ポスター（Poster）

Poster ポスター

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-001 神経突起再編成を制御する分子機構の解明 Eri Hasegawa（長谷川恵理）¹，Nagomi Tokumitsu（徳光和波）¹，Satoyoshi Yanagi（柳学理）¹，Rei K Morikawa（森川麗）¹，Williams R Claire（Claire R Williams）²，Parrish Z Jay（Jay Z Parrish）²，Kazuo Emoto（榎本和生）¹ ¹東京大院理生物科学 ²Dept. Biol., Univ. Washington, Washington, USA Neurons remodel their axons and dendrites during development to generate refined and matured neuronal circuits. Neurite remodeling should be strictly regulated, as defects in remodeling processes result in various neuronal disorders. It is known that axons and dendrites are remodeled at the similar time during development. For example, rat prefrontal cortex neurons undergo axon and dendrite refinement during adolescence. Due to the lack of a suitable model, little is known about how axon and dendrite remodeling in a single neuron is coordinated. The dendrites of Drosophila sensory C4da neurons undergo a large-scale pruning and regeneration during metamorphosis. For the past decade, we have studied the pruning process of C4da neuron dendrites, and identified multiple cellular and molecular mechanisms underlying the pruning (Dev Cell 2010; Science 2013; Nature Commun 2015). Here, we show that the axons of C4da neurons are also pruned at almost the same time as dendrites. C4da axons are pruned mostly at the synaptic sites, resulting in elimination of most synapses by 24 hours after puparium formation. Ecdysone signaling and phagocytic receptors, which are required for dendrite pruning, are also required for axon pruning, suggesting that at least some molecular mechanisms are shared between axon and dendrite pruning. To comprehensively understand the molecular and cellular mechanisms coordinating both types of pruning, we performed a large-scale genetic screen and identified several candidate genes including an RNA-binding protein. We are performing detailed analyses of candidate genes and would like to discuss about how axon and dendrite pruning are coordinated by shared and distinct mechanisms.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-002 マウス大脳新皮質におけるアストロサイトの多様性についての解析 Baekjun Pior（表伯俊），Darin Lanjakornsiripan（Lanjakornsiripan Darin），Daichi Kawaguchi（川口大地），Yukiko Gotoh（後藤由季子）東京大院薬分子 Astrocytes, abundant glial cells in the mammalian brain, play various roles in supporting neurons and regulating synaptic transmission. In the neocortex, various neuronal subtypes are organized into a laminar structure and display layer-specific heterogeneity in morphological features and functions. By contrast, neocortical astrocytes in layers II-VI have long been recognized as a non-diverse protoplasmic astrocyte population, and thus their diversity and properties remained largely unclear. In this study, we found that astrocytes of the mouse somatosensory cortex manifest layer-specific morphological and molecular differences. Three-dimensional observations revealed that astrocytes in different layers showed distinct morphology in terms of territorial volume, process arborization and cell orientation. Moreover, the extent of astrocytic ensheathment of synapses in layer II/III was found to be greater than that in layer VI. These findings suggest that astrocytes in different layers interact differently with neighboring cells. Importantly, layer-specific differences in astrocyte properties were also observed in terms of gene expression patterns based on an RNA-sequencing analysis. For example, Lef1 mRNA was expressed higher in upper-layer (layer II/III and IV) astrocytes (ULA) compared to deep-layer (layer V and VI) astrocytes (DLA). Lef1 protein was indeed expressed at a higher level in layer I-IV astrocytes compared to layer V-VI astrocytes. By contrast, Id1 mRNA was expressed at a higher level in DLA compared to ULA, which was again confirmed at the protein level. Interestingly, immunostaining experiments indicated that most immature astrocytes in the early postnatal cortex expressed Id1 proteins but they gradually lost Id1 expression depending on their laminar positions. We also examined reeler and Dab1 conditional knockout mice, in which neuronal layers are disturbed, and found that positioning of neuronal subtypes affected layer-specific astrocyte properties. These results together suggest that astrocyte diversification takes place during astrocyte maturation, probably through interaction with neuronal layers.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-003 脳室上衣細胞の発達とBMPシグナリング Kotaro Hiraoka（平岡宏太良）¹，Kazuhiko Yanai（谷内一彦）²，Noriko Osumi（大隅典子）³ ¹東北大学サイクロトロン・RIセンター　サイクロトロン核医学研究部 ²東北大学院医機能薬理学 ³東北大学院医発生発達神経科学 Surface of the ventricles in the brain is covered by a layer of the ependymal cells. The cells regulate transfer of various molecules such as water, glucose, and ions between the brain and cerebrospinal fluid (CSF), and thus are called "brain-CSF barrier". The cells also regulate dynamics of the CSF by movement of the multiple cilia that project from apical surface of the cells to ventricular lumens. Similar to the neurons, astrocytes, and oligodendrocytes, the ependymal cells differentiate from the radial glia. In development of the ependymal cells, mitosis from stem cells mostly occurs during embryonal day 14 to 16 in mice. Precursor cells mature to multi-ciliated cells during first postnatal week. Factors that regulate development of the ependymal cells have not been fully elucidated. Bone morphogenetic protein (BMP) signaling implicates in several processes during development and in adult tissue homeostasis. In a previous in-vitro study, it has been shown that ectopic expression of intracellular inhibitory factors of BMP signaling promotes differentiation of stem cells into ependymal-like cells (Nishimura, 2010). We hypothesized that inhibition of BMP signaling is critical for ependymal development. Initially, we investigated whether BMP-signaling factors are expressed in the ventricular wall during ependymal development by immunohistochemistry. Phosphorylated SMAD1/5/8, a marker showing that BMP signaling is transduced, was observed neither at postnatal day 0 nor at postnatal day 8, while expressions of BMP2 and BMP4 were observed at these stages. Expression of SMAD7, an intracellular inhibitory factors of BMP signaling, was also observed at these stages. Secondly, we investigated effects of BMPs on ependymal development in cell culture. We obtained progenitor cells from pups of postnatal day 0. Cells were cultured in medium with fetal bovine serum for a week, and in medium without serum for 2 weeks. We added BMP2, BMP4 or vehicle in medium without serum. Rates of cells with multiple cilia in total cells were 1.4%, 0.8%, and 52.1% in BMP2, BMP4, and vehicle conditions, respectively. The results of immunohistochemistry and cell culture support our hypothesis that inhibition of BMP signaling is critical for ependymal development.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-004 トリ蝸牛神経核の機能分化におけるKvチャネル発現とCa²⁺の連関 Ryota Adachi（安達良太），Rei Yamada（山田玲），Hiroshi Kuba（久場博司）名古屋大院医細胞生理 Precise temporal coding of sound requires proper adjustment of ion channel expression in auditory neurons. Avian nucleus magnocellularis (NM), a homolog of mammalian anteroventral cochlear nucleus, is involved in the temporal coding and well known for rich expression of two types of voltage-gated K⁺ (Kv) channels, Kv1.1 and Kv3.1. NM neurons are tuned to a specific frequency of sound (characteristic frequency, CF), and arranged tonotopically within the nucleus. Moreover, they are differentiated biophysically as well as morphologically along the tonotopic axis. One prominent example is the differentiation of Kv1.1; the expression level of this channel increases in neurons with higher CF, which is considered crucial in adjusting neuronal excitability to the CF-specific patterns of afferent input in NM. Recently, we reported that the differentiation of Kv1.1 was created, because afferent input augmented the expression to a larger extent in higher-CF neurons, showing the importance of afferent input in setting the level of Kv1.1 expression. Intriguingly, however, the auditory threshold is higher for higher-frequency sound, and therefore, the level of afferent input cannot solely explain the graded expression of Kv1.1 toward higher CF, raising a possibility that additional factors may contribute to the differentiation. In this study, we explored the mechanisms of tonotopic differentiation of Kv1.1 in NM, using organotypic culture of chicken brainstem. We found that chronic depolarization increased Kv1 current in a level-dependent manner, but the extent was larger at higher-CF regions, causing the tonotopic difference of the current in the culture. The depolarization increased Kv1 current via elevation of [Ca²⁺]_i, while it elevated basal [Ca²⁺]_i similarly irrespective of tonotopic regions. The results showed that the Ca²⁺-dependent process of Kv1.1 expression was more efficient at higher-CF regions, suggesting the importance of neuronal tonotopic identity as well as pattern of afferent input for the tonotopic differentiation of Kv1.1 in NM.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-005 Glial cells missing 1は成長因子分泌を促進し、細胞の分化・増殖および血管新生を誘導する Yoshitaka Hayashi（林義剛）¹，Satoshi Fuke（福家聡）¹，Yasuhiro Go（郷康広）²，Kazuhiko Nakabayashi（中林一彦）³，Takahiro Fuchigami（渕上孝裕）¹，Naoko Morimura（守村直子）¹，Natsu Koyama（小山なつ）¹，Seiji Hitoshi（等誠司）¹ ¹滋賀医大医統合生理 ²自然科学研究機構生命創成探究センター ³成育医療研究センター周産期ゲノミクス研究室 Glial cell missing (gcm) plays a critical role in the glial cell development in Drosophila. Overexpression of Gcm1 in the mammalian embryo brain was shown to promote the differentiation of neural precursor cells into astrocytes. However, the function of Gcm1 in the adult postnatal brain remains to be investigated because Gcm1-deficient mice are embryonic lethal. On the other hand, when the brain was injured, a lot of astrocytes proliferate and play a key role to repair. Here, we show that the Gcm1 was upregulated expression in the brain of 3 days after injury. To determine the function of Gcm1, we performed in utero electroporation studies to overexpress Gcm1 together with GFP in neural precursor cells at E14.5 and analyzed at E17.5. The Gcm1 significantly promoted the emergence of GFAP(+) and S100β(+) astrocytes, which is consistent with the precedent studies. Next, we investigated the differentiation into oligodendrocyte lineage cells by immunostaining the Gcm1 electroporated brains. The number of Olig2(+) cells, an oligodendrocyte lineage marker, were increased both in GFP(+) and in GFP(-) populations. Furthermore, we also noticed that Gcm1 overexpression resulted in more angiogenesis. Interestingly, the differentiation and angiogenesis were regulated by LIF, VEGFA, and VEGFC secretion. Thus, considering that brain injury requires gliogenesis and angiogenesis to repair the injury, our results suggest that Gcm1 plays an important role in the injured brain and have a possible association with injury treatment.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-006 小胞体膜分子 Derlin-1の成体海馬ニューロン新生における役割 Naoya Murao（村尾直哉），Hideki Nishitoh（西頭英起）宮崎大医機能制御・機能生化学 The endoplasmic reticulum (ER) has a quality control mechanism that ER resident stress sensors recognize unfolded or misfolded (malfolded) proteins and trigger the unfolded protein response (UPR). The UPR mediates the proper folding or degradation of malfolded proteins and the translational attenuation to inhibit the further production of ER proteins. A collapse of the ER quality control mechanism contributes to the onset and deterioration of several neurological disorders associated with impaired learning and memory. Adult neurogenesis is the process to generate new neurons from adult neural stem/precursor cells in restricted brain regions. Neurogenesis in the adult hippocampal dentate gyrus plays an important role in learning and memory formation, and its homeostasis is disrupted in several neurological disorders associated with memory impairment. ER quality control and adult neurogenesis are thought to be closely related to the mechanisms of learning and memory and neurological disorders, whereas the physiological relevance among them remains to be elucidated. The ER membranes protein Derlin-1 mediates ER-associated degradation of misfolded proteins (ERAD), and is known to be important for ER quality control mechanisms. Therefore, in the present study, we investigated the function of Derlin-1 in neural stem cell-specific deficient (Derl1^f/f;Nestin-Cre) mice in adult neurogenesis. We found that disruption of homeostatic maintenance mechanism of adult neural stem cells caused abnormalities of adult neurogenesis and depletion of stem cell pool in Derl1^f/f;Nestin-Cre mice. These results suggest that ER quality control mechanism in the CNS by Derlin-1 may play an important role in maintaining homeostasis of adult hippocampal neurogenesis. On the other hand, there was no change in major UPR-related genes in DNA microarray analysis of the hippocampal dentate gyrus region. Derlin-1 may contribute to the degradation of specific substrates which are related to the adult hippocampal neurogenesis. In this presentation, we will discuss the detail of homeostatic abnormalities of adult neural stem cells, and candidate factors causing depletion of stem cell pool due to deficiency of Derlin-1.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-007 胎児期の大脳皮質の神経細胞移動におけるATF5の機能解析 Kiko Shimahara（島原喜子），Mariko Umemura（梅村真理子），Rino Oshima（大嶋梨乃），Haruo Nakano（中野春男），Shigeru Takahashi（高橋滋），Yuji Takahashi（高橋勇二）東京薬科大院生命科学環境応用動物 Activating transcription factor 5 (ATF5), a member of the ATF/cAMP-responsive element-binding (CREB) transcription factor family, is reported to regulate cell differentiation, survival, and apoptosis. It is a stress responsive transcription factor, and increases expression levels of mRNA and protein in response to environmental stresses such as heavy metal exposure and amino acid deficiency. ATF5 is expression is obvious in the brain, olfactory tissue, testes and liver. Particularly, it is highly expressed in the ventricular zone (VZ) and in the subventricular zone (SVZ) where neurogenesis occurs during cortical development. We previously showed that ATF5-deficient (ATF5-/-) mice demonstrated abnormal olfactory bulb development, decreased survival rate at neonatal stage, and abnormal behavior. It was also revealed that the localization of upper layer neurons in the cerebral cortex of ATF5-/- mice was abnormal. Mammalian cerebral cortex has a six-layered structure that is formed by radial migration of neurons in an ""inside-out"" manner. Newly born neurons transform their neuronal migrate mode sequentially, multipolar migration, locomotion, and terminal translocation, during the radial migration of neurons. Thus, we predicted that ATF5 is involved in the neuronal migration of the cerebral cortex. In this study, to define the function of ATF5 in regulating neuronal migration, we performed live imaging experiments to track the migration of neurons expressing ATF5 shRNA. In embryonic day 14, knockdown of ATF5 impaired locomotion of neuronal migration in the cortex from VZ to surface. Our results suggest that ATF5 might have an important role for migration of neurons during cerebral cortex development.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-008 GFAP-GFPマウスを用いた発生過程における海馬歯状回形成細胞の移動解析 Hiroshi Shinohara（篠原広志），Tatsunori Seki（石龍徳）東京医科大学　組織･神経解剖学分野 In general, neurogenesis occurs during embryonic and early postnatal stages, and ceases at adult stage. However, the dentate gyrus (DG) continues neurogenesis from embryonic to adult stages. In the adult DG, granule neurons are generated in the subgranular zone, while during embryonic period, dentate neural progenitors are initially produced in the ventricular zone (VZ), and then migrate through the suprafimbrial region to the subpial region (SP) where a new proliferative zone is formed to develop the presumptive dentate gyrus. During the migration, the progenitors differentiate into granule neurons or maintain property of neural progenitors that further contribute to perinatal and postnatal neurogenesis. Although the migration of the neural precursors and relocation of the region of neurogenesis are key processes for the formation of the DG, the exact temporal and spatial patterns are still unknown. To address the problem, we performed cell-tracing analysis of the DG using Gfap-GFP Tg mice. GFP+ cells originated from the VZ migrated to the DG. In the early stage, most migrating cells differentiated to the neuron, whereas in the late stage, some migrating cells indicated proliferating state. Moreover, we performed time-lapse imaging in cultured hippocampal slices and found some types of cell migration in the DG: pia-touching cells, presumptive hippocampal fissure-touching and somal translocation-like cells. We will discuss possibility that the correlation between cell-type specification and modes of cell migration.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-009 小脳顆粒細胞の発生における転写因子Meis1の機能解析 Ryo Shiraishi（白石椋），Tomoo Owa（大輪智雄），Mikio Hoshino（星野幹雄）国立精神・神経セ神経研病態生化学 In cerebellar development, granule cell precursors (GCPs) actively proliferate in outer external granule cell layer (oEGL) just beneath the pial surface of cerebellum. Subsequently, GCPs exit cell cycle and differentiate in inner external granule cell layer (iEGL) and migrate along the fiber of Bergmann glia cell to form internal granule cell layer (iEGL). These events are tightly regulated by various interactions of molecules and numerous studies have tried to elucidate molecular events. However, comprehensive mechanism is still unknown. Meis1 is a member of the three amino acid loop extension (TALE) family of homeodomain containing transcriptional factors. Meis1 has been reported to be involved in hematopoiesis and leukemia. We previously reported that Meis1 is a key regulator of cerebellar granule cell differentiation via Pax6 transcription, BMP signaling and Atoh1 degradation (Owa et al. 2018). In this study, we observed many abnormalities of granule cell precursors in conditional knockout mice which delete Meis1 specifically in granule cell lineage. This phenotypic abnormalities included delay of cell cycle progression, delay of cell cycle exit and early migration to internal granule cell layer. This results suggested that Meis1 could have many roles in various aspects of GCP development and it is necessary to perform functional analysis of Meis1 in more detail. In this study, we performed functional analysis of Meis1 in GCP development mainly by in vivo electroporation of expression vector or shRNA knockdown vector to GCPs in EGL.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-010 Kcnab1はマウス大脳皮質サブプレートの早期マーカーであり、6b層の皮質内投射ニューロンに発現する。 Yuichiro Oka（岡雄一郎）^1,2，Sheena Y. X. Tiong（Tiong Y. X. Sheena）^1,2,3，Tatsuya Sasaki（佐々木達也）²，Miyuki Doi（土井美幸）²，Manabu Taniguchi（谷口学）²，Makoto Sato（佐藤真）^1,2 ¹大阪大院連合小児発達 ²大阪大院医神経機能形態学 ³Fac of Sci, Univ of Malaya, Kuala Lumpur, Malaysia Subplate (SP) neurons are among the earliest-born neurons in the cerebral cortex and heterogeneous in terms of cell morphology and gene expression. In addition, SP neurons also have multiple targets of axon projections including distant cortical areas, thalamus and other sub-cortical regions. However, the relationships between various axon projections and gene expression patterns of the SP neurons, and their remnant layer 6b (L6b) neurons, have not extensively been studied. In this study, we searched for the genes expressed in the L6b/SP neurons that has ipsilateral cortico-cortical projection, and analyzed their expression patterns and axon projection during cortical development. Among the candidate genes identified by our microarray analysis, we focused on the gene encoding a beta subunit of voltage-gated potassium channel (Kcnab1) and an established L6b marker gene, Ctgf, as they showed characteristic but distinct expression patterns. In situ hybridization analysis revealed that Ctgf expression gradually emerged as early as embryonic day (E) 16.5 and remained specific to SP/L6b throughout the developmental stages tested (up to postnatal day (P) 25), while Kcnab1 expression in SP was clearly detected already at E14.5 and expanded to the cortical plate postnatally. Double labeling with retrograde tracing and in situ hybridization demonstrated that Kcnab1 was expressed in more than half of the L6b/SP neurons that have ipsilateral cortico-cortical projection from the primary somatosensory cortex (S1) to the primary motor cortex (M1). Interestingly, retrograde tracing from the contralateral S1 did not give any labeling in L6b. This unilateral connection pattern was already observed at P2, suggesting sensory inputs are not necessary for establishing the gross axon projection patterns of SP/L6b neurons. Double immunostaining experiments revealed different degrees of co-expression of the protein product Kvβ1 with L6b/SP markers Ctgf, Cplx3, and Nurr1, suggesting molecular subdivision of L6b/SP association neurons. Together, our data suggest that Kcnab1 is a marker for embryonic SP with early onset and its expression defines a subpopulation of L6b/SP neurons that has ipsilateral cortico-cortical projection.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-011 高頻度チロシンリン酸化MAP1Bの神経発生過程における機能解析 Yasuyuki Ito（伊藤泰行），Atsuko Honda（本多敦子），Michihiro Igarashi（五十嵐道弘）新潟大学 (Background & Aims) Protein tyrosine phosphorylation has been recognized as an important mechanism in regulation of the neuronal functions, such as neuronal circuit formation and plasticity. It has been reported that activities of several protein kinases are the highest in the developing brain. To understand the comprehensive information on tyrosine phosphorylation signaling in neuronal development, we performed the phosphoproteomics of the embryonic mouse brain proteins and identified that Y1685 of the microtubule-associated protein 1B (MAP1B), which phosphorylation site is not characterized well, has been frequently phosphorylated. (Results) We confirmed that the peptide sequences including pY1685 has been highly detected using phosphoproteomics in the embryonic stage, namely, embryonic day (E) 12, E15 and E18. pY1685-MAP1B has been detected most frequently on E15. Immunohistochemistry using a phospho-specific antibody revealed that the endogenous pY1685-MAP1B was enriched into the filopodia of growth cones and was localized more distally to EB3, a marker of microtubule (+)-end tips. In addition, pY1685-MAP1B was co-localized with the F-actin rather than with microtubules in the filopodia. The neurons overexpressing Y1685F-MAP1B, mimicking its unphosphorylated form, showed the reduced number and the area of growth cones, compared with the neuron overexpressing the wild type MAP1B. In contrast, those overexpressing Y1685E-MAP1B, mimicking its phosphorylated form, induced the filopodial elongation and decreased the area of lamellipodia. (Discussion & Conclusion) Since pY1685-MAP1B was co-localized with F-actin and the genetic mutation at Y1685 affected the growth cone morphology, we suspected that pY1685-MAP1B is likely to regulate polymerization and/or reorganization of F-actin in the filopodia.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-012 バルプロ酸誘発性自閉症モデルラットの発達期小脳におけるグリア細胞性の制御 Midori Fukushima（福島翠）¹，Chihiro Nishikawa（西川ちひろ）¹，Yoko Nomura（野村洋子）³，Yukiko Fueta（笛田由紀子）⁴，Susumu Ueno（上野晋）⁴，Yuko Sekino（関野裕子）⁵，Yasunari Kanda（諫田泰成）²，Sachiko Yoshida（吉田祥子）¹ ¹豊橋技科大院工環境・生命工学 ²国立医薬品食品衛生研 ³Queens College, City University of New York, New York, USA ⁴産業医大 ⁵東京大学 The abnormality of interaction between neuron and glia is one of the causes of developmental disorders such as autism spectrum disorder (ASD). Valproate (VPA) is a widely used anti-epileptic drug, whereas it is known as an inducer of ASD. We observed some abnormalities of cerebellar cortex in ASD model rats maternally administered 600 mg/kg VPA p.o. at embryonic day 16 (E16). The irregular structure between the lobule V and VI was observed from P5 and maintained in adult. Epigenetic neuronal changes were expected to begin from the early developmental period. In this study, we examined early developmental alteration of neuron-glia interaction. We observed Reelin was more randomly expressed in the external granule layer of VPA-ad. animals than the control animals at P7. Reelin has been reported to be significantly reduced in human ASD brain and related some phosphorylation cascades. Reelin regulates the migration of neurons, suggesting that changes in the expression patterns of Reelin cause autism phenomena. Additionally, we examined the effect of glia and VPA-administered glia to the differentiation of neurosphere derived from E16 rat cerebrum. Glia-conditioned differentiation medium (GCM) was prepared with confluent glial culture. VPA-administered glia-conditioned differentiation medium (VPA-GCM) affected the differentiation of neurosphere, and this phenomena was modified with some kinase inhibitors. In both developing cerebellar cortex and glial culture, microglia was more decreased in VPA-administrated condition than control condition. Because Reelin was released from neuron, we suggest that a three-way relationships between neuron, astrocyte and microglia would modify the cerebellar development in VPA-administrated ASD-model rats.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-013 視神経オリゴデンドロサイトの起源領域の種間比較 Katsuhiko Ono（小野勝彦），Hitoshi Gotoh（後藤仁志），Tadashi Nomura（野村真）京都府立医大院神経発生生物学 Oligodendrocytes in the optic nerve originate in the rostral basal forebrain and migrate into the optic nerve during embryogenesis. We have previously reported a species difference of the embryonic origin of optic nerve oligodendrocyte precursor cells (ON-OPCs): the chiasmal region in the chick embryo and the preoptic area (POA) in the mouse fetus (Ono et al., Neuron, 1997; Brain Struct Funct, 2017). To further examine the species difference of ON-OPC development, we compared the expression of Olig2 and PDGFRa, evolutionary conserved markers for ON-OPCs, in embryos of various amniote species, such as mouse, chick and Chinese soft-shelled turtle. We used Hmx2/3 as a marker for POA in these species. Olig2+ and PDGFRa+ cells are detected in the POA but not in the chiasmal region of E12.5 mouse embryos. On the contrary, focal expression of Olig2 and PDGFRa was evident in the neuroepithelial layer of chiasma but not in the POA in HH28 chick embryos. Interestingly, in Chinese soft-shelled turtle embryos, Olig2+ cells were observed both in the chiasmal region and POA, while PDGFRa+ cells appeared only in the POA. Although lineage-tracing analysis of Olig2+ cells has not been carried out in the turtle embryos yet, the present histological data indicated that the embryonic origin of ON-OPCs in the rostral basal forebrain is comparable among amniotes. Furthermore, unique appearances of Olig2+ and PDGFRa+ cells in turtle embryos suggest that the POA is a common developmental origin of the ON-OPCs possibly shared in ancestral characteristics. We will discuss the functional relevance of species-specific origins of optic nerve OPCs and comparative anatomy of the visual system.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-014 マウス海馬CA1と大脳新皮質錐体細胞の放射状グリア線維依存的な移動様式の違いについての解析 Ayako Kitazawa（北澤彩子），Minkyung Shin（シンミンギョン），Kanehiro Hayashi（林周宏），Ken-ichiro Kubo（久保健一郎），Kazunori Nakajima（仲嶋一範）慶應大医解剖 Pyramidal neurons in the neocortex and hippocampus are born near the ventricle and migrate to their destinations during brain development. It has been known that neocortical neurons migrate in the mode called ""locomotion"", in which they migrate along with a single radial glial fiber for long distances in the cortical plate. On the other hand, hippocampal CA1 neurons migrate in the ""climbing"" mode, in which neurons migrate slowly using multiple radial glial fibers with multiple leading processes in the hippocampal plate. However, the precise mechanisms of the ""climbing"" mode remain unknown. In this study, we have investigated differences and similarities of the migration between the hippocampal CA1 region and neocortex using immunohistochemistry and electron microscopy. The leading processes of mouse CA1 neurons during migration were complex and have multiple sites of adhesion with other processes or cells in the hippocampal plate. Many CA1 neurons directly adhere to each other in the hippocampal plate, making the cell density higher than that of the cortical plate in the neocortex. On the other hand, knockdown of cell adhesion molecules using in utero electroporation suggested that CA1 neurons and neocortical neurons use common molecules for the migration. Based on these results, we hypothesized that the complexity of the shape of migrating neurons during climbing mode would be caused by the extracellular environment unique in the hippocampus. Thus, we attempted to transplant CA1 neurons into the neocortex. To investigate the direct effect of the neocortical radial glial fibers on the morphology of migrating CA1 neurons, we developed a novel culture system which allows the radial glial cells survive with their radial morphology in the absence of the surrounding cells like cortical plate neurons. When we placed neocortical and CA1 neurons onto the neocortical radial glial fibers, the ectopically transplanted CA1 neurons did not change their shape and did not migrate toward the cortical plate, while the transplanted neocortical neurons migrated normally into the cortical plate. These results indicate that the neocortical and hippocampal neurons might use distinct cues in their migrations.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-015 マウス海馬歯状回における自閉症感受性遺伝子AUTS2の機能解析 Saki Egusa（江草早紀）¹，Kei Hori（堀啓）¹，Asami Sakamoto（坂本亜沙美）¹，Manabu Abe（阿部学）²，Kenji Sakimura（崎村建司）²，Yasuhiro Go（郷康広）³ ¹国立精神・神経セ神経研病態生化学 ²新潟大脳研基礎神経科学細胞神経生物 ³生理研発達生理認知行動発達 The Dentate gyrus (DG) is a part of the limbic system crucial for higher brain functions, and malformation of the DG is associated with neuropsychiatric disorders. The molecular mechanisms underlying the pathogenesis are, however, largely unknown. Autism susceptibility candidate 2 (AUTS2) is a gene associated with a broad range of psychiatric illnesses. Auts2 is expressed at multiple brain regions as prefrontal cortex, hippocampus and DG as well as cerebellum. It has been previously reported that the cytoplasmic AUTS2 is involved in the regulation of neuronal migration and neuritogenesis in the developing cerebral cortex whereas the nuclear AUTS2 acts as a transcriptional activator. Although many evidences suggest that AUTS2 plays the crucial roles for the neurocognitive function, the physiological function of AUTS2 in brain development, however, remains to be elucidated. In this study, we investigated the role of AUTS2 in the DG development using the mutant mice that Auts2 gene is conditionally ablated at telencephalon. Immunohistochemistry shows that AUTS2 is expressed in the neural progenitor cells and the granule neurons, but is not detected in neural stem cells . We found that, the size of DG in the Auts2 mutant mice was drastically reduced in both the developing and mature brains. In the mutant mice, the number of granule neurons was decreased, and unexpectedly, the neural stem cells were also significantly reduced in total number and tertiary matrix, but in VZ of DG, the number of the neural stem cells were increased. It is suggested that AUTS2 may be involved in not only in the regulation of neuronal migration, but also the regulation of the granule cell differentiation. Based on these results, we demonstrate the crucial role of Auts2 for the DG development.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-016 CRISPR/Cas9によるゲノム編集によりHAタグラベルしたDab1発現マウスの開発 Takao Honda（本田岳夫），Kazunori Nakajima（仲嶋一範）慶應大医解剖 Reelin is a large glycoprotein secreted mainly from Cajal-Retzius cells in the marginal zone, and is critically important for regulation of neuronal migration especially in layered structure such as cerebral cortex and hippocampus. Reelin transmits the signal to a cytoplasmic adaptor protein Dab1 through its receptors ApoER2 and VLDLR, and induces Dab1 tyrosine phosphorylation. We previously showed that Dab1 has two nuclear localization signals (NLS1/2) and nuclear export signals (NES1/2), and shuttles between the cytoplasm and the nucleus. Although we observed nuclear translocation of Dab1 in Dab1-overexpressed HEK293T or Neuro2a cells and primary cultured neurons, it remains unclear whether Dab1 exists in the nucleus in cortical excitatory neurons in vivo due to its difficulty of detection by immunohistochemistry. Since the overexpressed Dab1 in cerebral cortex can be immunohistochemically detected, difficulty of Dab1 detection in vivo might be caused by its low protein amount. To overcome this difficulty, we have inserted a hemagglutinin HA and highly sensitive spaghetti monster tag just before the stop codon of the C-terminus of Dab1 using CRISPR/Cas9 genome editing system delivered by in utero electroporation. We could detect not only highly-sensitive spaghetti monster-tagged-Dab1 but also HA-tagged-Dab1 in excitatory cortical neurons. For further observation of Dab1 subcellular distribution in other brain regions, where it is difficult to introduce Cas9 genome editing system by electroporation, we are developing HA-tagged-Dab1-expressing mouse by editing fertilized-egg using CRISPR/Cas9 genome editing system.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-017 大脳皮質の神経幹細胞内における細胞周期因子Cyclin D2のmRNA輸送機構の解析 Takako Kikkawa（吉川貴子）¹，Yoshio Wakamatsu（若松義雄）¹，Yukiko U. Inoue（井上-上野由紀子）²，Yuji Tsunekawa（恒川雄二）³，Fumio Matsuzaki（松崎文雄）³，Kunihiro Suzuki（鈴木久仁博）⁴，Takayoshi Inoue（井上高良）²，Noriko Osumi（大隅典子）¹ ¹東北大院医発生発達神経 ²国立精神・神経セ神経研疾病６ ³理研CDB非対称細胞分裂 ⁴日大松戸歯生物 During corticogenesis, neural stem/progenitor cells need to adequately proliferate and differentiate. Neural stem/progenitor cells in the developing cortex are called radial glial (RG) cells, as they possess highly polarized morphology with long apical/basal processes. We have previously shown in the mouse that mRNAs of Cyclin D2 (Ccnd2), coding a cell cycle regulator, are transported to the basal end-foot of the RG cell by using a short 3'UTR sequence as a zip code (Tsunekawa et al., EMBO J, 2012). To examine the requirement of the Ccnd2 zip code in vivo, we applied a CRISPR/Cas9 genome editing system in mice to selectively remove the zip code by introducing two single guide RNAs targeting both ends of the zip code. In the zip code-deleted mutant cortex, Ccnd2 mRNA no longer localized in the basal end-feet of RG cells, yet remained around the RG cell soma. These results indicate that the zip code is essential for the basal transport of the Ccnd2 mRNA to the basal end-feet of RG cells. Interestingly, the zip code sequence is only conserved in placental mammals when we compared the sequences among various vertebrates. Consistently, Ccnd2 mRNAs were not detected in the basal end-feet of RG cells of opossum, a marsupial mammal. Furthermore, we electroporated a reporter construct containing opossum Ccnd2 3'UTR downstream to EGFP into mouse RG cells and found that the mRNA was not transported to their basal end-feet. Conversely, EGFP mRNA with mouse Ccnd2 3'UTR was basally transported in the opossum RG cells, suggesting that the transport machinery itself is conserved in mouse and opossum. Previous reports indicate that Tbr2-positive basal progenitors are proliferative in placental mammals but not in the developing opossum, suggesting the importance of the basal transport of Ccnd2 mRNA in producing proliferative basal progenitors to make a larger cortex of placental mammals.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-018 神経幹細胞におけるbHLH型転写因子のダイナミックな発現制御機構 Mayumi Yamada（山田真弓）^1,2,3，Shinji C Nagasaki（長崎真治）^1,2，Itaru Imayoshi（今吉格）^1,2,3 ¹京都大学大学院生命科学研究科 ²京都大学大学院生命科学研究科附属生命動態研究センター ³京都大学ウイルス再生医科学研究所 The mammalian brain consists of a complex ensemble of neurons and glial cells. Their production during development and remodelling is tightly controlled by various regulatory mechanisms in neural stem cells. Among such regulations, basic helix-loop-helix (bHLH) transcription factors have key functions in the self-renewal, multipotency, and fate determination of neural stem cells. Several bHLH transcription factors are coexpressed by neural stem cells and change its expression patterns uniquely during cell-fate determination. By time-lapse imaging, we found that these factors are expressed in an oscillatory manner by neural stem cells. Here, we focus on the importance of the expression dynamics of bHLH transcription factors. To analyze causal relationships between the dynamic gene expression changes of bHLH transcription factors and neural stem cell regulations, it is essential to manipulate bHLH transcription factors expressions with the fine temporal and spatial resolution. Recently, we developed a new photoactivatable (PA) tetracycline (Tet)-controlled gene expression system (PA-Tet system) for precise temporal and spatial control of gene expression at single-cell resolution. This system had a large dynamic range of downstream gene expression, rapid activation, and deactivation kinetics. We are planning to adapt this optogenetic system to control the expression dynamics of bHLH transcription factors to understand their functional roles in cell proliferation, fate-determination, and differentiation of neural stem cells.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-019 オリゴデンドロサイト前駆細胞維持に必須な新規Olig2結合因子の同定 Norihisa Bizen（備前典久）¹，Masato Yano（矢野真人）¹，Anna Simankova（Simankova Anna）¹，Li Zhou（周麗）^1,2，Manabu Abe（阿部学）²，Kenji Sakimura（﨑村建司）²，Hirohide Takebayashi（竹林浩秀）¹ ¹新潟大院医歯学総合研 ²新潟大学脳研究所 Oligodendrocytes form the myelin sheaths around axons and regulate the salutatory conduction in central nervous system (CNS). The failure of oligodendrocyte development leads to the misregulation of neuronal networks and thereby serious intractable neuronal disorders. Olig2, a basic loop helix loop transcription factor, is indispensable for the production and differentiation of oligodendrocytes in the CNS. However, it has remains unclear how Olig2 governs oligodendrocyte development. To elucidate the molecular basis underlying Olig2-mediated oligodendrocyte development, we have sought Olig2-binding factors, which cooperate with Olig2 for oligodendrogenesis. Yeast two hybrid screening using cDNA library prepared from mouse embryonic brains identified several candidate factors. Among them, Obp2 (Olig2-binding protein 2) has been known to be involved in snRNP biogenesis and transcriptional regulation. Nestin-Cre:Obp2 conditional knockout mice demonstrated that the number of Olig2- and PDGFRα-positive oligodendrocyte precursor cells started to decrease significantly in Obp2-deficient spinal cords from E13.5, which is oligodendrogenic stage. On the other hand, Obp2 deficiency did not affect motor neuron generation. Apoptosis and cell cycle arrest were significantly induced in oligodendrocyte precursor cells of Obp2-deficient mice. In addition, p53 pathway was also activated. Thus, these results suggest that Obp2 is a key factor for the maintenance of oligodendrocyte precursor cells in the CNS.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-020 Stk25とMST3による大脳皮質形成制御機構 Tohru Matsuki（松木亨），Akio Iio（飯尾明生），Masashi Ueda（上田昌史），Akie Toya（戸谷明恵），Atsuo Nakayama（中山敦雄）愛知県医療療育総合センター・発達障害研究所 Neuronal migration during cortical lamination is precisely regulated and necessary for development of functional brain. Stk25 has been identified as a modifier for Tau hyperphosphorylation regulated by Reelin-Dab1 signaling. Stk25 also have roles in Golgi morphological regulation, axonogenesis, neuronal polarization. However, we also found that although Stk25 has an additional role in neuronal migration during corticogenesis, Stk25 null mice show unclear histological aberrations compared to normal mice. In this study, we identified that MST3, a member of the GCKIII subgroup of the Ste20-like kinase family, plays similar roles in several biological functions, compensating for Stk25. In vivo, MST3 overexpression in Stk25 knockout neurons rescues neuronal migration and axonogenesis. Furthermore, defects in axonal formation induced by acute Stk25 knockout or knockdown recover within several days. This compensation system during neuronal migration is achieved by scaffolding abilities of Stk25 or MST3. These molecules recruit α -PIX or β -PIX, guanine exchange factors for Rac1. In addition, Stk25 and MST3 accelerate degradation of RhoA through the interaction. This study demonstrates that Stk25 and MST3 have similar roles in brain development through a mutual compensation mechanism by acting a hub protein for Rac1 activation and RhoA destabilization pathway. This study also provides insights not only for uncovering roles of GCKIII proteins, but also for understanding the basis of the cortical layer formation.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-021 大脳新皮質の腹側方向への拡張的形成には胎生初期プレプレートニューロンの腹側への流れが物理的に貢献する kanako Saito（齋藤加奈子），mayumi Okamoto（岡本麻友美），yuto watanabe（渡邊裕斗），Arata Nagasaka（長坂新），Takaki Miyata（宮田卓樹）名古屋大学医学研究科細胞生物学 Mammalian neocortical somatotopies (mapping body parts on the cortex) show a disproportionally large representation in the lateral filed (which is devoted for the fingers and lip in humans and the whiskers in mice), compared to the dorsal filed (representing the feet and trunk). Such "luxurious"; use of the lateral cortex depends on dorsal-to-ventral expansion of the pallium during development. Despite recent exploration in progenitor cells about molecular mechanisms underlying the cortical arealization/patterning, physical understanding of how the cortex expands through what force-generating events is missing. We hypothesized that neurons born very early in the dorsal pallium might ventrally migrate to physically increase the horizontal size/area of a superficial tissue (in a way analogous to pushing out of the Nile Delta into the Mediterranean sea). Formation of the neocortical neuronal layers in mice begins at E11 with the emergence of the preplate, the first and transient layer composed of the ealiest-generated cortical neurons. Preplate neurons' start to split into a superficial marginal zone containing Cajal-Retzius cells and a deeper subplate at E13. The importance of the preplate-constituting neurons has been known partly, only regarding their contribution to events in the mid- or late embryonic periods. We indeed found that neurons born earliest (at embryonic day 10 [E10]: dorsal pallial-derived and including subplate neuron) in the mouse pallium massively migrate ventrally during the E11-12 period, which was dependent on Rac1 and N-Cadherin, and then extend corticofugal axons to together form a ventral morphogenetic flow of the preplate, a transient layer of early-born neurons, until E13. Ablation of these E10-born neurons through in utero electroporation-based expression of diphtheria toxin to reduce the ventral "preplate stream" attenuated (1) the ventral deflection of radial glial fibers (by E13) and (2) the extension of the cortical plate at E14 (which was follow by immunohistochemistry for Ctip2, Satb2 and BrdU pulse at E12) that should occur ventrally through receiving later-born neurons. Weakening of the "preplate stream" also (3) shrank the postnatal neocortical map (including somatosensory barrels) dorsally. Thus, this previously unrecognized "preplate stream" physically primes the neocortical expansion and the somatotopic map formation.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-022 成長円錐のリン酸化プロテオミクスにより同定されたGAP-43のJNKリン酸化部位は伸長または再生する神経軸索で活性化される Asami Kawasaki（河嵜麻実）¹，Atsushi Tamada（玉田篤史）¹，Masayasu Okada（岡田正康）^1,3，Shujiro Okuda（奥田修二郎）²，Hirohumi Nishina（仁科博史）⁴，Tokiwa Yamasaki（山崎世和）⁴，Michihiro Igarashi（五十嵐道弘）¹ ¹新潟大学大学院　医歯学系総合研究科　分子細胞機能学 ²新潟大学大学院　医歯学総合研究科　バイオインフォーマティクス分野 ³新潟大学　脳研究所　脳神経外科学分野 ⁴東京医科歯科大学難治疾患研究所発生再生生物学分野 Axonal growth is a process to establish the functional neural circuits in brain. Toward the comprehensive understanding of the signaling pathways to regulate axonal growth, we took an unbiased phosphoproteomic approach to identify the whole phosphorylation sites involved in this event. Phosphoproteomic analysis of growth cone membranes prepared from postnatal day 1 rat forebrain identified more than 5,000 phosphorylation sites from ~1,200 proteins. The phosphorylation sites were highly proline directed and primarily mitogen-activated protein kinase dependent, owing to the activation of c-jun N-terminal kinase (JNK), suggesting that proteins that undergo proline-directed phosphorylation mediate nerve growth in the mammalian brain. Bioinformatics analysis revealed that phosphoproteins were enriched in microtubules and the cortical cytoskeleton. The most frequently phosphorylated site was Ser96 of growth-associated protein 43-kDa (GAP-43), a vertebrate-specific protein involved in axon growth. The S96 phosphorylation was specifically detected in growing and regenerating axons in mice by immunohistochemistry. Taken together, phosphorylated Ser96 of GAP-43 is a potential molecular marker for mammalian axonal growth. (Ref.) Kawasaki et al.: iScience 4: 190 [`18]

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-023 大脳原基皮質板におけるニューロン個性化完遂にはミクログリアの一時退出が必要である Yuki Hattori（服部祐季），Yu Naito（内藤裕），Ayano Kawaguchi（川口綾乃），Takaki Miyata（宮田卓樹）名古屋大院医細胞生物 Microglia, the resident macrophages in the central nervous system (CNS), have multiple functions in the embryonic brain such as phagocytotically regulating the number of neural progenitors and inducing neural stem-like cells to differentiate into intermediate progenitors. Although microglia account for only a minor population of the cells that constitute the cerebral wall, they extensively survey the entire structure, and are thus capable of providing the particular functions that are required in specific regions. Of note, these cells exhibit a mysterious behavior in the mid-embryonic cortex: they temporarily disappear from the cortical plate (CP), where post-migratory neurons accumulate and undergo subsequent specification programs that confer their projection subtype identity. To address the significance of microglial temporal absence from the mid-embryonic CP, we first investigated the molecular mechanism underlying their disappearance from the mid-embryonic CP. Using live imaging-based analyses, we found that microglia are bidirectionally expelled from the CP via attraction by CXCL12 produced from the meninges and subventricular zone. Next, we performed artificial delivery of microglia into the CP of cultured cerebral walls and thorough in vitro mixing of microglia and dissociated neurons homogeneously grown to an age corresponding to that of in vivo CP neurons. Both manipulations resulted in disturbed neuronal subtype specification (elevated expression of Tbr1, Satb2, and Cux1 and reduced expression of Ctip2). Further, We found that this experimental effect on CP neurons was primarily attributed to interleukin 6 (IL-6) produced by microglia. These results suggest that microglial temporal absence from the mid-embryonic CP is necessary for neurons to appropriately fine-tune the expression of transcription factors for proper cellular specialization, thus securing the well-balanced coexistence of functionally distinct neurons.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-024 転写因子の翻訳後修飾を介した小脳顆粒細胞の分化決定機構の解析 Satoshi Miyashita（宮下聡）¹，Yusuke Seto（瀬戸裕介）²，Tomoo Owa（大輪智雄）¹，Shinichiro Taya（田谷真一郎）¹，Yoshiya Kawaguchi（川口義弥）³，Mikio Hoshino（星野幹雄）¹ ¹国立精神・神経セ神経研病態生化学 ²京都大ウィルス研 ³京都大iCeMS The restrict regulation of the balance between proliferation and differentiation of neural precursors is essential for proper CNS organization. However, the underlining molecular mechanisms are still elusive. The development of cerebellar granule cells provides a good model to study this mechanism. During the development, granule cell precursors (GCPs) leave the rhombic rip, the origin of cerebellar excitatory neurons, migrate along the pial surface of the cerebellum and then form the external granule cell layer (EGL) that consists of two sublayers, outer EGL (oEGL) and inner EGL (iEGL). GCPs proliferate in the oEGL, and then exit from the cell cycle to move into iEGL. After staying in iEGL for a few days, GCPs leave iEGL for internal granule cell layer (IGL) to become mature granule cells(GCs). Although these processes for differentiation and maturation of GCs have been well described, its underlying machinery is poorly understood. Atonal homolog 1(Atoh1) is well-characterized bHLH transcriptional factor that has fundamental roles in the fate determination of GC lineage in embryonic rhombic lip and the proliferation of GCPs in EGL. Atoh1 is also suggested to have an oncogenic potential in sonic-hedgehog driven medulloblastoma derived from GC lineage. These features of Atoh1 indicate the requirement for the precise regulation of the expression level of ATOH1 protein. Although several studies have suggested the significant involvement of the proteasomal degradation in the ATOH1 protein level, it is still elusive how ATOH1 protein level is regulated in EGL during cerebellar development. In this study, we identified the post translational modification (PTM) of ATOH1 mediated by cell cycle related genes in neuronal cells. We also revealed that the PTM of ATOH1 were imporatant for the inhibition of the proteasomal degradation and the proper balance of the differentiation. These results suggested the novel mechanisms regulating the differentiation of GCs through the PTM of ATOH1.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-025 中枢神経系の髄鞘形成におけるLAMTOR1の役割 Takehiko Sunabori（砂堀毅彦），Masato Koike（小池正人）順大院神経生物形態 Myelination of axons by oligodendrocytes is crucial for efficient signal transduction in the central nervous system (CNS). Lines of reports have suggested that MAPK signaling pathway and mammalian target of rapamycin complex (mTORC) signaling pathway contributes in the differentiation of oligodendrocytes followed by axonal myelination. However, little is known about the underlining molecular mechanisms. LAMTOR1 is a lysosomal adaptor protein, which serves as an essential anchor for scaffolding both MAPK and mTORC pathways on the lysosomal membranes. To elucidate the in vivo function, we generated CNS specific LAMTOR1-deficient mice. Mice lacking LAMTOR1, specifically in the CNS, died around postnatal day 15 with severe epileptic seizures. Although we could not detect remarkable difference in neuronal positioning, axon projection and astrogenesis, dysmyelination was observed through the entire CNS. In situ hybridization and immunohistochemistry revealed that the generation and migration of oligodendrocyte precursor cells (OPCs) were normal. However, the proliferation of OPCs was significantly reduced from postnatal day 8 (P8) to P14. Since both mTORC and MAPK pathways are related to cell proliferation, we next asked the activation of both pathways by western blot analyses. Results showed that LAMTOR1-deficient mice showed inhibition only in the mTORC pathway, while the MAPK pathway remained normal. Taken together, these data suggests that LAMTOR1 has a crucial role for axonal myelination in the CNS by controlling OPC proliferation.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-026 ピロロキノリンキノンおよびポリフェノールにより惹起されるコンドロイチン硫酸の発現促進による神経機能制御 Hiroyuki Sasakura（笹倉寛之）¹，Masashi Ikeno（池野正史）¹，Yuki Morioka（森岡幸）¹，Yuka Takeuchi（武内由佳）¹，Ikue Mori（森郁恵）²，Kosei Takeuchi（武内恒成）¹ ¹愛知医科大学医学部　細胞生物学 ²名古屋大学大学院理学研究科生命理学専攻　生体構築論講座　分子神経生物学 Chodroitin sulfate (CS) proteoglycan is a major component of the extracellular matrix and plays an important role in neural development and maintenance. We recently showed that CS promotes neurogenesis in adult mouse hippocampus in response to enriched environment (Yamada et al., 2018, J.N.). This result revealed that CS functions as a critical regulator for the neurogenesis of mammalian brain in an activity dependent manner, however, the underlying mechanism is still unknown. The identification of chemicals that promote the expression of CS would be a powerful tool to dissect the CS-induced neurogenesis. Also, such chemicals enable us to test if the brain functions are improved in neural disordered or aged mouse by administration of them. We screened 271,040 natural organic compounds and identified hundreds of chemicals that up-regulated the expression of CS through in-cell western method. Pyrroloquinoline quinone (PQQ) and polyphenols were identified as chemicals that up-regulate CS. PQQ is an intriguing chemical which we previously identified as a lifespan extension drug for C. elegans (Sasakura et al., JCS, 2017). To elucidate the effects of PQQ and polyphenols for nervous system, we are conducting both in vitro and in vivo studies. We are studying the culture cells to clarify the mechanism of CS up-regulation. We are studying both C. elegans and mouse to evaluate the effects of chemicals for neural functions. The progress will be discussed in this presentation.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-027 分泌性糖タンパク質ReelinによるN-cadherinの制御機構 Kanehiro Hayashi（林周宏）¹，Seika Inoue（井上聖香）¹，Nobuhiko Nakao（仲尾信彦）²，Ken-ichiro Kubo（久保健一郎）¹，Taiji Adachi（安達泰治）²，Kazunori Nakajima（仲嶋一範）¹ ¹慶應義塾大学医学部解剖学 ²京都大学ウイルス・再生医科学研究所生命システム研究部門バイオメカニクス研究領域 The mammalian cerebral cortex has six layered structure, which enables higher cognitive functions. Extracellular glycoprotein, Reelin is critical for formation of this brain architecture because Reelin deficient mouse, reeler exhibits malformed six-layered structure. Reelin is secreted from mainly Cajal-Retzius cells in the marginal zone of the developing neocortex and affects migrating neurons to reach their final position. Recently, we reported that ectopic Reelin overexpression in the developing neocortex resulted in the formation of neuronal aggregation which is arranged in process-rich center area and cell body-rich peripheral region, and that formation of this aggregation is mediated by N-cadherin (Kubo et al., 2010, J. Neurosci.; Matsunaga, Noda et al., 2017, PNAS). In this study, we showed how Reelin regulate N-cadherin dependent cellular adhesion. The biochemical assay showed Reelin increases the amount of N-cadherin molecule on the cellular membrane of the primary cultured cortical neurons. Next, we measured the parameters of N-cadherin dependent adhesion on primary cultured cortical neurons after Reelin treatment by Atomic force microscopy (AFM). Reelin increased not only the number of N-cadherin on the plasma membrane but also adhesive strength of each N-cadherin molecule on the cellular membrane. Furthermore, we found Dbnl, which is actin-associated protein is the molecule to connect Reelin and N-cadherin. Dbnl was phosphorylated in the cultured neurons by Reelin treatment and retained N-cadherin on the plasma membrane in a Dbnl-phosphorylation dependent manner. These results suggest Reelin controls N-cadherin in both quality and quantity through Dbnl.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-028 低強度運動で高まる雌の海馬神経新生におけるアンドロゲン受容体の役割 Masahiro Okamoto（岡本正洋）¹，Takeru Shima（島孟留）¹，Yu-Fan Liu（劉宇帆）¹，Teresa A Milner（Milner A Teresa）³，Bruce S McEwen（McEwen S Bruce）²，Hideaki Soya（征矢英昭）¹ ¹筑波大　体育系ヒューマン・ハイ・パフォーマンス先端研究センター ²The Rockefeller University, New York, USA ³Weill Cornell Medicine, New York, USA Exercise has beneficial effects on hippocampal plasticity such as increasing adult neurogenesis. Our previous work revealed dihydrotestosterone (DHT) synthesized in hippocampus was potent mediator of mild exercise-induced neurogenesis in males. Although estrogens are considered a close relationship with neuronal plasticity and function in female, intriguingly, androgen receptor is distributed in female brain including hippocampus. This thing brings up a new question if androgens have a pivotal role on exercise effects in female as well as in male. To address this matter, we have investigated the effect of both sex steroid hormone, androgens and estrogens, on exercise-induced neurogenesis in female rats. Adult female rats were subjected to mild treadmill exercise for 2 weeks, which is a running speed of 15 m/min, 5 times per week for 30 min at a time. On the day before training, all the animals were given intraperitoneal injections of 5-Bromo-2-deoxyuridine (BrdU). Sedentary rats remained on the treadmill without running for the same amount of time. Ovariectomy and injection of estrogen receptor antagonist tamoxifen didn't affect AHN. On the other hand, androgen receptor (AR) antagonist flutamide blocked the mild exercise-induced neurogenesis. The inhibitory effects were notably the increase of DCX or BrdU/NeuN positive cells, not Ki67 positive cells. These data implied that androgen receptor regulates the differentiation and survival of hippocampal newborn cells, not proliferation. For genes expression analysis, hippocampus was separated into DG and CA regions. The validation of separation was confirmed with enrich expression genes in each region by conducting real time PCR. We found that mild exercise increased mRNA expression of androgen related to genes such as AR and srd5a2, which converts testosterone to DHT, in DG. These results provide evidence supporting our hypothesis that even in female rats, androgenic mediation plays a crucial role in mild exercise-enhanced hippocampal neurogenesis.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-029 小脳発達期における細胞間相互作用の計算モデル Mizuki Kato（加藤瑞己），Erik De Schutter（De Schutter Erik）沖縄科学技術大院計算脳科学ユニット The cerebellum is well known for its coordination of motor functions as well as its roles in non-motor functions such as cognition and emotion. Considering the complex information processing the cerebellum has to deal with, it is surprising that its output is given by a single type of neurons, Purkinje cells. In development of murine cortex, Purkinje cells undergo intensive dendritic remodeling stages to attain their mature morphology. In one of the remodeling stages, one or two primary dendritic trees are selected among several young trees. The selected one develops into a mature structure, whereas the unselected ones retract completely. The mechanism for this selection and definite rules for winning the selection have not been identified. However, there is indirect experimental evidence that intensive interactions with migrating granule cells in the vicinity are involved in this selection mechanism. In the real system, large variability of developmental stages of Purkinje cells even between neighbors during the same period make it difficult to capture the exact moment of the interest. As an alternative, computational modeling approaches offer greater control over such factors. However, most of the computational models have focused mainly on intracellular mechanisms. A new computational model is therefore needed for a better understanding of the interactions between different types of neurons. In the new model, the dendritic development of the Purkinje cells and the migration of granule cells are computationally simulated simultaneously in an imaginary 3D cube. The newest version of a computational framework, NeuroMaC, is chosen for its capability to generate models based on biological rules with consideration of various interactions with the environment. This model can produce a comprehensible representation of the early developmental dynamics of neurons, which are hard to track with existing models. Also, the results from this model are expected to bridge the gap in understanding of the early neonatal development of functional Purkinje cells with their typical morphology.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-030 ヘパラン硫酸エンドスルファターゼSulf1/2二重欠損マウスにおける皮質脊髄路の形成異常と巧緻運動障害 Satoshi Aizawa（相澤哲史）¹，Takuya Okada（岡田拓也）^1,2，Kazuko Keino-Masu（桝和子）^1,2，Akira Tamaoka（玉岡晃）^1,3，Masayuki Masu（桝正幸）^1,2 ¹筑波大　大学院人間総合科学研究科 ²筑波大　医学医療系　分子神経生物 ³筑波大　医学医療系　神経内科 The corticospinal tract (CST) plays a role in skilled voluntary movements. Accurate navigation of the CST axons from the motor cortex to the spinal cord requires coordinated regulation of multiple axon guidance molecules. Heparan sulfate (HS) is a linear carbohydrate bound to the core proteins of proteoglycans which exist in the extracellular matrix or on the cell surface. HS interacts with growth factors, morphogens, axon guidance molecules, and their receptors, thereby controlling their distribution and function. The specificity and affinity of the interactions beween HS and signaling molecules depend on the sulfation patterns of HS. HS endosulfatases, Sulf1 and Sulf2, desulfate specific sulfate groups from mature HS extracellularly, thereby controlling multiple signaling pathways. We previously reported that the Sulf1/2 double knockout (DKO) mice exhibited the defects in CST formation owing to the abnormal accumulation of Slit2 protein on the brain surface as the results of increases in the sulfated HS. However, the postnatal development of the CST and possible deficits in the skilled motor functions in the Sulf1/2 DKO mice remained unknown because the DKO mice in the C57BL/6 background died within a day of birth. Recently, we found Sulf1/2 DKO mice in a mixed genetic background could survive into adulthood. In this study, we investigated the trajectory of the CST using PKCγ immunostaining, BDA tracing, and 3D reconstruction. These studies revealed aberrant projection on the midbrain surface, midline crossing errors at the pyramidal decussation, and abnormal projection in the spinal cord. To investigate the possible deficits in motor function in the Sulf1/2 DKO mice as a consequence of the CST defects, we performed behavioral experiments including the grid walking test, double staircase test, and single pellet reaching test. These analyses revealed the impairment of skilled motor movements in the Sulf1/2 DKO mice. Taken together, our findings indicate that knockout of Sulf1/2 genes causes axon guidance defect of the CST and dysfunction of skilled voluntary movements.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-031 LIMKによるアクチン細胞骨格の制御を介した僧房細胞の樹状突起リモデリング Shuhei Aihara（藍原周平），Takeshi Imai（今井猛）九州大学大学院医学系学府 In the mammalian brain, neurons initially form excessive neurites, and then remodel their morphology during early postnatal development. It has been known that the dendrite remodeling is controlled by both secreted/transmembrane proteins and neuronal activity. However, it has remained unknown how these two pathways control mature neuronal morphology. Here we studied dendrite remodeling of mitral cells in the mouse olfactory bulb. Mitral cells are known to connect multiple dendrites to multiple glomeruli in the early development, and prune all but one primary dendrite during the first postnatal week. Using in utero electroporation, we performed CRISPR/Cas9 screening of receptor genes for secreted/transmembrane proteins that regulate dendrite remodeling in mitral cells. We found that the mitral cell-specific knockout of Bmpr2 impairs dendrite pruning. Deletion analyses revealed that C-terminal domain which inhibits LIMK activity, rather than the kinase domain necessary for the canonical Smad pathway, is critical for normal dendrite pruning. Active LIMK1 is known to inhibit cofilin and thereby stabilize actin cytoskeleton. Supporting this notion, LIMK1 overexpression also demonstrated a similar defect in dendrite pruning and the phenotype was rescued by BMPR2 overexpression. We also found that overexpression of Rac1, a candidate downstream effector of neuronal activity, impairs dendrite pruning, while this defect was rescued by Limk1 knockout. Taken together, these results suggest that regulation of actin cytoskeleton via LIMK1 is critical for remodeling of mitral cell dendrites mediated by BMP signaling and neuronal activity.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-032 内因性・外因性の軸索再生阻害因子の解除による皮質脊髄路再生の試み Yuka Nakamura（中村由香）^1,2，Masaki Ueno（上野将紀）^1,2,3，Jesse Niehaus（Niehaus Jesse）^2,3，Yi Zheng（Zheng Yi）⁴，Yutaka Yoshida（吉田冨）^2,5 ¹新潟大脳研システム脳病態学 ²Dept Dev Biol, Cincinnati Children's Hospital Medical Center, OH, USA ³JSTさきがけ ⁴Dept Exp Hematol and Cancer Biol, Cincinnati Children's Hospital Medical Center, OH, USA ⁵Burke Neurological Institute, Cornell Med School, NY, USA Axon regeneration is limited both by inhibitory molecules expressed around the lesion and less ability of adult neurons to grow axons after spinal cord injury. It remains unknown whether simultaneous suppression of both "extrinsic" and "intrinsic" factors liming axon regrowth has synergistic and sufficient effects to induce regeneration. Here we tested whether genetic blockade of extrinsic and intrinsic signals by deleting Rho, a small GTPase that mediates signals from extrinsic molecules to inhibit axon growth, and Pten, a phosphatase inhibiting the intrinsic mTOR signaling which is important for axon growth, induces axon regeneration robustly after spinal cord injury. RhoA and Pten deletion increased neurite outgrowth in the presence of myelin and glial scar-related inhibitory molecules in vitro. Cerebral cortex-specific deletion of RhoA and RhoC significantly suppressed dieback of transected corticospinal axons in mouse spinal cord injury model. In contrast, Pten deletion in the cortical neurons significantly increased the regrowth of transected axons in the lesion scar, whereas the axon dieback was not affected. Deletion of both RhoA and Pten further enhanced rewiring of corticospinal axons to form detour circuits connecting the cerebral cortex and hindlimb muscles, although the motor recovery was not significantly promoted. Our results indicate that targeting intrinsic and extrinsic barriers to axon regeneration has synergistic effects to reconstruct neural circuits after spinal cord injury.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-033 小脳プルキンエ細胞の発生における自閉症感受性遺伝子AUTS2の役割 Kunihiko Yamashiro（山城邦比古）¹，Ryo Aoki（青木瞭）^1,2，Asami Sakamoto（坂本亜沙美）²，Kei Hori（掘啓）²，Mikio Hoshino（星野幹雄）^1,2 ¹東京医科歯科大学医歯学総合研究科NCNP脳機能病態学分野 ²国立精神神経医療研究センター病態生化学研究部 Cerebellum has not only been a good model system to study the neural development, but also considered recently as one of the remarkable brain regions involved in higher brain functions such as the cognitions and sociability as well as various motor activities. Defects of cerebellar development have been associated with a variety of psychiatric disorders, but the underlying pathogenesis remains poorly understood. Autism susceptibility candidate 2 (AUTS2) has been implicated in a variety type of psychiatric disorders such as autism, intellectual disabilities and schizophrenia. In developing CNS, Auts2 is highly expressed at several brain regions such as frontal cortex, hippocampus and cerebellum. Previously studies have demonstrated that AUTS2 is involved in the regulation of neuronal migration and neuritogenesis, acting as an upstream factor for Rac1 and Cdc42 during the late embryonic stages. In addition, the nuclear AUTS2 has been reported to participate in the regulation of gene expression by interacting with the Polycomb repressor complex 1 (PRC1) in the developing cerebral cortex. The physiological functions for AUTS2 in the postnatal brain development, however, remain largely unknown. In this study, we investigated the role of AUTS2 in the postnatal cerebellar development. In the cerebellum, AUTS2 is specifically expressed in GABAergic neurons including Purkinje cells (PCs) and Golgi cells. The homozygotic mutant mice conditionally ablated Auts2 from the cerebellum exhibited the reduced size in the cerebellum. In the Auts2 mutant cerebellum, the number of PCs was significantly decreased. Loss of Auts2 also resulted in the morphological abnormalities of PC dendrites as well as the defects of extensions and arborization. Immunohistochemical analysis revealed that the Auts2-deficient mice displayed the delay of the VGluT2-positive climbing fiber synapse formation at postnatal 2 weeks whereas the number of VGluT1-positive parallel fiber synapses was significantly increased. Moreover, Auts2 mutant mice exhibited the defects of several motor skills that included a mild locomotor ataxia, a hindlimb clasping upon tail suspension, and a decreased latency of fall off an elevated platform. These results suggest that AUTS2 plays a critical role for Purkinje cell development and maturation to acquire motor functions of the cerebellum.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-034 ショウジョウバエの視神経軸索の最終安定化層を決定する分子コード Satoko Hakeda-Suzuki（羽毛田‐鈴木聡子）¹，Olena Trush（Trush Olena）²，Makoto Sato（佐藤純）²，Takashi Suzuki（鈴木崇之）¹ ¹東京工業大学　生命理工学院 ²金沢大学大学院医学系研究科 Formation of the functional neuronal network requires not only precise target recognition but also stabilization of the axonal contacts to the specific synaptic layer during development. However, little is known about the molecular mechanisms underlying the stabilization of the axonal connection after axons have reached their specific target layer. In the Drosophila visual system development, photoreceptors send their axons to distinct place in the optic lobe. Among eight photoreceptors, R7 and R8 axons extend through the first optic ganglion lamina, and terminate in the M6 and M3 layer respectively, in the second optic ganglion, medulla. Two protein tyrosine phosphatases Lar and Ptp69D are known to be required for R7 axons to target correct medulla layer. The simultaneous knock-down of the two RPTPs suggest that the depth of the final stabilizing layer relies primarily on the overall amount of the cytoplasmic activity. Further analysis revealed that among the neurons innervating the medulla, only neurons normally terminating between M3 and M6 were affected by the down-regulation of the two RPTPs. Although both RPTPs are widely expressed in optic lobes, not all of neurons were affected by down-regulation of two RPTPs. These results suggest that the existence of more sophisticated mechanisms which determine the final targeting layer of the axons. We assumed that cell adhesion molecules create molecular codes together with RPTPs which determine the final depth of axonal stabilizing layer. In this study, we down-regulate the molecules in combination with two cell adhesion molecules, N-cadherin and Capricious, and two RPTPs. All of these molecules are known to have function in the axon targeting of photoreceptor neurons. Our results will reveal the molecular mechanisms how widely expressed molecules determine the R7/R8 targeting layer in the Drosophila visual system.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-035 大脳皮質神経細胞の樹状突起形成におけるGαi1の役割 Nanako Hamada（浜田奈々子），Koh-ichi Nagata（永田浩一）愛知県心身障害者コロニー発達障害研究所 Heterotrimeric GTP-binding proteins (G proteins) function as molecular switches inside cells, and regulate transmitting signals from a variety of stimuli outside a cell to its interior. Gαi subfamily, composed of Gαi1, Gαi2, and Gαi3, are known to inhibit production of cAMP from ATP. Recently, GNAI1 encoding Gαi was identified as a responsible gene for developmental disorders from whole exome analysis with undiagnosed developmental disorder patients. While intensive biochemical analyses have been performed for the role of Gαi1 in the signal transduction, little is known about its physiological role during corticogenesis. We thus analyzed physiological significance of Gαi1 during mouse cortical development. We produced 3 RNAi vectors which efficiently knocked down Gαi1 overexpressed in COS7 cells. We also constructed expression vectors for disease-causative mutants, Gαi1-Q204R and Gαi1-K270R. Knockdown of Gαi1 or overexpression of the mutants using in utero electroporation did not affect neocortical neuronal migration and axon extension in the corpus callosum. On the other hand, dendrite development of cortical pyramidal neurons was impaired with Gαi1-knockdown as well as overexpression of the mutants in vivo; Gαi1-deficient neurons showed shorter and more elaborate dendrites compared with control ones. Taken together, disruption of Gαi1 function may result in the abnormal corticogenesis, leading to neurodevelopmental disorders with GNAI1 gene abnormalities.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-036 マウス海馬のパルブアルブミン陽性GABAニューロンに由来する有髄線維のトポグラフィー Jun Yamada（山田純），Shozo Jinno（神野尚三）九州大院医神経解剖 Axonal myelination in the brain plays an essential role for the fast and long-range propagation of action potentials. Although the majority of myelinated axons are derived from excitatory pyramidal neurons projecting to remote areas, recent studies have shown that some of them originate from GABAergic inhibitory neurons. To address this issue, here we aimed to elucidate the topography of myelinated axons derived from parvalbumin-positive (PV+) GABAergic inhibitory neurons in the mouse hippocampus. In developing mice (until postnatal day 28), the distributions of myelinated axons derived from PV+ neurons were less evident. In young adult mice (8-12 weeks old), the densities of myelinated axons originated from PV+ neurons were higher in the dorsal hippocampus than in the ventral hippocampus. A substantial population of axons originated from PV+ neurons were running along the longitudinal axis of the hippocampus. The immunofluorescent quadruple labeling for PV, myelin basic protein (a marker for axon), contactin-associated protein 1 (a marker for node of Ranvier), and SMI 31 (a marker for pan-axonal neurofilament) showed the potential differences in the topography of myelinated axons derived from PV+ neurons and pyramidal neurons. Our findings provide a key to understanding the role of PV+ GABAergic neurons that may innervate remote areas.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-037 LAR-RPTPs dimer formation: a potential key-step in the receptors activation control Mai Q Nguyen（Nguyen Mai Q）^1,2，Tokuichi Iguchi（Iguchi Tokuichi）²，Misato Yasumura（Yasumura Misato）²，Makoto Sato（Sato Makoto）^1,2,3 ¹Grad Sch of Front Bio, Osaka Univ, Osaka, Japan ²Dept Anat & Neurosci, Grad Sch Med, Osaka Univ, Osaka, Japan ³Div of Dev Neurosci, United Grad Sch of Child Dev, Osaka Univ, Osaka, Japan The leukocyte common antigen–related subfamily of receptor protein tyrosine phosphatases (LAR–RPTPs) plays essential roles in several key aspects of neural circuits establishment. It is reported to be involved in synapse formation, neurite growth, and nerve regeneration. Based on the general molecular structure of LAR–RPTPs, attentions have been paid to these receptors phosphatase activity as well as their ability to interact with membrane-associated proteins acting in cell–cell and cell–extracellular matrix interactions. The mechanism, by which these LAR–RPTPs interact with their ligands to activate downstream signals or by which these receptors phosphatase activities are modulated, however, remains unclear. Here we focus on the ability of LAR–RPTPs to form dimers suspecting it to be the intermediate step transducing ligand binding to switching phosphatase activity on or off. To address the role of LAR–RPTPs dimer formation, we established a system for the receptors dimerization detection in live cells. Were able to confirm LAR–RPTP dimerization, we interestingly found that its splicing variants possess different dimer formation potential. Further, to manipulate LAR–RPTP dimerization at will, the system was modified to allow dimerization induction and we successfully induced LAR–RPTP dimerization in live cells with drug application. Together with the fact that LAR–RPTP splicing variants showed distinct ligand selectivity, the observed difference in dimer formation potential of these variants is considered to have an essential role in regulating the receptor activity. The systems that we have established, thus, serve as a useful tool to elucidate the importance of receptor-dimerization in LAR–RPTPs function in neural development further.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-038 In vivo imaging of dendritic refinement in neonatal mouse barrel cortex with high time resolution Luwei Wang（Wang Luwei）^1,2，Shingo Nakazawa（Nakazawa Shingo）¹，Hidenobu Mizuno（Mizuno Hidenobu）³，Takuji Iwasato（Iwasato Takuji）^1,2 ¹Laboratory of Mammalian Neural Circuits, National Institute of Genetics, Mishima, Japan ²Department of Genetics, The Graduate University for Advanced Studies (SOKENDAI), Mishima, Japan ³International Research Center for Medical Sciences, Kumamoto University, Kumamoto, Japan Precise neural circuit formation is critical for the proper functioning of the mammalian neocortex. The development of dendritic projection, which is an important process for neural circuit formation, arises during postnatal stages in an activity-dependent manner. However, the mechanisms of how cortical neuron dendrites are refined during early postnatal development are still largely unexplored. The two-photon microscope in vivo time-lapse imaging could be a powerful strategy to investigate the dynamic nature of dendritic refinement. To study the dynamics of dendritic refinement in vivo, we used the mouse barrel cortex as a model. In layer 4 (L4) of the mouse barrel cortex, thalamocortical axons (TCAs) transmitting information corresponding to a single whisker forms a cluster, called "barrel", and L4 spiny stellate neurons (barrel cells) extend their dendrites asymmetrically toward the single barrel to make synapses with TCAs. The dendritic orientation of barrel cells is formed essentially during the first postnatal week. Previously, we developed the "Supernova" system, which enables sparse and bright labeling of cortical neurons, and the TCA-GFP transgenic mouse, which allows in vivo labeling of TCA termini. Then by using these systems, we performed 18 hour-long imaging of barrel cells at postnatal day 5 (P5) and provided the first in vivo observation of dendritic refinement process in the mammalian brain (Mizuno et al., Neuron 2014). Subsequently, we achieved long-term imaging from P3 to P6 (Nakazawa et al., Nature Commun. 2018) and revealed aspects of dendritic refinement dynamics of barrel cells, which could be regulated by the unbalanced inputs from TCAs at inside and outside of the barrel. In these experiments, imaging intervals were 4.5 to 9 hours, during which barrel cells changed their dendritic morphologies drastically. To understand the detailed dynamics and capture the possible transient development events of dendritic refinement more precisely, here we are performing higher time-resolution in vivo imaging. We observed the same neurons and their dendritic morphologies every 1 hour for 8 hours in the barrel cortex L4 of neonatal mouse. We found that the individual dendrites could change their morphologies even in 1 hour. In the meeting, we will show and discuss how individual dendrites refined their morphologies during neonatal stages.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-039 細胞膜剥離法によるラット脊髄後根神経節細胞の成長円錐の形態解析 Osamu Hoshi（星治）¹，Nobuyuki Takei（武井延之）² ¹東京医科歯科大学 ²新潟大学脳研究所 Growth cones play an important role in neuronal networking during nervous system development. Morphological changes associated with growth cone movement involve the cytoskeleton, but it is unclear how guidance of growth cone movement and local protein synthesis relate to the cytoskeleton. In this study, we observed growth cones using the unroofing technique involving ultra-sonication with atomic force microscopy (AFM). Rat dorsal root ganglia (DRG) were cultured in DMEM containing brain-derived neurotrophic factor. We then employed the unroofing method with ultra-sonication. After fixation, cells were labeled with an anti-α-tubulin antibody and Alexa Fluor 586 phalloidin to label actin filaments. Specimens were dried to a critical point in liquid CO2 after labeing and then the cytoplasmic surface and intracellular structures were observed by AFM. Immunofluorescence microscopic images revealed that actin filaments were distributed in the peripheral region of the growth cone and filopodia, and the α-tubulin-positive region was mainly the central domain of the growth cone. AFM images showed that high regions of DRG tended to be rich in actin filaments and tubulin compared with low regions of DRG. If ribosomal proteins or proteins involved in translation are also labeled, the relationship between local protein synthesis and the cytoskeleton can be clarified. A method to analyze the relationship between local protein synthesis and the cytoskeleton in growth cones is discussed.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-040 恐怖記憶の発達におけるOtx2発現ドーパミン作動性細胞の作用 Chika koyama（小山千佳），Tomoya Iigima（飯島友也），Xubin Hou（侯旭濱），Sayaka Sugiyama（杉山清佳）新潟大院医歯神経発達 Homeoprotein Otx2 is required for differentiation of dopaminergic cells during embryonic development. Otx2 is persistently expressed in dopaminergic cells in postnatal brain, however little is known how Otx2 affects dopaminergic function in emotional behavior. Ventral tegmental area containing Otx2-possitive dopaminergic cells responses to fear and reward stimulation and extends dopamine fibers to amygdala. It is widely accepted that amygdala is involved in fear memory and behavior. It prompted us to examine whether Otx2-possitive dopaminergic cells function in fear processing. In this study, we used knockout mice lacking Otx2 gene in dopaminergic cells by crossing Otx2-flox mice with DAT-Cre mice in which Cre recombinase is expressed under the control of the promoter of dopamine transporter. Previous study has indicated that Otx2 protein is localized in basolateral amygdala by delivering from outside (eg. CSF produced by choroid-plexus). Immunohistological analysis revealed that Otx2 immunoreactivity was decreased in basolateral amygdala by Otx2 deletion in dopaminergic cells. In addition, Parvalbumin(PV) expression, one of markers of inhibitory cells, was also decreased, suggesting that Otx2 might affect PV cell maturation. Then, in behavior analysis of tone-cued fear conditioning, we found that recall of fear memory were declined in Otx2 KO mice compared to control mice. Consequently, conditioned fear was erased by extinction learning, and 7 days later, spontaneous recovery of fear memory was weakened in KO mice. These results suggested that Otx2 expression in dopaminergic cells plays an important role in promoting function of amygdala and retaining fear memory. Hence, it would be important to elucidate a direct role of Otx2 in the dopaminergic function.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-041 大脳皮質神経細胞移動におけるSADキナーゼの役割 Keiko Nakanishi（中西圭子）^1,2，Hiroyuki Niida（丹伊田浩行）^3,4，Hidenori Tabata（田畑秀典）⁵，Yoshikazu Johmura（城村由和）^3,6，Takashi Ueda（植田高史）⁷，Kenichiro Yamada（山田憲一郎）⁸，Koh-ichi Nagata（永田浩一）⁵，Nobuaki Wakamatsu（若松延昭）⁸，Masashi Kishi（岸将史）⁹，Shinya Ugawa（鵜川眞也）⁷，Shoichi Shimada（島田昌一）^7,10，Yujiro Higashi（東雄二郎）¹，Makoto Nakanishi（中西真）^3,6 ¹愛知県心身障害者コロニー発達障害研周生期 ²愛知県心身障害者コロニー中央病院小児科 ³名市大医細胞生化学 ⁴浜松医大医分子生物 ⁵愛知県心身障害者コロニー発達障害研神経制御 ⁶東大医科研細胞生物学 ⁷名市大医機能組織 ⁸愛知県心身障害者コロニー発達障害研遺伝 ⁹野崎徳洲会病院神経科学研 ¹⁰大阪大医神経細胞生物学 SAD kinase, a serine/threonine kinase, is involved in presynaptic vesicle clustering and neuronal polarization. Mouse homologues SAD-A and SAD-B have been considered to be functionally redundant, because previous study reported Sada-/- and Sadb-/- double mutant mice showed perinatal lethality due to polarity defects while their single mutants did not show any apparent phenotype. To elucidate the non-redundant role, we generated Sada-/- mice and backcrossed them to a C57BL/6N background. The Sada-/- mice in C57BL/6N background died within a few days after birth. Their cortical lamination showed disorganized pattern. In utero electropolation technique using pCAG-EGFP vector revealed the aberrant radial migration in the mutant brains. Time-lapse imaging showed that migration velocity of Sada-/- bipolar neurons in the CP was slower than that of wild type. Double staining with Cux1(layer II-IV marker) plus EdU labeling confirmed that a considerable number of EdU-positive cells labeled at E14.5 (the time layer II-IV neurons were born) were Cux1-positive and located in the lower CP of Sada-/- mice at P0. In contrast, Sadb-/- mice did not show neonatal lethality and their radial migration appears to be normal. These results suggest that SAD-A and SAD-B have distinct roles in radial neuronal migration in the developing brain.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-042 EGFによる神経の軸索伸長とEphシグナルへの干渉 Yutaro Kobayashi（小林雄太朗），Daisaku Yokomaku（横幕大作），Yuriko Iwakura（岩倉百合子），Eiko Kitayama（北山栄子），Hiroyuki Nawa（那波宏之），Nobuyuki Takei（武井延之）新潟大学脳研究所分子神経生物学分野 Epidermal growth factor (EGF) and its derivative neuregulins are associated with schizophrenia neuropathology and/or genetics. We subcutaneously administered EGF to neonatal animals and employed those as a model for schizophrenia which shows impairments in several cognitive behaviors after puberty. However, the mechanism underlying the cognitive deficits has not been elucidated. In this experiment, we analyzed the effects of EGF on the axonal elongation of cerebral cortical neurons and subsequent influences on their synapse formation at the molecular and cellular levels. EGF enhanced the motility of growth cones and attenuated the synapse formation in culture. We found that the signal cross-talk between EGF receptor (ErbB1) and Ephrin-Eph appeared to be involved in this phenomenon. EGF induced EphA2 phosphorylation in cultured neurons as well as in the neonatal cerebral cortex. The chronic administration of EGF increased the expression of Eph molecules including EphA2 in cultured neurons. These facts raise the possibility that ErbB ligands including EGF disturb Ephrin-Eph signaling and perturbs inter-cortical synapse formation. All procedures adopted in this study were approved by and conducted under the control of the Niigata University Animal Care and Use Committee and the Ethics Committees of the authors' institutes. This study was supported by Grant-in-Aid for Scientific Research (C) (16K07053), Grant-in-Aid for Scientific Research on Innovative Areas (18H05429, Constructive Understanding of Multi-scale Dynamism of Neuropsychiatric Disorders), and a collaborative research grant for the Niigata University Brain Research Institute.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-043 ビトロネクチンによるPI3キナーゼ/GSK3β経路を介した小脳顆粒神経細胞の軸索決定 Yuko Oishi（大石佑子），Kei Hashimoto（橋本恵），Ayaka Abe（阿部彩香），Maho Kuroda（黒田真帆），Yasunori Miyamoto（宮本泰則）お茶の水女子大学院人間文化創成科学研究科ライフサイエンス専攻 Mouse cerebellar granule cells (CGCs) are the most numerous cells in cerebellum. During cerebellar development, the proliferation, differentiation, and axon elongation of CGCs is regulated by various factors such as sonic hedgehog and laminin, one of extracellular matrix proteins tempo-spatially. We previously revealed that vitronectin (VN), one of the extracellular matrix proteins, controls the maturation of CGCs through the promotion of the initial differentiation stage progress via αvβ5 integrin. During the progress of the initial differential stage, axon specification has been known to happen in CGCs. This fact motivated us to focus on the axon specification in CGCs. In this study, we examined the role of VN in the axon specification in CGCs. First, we analyzed the effects of VN on the number of axons during the axon specification of CGCs using VN knockout mice-derived CGCs. To identify the axons in CGCs, an axon marker, Tau1 was immune-stained with the antibody for Tau1. The immunostaining showed that the loss of VN increased the ratio of CGCs with no axon, but VN addition increased the ratio of CGCs with multiple-axon, indicating that VN controls the axon specification in CGCs. Next, to examine whether αvβ5 integrin is served as a receptor for VN and regulates the axon specification in CGCs, we analyzed the effect of β5 integrin knockdown on the axon specification in CGCs. Similar to VN knockout, the knockdown of β5 integrin using shRNA increased the ratio of CGCs with no axon. In addition, β5 integrin knockdown abolished the increase of the ratio of CGCs with multiple axon by VN, indicating that αvβ5 integrin is served as a receptor for VN and regulates the axon specification. Furthermore, we investigated the signal pathway of axon specification by VN. Previous study showed that the PI-3 kinase (PI3K)-GSK3β pathway regulates the axon specification in hippocampal neuron. So we checked the role of PI3K and GSK3β in CGCs using Wortmannin, a PI3K inhibitor and LiCl, a GSK3β inhibitor. Wortmannin suppressed the promotion of multiple-axon formation by VN, and LiCl, a GSK3β inhibitor, abolished the increase of the ratio of CGCs with no axon by VN knockout and β5 integrin knockdown. Taken together, our results indicate that VN promotes the axon specification of CGCs via the αvβ5 integrin and PI3K/GSK3β pathway.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-044 発生学的な筋分化コンパートメントに基づく脊髄神経分岐パターンの新しいモデル Shunsaku Homma（本間俊作），Shunsaku Homma（本間俊作），Takako Shimada（島田孝子），Hiroyuki Yaginuma（八木沼洋行）福島県立医科大学神経解剖・発生学講座 The branching pattern of the spinal nerves is essential information underlying the basic and clinical medicine. The spinal nerves, once exiting the vertebrae, bifurcate into the dorsal and ventral rami, which innervate the back muscles in the epaxial compartment and body wall and appendicular muscles in the hypaxial compartment, respectively. However, the epaxial-hypaxial distinction doses not reflect the embryonic somitic lineages. Intrinsic back muscles and proximal body wall muscles are derived from the medial portion of the somite, whereas the distal body wall muscles and appendicular muscles are derived from the lateral portion of the somite. Muscles from the two distinctive lineages differentiate exclusively either in screlotome-derived or the lateral plate-derived connective tissue environments and belong to respective embryonic compartments, namely the primaxial and abaxial compartments. Despite this fact, it has never been asked how the adult branching pattern corresponds to the embryonic muscle lineages. In this study, we first constructed an alternative model of the spinal nerve branching pattern that was based on the primaxial-abaxial distinction and yet was fully compatible to our current knowledge on the spinal nerve. In our model, a seemingly single cord of the ventral rami is composite: in embryonic periods, one primaxial-specific and two abaxial-specific branches innervate muscles in respective compartments, but later these distinctive branches are fasciculated into a single nerve like the adult form. To validate our model, we then analyzed the branching pattern of the thoracic spinal nerve in detail, using whole-mount transparent specimens of the body wall of mouse embryos. Contrary to a general perception, that is, the single thoracic spinal nerve runs in the intercostal space with sequentially sending out twigs to nearby muscles, costal nerves were divided into superficial and deep branches at the proximal segment close to the vertebra, traveling in the intercostal space until the anterior midline; but in later developmental stages, the two branches appeared to form a single nerve cord. This result supports our model and suggests that the adult branching pattern is modification of the embryonic form.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-045 小脳交連ニューロン回路の多様性形成におけるUNC5Cの役割 Naohiro Tsuji（辻尚宏），Yosuke Matsumoto（松本耀介），Ryuichi Shirasaki（白崎竜一）大阪大院生命機能細胞分子神経生物学 One of the fundamental issues in neural development is to understand the mechanisms that underlie the generation of neural circuit diversity within the nervous system. In the developing cerebellum, two distinct subtypes of commissural neurons, anatomically referred to as deep cerebellar nuclei (DCN) neurons, are generated from the common progenitor cells that selectively express bHLH transcription factor Atoh1: one is the interposed and lateral DCN (i/l-DCN) neurons, and the other is the medial DCN (m-DCN) neurons. Interestingly, axons derived from i/l-DCN extend ventrally, and cross the floor plate (FP) at the ventral midline. In contrast, axons from m-DCN cross the roof plate (RP) at the dorsal midline, instead of crossing the FP. Although many of the molecular programs downstream of the Atoh1 are shared among DCN subtypes, the critical determinant that generates the opposing identity of axonal navigation pattern remains unknown. To address this issue, here we focused on the role of the Netrin receptor UNC5C in mice, since it has previously been shown that the commissure formed by m-DCN axons is absent in Unc5c knockout mice (Kim & Ackerman, J. Neurosci. 2011). We first examined the expression pattern of UNC5C and another Netrin receptor DCC on DCN axons during the time when axons from m-DCN and i/l-DCN grow toward the RP and FP, respectively. Intriguingly, we found that, although i/l-DCN axons expressed DCC alone, m-DCN axons selectively expressed both UNC5C and DCC. Because UNC5C functions in long-range repulsive axon guidance through heterodimerization with DCC in response to Netrin, we next asked whether UNC5C defines the trajectory of m-DCN axons toward the RP in a dedicated manner. For this, we performed UNC5C loss- and gain-of-function experiments in vivo, using the Atoh1 enhancer-based conditional expression system that enables selective and sparse labeling of individual DCN axons. We found that the UNC5C loss-of-function caused m-DCN axons to behave like i/l-DCN axons (i.e., ventral growth toward the FP). Conversely, UNC5C gain-of-function in i/l-DCN axons elicited redirection of these axons toward the RP, resulting in an ectopic RP-crossing by i/l-DCN axons. Together, these results suggest that UNC5C has an essential function in assigning RP-crossing identity, thereby contributing crucially to the establishment of the neural circuit diversity of commissural neurons in the developing cerebellum.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-046 脊髄交連ニューロンの軸索ガイダンスにおける運動ニューロンの役割 Ryuichi Shirasaki（白崎竜一）¹，Takeshi Kaneyama（金山武司）¹，Katsuhiko Ono（小野勝彦）²，Hirohide Takebayashi（竹林浩秀）³ ¹大阪大院生命機能細胞分子神経生物学 ²京都府立医科大院神経発生生物学 ³新潟大院医歯学総合研 Axon guidance by commissural neurons in the developing spinal cord has been well documented, providing us with a valuable model for elucidating the logic and mechanisms of axon guidance. In the rodent spinal cord, axons of commissural neurons generated from the dorsal spinal cord initially extend ventrally toward the floor plate (FP) in response to FP-derived chemoattractants including Netrin-1 (Wu et al., Neuron 2019). After crossing the FP, these axons make a sharp turn to grow rostrally in close proximity to the FP without entering nearby motor columns. Notably, we have recently shown that the commissural neurons develop axonal projection to spinal motor neurons (MNs) via collateral branches arising later from the post-crossing segment of these axons. We have also demonstrated that these collateral branches further give rise to multiple secondary branches preferentially within the region where specific subtype of MNs (i.e., MMC subtype of MNs) resides (Kaneyama & Shirasaki, J. Comp. Neurol. 2018). These observations proposed a model in which spinal MNs play pivotal roles in several key aspects of commissural axon guidance in the developing spinal cord. For example, at the FP, commissural axons lose responsiveness to the attractants, and instead become responsive to the repellents expressed by MNs as well as by FP cells (Shirasaki et al., Science 1998; Zou et al., Cell 2000). This has long been considered to explain why post-crossing axons make a sharp turn to grow rostrally without invading nearby motor columns. However, it remains obscure to what extent MNs are actually involved in these processes of commissural axon guidance in vivo. In the current study, to address this issue, we analyzed in detail the behavior of commissural axons during the period from their ventral growth toward the FP until their generation of terminal branches in motor columns, using mutant mice that selectively lack MNs from the onset of their differentiation. Here, we define the essential contribution of spinal MNs to the guidance of commissural axons in the developing mouse spinal cord in vivo.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-047 A study of the function of DCLK1 short isoform CPG16 in the development of the murine cerebellum Emilia Bergoglio（Bergoglio Emilia）¹，Kazuya Togashi（Togashi Kazuya）¹，Hiroyuki Koizumi（Koizumi Hiroyuki）²，Kazuo Emoto（Emoto Kazuo）^1,3 ¹Graduate School of Science, the University of Tokyo ²School of Pharmaceutical Sciences, Ohu University ³International Research Center for Neurointelligence (WPI-IRCN) Doublecortin-like kinase 1 (DCLK1) is a neuronal serine-threonine protein kinase that is closely related to the Doublecortin protein family. DCLK1 is characterized by several splicing variants: the long, canonical variant with in which the kinase domain is preceded by a microtuble-binding one (DCLK1-L), a short variant which only possesses the kinase domain (CPG16), a Doublecortin-like isoform DCL, and others. Despite the fact that recent genome wide association and transcriptome studies (Le Hellard et al., 2009; Havik et al., 2012; Wu et al., 2012) have indicated that the expression levels of DCLK1-L and CPG16 are associated with psychiatric disorders such as attention-deficit hyperactive-disorder and schizophrenia, the precise differential roles of these isoforms are still poorly understood. In this study, we try to determine the localization and function of CPG16 in the murine brain development, with the aim of understanding the expression pattern and the roles exerted by this molecule during development and synaptogenesis. First, we examined the expression pattern of CPG16 by confronting wild-type and DCLK1-knockout brains. This mutant mouse is a particularly good model for this purpose, since it still retains CPG16, whereas DCLK1-L and DCL are knocked out. We observed different brain regions during different developmental time-points, namely the cortex, the hippocampus, and the cerebellum. Preliminary data suggests that the pattern of expression of CPG16 is remarkably different in cerebellar Purkinje cells, where only the main dendrite was stained by DCLK1 C-term antibody. Our next step is to look at the detailed expression of CPG16 in Purkinje cells using in-situ hybridization. In addition, we also aim to find out the precise cell localization of CPG16. Taken together, this data will give us a hint about CPG16 function in the developing brain and its potential link to disease.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-048 RhoA-GEFによる大脳皮質ニューロンの軸索分岐の制御 Kensuke Sasaki（佐々木健介）¹，Kei Arimoto（有本啓）¹，Kento Kankawa（寒川健斗）¹，Chikayo Terada（寺田睦世）¹，Tetsuo Yamamori（山森哲雄）²，Akiya Watakabe（渡我部昭哉）²，Nobuhiko Yamamoto（山本亘彦）¹ ¹大阪大院生命機能細胞分子神経生物学 ²理研CBS 高次脳機能分子解析 Axon branching is essential to form appropriate neuronal circuits. An indispensable process for branch formation is remodeling of cytoskeletons. We have previously shown that active RhoA promotes branch formation of horizontally elongating axons (horizontal axons), which originate from layer 2/3 neurons in the developing cortex. To further explore the upstream molecular pathway, we studied the role of Rho guanine nucleotide exchange factors that activate RhoA (RhoA-GEFs) in horizontal axon branching. In situ hybridization showed that about a half of 28 identified RhoA-GEFs was expressed in the developing rat cortex. These RhoA-GEFs were mostly expressed in the infant macaque cortex as well. The function in axon branching was studied in organotypic slice cultures by transfecting upper layer cells with the plasmids encoding enhanced green fluorescent protein and full-length RhoA-GEF genes, and horizontal axon morphology was analyzed at 14 days in vitro. An overexpression study demonstrated that horizontal axon branching was markedly promoted by overexpression of seven members among the RhoA-GEFs. Moreover, branching patterns were different between overexpressing RhoA-GEFs. In particular, ARHGEF18 increased the number of small branches in the terminal portion, while ABR made further smaller branches along the entire axonal shafts. The branch-regulating activity of the two RhoA-GEFs showed different sensitivity to the downstream inhibitor (Y-27632), suggesting the involvement of downstream RhoA signaling is different in the function of them. Finally, knockdown of either endogenous ARHGEF18 or ABR by introducing shRNA considerably suppressed axon branching. These results suggest that multiple RhoA-GEFs, which are expressed in the mammalian cortex, contribute to horizontal axon branching in different manners.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-049 Semaphorin3A-Fyn情報伝達におけるCRMP1リン酸化の樹状突起形態制御 Aoi Jitsuki-Takahashi（實木ー高橋葵）¹，Takeshi Kawashima（河嶌岳）²，Susumu Jitsuki（實木亨）³，Takuya Takahashi（高橋琢哉）³，Yoshio Goshima（五嶋良郎）²，Fumio Nakamura（中村史雄）¹ ¹東京女子医大院医生化 ²横浜市大医分子薬理神経生物 ³横浜市大医細胞生物・生理 Collapsin Response Mediator Protein 1 (CRMP1) is an intracellular phoshoprotein that mediates Semaphorin3A (Sema3A) intracellular signaling. Upon Sema3A stimulation, Fyn, a Src-type tyrosine kinase, phosphorylates and activates Cycline-dependent kinase 5 (Cdk5), which subsequently phosphorylates serine 522 of CRMP1. In addition, it has been shown that Fyn directly phosphorylates tyrosine 504 (Y504) of CRMP1 (Buel et al., 2010). Then, we investigated the functional role of this phosphorylation in Sema3A signaling. We found that Fyn phosphorylated Y504 but not other tyrosine residues of CRMP1 in HEK293T cells. Double immunostaining of phosphotyrosine and CRMP1 revealed that the co-localization signal was partially increased in the growth cones of primary cultured dorsal root ganglion (DRG) neurons after Sema3A stimulation. A dominant negative mutant of CRMP1 Y504F, substitution of tyrosine 504 to phenylalanine (F), suppressed Sema3A-induced growth cone collapse of chick E8 DRG neurons. We next tested the role of Fyn and CRMP1 in Sema3A-mediated dendritic growth in vivo. CRMP1-/- or Fyn -/- single-homozygous mice as well as Fyn+/-; Crmp1+/- double-heterozygous mice exhibited aberrant development of cortical layer V basal dendrites. Finally, we examined the dominant negative effect of CRMP1 Y504F on cortical dendritic morphogenesis. CRMP1 Y504F or CRMP1 WT with tdTomato was transfected in the mice cortical layer V neurons at E15 by in utero electroporation. CRMP1 Y504F-expressed layer V neurons showed poor development of basal dendrites compared with CRMP1 WT-expressed neurons at 5-weeks old mice. These results suggest that Fyn-induced phosphorylation of CRMP1 Y504 may participate in Sema3A-regulated axon pathfinding and cortical dendritic development.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-050 Coactosinによる皮質抑制性介在ニューロンの形態形成の決定 Xubin Hou（侯旭濱）¹，Mieko Morishima（森島美絵子）²，Naoki Matohara（眞戸原尚輝）¹，Junko Sugi（杉順子）¹，Kenji Sakimura（﨑村建司）³，Yasuo Kawaguchi（川口泰雄）²，Sayaka Sugiyama（杉山清佳）¹ ¹新潟大院・医歯学・神経発達 ²生理研大脳皮質機能大脳神経回路論 ³新潟大・脳研・細胞神経生物学 Cortical interneurons possess multiple properties on both morphological and physiological axes and inhibit distinct targets that vary depending on the cell-type. It remains unknown how cell-specific morphologies underlying multiple functional feature develop by sensory experience. Notably, there is scarce information about experience-dependent factors to promote cytoskeletal organization depending on cell-type of interneurons. Here, we addressed whether an actin-polymerizing factors were required for dendritic maturation of parvalbumin-positive cells (PV-cells) in the visual cortex. Among them, Coactosin was specifically localized in PV-cells, and it was dominantly detected at the dendritic base within PV-cells. The distribution attenuated in Otx2 depletion or dark raring, confirming that Coactosin served as a downstream effector of Otx2 which activates the critical period by strengthening PV-cell function. Then, we performed in vivo electrophysiological recording of visual-evoked potentials (VEPs) and revealed that the onset of critical period was impaired by conditional deletion of Coactosin in PV-cells. Although the visual acuity was not altered between groups, the stronger visual responses of Coactosin-deleted networks shifted toward the higher spatial frequency. Importantly, Coactosin deficiency in PV-cells caused a reduction in the number of primary dendrites of these cells. Remarkably, polar projection clustering analysis revealed that almost 62.5% of PV-cells was switched from multipolar shapes of their dendrites to bipolar-like morphology. In contrast, overexpression of Coactosin accelerated multipolar arborization of PV-cell dendrites and even converted bipolar dendrites of calretinin (CR)-expressing cells into multipolar morphology. These results indicated that Coactosin functions as a switch from bipolar shape to multipolar morphology in PV-cell maturation. As a second step, electrophysiological membrane properties of PV-cells were recorded to verify Coactosin requirement for physiological maturation. The characteristic properties such as low input resistance of PV-cells was changed in Coactosin deletion, suggesting a coupling between the morphometric and electrophysiological properties. Taken together, Otx2-inducible actin reorganization may operate the multipolar structure in maturation of PV circuits and provide insight into the mechanisms for the plasticity and processing of sensory information.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-051 Developmental roles of afadin in retinal laminar organization, synaptic choice, and circuit function Hannah Graham（Graham Hannah），Xin Duan（Duan Xin） University of California, San Francisco Various types of neurons are precisely wired up into circuits during development to support diverse physiological functions and neurological behaviors. During these processes neurons use cell-adhesion molecules (CAMs) to interact with the extracellular environment, prompting intracellular signaling cascades that guide neuronal polarity, neurite outgrowth, and synaptic partner recognition. Unique CAM expression patterns render neuronal subtypes with characteristic migration patterns, morphologies, and synaptic partner choices. Yet, how recognition at the cell-surface directs these cell behaviors is not well understood. We investigated the developmental role of afadin, a cytosolic adapter protein that links multiple CAM families to intracellular F-actin, during retinal circuit assembly and organization. Using Six3:Cre; Afadin F/F mice (cKORet), we genetically ablated afadin from the retina during development. Retinas from cKORet mutants showed significant aberrations in retinal layer organization. Significant fractions of all retinal neuron types and Muller glia ectopically localized to incorrect nuclear layers and exhibited aberrant morphologies. In contrast to radial disorganization, there were no changes in number and density of these neuron types, and levels of proliferation and apoptosis were unaffected. Afadin expression in retinal astrocytes is unaffected in cKORet mutants; however, we observed abnormal astrocyte patterning, suggesting a cell-non-autonomous effect of afadin loss. Next, we examined retinal output from retinal ganglion cells (RGCs) to two major retinorecipient areas, the dorsal lateral geniculate nucleus and superior colliculus. Although RGC axons of cKORet mutants generally targeted these areas correctly, the stereotyped patterning of axons within these structures was lost. Lastly, canonical synaptic pairings between subtypes of RGCs and amacrine cells persisted, despite both partners being mislocalized to the outer nuclear layer. These studies suggest that afadin plays a critical role during retinal development in terms of retinal lamination and nuclear organization, as well as RGC axon targeting. We reason that the primary defect concerns neuronal migration, but not proliferation or cell fate determination. Additionally, ectopic localization does not alter synaptic partner choice between amacrine cells and RGCs, suggesting that other molecular cues, independent of afadin, are required for synaptic specificity.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-052 Patch-RamDA-seq法によるクラスター型プロトカドヘリンのシングルセル発現様式と神経細胞間結合の関係 Fuko Masuda（増田風子）¹，Etsuko Tarusawa（足澤悦子）¹，Mana Umeda（梅田茉奈）²，Mika Yoshimura（芳村美佳）²，Tetsutaro Hayashi（林哲太郎）²，Takashi Kitsukawa（木津川尚史）¹，Makoto Sanbo（三宝誠）³，Masumi Hirabayashi（平林真澄）³，Itoshi Nikaido（二階堂愛）²，Takeshi Yagi（八木健）¹ ¹大阪大院生命機能 ²理化学研究所生命機能科学研究センターバイオインフォマティクス研究開発ユニット ³生理学研究所遺伝子改変動物作製室 Neural networks of the cerebral cortex are formed by specific intercellular connections. However, it is still unknown how the specificities of neural connections are determined. Previously, we revealed that neurons arising from a same single stem cell, called sister cells, form the synaptic connections more frequently than non-sister cells in the layer 4 of the mouse barrel cortex during developmental stage. Then the one-way synaptic connections are selectively removed so that the proportion of reciprocal connections are increased. Furthermore, we also showed that the cell-lineage dependent reciprocal connections are significantly decreased by deletion of clustered protocadherins (cPcdhs). cPcdhs are cell adhesion molecules which are encoded in their gene clusters. In mice, 58 Pcdh genes are organized into three gene clusters, Pcdh-a, Pcdh-b, and Pcdh-g, which have 14, 22, and 22 members, respectively. Pcdh-a and Pcdh-g additionally have constitutive expression isoforms, aC1, aC2, gC3, gC4, and gC5. Each neuron expresses a different combinational set of isoforms and determines the individuality of the neurons. It seems that cPcdhs form multimer on the cell surface and bind homophlically to the other cells. We assume that the neurons expressing the same kind of cPcdh isoforms can form a specific neural connection. Similarly to our previous study, chimeric mice were generated by transplanting the iPS cells which express GFP into blastocysts of wild-type mice, then we performed double whole-cell patch clamp recording from the visualized sister cells of the chimeric mice, and analyzed their connections. Then, we collected the cytoplasm from the recorded neuron and performed single-cell RNA sequence by RamDA-seq. RamDA-seq is a newly developed full-length total RNA-sequencing method for single cells. Compared with other methods, RamDA-seq shows high sensitivity to non-poly(A) RNAs and full-length coverage for extremely long transcripts by using random primed reverse transcription and a novel whole transcriptome amplification method, RT-RamDA. The relationship between the combinational expression of cPcdh isoforms and their neuronal connectivity was investigated. We found that sister cells tended to express the same type of variable cPcdh isoforms even without synaptic connections. On the other hand, cell lineage-dependent expression was not observed in constitutive cPcdh isoforms.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-053 グリピカンは非古典的BMP経路を介して経験依存的なシナプスと行動の可塑性を調節する Keisuke Kamimura（神村圭亮），Aiko Odajima（小田嶋愛子），Nobuaki Maeda（前田信明） (公財) 東京都医学総合研究所　脳発達・神経再生研究分野 Recent studies revealed that many synapses in the vertebrate brain are enwrapped with a proteoglycan-rich extracellular matrix (ECM) that plays important roles in synaptic plasticity. The synaptic boutons of Drosophila neuromuscular junctions (NMJs) are also surrounded by a rich ECM containing the heparan sulfate proteoglycans (HSPGs). These HSPGs play important roles in NMJ development by regulating Wnt/Wingless (Wg), BMP/glass bottom boat (Gbb), and the receptor protein tyrosine phosphatase, LAR. However, their roles in synaptic plasticity at the NMJ currently remain unclear. It has been shown that, under food deprivation conditions, larvae exhibit increases in locomotion speed and synaptic bouton numbers at NMJs. Octopamine, the invertebrate counterpart of noradrenaline, plays critical roles in this process; however, the underlying molecular mechanisms remain unclear. Here, we demonstrated that a glypican (Dally-like, Dlp) is required for this experience-induced behavioral and synaptic plasticity. Dlp is expressed by postsynaptic muscle cells, and suppresses the non-canonical BMP pathway, which is composed of the presynaptic BMP type II receptor Wishful thinking (Wit) and postsynaptic GluRIIA-containing ionotropic glutamate receptor. As a result, Dlp negatively regulates type I synaptic bouton formation, the postsynaptic expression of GluRIIA, and larval locomotion speed. Starvation-induced octopaminergic signaling decreases Dlp expression at NMJ, and, thus, its negative activities are repressed, leading to the stimulation of synapse formation and locomotion. The present results revealed that Dlp regulates starvation-induced neural plasticity as a downstream mediator of octopamine signaling, providing insights into proteoglycan-mediated behavioral and synaptic plasticity.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-054 神経細胞においてRNA修飾N⁶-メチルアデノシン (m⁶A) は樹状突起発達およびシナプス機能を制御する Momoe Sukegawa（助川桃枝）¹，Ikumi Oomoto（大本育美）^1,2，Rohini Roy（Roy Rohini）^1,2，Akane Hatsuda（初田茜）^1,2，Mineko Kengaku（見学美根子）^1,2，Dan Ohtan Wang（王丹）¹ ¹京都大学高等研究院物質－細胞統合システム拠点 ²京都大学生命科学研究科 Information processing and communication between the billions of neurons in our brain depend on elaborate structures of axons, dendrites, and synapses. In recent years, regulated local translation of specific messenger RNAs in these neuronal subcellular compartments has been shown to play an important role in triggering persistent local functional and structural changes, and thus remodeling of brain circuits at fine resolution; however, the regulatory mechanism is largely unclear. In this study, we tested the hypothesis that N⁶-methyladenosine (m⁶A) RNA modification is a regulatory component of local translation. m⁶A is the most prevalent internal modification of mammalian messenger RNAs, regulating multiple steps of RNA metabolism such as RNA splicing, stability, and translation. In mammalian central nervous system, m⁶A decorates one-third of the transcriptome, widely impacting brain functions such as neurogenesis, learning and memory, stress response and addiction. We hypothesized that localized mRNAs are subject to m⁶A modifications in neurons, allowing fine-tuning of activity-dependent local translation and plasticity. First, we used next-generation sequencing to identify 4,469 m⁶A sites in 2,921 RNA species localized to neuronal synapses. Gene ontology analysis predicted functional categories related to synapse function and neuropsychiatric disorders with m⁶A-mRNAs. We also found m⁶A binding proteins YTHDF1, YTHDF2, YTHDF3 (called m⁶A readers) in dendritic compartments both in vivo and in primary cultures. Knocking down either YTHDF1 or YTHDF3 in cultured hippocampal neurons led to shorter dendrites at an early developmental stage of neurons. In YTHDF1-KD neurons at later developmental stages, thin, filopodia-like immature spines were observed, reminiscent of spine abnormality in patients with mental retardation disorders. Reduced PSD-95 puncta, surface expression level of AMPA receptor subunit GluA1, and synaptic transmission indicate deficits in synaptogenesis of YTHDF1-KD neurons. Our results show a critical role of m⁶A-RNA signaling in regulating dendrite development and synaptogenesis, possibly in an activity-dependent manner.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-055 発達期小脳において、アルドラーゼC陽性プルキンエ細胞の登上線維シナプス刈り込みはアルドラーゼC陰性プルキンエ細胞よりも遅れ、ホスホリパーゼC β3を必要とする Takaki Watanabe（渡邉貴樹）^1,2，Yurie Rai（賴友梨恵）¹，Honoka Suzuki（鈴木穂香）¹，Tsubasa Akamatsu（赤松翼）¹，Naofumi Uesaka（上阪直史）^1,2，Masanobu Kano（狩野方伸）^1,2 ¹東京大院医神経生理 ²東京大学, 国際高等研究所, ニューロインテリジェンス国際研究機構 Neural circuits mature functionally during postnatal development by elimination of early-formed redundant synapses. In the cerebellum of neonatal mice, multiple climbing fibers (CFs) with similar input strengths form synapses on the soma of each Purkinje cell (PC). Then, single CF is strengthened in each PC from P3 to P7, and subsequently the other weaker CFs are eliminated from P8 to P17. Previous studies demonstrate that the metabotropic glutamate receptor subtype 1 (mGluR1) is indispensable for the elimination of weaker CFs from P12 to P17 (the late phase of CF elimination). Among downstream signaling molecules of mGluR1, phospholipase C β4 (PLCβ4) and β3 (PLCβ3) are expressed complementarily in PCs of aldolase C (Aldoc, also known as zebrin II)-negative and Aldoc-positive compartments, respectively. Our previous study demonstrates that PLCβ4 mediates the late phase of CF elimination in the rostral vermis that corresponds to Aldoc-negative regions. However, it remains unclear whether PLCβ3 mediates CF synapse elimination in the caudal vermis where PCs express Aldoc and PLCβ3, and whether there is any difference in developmental time course of CF synapse elimination between the Aldoc-positive and Aldoc-negative compartments. To address these questions, we used heterozygous Aldoc-tdTomato knock-in mice which enable us to identify PCs that express PLCβ3 and Aldoc in acute cerebellar slices by tdTomato fluorescence. We knocked down PLCβ3 in PCs by injecting lentivirus carrying miRNA against PLCβ3, and examined CF innervation patterns by recording CF-mediated excitatory synaptic currents in acute cerebellar slices. We found that significantly higher percentage of Aldoc-positive PCs remain multiply-innervated by CFs at P20 when compared to Aldoc-negative PCs. PCs with PLCβ3-knockdown remained multiply-innervated by two or more CFs after P12 in Aldoc-positive compartments. Thus, our results indicate that PLCβ3 is required for the late phase of CF synapse elimination in Aldoc-positive PCs, and that processes of CF synapse elimination are delayed in Aldoc-positive PCs when compared to Aldoc-negative PCs, resulting in higher degree of multiple CF innervation.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-056 発達期の大脳皮質におけるコヒーシンを介したクロマチン構造制御 Akiko Sakai（酒井晶子）¹，Ryuichiro Nakato（中戸隆一郎）²，Heiko Peters（Peters Heiko）³，Katsuhiko Shirahige（白髭克彦）²，Sayaka Sugiyama（杉山清佳）¹ ¹新潟大院医歯神経発達 ²東大定量研 ³Inst Genet Medicine, Newcastle Univ, Newcastle upon Tyne, UK Juvenile brain experiences a unique time window or critical period when neuronal circuits are remodeled by experience. Rewiring of circuits requires different classes of neurons develop and change their properties according to experience, and the underlying gene program is fundamental to proper brain function. Indeed, we have shown that experience-dependent transfer of Otx2 homeoprotein into Parvalbumin-positive interneuron (PV-cell) activates plasticity in mouse visual cortex, and that coordinated expression of Otx2 target genes promotes PV-cell maturation and maintains its function implicated in plasticity and neurodevelopmental disorders (Sugiyama et al. 2008, Sakai et al. 2017). As a transcription factor like Otx2 activates plasticity, it is implicated that the regulation occur through chromatin structure, but little is known about neuronal chromatin dynamics during the critical period. Here, we focus on cohesin, a protein complex regulating transcription via higher order structure of chromatin such as interaction between promoter and enhancer. Cohesin has been reported to be required for neuronal development and mutations in cohesin-related factors cause Cornelia de Lange syndrome which includes an autistic feature. We employed ChIP-seq (chromatin immunoprecipitation sequencing) to determine genome-wide binding sites of cohesin in juvenile mouse cortex. Comparison between P14 and P28 cortical cohesin binding sites revealed that categories of genes containing P28-enriched cohesin binding sites include synapse and ion transporter, consistent with developmental changes in neuronal functions. Cohesin binding sites in P28 cortex overlapped with nearly half of Otx2 binding sites, implying that Otx2 may affect chromatin structure through cohesin for juvenile-specific gene expression. On the other hand, genome-wide analysis of active enhancer-specific histone modification as well as cohesin binding sites in visual cortexes of normal- or dark-reared mice hardly revealed prominent sites of experience-dependent regulation. As one possibility is that cell-type specific difference is obscured by the large number of populations composing cortex, we are trying to establish a method to detect neuronal-type specific patterns of chromatin factors in postnatal cortex.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-057 Fluoro-Jade C 染色は神経前駆細胞の変性を検出できる Takuya Ikenari（池成拓哉）¹，Hirofumi Kurata（藏田洋文）^1,2,3，Tetsuji Mori（森徹自）¹ ¹鳥取大医生体制御学講座 ²鳥取大医脳神経小児科 ³国立病院機構熊本再春荘病院小児科 Neurons are vulnerable to many insults, including ischemia, traumatic injury, and excitotoxicity. Detecting neuronal death in tissue sections with precision is crucial in neuroscience research. Fluoro-Jade C (FJC) staining is one of the most widely used methods to detect degenerating neurons, regardless of cell death fashions, such as apoptosis and necrosis, even though the precise mechanism of FJC staining is remained to be clear. The biggest advantage of FJC staining is the specificity to the degenerating mature neurons. But, it is not still clear whether FJC staining can detect degenerating immature neurons and neuronal progenitors. First, we focused on subventricular zone (SVZ)-rostral migratory stream (RMS)- olfactory bulb (OB) system in the adult mouse brain. In this system, neural stem cell cells generate OB neurons via neuroblasts, and most neuroblasts and mature OB neurons die by apoptosis during differentiation. We detected small number of FJC positive (+) cells in the RMS (DCX+ neuroblasts) and OB (NeuN+ mature neurons), but almost no FJC+ cells in the SVZ. Then, we analyzed E14 embryonic brain in which massive apoptosis of neuronal progenitors was induced by administration of N-ethyl-N-nitrosourea (ENU). In the ENU treated embryonic brains, we detected FJC+/Sox2+ cells in the ventricular zone, namely proliferating radial glia, in addition to post-mitotic immature neurons. Furthermore, we analyzed E9-10 embryonic brain in which apoptosis of neuroepithelial cells by administration of toxic dose of ethanol. Next, we performed double staining with FJC and immunostaining with anti-activated Caspase-3 (Cas-3) antibody (early/mid phase apoptosis) or TUNEL staining (late phase apoptosis). In the apoptosis induced-embryonic brains, virtually all Cas-3+ cells and TUNEL+ cells were FJC+. On the other hand, in the normal adult RMS, there was considerable number of FJC negative cells among Cas-3+ cells. Present study indicates that FJC staining can detect degenerating neuronal lineage cells in wide range differentiation and apoptosis phases under the apoptosis induced experimental conditions. On the other hand, FJC staining can not detect all apoptotic neuronal cells under the normal condition. Our results could contribute to elucidate the mechanism of FJC staining.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-058 シトシンアラビノシドを暴露したマウス海馬神経細胞におけるヒストンH2AXリン酸化の新規メカニズム Saki Nakayama（中山紗希），Nobuyuki Fukushima（福嶋伸之）近畿大学総合理工学研究科 Cytosine arabinoside (Ara-C or cytarabine) is commonly used as an anticancer, genotoxic drug inhibiting DNA synthesis in mitotic cells. However, accumulating evidence has demonstrated that, in postmitotic neurons, Ara-C and other genotoxic drugs induce cell death by causing DNA damage or forcing cell cycle progression. We have also previously shown that Ara-C induces accumulation of phosphorylated histone H2AX (γH2AX) in nucleus as well as cell death in mouse hippocampal neurons. In general, γH2AX is generated by actions of phosphoinositol 3-kinase-related kinase (PIKK) family, which are activated by DNA double strand breaks resulted from errors in DNA replication or recombination. However, how Ara-C induces γH2AX formation in postmitotic neurons remains unclear. In this study, we first examine whether DNA polymerase is involved in Ara-C-induced cell death and γH2AX formation. RT-PCR analysis using single neurons showed that the expression of DNA polymerase α, δ, and ε was not observed in both control and Ara-C-treated neurons. Furthermore, Ara-C was not incorporated into DNA of neurons when neurons were cultured with [³H]-Ara-C. Thus, Ara-C was unlikely to target DNA polymerases to be incorporated into DNA in postmitotic neurons. Next we pharmacologically examine the mechanisms of Ara-C-induced γH2AX formation. When hippocampal neurons were co-treated with Ara-C and various inhibitors for PIKK family, there was no obvious inhibition in Ara-C-induced γH2AX formation, suggesting the involvement of non-canonical kinases in the Ara-C's actions. We then tested other kinase inhibitors and found that Ara-C-induced γH2AX formation was blocked by THZ-1, an inhibitor for cdk7 that is a critical kinase regulating not only cell cycle but also RNA transcription. Further study revealed that a transcription inhibitor actinomycin D (ActD) completely blocked Ara-C-induced γH2AX formation, whereas a direct RNA polymerase inhibitor α-amanitin failed to inhibit it. Interestingly, like Ara-C, a topoisomerase II inhibitor etoposide also generated γH2AX, which was blocked by THZ-1 and ActD. These results might imply a novel mechanism that Ara-C affects topoisomerase-related DNA reconfiguration and transcription, resulting in γH2AX formation in postmitotic neurons.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-059 X- irradiation of immature developing neurons causes delayed synaptopathy and decreases the number of bystander neurons Anggraeini Puspitasari（Puspitasari Anggraeini）¹，Hiroyuki Yamazaki（Yamazaki Hiroyuki）²，Kenji Hanamura（Hanamura Kenji）²，Kathryn D Held（Held Kathryn D）^1,3，Tomoaki Shirao（Shirao Tomoaki）² ¹Gunma University Initiative for Advanced Research ²Department of Neurobiology and Behavior, Gunma University Graduate School of Medicine ³Massachusetts General Hospital/ Harvard Medical School, Boston, MA USA Neurogenesis in adults is necessary to correct damage to the brain. However, the damaging effects of radiation on the synaptic function of immature neurons and surrounding, or bystander, neurons when the neurons reach maturity remain unclear. In this study, primary hippocampal neurons from embryonic day 16 C57BL/6 mice were seeded on poly-L-lysine-coated coverslips inside 12 wells plates. We have confirmed that more than 90 per cent of the cells at one day in vitro (DIV) were neurons using MAP2. The 1 DIV neurons were irradiated with 0.5 Gy and 1 Gy of X-rays, and for bystander study, the non-irradiated neurons were immediately suspended upon irradiated neurons then the cultures were maintained for three weeks. We analysed the apoptosis one day, one week, and two weeks after irradiation by examining the percentage of Caspase-3 positive cells, and the number of remaining neurons by analysing MAP2 /DAPI positive cells per region of interest. To elucidate the effects of irradiation on the synaptic function of the remaining neurons, at 14 DIV and 21 DIV the number of drebrin clusters along the dendrites were analysed by immunocytochemistry. Other synaptic proteins such as synapsin I and PSD95 were also determined using western blot analysis. We found that the percentage of activated Caspase-3 cells was increased a week after 1 Gy and two weeks for 0.5 Gy in parallel with a decrease in the number of MAP2/DAPI positive cells. The number of drebrin clusters along the dendrite was found to be significantly reduced at three weeks after irradiation but not two weeks after radiation. The western blotting analysis showed that there was no change of synapsin I expression two or three weeks after irradiation; however, decreased drebrin and PSD95 were found three weeks after radiation but not two weeks after radiation. Surprisingly, when we analysed the number of bystander neurons, the number of MAP2/DAPI positive cells in non-irradiated cells was decreased at three weeks after being co-cultured with irradiated neurons. These results suggest that the toxic effect of radiation on immature neurons does not occur immediately. The radiation-induced apoptosis occurs within one week while the synaptopathy of remaining neurons occurs three weeks after irradiation. Additionally, the irradiated neurons may cause cell death of bystander neurons.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-060 マウス海馬由来HT22細胞におけるブラジル産グリーンプロポリスの神経細胞保護作用 Madoka Takashima（高島麻都花）^1,2，Kenji Ichihara（市原賢二）¹，Yoko Hirata（平田洋子）^2,3 ¹アピ株式会社 ²岐阜大院連合創薬創薬科学 ³岐阜大工化学生命 Propolis is a resinous substance that honey bees produce by chewing buts and resins of various plants, and the components contained are characterized by botanical origins collected by honey bees. The botanical origin of Brazilian green propolis is Baccharis dracunclifolia, which contains various ingredients such as cinnamic acid derivatives, flavonoids and caffeoylquinic acids. It is also known that these chemical components and their amounts contained in the extracts of propolis differ depending on the extracting solvent. The functionality of the propolis extract has been widely used as a health food material, including antibacterial action and antiinflammatory action. Previously, a water extract of Brazilian green propolis has been reported to have protective effects against oxidative stress-induced cell damage in the rat retinal ganglion cell lines RGC-5. In this study, to further investigate the neuroprotective effects of Brazilian green propolis, we evaluated its extracts and their major components in in vitro tests using the mouse hippocampal cell lines HT22. We found that the water extract (WEP) and the ethanol extract (EEP) have protective effects on glutamate-induced oxidative stress in HT22 cells, and the effect of EEP was stronger than that of WEP. Among the main components of EEP, artepillin C, baccharin, drupanin and p-coumaric acid, only artepillin C exhibited neuroprotective effects. EEP and artepillin C also inhibited erastin-induced ferroptosis. On the other hand, EEP and these main components did not show protective effects on anticancer drug induced-apoptosis. EEP and artepillin C suppressed production of reactive oxidative species (ROS) and inhibited Ca²⁺ influx induced by glutamate. EEP activated the Nrf2-Keap1-ARE pathway, but artepillin C did not. In addition, EEP increased expression of heme oxygenase-1, as assessed by western blotting. From these results, we suggest that EEP and artepillin C have neuroprotective effects against oxidative stress-induced cell death and that the protection by EEP is due to the up-regulation of ROS scavenging enzymes. Interestingly, the mechanism of the neuroprotective effect of artepillin C is suggested to be different from that of EEP.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-061 成体および幼弱マウスで損傷した脊髄の1細胞解析を目的とした核の単離 Iyo Yorifuji（依藤依代）¹，Hiroshi Tsujioka（辻岡洋）¹，Yasushi Sakata（坂田泰史）²，Toshihide Yamashita（山下俊英）^1,3,4 ¹大阪大院医分子神経科学 ²大阪大院医循環器内科学 ³大阪大院生命機能 ⁴免疫学フロンティア研究センター In adult mammalian central nervous system, axon regeneration is blocked by glial scar formed at the lesion site. Moderate or complete depletion of pericyte, which is a component of glial scar, results in promotion or inhibition of axon regeneration after spinal cord injury (SCI) in mice, suggesting the heterogeneity of pericyte. In contrast, in neonatal mice, glial scar formation is almost absent, and axon regeneration is prominent after SCI, suggesting that regeneration-promoting subtype of pericyte might be enriched in neonate compared to adult. Here, to reveal the difference of subtype of pericyte between neonate and adult, and to identify the regeneration-promoting subtype of pericyte, we isolated single nucleus from injured spinal cord of adult and neonatal mice for single nucleus-RNA sequencing (snRNA-Seq). Dorsal half of the spinal cord was injured at Th10 level in adult (7 weeks old) female and neonatal (p1) mice. Comparison of lesion site at 1, 7 and 14 days post injury (dpi) by immunohistochemistry revealed that glial scar was formed at 7dpi or later in adult mice, whereas injury site was completely filled with cell population which did not express glial scar markers in neonatal mice. We compared the expression levels of 13 marker genes for major cell types in the spinal cord between the bulk tissue and isolated nuclei from injured spinal cord of adult and neonatal mice at 7dpi by quantitative RT-PCR, and found that the differences were less than 5 times in 85% (11) of them, suggesting that the isolated nuclei represent the cell population in the spinal cord. We performed a high-throughput nanogrid single nucleus isolation and sample preparation for snRNA-seq using ICELL8 system, and obtained 5448 single nuclei (1412, 1436, 1408 and 1192; nuclei from neonate sham, neonate SCI, adult sham and adult SCI, respectively). Average size of ~1000 bp DNA was observed after reverse transcription process, suggesting that high quality cDNA suitable for snRNA-seq was obtained from the isolated single nuclei. Sequencing of the cDNA libraries is now in progress.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-062 硫酸化糖鎖による神経軸索伸長阻害に対するラクトフェリン誘導体の軸索再生効果 Masao Nakamura（中村真男），Nao Tsutsumi（堤菜緒），Ami Iimori（飯森愛美），Atsushi Sato（佐藤淳）東京工科大院バイオ・情報メディア研究科 When central nervous tissues are damaged through spinal cord injury, reactive astrocytes accumulate in the vicinity of the injury and form glial scars. Injured nerve axons cannot regenerate across the glial scar. Chondroitin sulfate E (CS-E), a type of sulfated glycosaminoglycan, has recently attracted attention as an inhibitor of axon regeneration that is deposited in glial scars within the central nervous system. Receptor-type protein tyrosine phosphatase σ has been identified as a critical receptor for CS-E. Growth cones at the tip of damaged axons form inflated spherical structures called dystrophic endballs, and the mechanism preventing their forward movement is being elucidated. A recent report has shown that when CS-E present in the vicinity of the injury is degraded by the CS-degrading enzyme, neural regeneration is promoted, which in turn promotes recovery of motor function. Furthermore, In In vitro experiments using primary cultured neurons, addition of CS-E to the media inhibited the axon growth. When the added CS was enzymatically digested, the observed inhibition was suppressed. Based on these in vivo and in vitro studies, CS-E is expected to be a therapeutic target in the treatment of spinal cord injury. We aim to develop novel therapetic drugs that would facilitate patient recovery from spinal cord injury. We recently discovered that lactoferrin (LF) has high affinity for the axon growth inhibiting factor CS. A neural regeneration effect of LF has been previously reported. We expect it will also serve as a lead in the discovery and development of CS-targeting drugs promoting neural regeneration. In this study, we evaluated the neural regeneration effect of an LF derivative capable of binding CS. First, we developed an in vitro spinal cord injury model of CS-E using the rat-derived pheochromocytoma PC-12 cell line. We used NGF-induced protrusive elongation and sensitivity of growth cones of PC-12 cell to CS-E as indicators of efficacy. We used this system to examine the ability of LF to block CS-E-mediated inhibition of axon growth and growth cone collapse. We observed that CS-mediated inhibition of protrusive elongation in NGF-differentiated PC-12 cells was weaker in the presence of the LF derivative, which therefore promoted protrusive elongation. CS-induced growth cone collapse of PC-12 cell was also suppressed in the presence of LF. These findings suggest that the LF derivative can suppress inhibition of axon growth by CS.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-063 細胞低接着性コラーゲンを足場とした神経細胞の培養 Norihiko Nakano（中野法彦）¹，Kenji Kanekiyo（兼清健志）¹，Saori Kunii（國井沙織）²，Koichi Morimoto（森本康一）²，Kaoru Omae（尾前薫）³，Chizuka Ide（井出千束）¹ ¹藍野大中央研 ²近畿大生物理工 ³神戸医療産業都市推進機構 Extracellular matrix proteins such as type I collagen or gelatin are known as scaffolds for cells adhesion and proliferation, and are utilized in regenerative medicine and stem cell research. Recently, we succeeded in developing low adhesive scaffold collagen (LASCol) from porcine skin. Cells cultured on LASCol coated-dish collide each other one after another and get to form cell aggregates (spheroids) larger. However, the behavior of neuronal cells remains still unknown. In this study, we report that the morphology and behavior of nerve cells and astrocytes cultured on LASCol. We observed an influence of LASCol on neuronal cells in culture system. At first, neurons derived from newborn rat hippocampus were seeded on a culture dish coated with LASCol, atelocollagen or poly-L-lysine (PLL). As a result, it was revealed that neurons on LASCol have increased cell viability and remarkably elongated neurites as compared with cells on atelocollagen or PLL. On the other hand, astrocytes derived from rat brain rarely adhered on LASCol and the growth was suppressed compared with astrocytes on other coated dishes. Therefore, it was suggested that LASCol is a suitable scaffold that acts specifically on nerve cells. Acknowledgments; The content of this research is the result obtained by research support of the AMED Translational Research Network Program.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-064 終糸の中心管上衣細胞の特徴と脊髄損傷治療への可能性 Kenji Kanekiyo（兼清健志）¹，Norihiko Nakano（中野法彦）¹，Chihiro Tsukagoshi（塚越千尋）²，Chimi Miyamoto（宮本陳敏）²，Seiya Abe（安部征哉）²，Chizuka Ide（井出千束）¹ ¹藍野大・中央研 ²藍野大・医療保健・作業療法 We have reported that the cell transplantation, such as bone marrow stromal cells, bone marrow-derived mononuculear cells, and choroid plexus epitherial cells, was effective for spinal cord injury. Regarding the endogenous repair, ependymal cells in the central canal of the spinal cord are known to play a pivotal role in axon regeneration after spinal cord injury. The filum terminale (FT) is a thin strand continuous with the conus medullaris, a caudal end of the main spinal cord. The parenchymal tissue of spinal cord contains numerous glial cells other than the ependymal cells. On the other hands, the FT consists of the central canal associated with no, or only a negligible amount of spinal cord parenchymal tissue. The FT is an ependymal cell rich source therefore is a possible candidate for transplantation. In this study, we first clarified the characteristics of ependymal cells of the central canal of the FT in rats. Immunohistochemical analysis revealed that almost all ependymal cells were positive for Sox2, Sox9, FoxJ1, and CD133, markers for ependymal cells of the central canal in the spinal cord. Thus, the ependymal cells of the central canal of the FT have same characteristics as those of the main spinal cord. Nest, we examined the fate of FT ependymal cells after transplantation in the lesion of spinal cord. The FT from GFP-transgenic rat was minced and transplanted into the lesion caused by contusion. Engrafted FT cells were remined 1 week after lesion site in the spinal cord and tightly interacted with regenerating axons. This indicates that engrafted ependymal cells from FT promotes axon regeneration. Now we are further investigating the usefulness of the FT ependymal cells for spinal cord injury.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-065 若年性lncRNA (JALNC) による細胞若年性の維持 Masaki Mori（森雅樹）¹，Takao Morimune（森宗孝夫）^1,2，Faidruz Azura Jam（Jam Faidruz Azura）¹，Yosuke Kadota（門田陽介）¹，Ayami Tano（田埜郁実）¹，Yuya Tanaka（田中雄也）¹，Sayumi Akahane（赤羽紗由美）¹，Mayu Fukumura（福村真優）¹，Haruka Yukiue（雪上晴加）¹ ¹滋賀医科大学　神経難病研究センター ²滋賀医科大学　小児科学講座 Young individuals possess physiological advantages that adults do not possess. These features include the abilities to grow in size, maturate, heal faster, learn, and so on. It is not still understood how only the younger individuals have these characteristics, which are lost when the ones fully maturate or grow older. The molecular basis for the differences has not been well elucidated. Aiming at discovering the mechanisms behind these strong points of juvenile animals, we performed next-gen sequencing and identified young cell-specific genes in mice. We call these genes as "juvenility-associated genes (JAGs)". As a result of further biological and bioinformatics analysis of the JAGs, we found a non-coding element of JAGs had significant functional implications. Long non-coding RNA (lncRNA) consists an unexplored part of the transcriptome, though they have a high therapeutic potential via targeting with antisense strategies such as siRNA or antisense oligonucleotide. We identified the young cell-specific lncRNAs as "juvenility-associated long non-coding RNAs (JALNCs)", and found they possess essential functions in a cell. Among the JALNCs, a novel lncRNA Gm14230 is expressed in juvenile cells and is essential for cell growth. Gm14230 is conserved across species in terms of a nucleotide sequence and an RNA secondary structure. Loss of Gm14230 via RNA interference causes cell cycle exit and cellular senescence, indicating Gm14230 maintains the cellular juvenescence. Thus, Gm14230 has a potential as a therapeutic target for the neurological diseases, shedding light on the functional relevance for the noncoding element of the transcriptome.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-066 新生仔マーモセット脳における領域特異的な遺伝子発現の探索 Yoshiaki Kita（喜多善亮），Aya C Yoshida（吉田彩），Satomi S Kikuchi（菊池里美），Chihiro Yoshida（吉田千尋），Mami U（禹麻美），Karl J Windak（ウィンダックカール），Yan Wang（王燕），Hirozumi Nishibe（西部弘純），Tomomi Shimogori（下郡智美）理研CBS 脳発達分子メカニズム The common marmoset (Callithrix jacchus, a small New World monkey) as a non-human primate model has been attracting many neuroscientists because of its unique behavior which has a resemblance to human behavior. For better understanding of primate-specific circuit and its formation mechanism, it is crucial to visualize genes that are expressed in the marmoset brain with spatiotemporal resolution. The Marmoset Gene Atlas (MGA) (https://gene-atlas.brainminds.riken.jp/) provides a whole brain gene expression atlas obtained by in situ hybridization (ISH) of the marmoset brain. The MGA enables researchers to uncover connections between structure, circuit function, and underlying molecular mechanisms of circuit formation. Here we introduce the new function, "Structural search", which allows us to search for genes expressed in target areas systematically. First, development of the grid system that provides positional information on each ISH section enables us to convert ISH signal to gene expression data on the grid. Next, defining each grid for corresponding brain area provides spatial information of gene expression in different brain region. Lastly, registration of 1,342 genes to the grid system allows us to analyze gene expression systematically. Using this function, we can identify genes that are expressed in specific regions in cortex, thalamus, hypothalamus, midbrain and hindbrain. By comparing gene expression in marmoset and those in other species, we can reveal primate-specific gene expression patterns. We anticipate that this search function will accelerate the scientific understanding of primate-specific circuit and its formation mechanism.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-067 MIFによるグリオーマ幹細胞増殖制御機構の解明 Shigeki Ohta（大多茂樹）¹，Yukina Morimoto（森本佑紀奈）²，Ayako Tokumitsu（徳光綾子）³，Takafumi Sone（曽根岳史）⁴，Ayuna Sakamoto（坂本鮎菜）¹，Masahiro Toda（戸田正博）²，Yutaka Kawakami（河上裕）¹，Hideyuki Okano（岡野栄之）⁴ ¹慶應大医先端研 ²慶應大医脳神経外科 ³慶應義塾大学病院臨床研究推進センター ⁴慶應大医生理 In the previous study, we identified macrophage migration inhibitory factor (MIF) as a novel factor that can support the proliferation and/or survival of NSPCs (Neural stem cell/progenitor cells) based on functional cloning strategy in vitro (Ohta S et al., JCS, 2016). In the present study, we identify TPT1 (Tumor Protein, Translationally-Controlled 1) as a downstream target of MIF signaling in NSPCs. Lentivirus-mediated gene silencing of Mif decreased the gene expression of Tpt1 in mouse NSPCs in vitro. Overexpression of Tpt1 in mouse NSPCs increased cell proliferation, whereas gene silencing of Tpt1 decreased NSPC proliferation. We also show that Zfhx4 (zinc finger homeobox 4) gene expression is downregulated by silencing of the Mif or Tpt1 gene in mouse NSPCs. In human ES cell-derived NSPCs (hES-NSPCs), lentivirus-mediated gene silencing of MIF decreased the expression of Tpt1. Overexpression of TPT1 increased cell proliferation, while lentivirus-mediated gene silencing of Tpt1 decreased both cell proliferation and neurogenesis in hES-NSPCs. Importantly, the same decreases in cell proliferation and defect of neurogenesis were observed in human NSPCs generated from iPSCs harboring CRISPR/Cas9-mediated heterozygous disruption of the Tpt1 compared with wild-type cells. In a previous study, we reported the functions of MIF in regulating the proliferation of brain tumor-initiating cells (BTICs) from primary glioblastoma (GBM) tissues (Fukaya R et al., Cancer Res., 2016). We also identified the role of the MIF-CHD7 (Chromodomain-Helicase-DNA-binding protein 7) signaling axis in regulating BTIC proliferation (Ohta S et al., Molecular Brain, 2017). The Tpt1 gene is highly expressed in GBM and a correlation in gene expression was observed between MIF and TPT1 in a GBM cohort based on in silico data base analysis. As seen in mouse and hES-derived NSPCs, lentivirus-mediated gene silencing of Mif caused the downregulation of Chd7 andTpt1 gene expression in BTICs. Moreover, Tpt1 gene silencing showed the downregulation of Bmi1, Aspm and Mycn, and decreased the proliferation of BTICs. Finally, we examined the therapeutic effect of TPT1 in a nude mouse model. BTICs infected with either lenti-shTPT1 or lenti-shControl were implanted into the brains of nude mice. Mice implanted with lenti-shTPT1infected BTICs showed longer overall survival than the control group. These results open new avenues for the development of glioma therapies.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-068 マーモセット脳における発達〜老化の多角的MRI解析 Fumiko Seki（関布美子）¹，Keigo Hikishima（疋島啓吾）^1,2,4，Yuji Komaki（小牧裕司）^1,2，Marin Nishio（西尾まりん）^1,2,5，Junichi Hata（畑純一）^1,2,3，Akiko Uematsu（植松明子）^1,2,3，Norio Okahara（岡原則夫）¹，Erika Sasaki（佐々木えりか）^1,2，Hideyuki Okano（岡野栄之）^2,3 ¹実験動物中央研究所 ²慶應大医生理 ³理研CBS マーモセット神経構造 ⁴OIST ⁵首都大院人間健康放射線 INTRODUCTION This study investigated developmental patterns of white matter structures in common marmosets (marmosets), for the purpose of characterization the growth patterns of white matter tracts in marmoset brain using multiple parameters (MTR; magnetic transfer ratio, RD, radial diffusivity, FA, fractional anisotropy). They indirectly measure the maturation of tissue microstructure. METHODS Sequential MRI was performed to 33 marmosets at the age of 1 - 190 months, ranged from 1 to 8 times on 7.0 T Biospec 70/16 scanner. Tract-based ROIs were created for assessment of major fiber bundles. The population trajectories for respective regions were estimated using nonlinear mixed model, given the previous study found that the developmental pattern of global white matter development was rapid increase and stabilized thereafter. RESULTS Population growth trajectories of association fibers estimated using Gompertz function showed different developmental patterns. Inferior longitudinal fasciculus (ILF) showed earlier maturation with slow speed, whereas inferior fronto-occipital fasciculus (IFOF) showed slower maturation with fast speed observed in MTR, RD and FA. The fornix showed the earlier maturation among MTR, RD and FA. However, the growth speed was different between MTR and FA. DISCUSSION We could characterize the maturation/degeneration patterns of different white matter structures ranged from 1 month to 90 months, which covered the whole developmental process. The timing of maturation and accelerated speed were varied among regions. The maturation patterns of investigated regions were consistent with human studies previously reported. This study revealed that marmosets also showed normal growth patterns of major white matter structures were region-specific. By detailed comparison of growth trajectories across the species, it allows us to characterize what may be marmoset-specific features. Such evaluation is crucial to investigate whether brain disease models not only neurodevelopmental disorders but neurodegenerative diseases showed abnormal time course of brain development.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-069 Evidence of Chemically Modified Transcriptome at Neuronal Synapses Rohini Roy（Roy Rohini）^1,2，Yoshie Fujiwara（Fujiwara Yoshie）¹，Masatsugu Denawa（Denawa Masatsugu）³，Kei Iida（Iida Kei）³，Dan Ohtan Wang（Wang Dan Ohtan）¹ ¹INSTITUTE OF INTEGRATED CELL-MATERIAL SCIENCES ²Graduate School of Biostudies, Kyoto University, Kyoto, Japan ³Medical Research Support Centre, Graduate School of Medicine, Kyoto University, Kyoto, Japan Higher Order cognitive functions emerge through the development and maintenance of complex neuronal circuits. Synapses, being the functional sites of activity sensing and propagation of neurons, play a crucial role in connecting and fine-tuning the circuits; thus understanding the molecular mechanisms regulating synapse function is of central interest in neuroscience. One potent functional regulation pathway of the synapse is through proteostasis, a cellular pathway that involves dynamic biogenesis, modification, folding, trafficking, assembly, and degradation of proteins. Previous studies have shown that in addition to polyribosomes, a large number of RNA species are detected at synaptic compartments, serving as a versatile template pool for local translation. However, except for a handful of examples, the coding potential of the synaptic transcriptome and the spatiotemporal regulation mechanisms have not been fully addressed. Recently emerged RNA epigenetics such as N6-methyl-adenosine (m⁶A) modification in mRNAs, has been shown to be a new powerful regulatory layer of complex gene expression underlying brain functions such as cortical neurogenesis, rewarding behaviors, circadian clocks, stress response, and injury-related regeneration. Using next-generation sequencing techniques, we have characterized m⁶A modified transcripts in synaptic compartments. We found over one thousand differentially methylated RNA species preferentially localized to the synapse. We also demonstrated an indispensable role of m⁶A signal for synapse formation, maintenance, and transmission. On the verge of characterizing the full complement of synaptic epitranscriptome, we now extend our study from mRNA to other synaptically localized RNA species and their chemical modifications. For example, N1-methyl-adenosine (m¹A) is another reversible RNA chemical modification that can induce changes in RNA secondary structures, stabilizing and recruiting tRNAs for translation. We show evidence suggesting the presence of m¹A at neuronal synapse and RNA species carrying this modification. Our study suggests that the expanded coding potential through RNA epigenetics at neuronal synapses is an essential component of a highly flexible and responsive local genetic network underlying synapse function.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-070 D1ドーパミン受容体コンディショナル発現マウスを用いたドーパミンによる運動制御機構の解明 Nae Saito（齊藤奈英）¹，Satoshi Hara（原怜）²，Kazuki Tainaka（田井中一貴）¹，Asako Sato（佐藤朝子）³，Manabu Abe（阿部学）¹，Meiko Kawamura（川村名子）¹，Shun Yamaguchi（山口瞬）⁴，Satomi Chiken（知見聡美）⁵，Hiroshi Ichinose（一瀬宏）²，Kenji Sakimura（﨑村建司）¹，Atsushi Nambu（南部篤）⁵，Toshikuni Sasaoka（笹岡俊邦）¹ ¹新潟大学　脳研究所 ²東京工業大学生命理工学院 ³北里大学医学部 ⁴岐阜大院　医学系研究科 ⁵生理学研究所 Parkinson's disease (PD) is a neurological disorder that exhibits tremor, dyskinesia, rigidity and non-motor symptoms such as cognitive impairment and depression. PD is caused by impairment of dopaminergic neurotransmission due to degenerative loss of midbrain dopaminergic neurons. PD as well as Alzheimer's disease are fairly common in the elderly, and thus, elucidation of pathophysiology underlying symptoms of PD is urgent to develop better treatments and preventive measures. Midbrain dopaminergic neurons mainly project to the striatum, a main input station of the basal ganglia. In the striatum, there are two types of projection neurons: ""direct pathway"" neurons that express dopamine D1 receptors (D1R) and project to the internal segment of the globus pallidus and ""indirect pathway"" neurons that express dopamine D2 receptors (D2R) and project to the external segment of the globus pallidus. However, details of dopamine functions via D1R and D2R in information processing through the basal ganglia remain unclear. To understand neural mechanisms of motor and non-motor symptoms of PD and the roles of dopamine in information processing through the basal ganglia, we developed novel D1R knockdown mice in which D1R expression can be reversibly regulated. Suppression of D1R expression using doxycycline (Dox) administration decreased their spontaneous motor activity and impaired their motor ability. It depressed neurotransmission through the direct pathway. A few days after termination of Dox administration, spontaneous motor activity temporarily increased by several times, and then returned to normal levels in a few days. To investigate the mechanism of such rebound motor activity, we analyzed contents of dopamine and various metabolites at several time points: before Dox administration, and just after and three days after termination of Dox administration for four weeks. In addition, we performed several behavior tests and observed the neural activity after passive avoidance test using the whole brain imaging technique.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-071 GAD65欠損ラットは脳内GABA含量の減少とてんかん重積状態を示す Toshikazu Kakizaki（柿崎利和）¹，Weiru Jiang（姜玮茹）¹，Takashi Suto（須藤貴史）²，Shigeru Saito（齋藤繁）²，Makoto Itakura（板倉誠）³，Masahiko Watanabe（渡辺雅彦）⁴，Yuchio Yanagawa（柳川右千夫）¹ ¹群馬大院医遺伝発達行動 ²群馬大院医麻酔神経 ³北里大医生化学 ⁴北海道大院医解剖発生 γ-Aminobutyric acid (GABA) is a major inhibitory neurotransmitter in the mammalian CNS. GABA is synthesized by two types of glutamate decarboxylase (GAD), GAD65 and GAD67, which are encoded by independent genes. The GAD65 knockout mice survive after weaning and display several behavioral alterations including spontaneous seizures and increased anxiety in adult. GABA content is reduced by half in the adult GAD65 knockout mouse brain. The GAD65/GAD67 protein ratio (GAD65/67 ratio) has been reported to differ between adult mouse and rat brains. We examined whether this ratio is changed during development in both species. Our western blot analyses with anti-GAD65/67 antibody revealed that the mouse GAD65/67 ratio was about 1 in cerebral cortex at postnatal week 2-8 (PW2-8). On the other hand, the ratio of the rat cerebral cortex was about 2 at PW2 and PW4, and 3 at PW8. This result implies that the contributions of GAD65 to GABA synthesis and brain function are bigger in rats than those in mice. In this study, to investigate the physiological significance of GAD65 or GABA in rats, we generated GAD65 knockout rats using TALEN-mediated genome editing. Western blot analysis demonstrated that the GAD65 protein was not detected in whole brain crude extracts of GAD65 knockout rats. On the other hand, the GAD67 protein amounts in the knockout rats were similar to those in their wild-type littermates. These results suggest that GAD67 does not compensate for a loss of GAD65 at the protein level. More than 80% of the GAD65 knockout rats died until PW4. The GAD65 knockout rats displayed wild running/jumping and clonic convulsion, and repeated them (status epilepticus) around PW3 before they died. GABA contents in the GAD65 knockout rats were reduced to 64-72% in the cerebral cortex, hippocampus and cellebellum of their wild-type littermates at postnatal day 17-19. On the other hand, those were reduced to 77-82% at postnatal month 3-7. These results are inconsistent with the fact that the GAD65/67 ratio increases along with ages. In patients with temporal lobe epilepsy and epileptic animals, mossy fiber sprouting can be occurred in the molecular layer of the hippocampal dentate gyrus. In order to determine whether such the reorganization is induced in the GAD65 KO rat hippocampus, we are performing an immunohistochemistry of Zn transporter 3 that is a marker of hippocampal mossy fiber terminal.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-072 Animal Models of Treatment-Resistant Depression: Building on the Insights from Bedside to Bench to Improve Translatability Srikumar Bettadapura Narayanaswamy（Bettadapura Narayanaswamy Srikumar） Natl Inst of Mental Hlth Neuro Sci (NIMHANS) Compound attrition during drug development is perhaps highest in clinical trials of psychiatric illnesses. This can be partly attributed to the failure of accurate prediction of therapeutic activity by animal models. In this context, the quest to develop animal models with better face, construct and predictive validity is more important now than ever before. Traditional tests of depression such as forced swim or tail suspension tests lack construct validity. Accordingly, adopting approaches utilizing clinical mechanisms involved in depression or treatment-resistant depression (TRD) is necessary. In this direction, we developed the ACTH-induced model of depression in mice, which showed good predictive and construct validity but lacked face validity. Accordingly, we explored the MRL/lpr mouse, a model for lupus, as a potential TRD model. These mice showed a depression-like phenotype, construct and predictive validity profile similar to the ACTH model. Thus, while the MRL/lpr mouse model satisfies the criteria for all the three validities, the pathophysiology seen in these mice might be applicable only in a sub-population of patients with immune-mediated depression. Accordingly, currently we are developing a potential TRD model based on neurosteroids. In the clinic, patients on finasteride treatment (for benign prostatic hyperplasia and androgenic alopecia) develop depression that persists beyond the treatment regimen. The neurobiological mechanisms underlying this phenomenon is not understood and has the potential to be developed as a model of TRD. Pursuing this, we found that repeated finasteride (5-alpha reductase inhibitor used to treat benign prostatic hyperplasia) administration induced depression-like phenotype in the FST and splash test and anxiety in the elevated plus maze. Interestingly, antidepressant effects of fluoxetine were blocked by finasteride. Further pharmacological, electrophysiological and neurochemical characterization is underway. In summary, our preliminary results indicate that the finasteride model could represent an animal model with a good construct validity and with a potential for the discovery of novel antidepressants.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-073 オレキシンが縫線核セロトニン作動性ニューロンの発火様式に及ぼす影響 Masaru Ishibashi（石橋賢）^1,2，Atsuo Fukuda（福田敦夫）¹，Christopher S Leonard（Leonard S Christopher）² ¹浜松医科大医学部神経生理 ²Dept of Physiology, New York Medical college, Valhalla, USA Serotonergic (5-HT) dorsal raphe (DR) neurons regulate numerous brain functions including sleep-wake states, circadian phase, reward and mood. It has been well established that the wake-promoting neuropeptide orexin depolarizes and increases the firing rate of 5-HT DR neurons. However, how orexins influence the response of 5-HT DR neurons to their inputs has not been well explored. This is important because 5-HT DR neurons are known to fire at slow tonic rates in relation to behavioral state but also to encode sensory/motor and reward events with phasic firing. To investigate this question, we used whole cell recording methods in mouse brain slices. We found that, in addition to depolarizing these neurons, orexin-A also strongly increased the amplitude and duration of the Ca²⁺-dependent post-spike afterhyperpolarization (AHP). This orexin-enhanced AHP (oeAHP) was mediated by both orexin 1 and orexin 2 receptors involved two distinct molecular mechanisms. The first was of medium-duration and involved apamin-sensitive SK Ca²⁺-activated K⁺ channels. The second was of longer duration, was apamin-insensitive (termed the ai-oeAHP) and appeared similar to a slow AHP (sAHP). Additional experiments exploring the extracellular Ca²⁺ dependence ([Ca²⁺]_o) of both orexin-induced excitation and the oeAHP revealed they are both nearly blocked by 10 mM [Ca²⁺]_o and are near-maximal at more physiological (~ 0.5 mM) [Ca²⁺]_o. Surprisingly, we found that the ai-oeAHP was not attenuated by a cesium-based patch solution as expected for a K+ currents, but rather was blocked by substituting NMDG for Na+ in the ACSF or by application of flufenamic acid (FFA), both of which attenuated the orexin-induced inward current. Functionally, the oeAHP reduced both the steady-state firing rate and firing fidelity for a given input, without attenuating the initial firing rate or firing fidelity. Thus, orexin-A increases spike-frequency adaptation and makes these 5-HT DR neurons respond more phasically, which could influence 5-HT release. These results reveal new orexin actions on 5-HT DR neurons and suggest that, orexin functions to enhance phasic responding of these neurons to their inputs. Moreover, these findings are relevant to the pathophysiology of hypercalcemia which would attenuate these orexin actions and might contribute to the associated clinical sign of drowsiness.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-074 次世代型アルツハイマー病モデルマウスを用いた神経細胞1次繊毛動態の解析 Shougo Koubuchi（河渕省吾）¹，Takashi Saitoh（斉藤貴志）²，Takaomi C Saido（西道隆臣）²，Yuki Kobayashi（小林勇喜）¹，Yumiko Saitoh（斎藤祐見子）¹ ¹広島大学総合科学研究科 ²理研脳神経科学研究センター・神経老化制御 Primary cilia are small non-motile microtubule and cell membrane protrusions appeared on most mammalian cell types. These organelles serve as sensory structures receiving the extracellular milieu and reprogramming the transcriptional machinery in response to environmental change. Neuronal ciliary membrane is highly enriched for receptors, in particular a set of certain G protein-coupled receptors including melanin-concentrating hormone receptor 1 (MCHR1), determine a host of crucial physiologies. Disrupting cilia structure or function results in a spectrum of diseases collectively referred to as ciliopathies. Common to human ciliopathies is cognitive impairment, a symptom also observed in Alzheimer's disease (AD). AD is associated with impairments in memory, cognition, language, behavior, and personality. One hallmark of AD is accumulation of senile plaques composed of neurotoxic amyloid-β (Aβ) peptide. The Aβ peptide is generated by the proteolytic cleavage of the amyloid precursor protein (APP). Transgenic mouse models of AD with nonphysiologic overexpression of amyloid precursor protein exhibit various unnatural symptoms/dysfunctions. To overcome this issue, mice with single humanized APP knock-in carrying Swedish (NL), Arctic (G) and Beyreuther/Iberian (F) mutations in different combinations were recently developed. Here, we report that the hippocampal CA1 and the CA3 neuronal cells in APP^NL-G-F knock-in mice (NLGF) still had canonical cilia molecule adenylyl cyclase 3- and MCHR1-positive cilia but their length were significantly changed as compared to the corresponding wild-type mice. In addition, a significant difference of cilia prevalence between NLGF- and wild-mice also observed in selective hippocampal regions. There is growing evidence that primary cilia dynamics regulate interneuronal morphology, synaptic connectivity and formation of neuronal dendrites. Thus, our observations raise the important possibility that structural and functional alterations in neuronal cilia might have a role in AD development.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-075 顔面運動ニューロン傷害後の機能低下に関連するシグナル分子 Takashi Ishijima（石嶋貴志），Kazuyuki Nakajima（中嶋一行）創価大学、理工学部、共生創造理工学科 Transection of adult rat facial nerve led to the functional down-regulations of m2 muscarinic acetylcholine receptor (m2MAchR) and GABA_A receptorα1(GABA_ARα1) in the ipsilateral motoneurons. However, it was not clear how these changes were caused and/or what kinds of signaling molecules are related to the phenomenon. Thus, in the present study, we surveyed the signaling molecules associated with down-regulation of motoneuronal function in injured nucleus. The axotomized facial nucleus and the control nucleus were cut out from the brainstem of rats whose facial nerve was previously transected, and the tissue extracts were analyzed for various signaling molecules such as transcription factors, mitogen-activated protein kinases (MAPKs) and protein kinase C(PKC). We first analyzed activation of cAMP-response element binding protein (CREB) in injured nucleus. Immunoblotting revealed that the levels of phosphorylated CREB in axotomized facial nucleus were largely decreased at 1.5-6 h post-insult. The levels of phosphorylated extracellular signal-regulated kinase were not significantly changed, but those of phosphorylated p-38 were down-regulated at 3-12 h post-insult. We further analyzed the levels of classical protein kinase C (PKCα, PKCβ, PKCγ) in axotomized facial nucleus. As the result, the levels of PKCα and PKCβ were significantly suppressed at 1.5-12 h after transection. On the other hand, the levels of PKCγ did not change at any time points. Together all, these results suggested that the functional down-regulation of motoneurons in axotomized facial neucleus was caused by inactivation of p-38 MAPK and CREB and/or by decrease of PKCα/β.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-076 光遺伝学を用いた延髄孤束核領域における L-DOPA 性神経伝達の機能解析 Daiki Masukawa（増川太輝）¹，Yuka Nakao（中尾友香）¹，Hiroki Muneto（宗藤大樹）¹，Ryang Kim（金亮）²，Haruhiko Bito（尾藤晴彦）²，Yoshio Goshima（五嶋良郎）¹ ¹横浜市大医分子薬理神経生物 ²東京大院医神経生化学 We have proposed that L-DOPA is a neurotransmitter. L-DOPA when exogenously microinjected into the nucleus tractus solitarii (NTS) induces depressor and bradycardic response. Electro-stimulation of aortic depressor nerves (ADN) induces L-DOPA release from the NTS, and induces depressor and bradycardic response. These our findings suggested that L-DOPA plays a neurotransmitter role in the primary baroreceptor afferents terminating in the NTS. To further determine the role of L-DOPA as a neurotransmitter in the ADN, we investigated whether depressor and bradycardic responses to L-DOPA was mimicked by stimulating ADN using optogenetic procedure in rats. We injected adeno-associated virus encoding ChR2-EYFP or EYFP into the ganglion of ADN. Four to five weeks after injection, ChR2-EYFP and EYFP signals were partially localized with tyrosine hydroxylase-positive neurons in the NTS of the fixed brain tissues. Photostimulation (473 nm, 40 mW, 20 Hz, 20s) from the surface of the NTS induces depressor, bradycardic response in the animals expressing ChR2-EYFP, but not EYFP in the ADN. The effects of photostimulation were attenuated by treatment with L-DOPA cyclohexylester (1 μg), an antagonist for L-DOPA, in the NTS, as were those of exogenously applied L-DOPA. Our preliminary data show that the photostimulation induces the release of L-DOPA detected by microdialysis in vivo at the NTS. These results further provide the evidence that L-DOPA is a neurotransmitter in the ADN terminating into the NTS.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-077 末梢神経損傷後の脊髄後角におけるBDNF mRNA 発現変化 Kimiko kobayashi（小林希実子），Hiroki Yamanaka（山中博樹），Masamichi Okubo（大久保正道），Koichi Noguchi（野口光一）兵庫医大医解剖（神経） Brain derived neurotrophic factor (BDNF) is known to be a neuromodulator in the dorsal horn circuit. In the neuropathic pain model, BDNF in the spinal dorsal horn has been recently considered to play a crucial role in the sensitization of dorsal horn neurons leading to mechanical allodynia (Coull et. al., Nature. 2005). However, the cellular distribution of BDNF in the spinal cord has remains unclear. The aim of this study was to detect the BDNF mRNA expressing cells in the spinal cord and evaluate the effect of nerve injury on the transcriptional activity of BDNF mRNA. In this study, we examined the expression of the BDNF mRNAs in the spinal cord after spared nerve injury (SNI). Male Sprague-Dawley rats (weighing 200-250 g) were sacrificed at several time points (1 day to 2 weeks) and the expression of BDNF mRNA in spinal cord was examined by reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization histochemistry (ISHH). RT-PCR showed that the nerve injury increased the BDNF mRNA in dorsal horn. Quantitative analysis revealed that BDNF mRNA expression was significantly increased from 1 to 3 days and returned to normal level at 14 days after injury. We could detect the expression of BDNF mRNA in naive spinal dorsal horn by ISHH. Spinal cord section of SNI model showed the increase of the intensity of positive signals but not the number of BDNF mRNA labelled cells. Upregulated BDNF mRNA expressing cells are located mainly in lamina I and III-IV of dorsal horn. Double labeling ISHH of BDNF with several marker proteins showed that the upregulated signal of BDNF heavily co-localized with a subsets of NeuN (neuronal marker) and Lmx1B (excitatory neuronal marker ) immunoreactive cells in dorsal horn of SNI model rats. Signals for BDNF mRNA were exclusively localized in excitatory dorsal horn neuron. These results indicate that the specific population of spinal neurons have high level BDNF expression and that limited subset of spinal neurons up-regulate BDNF expression following nerve injury. These results indicate that a subset of spinal neurons use BDNF as a modulator of spinal local circuit or as a signaling molecule from spinal cord to brain that integrates sensory inputs.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-078 神経細胞が持つGPCR局在型一次繊毛動態とエネルギー代謝の関連 Yuki Kobayashi（小林勇喜），Tomoya Okada（岡田智哉），Daisuke Miki（三木大輔），Shogo Kohbuchi（河渕省吾），Yumiko Saito（斎藤祐見子）広島大学総合科学研究科 Most mammalian cells have single nonmotile primary cilia, which are considered to be important organelles that function as hubs for selective signaling. Accumulating evidence suggests a strong association between genetic ciliopathies and obesity in humans and animals. Indeed, obesity is a common manifestation observed in human genetic ciliopathies such as Bardet-Biedl syndrome (BBS). Many neurons in the mammalian brain possess primary cilia that are enriched for certain G protein-coupled receptors (GPCRs), including melanin-concentrating hormone (MCH) receptor 1 (MCHR1). The MCH-MCHR1 system is known to be mainly involved in feeding behavior and energy homeostasis. Recently, we reported that MCH causes a 30-40% reduction in primary cilia length via ciliary MCHR1-expressing epithelial RPE1 cells. Mice that lack BBS proteins (BBS4) is obese with short cilia length in selected brain regions such as hippocampus. However, a possible correlation between MCHR1-positive neuronal cilia length and energy metabolism has not been characterized. Here, we report that 48 h-starved mice displayed shorter MCHR1 positive cilia than did normal diet-fed mice in the hippocampal CA1 region. Quantitative real-time PCR revealed that preproMCH mRNA expression was highly upregulated in the lateral hypothalamus after 48 h-starved mice. Further, in rat-derived hippocampal slice culture, MCH treatment at the nanomolar level caused cilia length shortening of MCHR1 positive cilia in the CA1 region. Our results suggest the involvement of MCH-MCHR1 axis in regulation of MCHR1-positive neuronal cilia length in vivo. It is currently in progress for the long-term effect of high-fat diets on the dynamics of MCHR1-positive neuronal cilia.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-079 嫌悪刺激によるドーパミン放出の抑制が側坐核D2R-中型有棘神経細胞内のPKA-Rap1経路を活性化する You-Hsin Lin（林祐新）¹，Taku Nagai（永井拓）²，Yukie Yamahashi（山橋幸恵）¹，Keisuke Kuroda（黒田啓介）¹，Kozo Kaibuchi（貝淵弘三）¹ ¹名古屋大学大学院医学系研究科神経情報薬理学講座 ²名古屋大院医医療薬学 Aversive behavior is one of the emotional behaviors that is critical in avoiding dangerous situation or unpleasant/harmful stimuli. Its deficiency is known to be associated with various neuropsychological diseases, including Alzheimer's disease and Parkinson's disease. To date, accumulated studies have shown that the output of aversive behavior is regulated by the nucleus accumbens (NAc). NAc consists mainly of medium spiny neurons that express dopamine D1 receptor (D1R-MSN) and dopamine D2 receptor (D2R-MSN). Receiving various neuromodulator inputs, such as dopaminergic input, it determines the output of aversive behavior and other emotional behaviors by controlling the balance between D1R-MSN activation and D2R-MSN activation. Aversive stimuli are thought to suppress dopamine release in the NAc and activate specifically accumbal D2R-MSN through protein kinase A (PKA). However, the linkage between the suppressed dopamine release in the NAc and PKA activation in accumbal D2R-MSN is not well understood yet. Our lab recently has shown that PKA-Rap1 signaling is activated in accumbal D2R-MSN at low dopamine concentration in NAc. In this study, we found that PKA-Rap1 signaling activation is involved in aversive behavior by using adeno-associated virus (Flex-AAV) that express PKA and Rap1 mutants in D2R-MSN specific manner. Furthermore, we found that electric shock used as aversive stimulus activated PKA-Rap1 signaling specifically in accumbal D2R-MSN. We also found that the inhibited dopamine release by optogenetics induced aversive behavior through PKA-Rap1 signaling in accumbal D2R-MSN. How aversive stimulus activates PKA-Rap1 signaling in D2R-MSN would need to be investigated in the future studies.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-080 線条体ニューロンにおけるLrtm2の役割 Misato Ichise（市瀬実里），Minoru Hatayama（畑山実），Jun Aruga（有賀純）長崎大院 Recent studies have shown critical roles of neural leucine-rich-repeat (LRR)-containing transmembrane superfamily proteins in health and disease. Among them, proteins with LRR with no other recognizable extracellular domains (`'LRR-only'' proteins such as Lrrtm1-4 and Slitrk1-6) are known to be essential for synaptogenesis or maintaining synaptic functions. Lrtm2, a member of the LRR-only proteins, was strongly detected in striatum, globus pallidus and substantia nigra pars reticulata in mouse brain, and the protein content increased in the postnatal development. In previous studies, Lrtm2-deficient mice exhibited altered motor function and altered dopamine metabolites content in striatum and other regions. To clarify the role of Lrtm2 in the striatal medium spiny neurons, we measured the expression level of various molecular markers, and found that distribution of proteins involved in controlling neurotransmitter dynamics were altered in the of striatonigral GABAergic pathway of the Lrtm2-deficient mice. Based on these results, we hypothesized a role of Lrtm2 in macromolecular trafficking in the striatal neurons. To address this possibility, we generated a series of Lrtm2 mutant proteins that show impaired subcellular localization. The role of Lrtm2 was investigated by expressing these mutant proteins in vitro and in vivo.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-081 部位特異的蛍光標識を用いたドーパミンに応答するチロシン水酸化酵素の構造変化の解析 Shunya Inukai（犬飼駿弥），Satoshi Hara（原怜），Hiroshi Ichinose（一瀬宏）東工大院生命理工 Tyrosine hydroxylase (TH) is an enzyme that synthesizes L-DOPA from L-Tyrosine. The reaction is the rate-limiting step of the biosynthetic pathway for dopamine (DA). The TH protein structure consists of three domains; the regulatory domain at the N-terminal region (1-165 a.a.), the catalytic domain at the central region (165-477 a.a.), and the tetramerization domain at the C-terminal region (477-497 a.a.). The TH activity is inhibited by DA through the binding to iron at the active site, and the inhibition is canceled by phosphorylation of the 40th serine residue. Although the conformational changes of the N-terminal region are deeply involved in the regulatory mechanism, the crystal structure of the full-length TH containing the N-terminal region has not been solved. In order to understand the structural information in the regulation of TH, we prepared the fluorescence-labeled TH proteins at the desired residues and measured alterations in the fluorescence intensity caused by the DA binding. We labeled the fluorescent dye to a cysteine residue (Cys) on TH via a maleimide group. We substituted four out of seven Cys residues on the surface of TH to alanine, and prepared mutant THs in which Cys was introduced into four different sites A59, C176, A230, and S295. Each mutant TH was expressed in E. coli, purified, and labeled with a fluorescent dye. The fluorescence intensity of each fluorescence-labeled mutant TH was measured, and the changes of the fluorescence intensity after the addition of DA were recorded. The fluorescence intensities were decreased by the addition of DA in those mutants at the sites of A59, C176, A230 and S295. These results suggested that A59, C176, A230, and S295 are the sites affected by the conformational changes caused by the DA binding to the active site. The results of A59 and S295 corresponded to the study obtained from hydrogen-deuterium exchange (Wang et al., 2009). In addition, we presumed the sites affected by the conformational change, which have not been identified.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-082 ドーパミンシグナルとアデノシンシグナルとのバランスは、線条体中型有棘神経細胞におけるPKA/Rap1経路を制御する Xinjian Zhang（張心健）¹，Taku Nagai（永井拓）²，Rijwan Uddin Ahammad（Ahammad Rijwan Uddin）¹，Keisuke Kuroda（黒田啓介）¹，Takashi Nakano（中野高志）³，Junichiro Yoshimoto（吉本潤一郎）³，Kiyofumi Yamada（山田清文）²，Kozo Kaibuchi（貝淵弘三）¹ ¹名古屋大院医神経情報薬理 ²名古屋大院医医療薬学 ³奈良先端大情報 Medium spiny neurons (MSNs) expressing dopamine D1 receptor (D1R) or D2 receptor (D2R) are major components of the striatum. In the ventral striatum, D1R-MSNs are involved in rewarding behavior, while D2R-MSNs are involved in aversive behavior. Stimulation of D1R activates protein kinase A (PKA) through Golf to increase neuronal activity, while D2R stimulation inhibits PKA through Gi. Adenosine A2A receptor (A2AR) coupled to Golf is highly expressed in D2R-MSNs within the striatum. However, how dopamine and adenosine co-operatively regulate PKA activity remains largely unknown. Here, we measured Rap1gap serine 563 phosphorylation to monitor PKA activity and examined dopamine and adenosine signals in MSNs. We found that a D1R agonist increased Rap1gap phosphorylation in striatal slices and in D1R-MSNs in vivo. A2AR agonist CGS21680 increased Rap1gap phosphorylation, and pretreatment with the D2R agonist quinpirole blocked this effect in striatal slices. The A2AR agonist showed minimal effects on Rap1gap phosphorylation in vivo whereas D2R antagonist eticlopride increased Rap1gap phosphorylation in D2R-MSNs in vivo, and the effect of eticlopride was blocked by the pretreatment with the A2AR antagonist SCH58261. These results suggest that adenosine positively regulates PKA in D2R-MSNs through A2AR, while this effect is blocked by basal dopamine in vivo. Incorporating computational model analysis, we propose that the shift from D1R-MSNs to D2R-MSNs or vice versa appears to depend predominantly on a change in dopamine concentration.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-083 代謝型グルタミン酸受容体1型とGABA_B受容体の複合体形成と機能的相互作用 Hakushun Sakairi（坂入伯駿）¹，Yuji Kamikubo（上窪裕二）²，Masayoshi Abe（阿部匡良）³，Keisuke Ikeda（池田圭祐）³，Arata Ichiki（一木新）³，Toshihide Tabata（田端俊英）³，Takashi Sakurai（櫻井隆）^1,2 ¹順天大院　医　細胞・分子薬理 ²順天大医　薬理 ³富山大院理工 G-protein coupled receptors (GPCRs) mediate diverse modalities of cellular responses including synaptic transmission. Recently, many studies have suggested that complexed GPCRs may mediate more diverse modalities of cellular responses. We pursue the physiological significance of complex formation between GPCRs for neurotransmitters, focusing on the metabotropic glutamate receptors and gamma aminobutyric acid B receptor (mGluRs and GABA_BR, respectively). Type-1 mGluR (mGluR1) is localized to the post-synaptic membrane and mediates long-term depression (LTD), a form of synaptic plasticity crucial for learning and memory. GABA_BR reduces neurotransmitter release and postsynaptic excitation by activating the K⁺ channels, and inhibiting the activities of adenylyl cyclase and Ca²⁺ channels. In the previous reports, we showed that GABA_BR activation augments mGluR1 signaling and LTD in cerebellar Purkinje cells. These results suggest the possibility of heteromeric complex formation between these GPCRs. In this report, we aimed to clarify the details of physical and functional interaction between mGluR1 and GABA_BR at the plasma membrane in a transient or stable heterologous expression system of HEK293 cells. Our fluorescence imaging and biochemical assay uncovered that mGluR1 and GABA_BR formed heteromeric complex at cell surface. Moreover, Our Ca²⁺ imaging and homogenous time-resolved Förster resonance energy transfer (TR-FRET) cAMP assay revealed that GABA_BR activation augmented mGluR1 signal transduction, while mGluR1 activation suppressed GABA_BR signaling. These results indicate that mGluR1 and GABA_BR regulate their function mutually. Our findings provide a novel insight into the regulatory mechanism of synaptic transmission.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-084 β-アレスチンを介する PACAP-PAC1受容体シグナルの解析 Yusuke Shintani（新谷勇介）¹，Atsuko Hayata-Takano（早田敦子）^1,2，Keita Moriguchi（森口啓太）¹，Hitoshi Hashimoto（橋本均）^1,2,3,4 ¹大阪大学大学院薬学研究科神経薬理学分野 ²大阪大学・金沢大学・浜松医科大学・千葉大学・福井大学連合小児発達学研究科附属子どものこころの分子統御機構研究センター ³大阪大学データビリティフロンティア機構バイオサイエンス部門 ⁴大阪大学先導的学際研究機構超次元ライフイメージング研究部門 Pituitary adenylate cyclase-activating polypeptide (PACAP) is a multifunctional neuropeptide widely expressed in the nervous system and acts as a neurotransmitter and neurotrophic factor. Type 1 PACAP receptor (PAC1R) is highly expressed in the central peripheral nervous system and involved in the regulation of, e.g., emotion and stress response, pain transmission. Two nonvisual β-arrestins, β-arrestin1 and β-arrestin2 are functional adaptor proteins that interact with activated GPCRs and mediate several signaling pathways in G-protein-independent manner. Recently, several studies have demonstrated that the β-arrestin-mediated signaling can be therapeutic targets for several disorders with improved therapeutic potential. However, the precise roles of the two β-arrestin isoforms in PACAP-PAC1R signaling remain unclear. In this study, we examined PACAP-induced interaction between PAC1R and the two β-arrestin isoforms, β-arrestin-dependent PAC1R subcellular trafficking and ERK1/2 activation using the NanoBiT system, HaloTag technology and siRNA-mediated silencing in HEK293T cells and primary cultured cortical neurons. PAC1R interacted with both β-arrestins with similar PACAP dose- and time-dependency. Upon PACAP stimulation, the complex of PAC1R and β-arrestin2 translocated from the cell surface into cytosol, but that of β-arrestin1 remained at the cell surface regions. siRNA-mediated silencing of β-arrestin2 significantly reduced PACAP-induced PAC1R internalization and prolonged ERK1/2 activation, but silencing of β-arrestin1 showed no significant effect on PAC1R internalization and rather increased ERK1/2 phosphorylation. Taken together, these results suggest that the fine and precise tuning of the PAC1R signaling pathways is mediated by the differential actions of β-arrestin1 and β-arrestin2. Because altered PACAP-PAC1R signaling might be related to psychiatric and neurological disorders, the present observations are expected to contribute to elucidation of molecular mechanisms of these disorders.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-085 癲癇発作中の脳神経活動イメージング Leona Shigemitsu（重光玲於奈），Kazutoyo Ogino（荻野一豊），Hiromi Hirata（平田普三）青山学院大学大学院理工学研究科 Epilepsy, a chronic brain disorder, is pathologically marked by spasm and/or loss of consciousness and is caused by hyperexcitability of the CNS. The excess excitability of the CNS can be induced by efflux of Cl- from the cells. One of the causes of epilepsy is upregulation of chloride ion reversal potential, which can be induced by the loss of potassium-chloride cotransporter KCC2, resulting in the excess activation of neural networks. Zebrafish have two paralogs of KCC2 gene (KCC2a and KCC2b) generated by the ancestral gene duplication and are expressed by the CNS. Here, we generated KCC2a-KCC2b double knockout mutant (DKO) zebrafish by genome editing and found that DKO larvae exhibit the epileptic phenotype when exposed to LED flash light. However, how this epilepsy is triggered by external optical stimuli remains unclear. We here aim to elucidate the focal area of seizured brain in larval zebrafish. Firstly, we established a motor assay to induce epilepsy following exposure to red flash light at 20 Hz. Secondly, to visualize brain neural activity during epileptic seizure, we prepared DKO zebrafish larvae (5 dpf) expressing GCaMP6s for visualization of whole brain activity in the brain. Lastly, we performed in vivo calcium imaging and analyzed the result of a fluorescence intensity and kinetics divided by three of brain regions; telencephalon, optic tectum and hindbrain. Surprisingly, we found that the fluorescence intensity in each region of the DKO larval brain was more persistent and higher compared to wild type. Our temporal analysis revealed that excitability during epileptic seizure is prolonged in DKO larval brain compared to wild type brain. Taken together, we established a new pathologically relevant seizured model in zebrafish.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-086 リン酸化によるKCC2の機能制御はGABAによる抑制性伝達および生存に必須である Miho Watanabe（渡部美穂）¹，Jinwei Zhang（Zhang Jinwei）²，Mohammad Mansuri（Mansuri Mohammad）³，Jingjing Duan（Duan Jingjing）³，Kristopher T Kahle（Kahle T Kristopher）^3,4，Atsuo Fukuda（福田敦夫）¹ ¹浜松医大神経生理 ²Inst Biomed Clinical Sci, Univ Exeter Med Sch, Exeter, UK ³Dept Neurosurgery, Yale Sch Med, New Haven, CT ⁴Depts Neurosurgery, Pediatrics, Cell and Mol Physiol; Centers for Mendelian Genomics, Yale Sch Med, New Haven, CT The Cl^- extruding transporter KCC2 (SLC12A5) critically modulates GABA_A receptor signaling via its effect on neuronal Cl^- homeostasis. A postnatal decrease in neuronal Cl^- concentration mediated by KCC2 establishes GABA-mediated fast synaptic inhibition in the central nervous system. However, the mechanisms that regulate the functional expression of KCC2 during development are poorly understood. Here, we showed that in vivo KCC2 phosphorylation at two carboxyl-terminal threonine (Thr) residues 906 and 1007, essential for in vitro inhibition of KCC2 activity, decreased in parallel with the development onset of GABA inhibition. Mice engineered with homozygous glutamate (E) substitution at Thr906/Thr1007 (Kcc2^E/E), modeling constitutive phospho-mimetic inhibition of KCC2, died shortly after birth due to apparent spontaneous respiratory arrest. Kcc2^E/E mice exhibited diminished GABA-dependent neuronal Cl^- extrusion capacity, lacked spontaneous respiratory discharges from cervical spinal cord neurons (C4), and displayed severely altered locomotor rhythm recordings from lumbar spinal cord neurons (L2). Kcc2^E/E mice exhibited touch-evoked generalized seizures and an anomalous neuronal distribution but normal dendritic spine morphology. These data demonstrated that regulated KCC2 phosphorylation at Thr906/Thr1007 is essential for postnatal survival through its control of Cl^- homeostasis governing postnatal developmental changes in GABA signaling.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-087 Slitrk1によるPC12細胞のエンドサイトーシス制御 Minoru Hatayama（畑山実）^1,2，Kei-ichi Katayama（片山圭一）²，Jun Aruga（有賀純）^1,2 ¹長崎大 ²理研CBS Slitrk family encodes transmembrane protein with extracellular leucine-rich repeat domain and intracellular domain. This family consists of six members that are expressed in CNS. A member of Slitrk family, Slitrk1, is involved in synapse formation, neurite extension, and its knockout mice show increased anxiety- and depression-like behavior. However, the molecular mechanisms underlying Slitrk1-mediated control of the higher brain function are poorly understood. Here we show Dynamin1 is a binding partner for Slitrk1, and Slitrk1-Dynamin1 complex regulated membrane internalization of PC12 cell. Dynamin1 was identified by mass-spectrometry analysis of the Slitrk1 co-precipitates in an immunoprecipitation assay using anti-Slitrk1 and embryonic brain lysate. Analysis of PC12 cells cotransfected with HA-tagged Slitrk1 and mCherry-tagged Clathrin revealed that Slitrk1 increased Clathrin-positive vesicle number 24 hours after transfection. In addition, Slitrk1 transfected cell decreased membrane internalization indicated by FM 4-64 dye uptake. To investigate membrane dynamics in living cell, time-lapse imaging was performed for mCherry-Clathrin in PC12 cell stimulated with nerve growth factor. This assay revealed that Slitrk1 extended duration of Clathrin-positive vesicle, and that Slitrk1 was one of the regulatory factors of endocytosis. These findings suggest that Slitrk1 inhibits Clathrin-dependent endocytosis by interacting with Dynamin1 in PC12 cell. To address if Slitrk1 is similarly involved in the regulation of endocytosis in CNS neurons. We also analyzed Clathrin and Dynamin1 dynamics in Slitrk1-overexpressing primary cultured neurons under various neurotransmitter stimuli.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-088 マウス脳室周囲器官に局在するToll様受容体4の発現量はLPS濃度依存的に減少する Yousuke Nakano（中野洋輔）^1,2，Masato Maruyama（丸山正人）¹，Masahiko Kase（加瀬政彦）¹，Seiji Miyata（宮田清司）²，Tetsuo Sugimoto（杉本哲夫）¹ ¹関西医科大学医学部解剖学第二講座 ²京都工芸繊維大学生体機能学研究室 The immunoreaction induced by pyrogen lipopolysaccharide (LPS), a component of gram-negative bacteria, is a well-characterized example of innate recognition which leads to a robust inflammatory response. It has been reported that LPS-induced inflammation led to different two systemic responses, low-dose LPS-induced fever and high-dose LPS-induced hypothermia. The hypothermia induced by LPS is often used as a model of sepsis. In addition, previous reports had shown that Toll-like receptor 4 (TLR4) mRNA was reduced in septic rat brain. However, in mammalian brain, the mechanism of these opposite immunoreactions has still been unclear. The sensory circumventricular organs (CVOs), comprised of the organum vasculosum lamina terminalis(OVLT), subfornical organ(SFO) and area postrema(AP), are lack the blood-brain barrier, and the sensory CVOs expressed various receptors to recognize molecules in the blood, pro-inflammatory cytokines, pathogen and more. In our previous study in vivo, TLR4 was expressed in astrocytes throughout the sensory CVOs and in microglia of the AP. In addition, the peripheral and central administration of LPS induced a nuclear translocation of STAT3 in astrocytes, respectively. These results indicated that the sensory CVOs can directly recognize LPS through TLR4. To investigate the immunoreaction in the sensory CVOs, LPS was administered intraperitoneally at the low (50 ug/kg) or high (1 mg/kg) dose into ICR adult male mice. Twenty-four hours after administration of LPS, brain slices were immunostained with antibodies against TLR4, GFAP (astrocyte marker), and CX3CR1 (microglia marker), and the number of TLR4 positive cells was counted. As a result, low dose of LPS reduced TLR4 positive cells in AP, but not in OVLT and SFO. On the other hand, high dose of LPS reduced TLR4 positive cells in all sensory CVOs. Taken together, our results showed that LPS administration reduced the TLR4 positive cells in CVOs and AP was most sensitive to LPS among CVOs.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-089 Gタンパク質共役型受容体とβアレスチンの相互作用が1次繊毛局在に与える影響 Sho Shikada（鹿田星）¹，Ko Miyoshi（三好耕）¹，Atsuko Hayata（早田敦子）^2,3，Takeshi Yoshimura（吉村武）¹，Sarina Han（韓薩日娜）¹，Genki Amano（天野元揮）¹，Hironori Takamura（高村明孝）¹，Taiichi Katayama（片山泰一）¹ ¹大阪大院・連合小児・分子生物遺伝学 ²大阪大院・薬・神経薬理 ³大阪大院・連合小児・子どものこころセンター On the surface of neurons in most vertebrates there is a non-motile protrusion of the cell membrane called primary cilium. In recent years, primary cilia have been studied in roles of a non-synaptic sensor that transmits extracellular stimuli into the cell body. Defective formation or function of primary cilia is implicated in the pathogenesis of many human developmental diseases, termed ciliopathies. In ciliopathies several symptoms of the central nervous system such as neurodevelopmental disorders are accompanied, pointing out the importance of cell function at primary cilia on neurons. Several G-protein-coupled receptors (GPCRs) have been revealed to localize at primary cilia as signaling factors, and it has been clarified that many of them are essential for activating signaling pathways responsible for multipleneural functions. It has been suggested that among the ciliary GPCRs serotonin receptor 6 (HTR6) is implicated in social cognition. On the other hand, serotonin receptor 7 (HTR7), highly homologous to HTR6, does not localize to primary cilia. Therefore, localization of GPCRs in primary cilia is likely to be specifically regulated by other factors. In this study, in order to elucidate regulatory mechanisms of GPCR localization in primary cilia, we performed cell biological or pharmacological analysis, and attempted to identify factors that specifically bind to and regulate HTR6. As a result, we found that HTR6 interacts with β-arrestin while HTR7 does not, and that HTR6-β-arrestin interaction is likely to important for the cilia localization of HTR6, where this interaction is regulated by activity of HTR6.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-090 PC12細胞分化により発現誘導するGPR3はシナプシンリン酸化を修飾する Humiaki Ikawa（猪川文朗），Shigeru Tanaka（田中茂），Kenta Sasaki（佐々木健太），Tomohiro Noguchi（野口智裕），Kana Harada（原田佳奈），Izumi Hide（秀秀和泉），Norio Sakai（酒井規雄）広島大院医歯薬神経薬理 The G protein-coupled receptor 3 (GPR3), which is widely expressed in various neuron types, is a member of the class A rhodopsin-like GPCR family. GPR3 is unique in its ability to constitutively activate Gαs protein without the addition of ligands, which elevates the basal level of intracellular cyclic adenosine monophosphate (cAMP). In a previous study, we reported that the neuronal expression of GPR3 enhances neurite outgrowth and neuronal survival. Recent our time-lapse observations of GPR3-transfected cerebellar granular neurons revealed that GPR3 is transported along the neurite and predominantly distributed at the tip of neurite, where the expression of GPR3 is highly correlated with local elevation of protein kinase A (PKA). Synapsin is a major endogenous substrate for cAMP-dependent protein kinase and is known to be highly concentrated in the presynaptic axon terminal. These findings suggest that GPR3 may interact with synapsin to modulate synaptic functions. To address this question, PC12 cells were utilized to explore presynaptic function. When the differentiation of PC12 cells was induced by serum deprivation and addition of a nerve growth factor, GPR3 mRNA was highly upregulated from 12-48 hours after the stimulation. Similarly, synapsin 2 mRNA was increasingly upregulated until 48 hours after the stimulation, whereas synapsin 1 and synapsin 3 decreased during differentiation of PC12 cells. In PC12 cells transfected with a GFP-tagged GPR3 expressing vector, fluorescent GPR3 was transported along the neurites and concentrated in the presynaptic terminal. We then investigated if the induction of GPR3 affected the phosphorylation of synapsin. When the differential induction of GPR3 was decreased by shRNA, the phosphorylation of synapsin tended to decrease 28 hours after transfection, as shown by Western blotting analysis. These results suggest a potential role of GPR3 in the phosphorylation of synapsin that may serve as a potential modulator of presynaptic functions.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-091 電位依存性ナトリウムチャネルNav1.1と相互作用する脳タンパクの同定 Ikuo Ogiwara（荻原郁夫）日本医科大医生理（システム生理） Nav1.1 is one of four brain-types of voltage-gated sodium channel alpha subunit that co-assemble with beta subunits to form heteromultimeric protein complexes. In juvenile mouse brain, Nav1.1 is localized predominantly at the axon initial segments (AISs) in parvalbumin-expressing (PV) cells, and plays critical roles in maintaining sustained fast-spiking from these inhibitory neurons. Selective removal of Nav1.1 in PV cells in mice causes epileptic seizures and autism-like behaviors resembling clinical features of human Dravet syndrome. Meanwhile, Nav1.1 is obviously detectable in some excitatory cells, but is indiscernible at the AISs in these neurons. Although Nav1.1 interacts with ubiquitously-expressed calmodulin through the IQ domain in its C-terminal domain, we wanted to identify other proteins that would interact and contribute to subcellular localization of Nav1.1. In the present study, we first screened candidate proteins using immunoprecipitation with anti-Nav1.1 antibody and juvenile mouse brain extract, followed by tandem mass spectrometry. The list of proteins included previously known Nav1.1-interacting proteins, such as sodium channel beta subunits and calmodulin. We selected some candidates from the list, which were seemingly expressed in subsets of neurons on the basis of the Allen brain atlas database, and verified that two candidates, namely, A and B, were co-immunoprecipitated with Nav1.1, and vice versa. Previous studies showed that proteins A and B are localized in the AISs and post-synapses of neurons, respectively. We finally found that proteins A and B both interacted with the intracellular loop I, but not the C-terminal domain, of Nav1.1 using heterologous expressing systems with HEK293 cells. Our findings may suggest that these Nav1.1-interacting protein candidates can be involved in subcellular localization of Nav1.1.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-092 静磁場刺激は細胞膜Cl−透過性を高めることでマウス大脳皮質錐体神経細胞の興奮性を低下させる Adya Saran Sinha（Sinha Saran Adya）¹，Yasuyuki Takamatsu（高松泰行）^2,3，Tenpei Akita（秋田天平）¹，Tatsuya Mima（美馬達哉）²，Atsuo Fukuda（福田敦夫）¹ ¹浜松医大・医・神経生理学 ²立命館大・先端総合学術研究科 ³北海道大・保健科学研究院・機能回復学 Electromagnetic fields of varying intensities are commonplace in everyday life. With the advent of technology using stronger magnetic fields like 7 Tesla (T) for higher resolution brain imaging and using transcranial magnetic stimulation as a therapeutic strategy for neurological diseases, it becomes essential to understand their effects on normal neuronal function in detail. Previous reports suggested that exposure of the brains of normal human subjects to static magnetic fields (SMF) reduced neuronal excitability in the motor cortex. This effect persisted several minutes after termination of the exposure. Using 350 μm thick acute brain slices of mice including the motor cortex, we examined in detail the effect of SMF exposure on the excitability of layer II/III pyramidal neurons in the motor cortex with the whole-cell patch-clamp technique. The slices were obtained from 3 weeks old C57BL/6J male mice, and 0.3 T SMF was applied with a neodymium (NdFeB) magnet for 30 min. Using a standard low Cl^- (14 mM) patch pipette solution, we found that SMF exposure reduced membrane input resistance without changes in resting membrane potential (RMP) at 10 min after the exposure in pyramidal neurons. This caused the increase in minimum threshold current for action potential (AP) generation, i.e. rheobase, and the decrease in frequency of AP spikes in response to a given current stimulus. In addition, some neurons appeared to swell after the exposure. These effects were attenuated at 20 min. The AP waveform was, however, unchanged by the exposure. When using a high Cl^- (154 mM) pipette solution, we found that RMP was significantly depolarized at 10 min after SMF exposure. The increased rheobase and the decreased AP frequency were reproduced in this condition. However, the amplitude and the slope of upstroke of APs were reduced, while the downstroke and half-width of APs were unchanged, in this condition. These results strongly suggest that SMF temporarily suppresses the excitability of pyramidal neurons by enhancing the membrane Cl^- conductance of these neurons.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-093 脊椎動物の電位依存性Naチャネルの集積機構の起源 Yasushi Okamura（岡村康司）¹，Yuka Jinno（神野有香）¹，Masaki Hashimoto（橋本真宜）¹，Junko Nishino（西野純子）²，Yasunori Sasakura（笹倉靖徳）³，Atsuo Nishino（西野敦雄）²，Takafumi Kawai（河合喬文）¹ ¹大阪大学院医生理 ²弘前大学農学生命科学生物学科 ³筑波大学下田臨海実験センター Voltage-gated sodium channel, Nav, plays fundamental roles in membrane excitabilities in neuron, muscle and endocrine cell. Totally ten ortholog genes encoding Nav alpha subunit exist in mammalian genome. It has been suggested that diversity of Nav alpha subunit genes resulted from gene duplications of a single ancestral gene (Nav1), whereas Nav2 is only found in invertebrates and in invertebrates as well as in cartilaginous and sarcopterygian fish. Mammalian Nav1 forms clusters through interacting with an anchoring protein, ankyrin, at specialized sites such as the node of Ranvier or the axon initial segment in neurons and intercalated discs of cardiac muscles, playing key roles in regulation of electrical signals. Clustering of Nav1 at these sites accelerate conduction velocity through forming discrete regions (node and internode) in myelinated fibers and clustered region and unclustered region in nonmyelinated fibers. Tuning of clustering of Nav1 in the axon initial segment is important for adaptation of neuronal excitability in response to changes of input signals. Despite such functional importance of Nav1 cluster, it remains unclear how machinery for such Nav1 clustering was established during animal evolution. The simple nervous system of a sea squirt, Ciona intestinalis, which belongs to chordates, exhibits structural and evolutional relationship to vertebrate CNS, such as neural tube formation as well as neural crest or placode-related cells. In a previous study, we reported that sea squirt Nav1 gene, CiNav1a, exhibited rapidly activating Na⁺ currents upon heterologous expression in Xenopus oocyte. In situ hybridization analysis showed that CiNav1a is specifically expressed in neurons. In this study, we examined pattern of protein expression of CiNav1a in neurons by immunostaining using polyclonal antibodies to CiNav1a. Interestingly, we observed some discontinuous patterns of clusters on sea squirt neurons which are reminiscent of patterns of Nav1 clusters on mammalian unmyelinated axons. We are currently studying detailed protein distribution of CiNav1a and its relationship with neuronal cell types.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-094 報酬行動を規定するKCNQチャネルの分子メカニズム Daisuke Tsuboi（坪井大輔）¹，Tomoki Nishioka（西岡朋生）¹，Takushi Shimomura（下村拓史）²，Takeshi Otsuka（大塚岳）³，Yoshihiro Kubo（久保義弘）²，Yasuo Kawaguchi（川口泰雄）³，Kozo Kaibuchi（貝淵弘三）¹ ¹名古屋大医神経情報薬理 ²生理研分子生理神経機能素子 ³生理研大脳皮質機能大脳神経回路論 Dopamine plays a key role in the modulation of the circuit activity in striatum for reward behavior. We have reported that dopamine type 1 receptor (D1R) signaling in the striatum presumably regulates neuronal excitability and reward-related behaviors through PKA/Rap1/MAPK pathway. However, how D1Rs and its downstream signaling regulate neuronal excitability and behavior remain largely unknown. We focus on the post-modification of ion channels for neuronal excitability and reward behavior because protein phosphorylation of ion channels is vital for neuronal function. In this study, we identified a voltage-gated potassium channel, KCNQ2, as a phospho-candidate that is regulated by D1R signaling. Phosphorylation of KCNQ2 by MAPK cascade altered the open probability of KCNQ2/3 channels in Xenopus oocyte. The expression of phospho-defective mutants of KCNQ2 suppressed the functional modulation of KCNQ channel by MAPK. D1R agonist, SKF38393 caused a decrease in KCNQ-sensitive current in striatal slices, whereas D2R agonist, Quinpirole did not cause the effect. These results suggest that D1R signaling controls the channel activity of KCNQ via its phosphorylation for neuronal excitability and reward behavior.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-095 大脳皮質第５層におけるシナプス特異性形成の時系列解析 Nao Nakagawa（中川直）^1,2，Yoshiaki Tagawa（田川義晃）¹，Toshihiko Hosoya（細谷俊彦）² ¹鹿児島大学大学院医歯学総合研究科神経筋生理学分野 ²理化学研究所脳神経科学研究センター局所神経回路研究チーム The developmental processes that generate the specificity of neocortical synaptic connections are not well understood. In particular, many neocortical neurons initially generate excessive synapses and later prune unnecessary ones, and the specificity of these processes is unclear. Neocortical layer 5 contains two major types of pyramidal neurons; subcerebral projection neurons (SCPNs) innervating subcortical targets, and callosal projection neurons (CPNs) innervating the contralateral cortex. In adults, mutual connections between these pyramidal neurons exhibit highly specific patterns. For example, SCPNs have significantly frequent bidirectional connections (Morishima et al., 2011). In addition, connections between SCPNs and CPNs are biased in the direction from CPNs to SCPNs (Morishima and Kawaguchi, 2006; Brown and Hestrin, 2009). One hypothesis regarding the development of these specific connections is that excessive synapses are initially generated randomly and later deleted in a specific manner. Another possibility is that the initial excessive synapse formation is already specific. To test these hypotheses, we examined the time course of synapse formation after the beginning of the second postnatal week, when the formation of mutual connections initiates. We found that SCPNs temporarily make strong and excessive synapses on SCPNs; the connection probability between SCPNs rapidly increased during the second postnatal week and reached to >30%, and then decreased to <10% in adults. In contrast, SCPNs made synapses on CPNs only with the probability of ~10%, and the probability of CPN-CPN connection also remained low (~10%). Moreover, SCPNs had significantly high probability of bidirectional connections at the latter half of the second postnatal week. In contrast to the adult stage, during the second postnatal week, the connection probability between SCPNs and CPNs was similarly low in both directions. These results suggest that excessive synapse formation occurs in a cell type-specific manner, that preferential bidirectional connections develop before pruning of excessive synapses, and that the bias in connection between different cell types is created at stages later than excessive synapse formation. Thus, different specificity in neocortical synaptic connections may be generated by different processes in excessive formation and pruning of synapses.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-096 超高解像度顕微鏡を用いた神経筋接合部シナプス小胞放出部位、アクティブゾーン、の解析 Hiroshi Nishimune（西宗裕史）¹，Yomna Badawi（Badawi Yomna）¹，Toshihisa Ohtsuka（大塚稔久）²，Kazuhiro Shigemoto（重本和宏）³ ¹Dept. Anat. & Cell Biol, Univ of Kansas Sch of Med, Kansas City, USA ²山梨大院医生化学一 ³都健康長寿医療セ研老年病・運動器医 Synaptic transmission requires the release of neurotransmitters by fusing synaptic vesicles at presynaptic active zones. We used stimulated emission depletion (STED) microscopy to reveal the molecular architecture of active zones at sub-diffraction limited resolution. We previously reported a molecular mechanism for organizing the active zones at mouse neuromuscular junctions (NMJs) involving, synapse organizer laminin β2, presynaptic voltage-gated calcium channels (VGCCs), and active zone-specific protein bassoon. Laminin β2 is secreted from muscle, binds specifically to P/Q-type VGCCs, and VGCCs bind directly to active zone-specific proteins called cytomatrix of active zones (CAZ). Here, the new STED images indicated that these molecular interactions are likely taking place in vivo at NMJs. The data suggest that laminin β2 anchors VGCCs in front of postsynaptic junctional folds, and VGCCs function as scaffolding proteins for accumulating CAZ proteins, which together organize active zones of NMJs. Furthermore, STED microscopy data revealed the organization of CAZ proteins at sub-diffraction limited resolution. An unexpected finding was non-overlapping localization of the active zone proteins Bassoon and Piccolo in NMJs. Piccolo puncta sandwiched a Bassoon punctum in a side-by-side pattern, which were not resolved previously using confocal microscopy. P/Q-type VGCCs localized as a small punctum in between Piccolo and colocalized with Bassoon. We quantified the distribution patterns of other CAZ proteins, including ELKS, Munc13-1/2, Rim2, and Rim-BP. These CAZ proteins showed unique distribution patterns and differed from the three proteins mentioned above. These structural analyses suggested a unit structure of active zone organized by the CAZ proteins, which will be shown using an active zone model.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-097 視床体性感覚野における求心性シナプス前終末からの伝達物質放出機構 Mitsuharu Midorikawa（緑川光春），Mariko Miyata（宮田麻理子）東京女子医大院医第一生理 Somatosensory information from the maxillary region of rodent is conveyed to the ipsilateral trigeminal nuclei via the infraorbital nerve and then to relay neurons in the contralateral ventral posteromedial thalamic nucleus (VPM) via medial lemniscal fibers. During early postnatal development, the lemniscal fibers undergoes a number of morphological and functional changes. Previous studies including reports from our lab have shown that VPM relay neurons are innervated by multiple afferent fibers before the maturation, but synapses are eliminated upon the developmental maturation and eventually dominated by a single strong lemniscal fiber. During the maturation, it is also shown that the properties of the postsynaptic receptors takes place. However, our knowledge is largely limited to the pruning of the lemniscal fibers and the changes of postsynaptic receptors during the development. The kinetics of the transmitter release from the presynaptic lemniscal terminals have been estimated by postsynaptic excitatory currents recorded from the postsynaptic VPM relay neurons, but it remains largely enigmatic mainly because of the technical difficulty to apply direct measurement technique from the presynaptic terminal just as most of the neuronal presynaptic terminals. Here, we managed to measure the kinetics of transmitter release from medial lemniscal fiber presynaptic terminal by applying patch-clamp recording to the terminal. We applied capacitance measurement technique and paired recordings from a medial lemniscal fiber terminal-VPM relay neuron, and dissect the kinetics of transmitter release at the synapse as well as their developmental changes.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-098 Hapln4/Bral2欠損マウスの台形体内側核シナプスにおける形態学的および組織学的解析 Midori Edamatsu（枝松緑）¹，Tetsuya Hori（堀哲也）²，Toshitaka Oohashi（大橋俊孝）¹ ¹岡山大院医歯薬分子医化学 ²同志社大生命医科学神経生理 Perineuronal nets (PNN) are the reticular structure that surrounds the cell body and dendrites of many adult central nervous system neurons. PNN are formed during late development and known to play important roles in regulation of neuronal plasticity and protection. PNN are further hypothesized to create a charged milieu around the neurons and thus, might directly modulate synaptic activity. The scaffold of PNN is based on hyaluronan polymer chains which are anchored via hyaluronan synthase in the membrane of neuron somata and proximal dendrites. PNN are composed of hyaluronan, the chondroitin sulfate proteoglycans (CSPG) and the hyaluronan and proteoglycan binding link proteins (Hapln). Hapln is a key molecule in the formation and control of hyaluronan-based condensed perineuronal matrix in the adult brain. Previously, we showed that Hapln4/Bral2 is expressed mainly in the PNN of brainstem and cerebellum (Bekku et al., Mol Cell Neurosci, 2003). Hapln4/Bral2-deficient mice have attenuated PNN, but the overall levels of CSPGs are unchanged except for neurocan. We show that Hapln4/Bral2 deficiency markedly affected the localization of brevican in PNN, whereas no effect was seen on aggrecan (Bekku et al., J Comp Neurol, 2012). This is a genetic model of disorganized PNN via deletion of link protein gene. To investigate whether the lack of Hapln4/Bral2 induced changes in presynaptic terminals, we performed morphometric measurements of presynaptic terminal by fluorescent tracer. Furthermore, we performed whole-cell recording of postsynaptic responses from principal cells of the medial nucleus of the trapezoid body in mice brainstem slices. Our results may provide a clue to understand the functional significance of Hapln4/Bral2 in the calyx of Held synapse.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-099 前脳基底核コリン作動性ニューロンへのグルタミン酸性伝達に対するセロトニンによるP/Q型カルシウムチャネル依存的な調節 Takuma Nishijo（西條琢真），Toshihiko Momiyama（籾山俊彦）東京慈恵医大・薬理 Basal forebrain (BF) is an origin of cholinergic neurons which project to various brain regions including cortex and hippocampus. BF has been shown to be involved in attention, arousal, learning, and memory, as well as the related disorders, including dementia and Alzheimer's disease. Serotonergic fibers in the dorsal raphe nuclei project and innervate various brain areas including BF nuclei. However, serotonin (5-HT)-induced modulatory effects on the excitatory synaptic transmission in the BF are unknown. Therefore, this study was aimed to elucidate 5-HT-induced modulation of glutamatergic synaptic transmission onto BF cholinergic neurons. BF cholinergic neurons were identified with Cy3-192IgG injected into lateral ventricles at P8-9 and investigated in P12-20 rat brain slices using whole-cell patch-clamp technique. Pharmacologically isolated excitatory postsynaptic currents (EPSCs) were evoked by focal electrical stimulation. Bath application of 5-HT inhibited the amplitude of the evoked EPSCs. Both a 5-HT_1A receptor agonist and a 5-HT_1B receptor agonist inhibited the amplitude of EPSCs. A 5-HT_1A receptor antagonist or a 5-HT_1B receptor antagonist partially antagonized 5-HT-induced inhibition of EPSCs. In the presence of both antagonists, most of 5-HT-induced effect disappeared. 5-HT could still inhibit the EPSCs in the presence of ω-conotoxin GVIA, whereas 5-HT-induced inhibition was significantly smaller in the presence of ω-agatoxin TK than that without ω-agatoxin TK. While 5-HT had no significantly effect on paired-pulse ratio, 5-HT reduced synaptic strength by changing AMPA/ NMDA ratio. These results suggest that 5-HT inhibits glutamatergic transmission onto BF cholinergic neurons and that both 5-HT_1A and 5-HT_1B receptors contribute to 5-HT-induced inhibition. They also suggest that 5-HT inhibits glutamatergic transmission by selectively blocking P/Q-type calcium channels.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-100 ゼブラフィッシュ幼生のマウスナー神経細胞を制御するGABA作動性シナプスと回路の解析 Mio Aoki（青木澪）¹，Shunpei Baba（馬場俊平）²，Tomohiro Inoue（井上智裕）^1,2，Tsuyoshi Higashi（東毅）^1,2，Takayuki Sumimoto（角本貴進）^1,2，Takanori Ikenaga（池永隆徳）^1,2，Masataka Nikaido（二階堂昌孝）^1,2，Kohei Hatta（八田公平）^1,2 ¹兵庫県立大学大学院生命理学研究科 ²兵庫県立大学理学部 Mauthner cells (M-cells) are a pair of largest neurons of the fish brain, bilaterally located in the forth rhombomere. They gather information from somatic, auditory and visual sensations, to decide the timing and direction of the escape behavior. The axons of the M-cells cross the midline, descending the spinal cord to the tail to control the spinal motoneurons to perform C-shaped escape behavior. M-cell has been considered as an important model to analyze inhibitory circuits in vertebrates. Feedback and feed forward glycinergic inhibitory neurons have been well characterized. On the other hand, electrophysiological evidence suggested that M-cell are also strongly inhibited by tonic activity of GABAergic neurons and the characteristics of their IPSPs differ from the glycinergic ones (Hatta et al. 1999, 2001). However, little is known on the actual GABAnergic innervations on the M-cells. Here we visualized synapse boutons of GABAergic neuron by immunohistology as well as by using a glutamic acid decarboxylase 1b (gad1b) reporter line. By live-imaging Cre-induced red-green mosaic embryos, we find that there are nerve endings derived from, at least, three distinct GABAergic neurons on the lateral dendrite and the cell body of the M-cell. In addition, we have succeeded in tracing the neuronal soma from a single synaptic bouton, using photoconvertible fluorescent proteins, such as Kaede. We are currently using this technique to identify the neuronal circuits controlling the activity of the M-neurons and the animal behavior.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-101 小胞型グルタミン酸輸送体の発現量が中枢神経系シナプス伝達に与える影響 Yutaro Nakakubo（中久保佑太郎）¹，Saeka Abe（阿部冴華）¹，Masayuki Isa（伊佐真幸）²，Shigeo Takamori（高森茂雄）²，Tetsuya Hori（堀哲也）¹ ¹同志社大院生命医科学 ²同志社大・院・脳科学 After releasing neurotransmitter, synaptic vesicles are retrieved by endocytosis and recycled via fast or slow pathways to be reused for synaptic transmission. To maintain the synaptic efficacy, vesicles must be refilled with neurotransmitter during recycling. The vesicular glutamate transporters (VGLUTs) play an essential role in synaptic transmission by filling vesicles with glutamate. At the synapse in mammalian central nervous system, the expression level of VGLUTs may determine the speed of filling with glutamate into synaptic vesicles, thereby affecting the efficacy of neurotransmission. To address this question, we first examined the vesicle refilling rate directly at the calyx of Held-MNTB glutamatergic synapses in brain slice preparation derived from VGLUT1 knock-out (KO) mice using simultaneous presynaptic and postsynaptic whole-cell recording combined with caged glutamate photolysis. We found that the recovery of EPSC after uncaging was remarkably slower in VGLUT1 KO mice than in WT mice. However, several parameters including the mean amplitudes or shape of EPSCs evoked by single action potential (AP) as well as the mean amplitudes and frequency of spontaneous EPSCs remained unchanged. Next, we tested the effect of slowing of filling speed of neurotransmitter on the fidelity of high-frequency neurotransmission by recording postsynaptic APs in response to presynaptic APs elicited by 100 Hz stimulation. During a sustained 100 Hz stimulation, postsynaptic AP firing rates remained constant at least for 10 s in WT mice, whereas at synapse in VGLUT1 KO mice, AP generation started to fail within 3 s after the onset of stimulation. These results suggest that the sufficient amount of VGLUT expression is required for the maintenance of the high-frequency neurotransmission at the fast glutamatergic synapse.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-102 マウス脳におけるNMDA型グルタミン酸受容体サブユニットの定量的解析 Yasuhiro Suzuki（鈴木康浩）^1,2，Chihiro Nakamoto（中本千尋）^1,3，Manabu Abe（阿部学）^1,3，Toshikuni Sasaoka（笹岡俊邦）²，Kenji Sakimura（﨑村建司）^1,3 ¹新潟大脳研基礎神経科学細胞神経生物 ²新潟大脳研動物資源研究開発分野 ³新潟大脳研モデル動物開発分野 The NMDA-type glutamate receptors (NMDARs) are involved in activity-dependent changes of synaptic efficacy. The main NMDARs are composed of GluN1 and GluN2 subunits. The GluN1 subunit is encoded by a single gene, but exists as several splice variants. Focusing on these C-terminal side patterns, two types of GluN1, C2 and C2', have been identified. On the other hand, four GluN2A-2D subunits are encoded by separate genes. Although it is very important to know the quantity of each subunit in various brain regions for the understanding of NMDAR function, no quantitative analysis has been made. We newly developed a quantitative western blot method to determine the amount of two main variants GluN1-C2 and GluN1-C2' and four GluN2 subunits in the cortex, hippocampus and cerebellum. In the crude fractions, there were abundant GluN1-C2 and GluN1-C2' subunits in the cortex and hippocampus (1-C2 : 1-C2' : 2A : 2B : 2C : 2D = 1.0 : 2.38 : 0.82: 1.24 : 0 : 0.05 in the cortex, 1.0 : 3.8 : 1.1 : 1.78 : 0 : 0.09 in the hippocampus, respectively), whereas GluN1-C2' subunits were abundant in the cerebellum (1-C2 : 1-C2' : 2A : 2B : 2C : 2D = 1.0 : 10.3 : 1.5 : 0 : 3.0 : 0.63). On the other hand, in the synaptosomal fractions, the ratio of GluN1 to GluN2 subunits was almost the same in all the regions, showing that relative amounts of GluN1 subunits decreased during trafficking to synaptic membranes and resulted in the formation of functional GluN1/N2 (1:1). It still remains to be clarified why unnecessarily abundant GluN1 is produced in these regions. The amount of NMDA-type receptor subunit protein in the developmental stage was almost equal to that of mRNA calculated by in situ hybridization, but there were some differences.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-103 皮質-線条体投射刺激における線条体投射ニューロンの入出力特性 Ryo Inagaki（稲垣良）¹，Masato Sasagawa（笹川正人）¹，Makoto Osanai（小山内実）² ¹東北大院医 ²大阪大院医 It is known that striatum, a subcortical nucleus in the forebrain, regulates the higher brain functions including voluntary movement and reward learning. In the striatum, the projection neurons are divided into the two subtypes, dopamine D1 receptor type medium spiny neuron (D1-MSN) and the dopamine D2 receptor type MSN (D2-MSN). These two subtypes of MSN receive excitatory inputs from the cerebral cortex, which may in turn affect the activity of the basal ganglia. However, the properties of the responses in D1- and D2- MSNs to the cortico-striatal input still remain unclear. On the other hand, the cortical beta waves are observed at the time of planning and/or execution of a motion, and the striatal D1-MSN and D2-MSN show a contribution to the initiation and the termination of the motion, respectively. Therefore, we try to elucidate the responses of D1- and D2- MSN to the various frequency inputs from the cortex. For assessing the transient response of numerous D1- or D2- MSNs in striatal slices, we applied in vitro Ca2+ imaging method which widely used for probing action potentials. In addition, to characterize the D1 and D2-MSNs, we prepared transgenic mice expressing yellow fluorescent protein in D1 and D2- MSNs, respectively. As a result, the amplitude of the intracellular Ca2+ concentration ([Ca2+]i) elevation become larger depends on the stimulus frequency in both MSN. The D1- MSN showed significant increase [Ca2+]i transient rates in the stimulation of beta frequency band compared with that of D2-type MSN. Furthermore, application of the dopamine enhanced the amplitude of [Ca2+]i transients of D2-type MSN in beta frequency band stimulation. Thus, these results suggested that the two types of MSNs have different properties in the response to the cortico-striatal inputs, which modulated not only by the cortical excitatory inputs but also by the dopaminergic inputs.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-104 Direct recruitment of retrieved vesicle to the readily releasable pool is scaled with exocytic mode Yujin Kim（Kim Yujin），Unghwi Lee（Lee Unghwi），Sunghoe Chang（Chang Sunghoe） Seoul National University College of Medicine Sustained release of synaptic vesicles is limited by the rate of reloading of new vesicles to the readily-releasable pool (RRP). How neurons accommodate the demand of releasable vesicles, however, has not been answered. Here, using an axon-targeting glutamate sensor, iGluSnFRpre, we imaged the amount of glutamate released from individual hippocampal presynaptic terminals in response to a single action potential. We found that synapses changed their predominant exocytic mode from univesicular release (UVR) to multivesicular release (MVR) at high extracellular calcium levels. After vesicle release, the RRP was replenished on sub-second timescales through dynamin-dependent endocytosis. Evidently, MVR sped up RRP depletion, yet this is compensated by direct recruitment of recycled vesicles to the RRP. Indeed, glutamate depletion experiments followed by computational modeling suggested that ~70 % of retrieved vesicles are directly recruited to the RRP during MVR compared with only 7% during UVR. Together, our results indicate that presynaptic short-term plasticity plays a crucial role on the RRP refilling mechanism depending on the mode of exocytosis to meet the requirement that fusion-competent vesicles are immediately released.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-105 Presynaptic mitochondrial regulation of microdomain calcium and short-term plasticity Suk Ho Lee（Lee Suk Ho），Che Ho Yang（Yang Che Ho），Won Kyung Ho（Ho Won Kyung） Seoul National University College of Medicine. Mitochondria play important roles in generating ATP and calcium sequestration. However, little is known about the role of mitochondrial calcium dynamics in synaptic transmission. We have previously shown that 2 μM tetraphenylphosphonium (TPP+) specifically blocks mitochondrial Na+/Ca2+ exchanger (mNCX) without affecting the peaks and fast decaying phase of cytosolic calcium transients at the calyx of Held. Nevertheless, we found that TPP+ reduced short-term facilitation (STF) and steady-state EPSCs (EPSCss) evoked by a brief high frequency stimulation (HFS, 30 pulses at 100-300 Hz) at mature calyx of Held and hippocampal mossy fiber synapses under physiological extracellular [Ca2+] (1.2 mM). TPP+ had no effect on STP at immature calyces. When 50 μM EGTA-AM was pre-treated, however, TPP+ reduced STF and EPSCss, suggesting that low cytosolic [Ca2+] may be required for mitochondrial Ca2+ release via mNCX during HFS to modulate STP. The fast releasing vesicle pool (FRP) size, probed by a 3 ms depolarizing pulse, at the end of HFS was reduced by TPP+ in immature but not in mature calyx, indicating that low EPSCss in the presence of TPP+ is caused by low release probability at mature calyx. TPP+ had no effect on global cytosolic calcium ([Ca2+]i) during and after HFS. Monitoring mitochondrial [Ca2+] ([Ca2+]m) using 10 μM rhodFF-AM, we found that TPP+ enhanced the peak [Ca2+]m level caused by HFS under 0.5 mM EGTA, but not under 0.1 mM EGTA (mimicking physiological buffer at immature calyx). Moreover, TPP+ slowed down the decay of [Ca2+]m after a HFS, indicative of mitochondrial Ca2+ release inhibition. We investigated whether mitochondrial Ca2+ release regulates the recovery of FRP. TPP+ suppressed the recovery of FRP, which was occluded by calmodulin binding domain. To further study the role of mitochondrial Ca2+ release in Ca2+-dependent recovery, we measured capacitance (Cm) decay that reflects endocytosis. TPP+ significantly slowed down the decay of Cm. Our results suggest that mitochondrial calcium release under physiological [Ca2+]o enhances not only release probability but also recovery of FRP during and after a brief HFS by the increase of microdomain calcium.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-106 Disparities in Short-Term Depression Among Prefrontal Cortex Synapses Sustain Persistent Activity in a Balanced Network Jae Young Yoon（Yoon Jae Young），Hyoung Ro Lee（Lee Hyoung Ro），Won-Kyung Ho（Ho Won-Kyung），Suk-Ho Lee（Lee Suk-Ho） Seoul National University The prefrontal cortex (PFC) is a key cortical area for working memory. Activity of neuronal subsets in the PFC that represent cue information persists in the absence of cue during the delay period in working memory tasks. Such persistent activity has been regarded as a neural correlate of working memory. High temporal irregularity in the spiking patterns observed during delay activity suggests that persistent activity emerges as a network property of the PFC, rather than from intrinsic properties of individual neurons. Multiple network properties have been suggested from computational studies that may cooperatively support persistent activity. Nevertheless, it has not been tested whether short-term synaptic plasticity (STP) as observed experimentally from recurrent excitatory synapses of the PFC can sustain persistent activity in a cortical network model. Moreover, diverse forms of STP are exhibited by neocortical synapses of different presynaptic and postsynaptic identities, but it has not been studied whether STP at synapses other than excitatory recurrent ones contribute to persistent activity. In the current study, we characterized STP from synapses in layer 5 of the medial prefrontal cortex (mPFC) with identified presynaptic and postsynaptic cell types by optogenetically expressing the channelrhodopsin variant ChIEF which has fast and stable light activation kinetics, in conjunction with morphological and electrophysiological characterization of postsynaptic cells. Incorporation of STP at major synaptic types of the mPFC into a cortical network model revealed that the unique combination of little depression at recurrent excitatory synapses and stronger depression at other synapses in the mPFC could support the maintenance of persistent activity in a balanced network.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-107 マウス海馬長期増強（LTP)の光学測定 Yoko Tominaga（冨永洋子），Makiko Taketoshi（竹歳麻紀子），Takashi Tominaga（冨永貴志）徳島文理大神経研 The long-term potentiation (LTP) at the hippocampal circuit is the model of the activity-dependent changes in the input-output (I-O) relationship of a neural circuit and the cellular model of the learning and memory functions of the brain. It is crucial to capture and detect the modification of the LTP over the circuit to understand the higher functions of the brain, which is often the key to certain types of pathology. Optical imaging techniques to probe the membrane potential at every component of neurons, such as dendrites, axons, and somas, in the circuit are essential. We have developed fast single-photon wide-field optical imaging with a voltage-sensitive dye (VSD) for quantitative measurements. Here, we show that VSD imaging (VSDI) can be used to record changes in circuit activity in association with the LTP in the CA1 area. We achieved LTP measurements over 12 hrs (every 30 sec) with 0.1 ms/frame recordings. We also developed a method to evaluate circuit activity changes by mapping the variation in recordings from the CA1 area to coordinates defined by the morphology of CA1 pyramidal cells. The results clearly showed two types of spatial heterogeneity in LTP induction. The first heterogeneity is that LTP increased with distance from the stimulation site. The other heterogeneity is that LTP is higher in the stratum pyramidal-oriens region than in the stratum radiatum. We also showed that the pattern of heterogeneity changed according to the induction protocol, such as induction by theta burst stimulation (TBS) or high-frequency stimulation (HFS). We further demonstrated that part of the heterogeneity depends on the I-O response of the circuit elements. The LTP patterns are modified by other factors, such as the difference in the strain of the mouse (B6) and (DBA2). The robust evaluation of LTP should be useful in many applications.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-108 Homeostatic and heterosynaptic plasticity in hippocampal pyramidal neurons Thomas Chater（Chater Thomas），Yukiko Goda（Goda Yukiko） RIKEN Center for Brain Science Individual neurons must be able to respond to strong inputs in order to encode new information, and concurrently maintain their firing rates within appropriate boundaries. These two demands can be epitomized by input-specific Hebbian plasticity and cell-wide homeostatic plasticity. How these multiple mechanisms operate in single neurons is not well understood. In order to probe this question, here we imaged single dendrites on CA1 and CA3 rat hippocampal neurons, whilst subjecting them to both Hebbian and homeostatic plasticity. Single synapses were potentiated via glutamate uncaging, and concurrently synaptic scaling was triggered by chronic TTX or picrotoxin treatment. Remarkably, CA1 and CA3 spines responded differently to activity manipulation. During activity blockade with TTX basal spine size in CA1 was unaffected, while CA3 spines shrank. These changes in spine dynamics appear to be developmentally regulated, as CA3 neurons from older slices did not change size after TTX treatment. In both CA1 and CA3 neurons, glutamate-uncaging induced robust structural LTP, with an enhancement of the magnitude of early LTP in CA1, but not CA3 neurons. Finally, we have identified a subtle but consistent heterosynaptic spreading of plasticity following single-spine potentiation that leads to spine enlargement along the target dendrite. This occurs in both CA1 and CA3 neurons, and appears to be independent of previous network scaling manipulations. These results point to a set of interacting plasticity mechanisms that are determined by the cell-type and developmental stage in the hippocampal circuitry.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-109 ショウジョウバエの視神経におけるシナプスの安定化・不安定化シグナルのRNAiスクリーニング Jiro Osaka（小坂二郎），Yuya Kato（加藤由野），Yusuke Watanuki（綿貫雄介），Tomohiro Araki（荒木智裕），Satoko Hakeda-Suzuki（羽毛田-鈴木聡子），Takashi Suzuki（鈴木崇之）東工大院生命理工 In our previous research, it was found that the synaptic number of Drosophila photoreceptor decreases after their prolonged ambient light exposure (Sugie et.al. 2015). The number recovers after stopping the light exposure. We consider this reversible change as the indicator of synaptic modulations with natural stimuli. Moreover, it turned out that postsynaptic neurons send WNT signal to presynaptic neurons as a feedback signal and contribute to synaptic stabilization. We assumed that synaptic modulation is achieved by regulating not only synaptic stabilizing signals but also synaptic destabilizing ones. However, little is known about synaptic destabilizing signals. Furthermore, there may exist other synaptic stabilizing signals than WNT signal. We attempt to research these two kinds of signals by utilizing the phenomenon of synaptic number change we found. In this study, we will show the RNAi screening for searching synaptic stabilizing and destabilizing signals. We first conducted the screening for synaptic destabilizing signals. Since WNT signals transduce a feedback signal, cell-cell communication seemed to play an important role in synaptic modulations. Therefore, we consider transmembrane protein as important and conducted the screening for 209 genes. Six genes were selected as synaptic destabilizing genes. In the screening for the synaptic stabilizing signals, we performed the screening for synapse related genes as well as transmembrane genes. We conducted the screening in two steps. For the first screening, we utilized localization pattern of brp-short-mCherry as a simple indicator of synaptic stability since they make aggregation when flies are put on light condition (LL). Two Golgi body related genes, OstΔ(Oligosaccharide transferase Δ subunit) and KdelR (KDEL receptor) showed strong synaptic destabilization phenotype when they are knocked down. Furthermore, we will show the strategy of the screening and consideration for other genes we selected as candidates.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-110 脳出血モデルラットにおける運動機能回復に伴うシナプス変化 Chihiro Sato（佐藤ちひろ）¹，Kunikazu Tanji（丹治邦和）²，Shuji Shimoyama（下山修二）³，Mitsuru Chiba（千葉満）⁴，Kazuki Akahira（赤平一樹）¹，Junko Yamada（山田順子）¹ ¹弘前大学大学院保健学研究科総合リハビリテーション科学領域 ²弘前大学大学院医学研究科神経病理学講座 ³弘前大学大学院医学研究科脳神経生理学講座 ⁴弘前大学大学院保健学研究科生体科学領域 We previously reported that the exercise promotes the motor recovery in the intracerebral hemorrhage (ICH) model rats. This study aimed at further examining synaptic plasticity underlying the motor recovery with rehabilitative training. To clarify the synaptic function on the corticospinal tract of the injury side, DiI, the retrograde tracer, was injected into the right striatum. After 10 days training, coronal brain slices were obtained from ICH rats. Whole cell patch clamp recordings were performed from DiI labeled layer V cortical neurons in motor cortex (M1) using voltage -clamp mode (V hold = -70 mV). The amplitude and frequency of miniature excitatory postsynaptic currents (mEPSC) were recorded in the presence of TTX at 32 degrees. The amplitude and frequency of mEPSC were increased in both voluntary exercise (V-Ex.) and forced exercise (F-Ex.) groups than the non-exercise group. However, there were no differences in the F-Ex. and V-Ex. groups. To assess the motivation and stress levels with the training, the expression of ΔFosB in nucleus accumbens and the concentration of the plasma corticosterone were confirmed after training. The expression of ΔFosB was increased in the V-Ex. group than the F-Ex. and Non-Ex. groups. The plasma corticosterone was higher in the F-Ex. group. than the other groups. These data suggested that the recovery of motor function due to the synaptic plasticity in the motor cortex that project to the striatum. The promotion of synaptic functions might be affected by the motivation and stress with exercise.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-111 運動皮質損傷後機能回復における線条体のAMPA受容体の役割 Susumu Jitsuki（実木亨）¹，Yutaka Kawakami（川上裕）¹，Sayaka Kogami（湖上爽）¹，Aoi Jitsuki-Takahashi（實木-高橋葵）²，Takuya Takahashi（高橋琢哉）¹ ¹横浜市大院医生理 ²東京女子医大院医生化学 Brain damage such as stroke is a devastating neurological condition that may severely compromise patient quality of life. Restoration of motor impairment after brain damage is considered to be the result of compensative neural plasticity in intact brain regions, mediated by the reorganization of cortical motor maps. Experience-dependent synaptic AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic-acid) receptor (AMPAR) delivery underlies behaviors that require neural plasticity such as learning. We have previously found that rehabilitation-dependent synaptic delivery of AMPAR in the peri-injured area facilitate functional recovery after cortical damage. For execution of appropriate motor function, it requires involvement of motor-related brain regions such as striatum, thalamus or brain stem. However, it remains unclear that the role of other brain region in recovery after brain damage. Here, using cortical injury rat model, we found that AMPAR-mediated miniature EPSC (mEPSC) of the layer 5 pyramidal neurons in the periinjured cortex were positively correlate with recovery rate of forelimb reaching motor performance after rehabilitative training. On the other hand, mEPSC of the medium spiny neurons in the periinjured striatum were negatively correlate with recovery rate of motor performance. Furthermore, pharmacologically blockade of AMPAR in the peri-injured region inhibited recovery of forelimb reaching performance. These results suggest that AMPAR accumulation level in the peri-injured region could contribute functional recovery after cortical damage.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-112 海馬シナプス可塑性の時間枠を同定する新規光学技術 Akihiro Goto（後藤明弘）^1,2，Ken Miya（宮顕）^2,3，Tomoki Matsuda（松田知己）⁴，Takeharu Nagai（永井健治）⁴，Yasunori Hayashi（林康紀）^1,2,5 ¹京都大学大学院医学研究科 ²理化学研究所脳神経科学研究センター ³筑波大学　医学医療系 ⁴大阪大学　産業科学研究所 ⁵埼玉大学　脳末梢科学研究センター LTP is a cellular model of memory and plays a critical role in behavioral memory. To find out when and where LTP takes place and contributes to the memory formation in vivo, we developed a new optical method for erasing LTP. For this purpose, we introduced CALI (chromophore-assisted light inactivation) system. In this system, cofilin was fused with a photosensitizer protein SuperNova (CFL-SN) and light was illuminated to inactivate cofilin. Cofilin is one of actin binding proteins and we previously found that cofilin is transported to the spine in the initial phase of LTP and consolidates spine expansion. SuperNova (SN) is a protein of GFP family that generates reactive oxygen upon light illumination. To validate this system, a persistent enlargement of the spine (structural LTP) was induced with two-photon (2P) uncaging of MNI-glutamate in the single dendritic spine in the hippocampal slice culture, which expresses SN-fused cofilin (CFL-SN), and 559 nm laser light was subsequently irradiated. Followed by light illumination on the single spine 10 min after LTP induction, the spine volume gradually decreased. However, decrease in the spine volume was no longer observed when light was illuminated 50 min after LTP induction. Furthermore, the light illumination on the spine 1 min before sLTP had no effect on the spine enlargement. The light illumination on the spine without LTP induction had no effect on the spine volume. These results indicate cofilin is critical for maintenance of LTP up to 40 min after induction, and the inactivation of cofilin had negligible effect on LTP induction and the volume of unstimulated spine . To test system in memory formation, we expressed CFL-SN in hippocampal neurons by injecting AAV-floxed-CFL-SN into CA1 region of CaMKII-Cre mouse, then implanted an optical fiber above the virus injection site. Memory was examined by Inhibitory Avoidance (IA) task. Memory was significantly impaired when 593 nm light was illuminated on CA1 neurons expressing CFL-SN 2 min after an electrical shock at day 1. On the other hand, memory was not impaired when the light was illuminated either 1 min before or 1 hours after the electrical shock. These results indicate that this new method can specifically erase memory shortly (2 min) after memory formation without any effect on subsequent induction of memory.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-113 妊娠、出産、子育て経験による視床下部TIDAニューロンへの興奮性シナプス入力の影響 Miyako Furuta（古田都），Chiaki Kakehashi（掛橋千彰），Toshiya Funabashi（舩橋利也）聖マリアンナ医大医生理 Dopaminergic neurons located in the arcuate nucleus and adjacent periventricular region of the mediobasal hypothalamus, are called A12 tuberoinfundibular dopaminergic (TIDA) neurons which control prolactin (PRL) release from the anterior pituitary as an inhibitory factor. Intracerebroventricular administration of PRL induces maternal behavior in virgin female rats, suggesting that TIDA neurons are involved in controlling maternal behavior through PRL regulation. However, whether TIDA neurons per se play a role on maternal behavior remains unclear. We examined in the present study whether reproductive experiences affect the excitatory synaptic inputs to TIDA neurons which might be involved in maternal behavior in female mice. We previously determined that green fluorescent protein (GFP) was a reliable marker of TIDA neurons in transgenic mice expressed GFP under the control of the rat tyrosine hydroxylase gene (RBRC02095). In 16-weeks old primiparous and nulliparous transgenic mice (N=8-9), brain slices including TIDA neurons were made and trunk blood were collected in proestrous mice. They were subjected to whole-cell voltage- and current-clamp study, and PRL assay, respectively. TIDA neurons were identified by fluorescence microscopy. The frequency, but not amplitude, of miniature excitatory post synaptic potential (mEPSP) was augmented in primiparous mice compared to nulliparous counterpart. Plasma PRL concentrations of primiparous mice were significantly lower than those of nulliparous mice. Resting membrane potentials and the number of action potentials induced by current injection were not significantly different between the group. These results suggest that the excitatory presynaptic inputs to TIDA neurons may be changed by reproductive experiences and consequently be involved in the regulation of maternal behavior. Immunohistochemical analysis revealed that excitatory synapses on TIDA neurons expressed vesicular glutamate transporter 2 (vGlut 2). We are currently examining the number of synapses on TIDA neurons whether or not changed by reproductive experiences with confocal laser scanning microscopy.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-115 神経変性疾患家系における新しいKIF1Bβ変異はIGF1R輸送と軸索伸長を障害する Fang Xu（徐方）¹，Hironori Takahashi（高橋光規）¹，Yosuke Tanaka（田中庸介）¹，Sotaro Ichinose（一ノ瀬聡太郎）¹，Shinsuke Niwa（丹羽伸介）¹，Matthew P Wicklund（Wicklund P Matthew）²，Nobutaka Hirokawa（廣川信隆）^1,3 ¹東京大院医細胞生物・解剖 ²Dept Neurology, Penn State Hershey Medical Center, Hershey, USA ³CEGMR, King Abdulaziz Univ, Jeddah, Saudi Arabia Kinesin superfamily proteins (KIFs) largely serve as microtubule molecular motors that transport essential cellular materials to specific destinations. KIF1Bβ is a kinesin-3 family member that plays an indispensable role in neuronal survival, morphogenesis, and function (Zhao et al., 2001). Kif1b^-/- mice die soon after birth due to apnea. The brains of Kif1b^-/- mice are reduced in volume by ca. 10% compared with the brains of WT littermates. The cellularity, organization, and the development of the brain stem nuclei and commissural fibers are also significantly decreased. The dissociated hippocampal neuron culture shows severe neuronal death by day in vitro (DIV) 3, and KIF1Bβ expression restores neuron number. However, a distinct splicing isoform with a totally different cargo-binding domain, KIF1Bα, cannot restore the neuron number. In contrast, KIF1Bβ and its closely related motor protein KIF1A both transport synaptic vesicle precursors including synaptophysin, SV2, and synaptotagmin down the neuronal axons by binding to the dynamic adaptors Rab3A-GTP and differentially expressed in neoplastic versus normal cells protein (DENN)/MAPK-activating death domain (MADD). However, deficiency of these cargoes cannot sufficiently the severe neurodegeneration in Kif1b^-/- neurons from previous knockout phenotypes. KIF1Bβ mutations have previously been reported in a limited number of pedigrees of Charcot-Marie-Tooth disease type 2A (CMT2A) neuropathy. However, the gene responsible for CMT2A is still controversial, and the mechanism of pathogenesis remains elusive. In this study, we show that the receptor tyrosine kinase IGF1R is a new direct binding partner of KIF1Bβ, and its binding and transport is specifically impaired by the Y1087C mutation of KIF1Bβ, which we detected in hereditary neuropathic patients. The axonal outgrowth and IGF-I signaling of Kif1b^-/- neurons were significantly impaired, consistent with decreased surface IGF1R expression. The complementary capacity of KIF1Bβ-Y1087C of these phenotypes was significantly impaired, but the binding capacity to synaptic vesicle precursors was not affected. These data have supported the relevance of KIF1Bβ in IGF1R transport, which may give new clue to the neuropathic pathogenesis.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-116 軸索内の停止型ミトコンドリアの時間的・空間的制御 Yoshiyuki Konishi（小西慶幸）^1,2，Ikuma Hori（堀生実）¹，Nozomu Matsumoto（松本望）¹，Seiji Miyake（三宅誠司）^2,3 ¹福井大工生物応用化学 ²福井大ライフセ ³福井大医眼科 Neurons are capable to establish branched axonal arbors, and maintain their morphology even in culture without particular extrinsic cues. Molecular systems that control this cell-autonomous process remain to be elucidated. Axonal transport of mitochondria mediated by microtubules and their morters is crucial to deliver mitochondria throughout axonal arbors. In mature cortical neurons, only a part of axonal mitochondria are being transported via axonal transport, whereas majority of mitochondria are anchored to the microtubules via the function of anchoring protein, syntaphilin. Intriguingly, inhibition of syntaphilin results in the loss of axonal branches, suggesting that these stationary mitochondria play critical roles to maintain the axonal arbor shape presumably by locally providing the ATP. The intracellular systems that control corrective behavior of axonal mitochondria remain to be elucidated. We aimed to understand the system regulating the distribution of stationary mitochondria in axons. In cerebellar granule neurons, about 90 % of mitochondria were stationary at 7 DIV, but they constantly swithced to motile mitochondria with 90 min of half life. Compartmentalization analysis revealed that their spatial distribution is maintained as relatively uniform, whereas the position of each mitochondria is altered over the time. Local inactivation of mitochondria by mito-KillerRed affected the positioning of other mitochondria. These results raised the possibility that position of mitochondria in axon is regulated by inter-mitochondrial interactions. We will also discuss about the relationship between the mitochondrial position and presinaptic sites.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-117 成長円錐におけるF-アクチンは局所的エンドサイトーシス及び軸索ガイダンス受容体の局在に関係する Motohiro Nozumi（野住素広），Michihiro Igarashi（五十嵐道弘）新潟大院医歯分子細胞機能 Growth cones are the highly motile structures at the tip of extending axons in the developing or the regenerating neurons. The highly organized actin structure in the peripheral domain is important to the growth cone motility and its guidance. We recently revealed, using superresolution microscopy (SIM), that local endocytosis is associated with the F-actin-bundling at the leading edge of growth cones. The F-actin-dependent endocytosis most likely contributes to the membrane retrieval, containing the lipid rafts from the nonadherent/apical surface in the growth cone [Cell Rep 2017]. Using 3D-SIM, we also found that F-actin bundles were distributed along the apical surface, and some of them were protruding, not only towards the leading edge, but also the apical side. An axon guidance receptor neuropilin-1 was transiently concentrated in the nonadherent apical filopodia. Interestingly, synaptophysin and two endocytic components, endophilin A3 and dynamin 1, also were accumulated in the apical filopodia. These results suggest that the formation of apical filopodia promotes the neuropilin-1-associated local endocytosis effectively. Our model indicates that the growth cone has a cooperating, sensing mechanism as it physically pushes itself forward by actin polymerization.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-118 ドレブリンノックアウトによるNMDA受容体機能変化と微小管への影響 Noriko Koganezawa（小金澤紀子），Hiroyuki Yamazaki（山崎博幸），Tomoaki Shriao（白尾智明）群馬大院医神経薬理 Dynamic microtubule has an important role in the maintenance of dendritic spines and inhibition of microtubule growth affects spine morphology. It has known that depletion of EB3, a microtubule plus-end binding protein, also affects spine morphology. Drebrin regulates actin dynamics and has a critical role in synaptic plasticity. Microtubules are known to enter into spines in response to Ca2+ influx through synaptic NMDA receptors (NMDARs) and drebrin which directly binds to EB3 regulates this entry. Recently an interaction between drebrin and mDia2 fromin has been shown and some of the formins can bind microtubules and actin filaments. These facts suggest that microtubule dynamics couple with actin dynamics which is regulated by drebrin. Here, to investigate if drebrin regulates microtubule dynamics in dendrites, we used cultured neurons derived from drebrin knockout (DXKO) mice. Microtubule-associated protein 2 (MAP2) binds to microtubule and modulates microtubule stability. As EB3 binds directly to MAP2, we first observed MAP2 immunoreactivity using wild-type (WT) neurons and DXKO neurons. We detected less MAP2 positive neurons in DXKO neurons and also found that application of AP-5, an NMDAR antagonist, induced less immunoreactivity of MAP2 in WT neurons. This result suggests low functionality of NMDARs in DXKO neurons. We therefore examined the accumulation and expression level of NMDAR subunits and found that DXKO neurons had different pattern of their accumulation and expression level compared to WT neurons. Taken together, drebrin depletion affects NMDAR function and changes MAP2 immunoreactivity which might affect stability of microtubules. Further biochemical analysis has been in progress to reveal drebrin function in microtubule dynamics.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-119 髄鞘形成過程におけるミクログリアの関与 Tomoko Ishibashi（石橋智子），Kaori Nagasaka（長阪香），Takumi Kusayanagi（草柳匠美），Hiroko Baba（馬場広子）東薬大薬機能形態学 Microglia are resident macrophages in the brain and are the main active immune cells in the CNS. There is growing evidence that microglia also play a key role in myelinogenesis in the developing brain. Activated amoeboid microglia -and especially microglia that express integrin complement receptor CD11c - transiently accumulate in white matter during postnatal development. These early postnatal CD11c⁺ microglia regulate CNS myelination through insulin-like growth factor 1 (IGF-1) secretion (Wlodarczyk et al., 2017), but how exactly microglia-released IGF-1 promotes myelination - by direct action on oligodendrocytes or indirectly by promoting neuron or axonal survival - is not completely understood. Furthermore, it is also still unclear why and how microglia migrate to white matter before oligodendrocytes as well as how the timing of their disappearance from white matter after oligodendrocyte myelination is regulated. The present study aims to contribute to a better understanding of the physiological role of microglia in myelination by investigating the state of microglia in cerebella and optic nerves of developing animals. Activated CD68/Iba1-positive cells were observed in the myelinating distal white matter but not in the already myelinated area at P7 to P10 mouse cerebella. Iba1-positive microglial processes touched or surrounded MBP-positive myelinated axons in the myelinating area. Macrophage antigen complex (Mac2, also known as Galectin-3, a myelin phagocytic marker) positive cells were also observed in the myelinating distal area, but not in the myelinated white matter, and some Mac2-positive cells contained MAG-positive oligodendrocytes. In myelinating optic nerves, many Iba1-positive microglia were also transiently observed. Additionaly, mice with conditionally deleted Csf1r from microglia showed a significant loss of microglia in comparison to control mice (Nakayama et al., 2018), but no significant alterations in myelination, which suggests that other cells compensate for the lack of microglia. While these results suggest that microglia are not essential contributors in myelinogenesis, they may play an important role in creating the right environment for oligodendrocytes to make myelin.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-120 ヒトアストロサイトにおいてアラキドン酸はBDNFの発現を誘導する Satoshi Morita（森田賢），Toshiaki Sueyasu（末安俊明），Hisanori Tokuda（得田久敬），Yoshihisa Kaneda（金田喜久），Tomohiro Rogi（櫓木智裕），Hiroshi Shibata（柴田浩志）サントリーウエルネス株式会社　健康科学研究所 Arachidonic acid (ARA) and docosahexaenoic acid (DHA) are essential fatty acids. They are major polyunsaturated fatty acids (PUFA) in the brain phospholipids that decrease with age. Supplementation of ARA and DHA has been reported to improve the age-related deficits such as cognitive function. However, the mechanisms underlying this phenomenon has not been elucidated. Brain-derived neurotrophic factor (BDNF) is a protein that regulates memory formation, synaptic plasticity, and neuron survival. Interestingly, previous studies have reported relationships between BDNF and DHA. DHA ingestion was found to increase the expression of BDNF in the brain. Additionally, in cultured astrocytes, DHA increased the expression of BDNF via p38 MAPK. In the present study, we examined the possible roles of ARA and DHA in the expression of BDNF using cultured astrocytes derived from human cortex. Cells were cultured in astrocyte medium containing fetal bovine serum and astrocyte growth solution. Each fatty acids dissolved in ethanol was conjugated with bovine serum albumin (BSA). The purity of the cultured human astrocytes was more than 90%, which was examined using a well-known astrocyte marker protein. The composition of ARA and DHA in the phospholipid fatty acids was increased by the addition of each fatty acids in the culture medium. We found that DHA increased the expression of BDNF in human astrocytes. Besides, that was impaired by co-treatment of DHA with the p38 MAPK inhibitor. Furthermore, ARA also increased the expression of BDNF. The two fatty acids increased BDNF expression to approximately the same extent, which suggested important roles of ARA and DHA in the brain. The activity of p38 MAPK and CREB are expected to be the molecular mechanisms underlying the increased expression of BDNF. It is an interesting point how ARA induces the expression of BDNF and whether the molecular mechanisms are different compared to that of DHA. Further biochemical and molecular biological experiments are currently underway.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-121 脊髄の発達過程におけるグリシントランスポーター１(GlyT1)の発現変化 Chigusa Shimizu-Okabe（清水千草），Ryuji Tomoyose（友寄竜司），Kie Henman（平安山貴江），Shiori Kobayashi（小林しおり），Chitoshi Takayama（高山千利）琉球大学医学研究科分子解剖学講座 In the spinal cord, glycine and Gamma-amino butyric acid (GABA) are inhibitory neurotransmitters. Released glycine and GABA is removed from the synaptic cleft by glycine transporter1 (GlyT1) and GABA transporter 3 (GAT-3). We have previously investigated the developmental changes in GABA-removal system (Kim et al. 2014). However, the development of glycine-removal system in astrocytes remains unclear. The present study aimed to reveal the ontogeny of the glycine-removal system in the astrocytes by examining the immunohistochemical localization of GlyT1 in the embryonic and postnatal mouse cervical spinal cord. On embryonic day 12 (E12), GlyT1 was expressed in the mantle layer. GlyT1 was localized in the process of radial glia. On E14, GlyT1 was localized in the ventral horn. This expression pattern was different from GAT-3. Weak GAT-3 immunolabeling was localized to several radial fibers extending from the central canal to the pial surface on E12. On E14, GAT-3 was detected in the radial processes. After E18, GlyT1 was colocalized with GAT-3 in astrocytes. In our previous study, GlyT2, which uptakes glycine in presynaptic terminal, was first detected in the ventral horn on E16 after GlyT1 was expressed (Sunagawa et al. 2017). These results suggested that glycine and GABA were uptaken in different region of the radial glia in E12-14, but after that, both neurotransmitters were removed by the same astrocytes.Before the formation of glycinergic terminal, glycine-removal system in astrocytes might be ready.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-122 アストログリア由来MHCIは病理的変化により社会性および認知機能障害を誘発した Kazuhiro Hada（羽田和弘）¹，Akira Sobue（祖父江顕）¹，Wulaer Bolati（Bolati Wulaer）¹，Ito Norimichi（伊藤教道）¹，Akira Nakajima（中島晶）¹，Toshitaka Nabeshima（鍋島俊隆）^2,3，Taku Nagai（永井拓）¹，Kiyofumi Yamada（山田清文）¹ ¹名古屋大学大学院医療薬学 ²藤田医科大学先進診断システム探索研究部門 ³藍野大学 Major histocompatibility complex (MHC) plays an important role in the adaptive immune response. The MHC genomic region in the mouse, located on chromosome 17, is named H-2 and the genes within this region are classified into three distinct classes (I to III). The MHC class I (MHCI) genes, such as H-2K and H-2D in the mouse, are highly polymorphic, and the unique roles of MHCI molecules have been demonstrated in the central nervous system. Association studies have also implicated MHCI genes in several neuropsychiatric disorders such as schizophrenia. We have previously reported that polyriboinosinic-polyribocytidylic acid (polyI:C) treatment in neonatal mice results in an impairment of neurodevelopment, which is accompanied by the development of schizophrenia-like behaviors in adulthood. Neonatal polyI:C treatment increased expression of MHCI H-2K and H-2D mRNA levels in the medial prefrontal cortex (mPFC). In order to clarify the role of MHCI in astrocytes, we developed a mouse model expressing MHCI molecules in astrocytes of mPFC, using an adeno-associated virus (AAV) vector that expresses the transgene under the control of glial fibrillary acidic protein (GFAP) promoter. MHCI / H-2D or its soluble form MHCI (sMHCI) was secreted into the exosomes from astrocytes. The MHCI-expressing mice showed impairments in social interaction and novel object recognition tests, as well as in a touchscreen-based visual discrimination learning. The number of Iba1-positive microglial cells was significantly increased in the mPFC of MHCI-expressing mice compared with control mice. Astroglial expression of MHCI decreased the number of parvalbumin-positive cells and reduced the dendritic spine density of pyramidal neurons in the mPFC. Furthermore, a single treatment of atypical antipsychotic drug clozapine or repeated treatment of neutral-sphingomyelinase inhibitor GW4869 ameliorated these abnormal behaviors. Repeated treatment of GW4869 also improved neuropathological changes observed in MHCI-expressing mice. These results suggest that the exosomal MHCI derived from astrocytes affects the microglial proliferation as well as the neuronal number and spine density, thereby leading to behavioral dysfunctions in mice.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-123 ミクログリア選択的Cre発現マウスの作製と解析 Manabu Abe（阿部学）^1,2，Keisuke Watanabe（渡辺啓介）³，Fei Peng（彭菲）²，Keiji Sakimura（崎村建司）² ¹新潟大脳研モデル動物開発 ²新潟大脳研細胞神経生物 ³新潟大院医肉眼解剖学 Microglia, the resident immune cells of the brain, are known to play important roles in pathological and physiological conditions. Their main function, as immune cells, is to detect the signs of pathogenic invasion or tissue damage, and moderate the potential damage to the CNS and support tissue repair. In addition, recent findings have revealed that microglia are also involved in maintaining the neurodevelopment process and synaptic plasticity and controlling other neural functions. In order to investigate detailed functions of microglia, we have undertaken development of a conditional gene manipulation system in microglia using Cre/loxP recombination. We generated a knock-in mouse line expressing codon-improved Cre recombinase (iCre) under the control of the Aif1 (Allograft inflammatory factor 1, also known as Iba1) promotor. This Aif1-iCre line was established by insertion of iCre gene into the translation initiation site of the Aif1 in frame using C57BL/6N ES cells via homologous recombination. Resulting Aif1-iCre mice were viable, grew normally, and were fertile. By crossing the Aif1-iCre line with the tdTomato reporter mouse line, we investigated its recombination pattern in the brain. We found that tdTomato fluorescence was detected in the almost cells expressing Iba1 protein, indicating that conditional recombination was performed in a very large population of microglia. Although the recombination seemed to be highly microglia-selective, we also detected fluorescence in a minor population of excitatory neurons, which express a neuronal marker NeuN but contain no inhibitory transmitter GABA, in the forebrain and cells in capillary vessel throughout the brain. It seems that the recombination in these cells was due to transient expression of Cre recombinase at an early stage of development because no immunoreactivity for Iba1 was detected in the adult brain. However, no tdTomato fluorescence was overlapped with the immunofluorescence for astrocyte marker GFAP, glial fibrillary acidic protein, or oligodendrocyte marker APC, adenamatous polyposis coli. Thus, the Aif1-iCre line on the C57BL/6N background will be a useful genetic tool to manipulate gene expression in microglia without affecting the functions of other glial cells.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-124 Ensheathing Gliaからの小胞性グルタミン酸放出が、連合学習における嫌悪情報を伝達する。 Tomoyuki Miyashita（宮下知之）¹，Kyohei Mikami（三上恭平）²，Kentaro Endo（遠藤堅太郎）²，Takaaki Miyaji（宮地孝明）³，Sawako Moriyama（森山佐和子）^3,4，Kotaro Konno（今野幸太郎）⁵，Yoshinori Moriyama（森山芳則）⁴，Masahiko Watanabe（渡辺雅彦）⁵，Minoru Saitoe（齊藤実）¹ ¹東京都医学総合研究所　学習記憶プロジェクト ²東京都医学総合研究所　基盤技術研究センター ³岡山大学　自然生命科学研究支援センター ⁴松本歯科大学　口腔生化学講座 ⁵北海道大学大学院医学研究院　解剖学分野　解剖発生学教室 Recent studies demonstrate that glial cells release gliotransmitters to modify synaptic transmission. However, release mode and function of gliotransmitters are still largely unknown. We here identified a novel Drosophila vesicular glutamate transporter (DVGLUT2) specifically expressed in the Drosophila ensheathing glia, which surround neuronal structures including mushroom body (MB), a neuronal center for association of CS and US information. Electron microscopic study shows synaptic vesicle-like structure in ensheathing glia, and invasion of tentacles of ensheathing glia into the MB neuropils. Acute knocking down of DVGLUT2 in glial cells but not in neuronal cells impaired aversive olfactory conditioning, pairing an odor with electrical shocks. Synaptic-GRASP analysis demonstrated significant trace of vesicular exocytosis from ensheathing glia to the MB compartments, which form synapses with mushroom body output neurons (MBON) that mediate escape behaviors. Using glutamatergic vesicular exocytosis probe, dvGluT2::pHluorin, and glutamate sensor probe, iGlusnFR, we found that electrical shock presentation induces vesicular exocytosis in ensheathing glia and increases extracellular glutamate level in the MBs that is blocked by inhibiting vesicular exocytosis in ensheathing glia. Notably, we observed Ca²⁺ responses in the MB upon US electrical shock presentation alone only under low extracellular (0.5 mM) Mg²⁺ condition and these responses were blocked by NMDA receptor (NR) blocker as well as inhibiting vesicular exocytosis in ensheathing glia. We also observed shock-evoked Ca²⁺ responses in the MB in combination with CS odor presentation. Given that Mg²⁺ blockade of NR in the MB is removed by depolarization of MB neurons, we propose a following model; while US electrical shock evokes vesicular exocytosis of glutamate from ensheathing glia to the MBs, released glutamate induces Ca²⁺ influx via NR only when postsynaptic MB neurons are depolarized by presentation of CS odor stimuli. Our data suggest that glial cells not only modify synaptic transmission but also play primary role in sensory transmission.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-125 酪酸はアストロサイトのLPS誘導性NO産生を増強する Mitsuaki Moriyama（森山光章），Hideyo Satoh（佐藤秀世），Katsura Takano（高野桂），Yoichi Nakamura（中村洋一）大阪府立大学　獣医　統合生理学 Cumulative evidence indicates that short-chain fatty acid attenuates inflammatory response in macrophages and adipocytes. Among them, butyrate, a four-carbon short-chain fatty acid produced by microbial fermentation, has been reported to suppress microglial inflammation through inhibition of histone deacetylase. Since the effect of butyrate on inflammatory response in astrocytes has not been fully understood, we examined the effect of butyrate on nitric oxide (NO) production in cultured astrocytes, which is experimentally stimulated by lipopolysaccharide (LPS). Butyrate augmented LPS-induced NO production with a concomitant increase in expression of inducible NO synthase (iNOS), although cell viability was not affected. Butyrate also increased LPS-induced production of intracellular reactive oxygen species (ROS) and proinflammatory cytokines. Butyrate did not affect LPS-induced activation of MAPKs. Addition of AR420626, agonist of G protein-coupled receptor (GPR) 41 which is a butyrate receptor on the plasma membrane, also potentiated LPS-induced expression of iNOS and production of NO and ROS. Butyrate-induced increase in NO production was partially attenuated in the presence of β-hyrdroxybutyrate, GPR41 antagonist. These findings suggest that augmentation of NO production by butyrate may exacerbate glial damage during neuroinflammation. Such changes may occur partially via GPR41-dependent pathway.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-126 アストロサイト特異的Tsc1ノックアウトマウスの社会行動異常はRheb阻害により回復する Tadayuki Shimada（島田忠之），Hiroko Sugiura（杉浦弘子），Kanato Yamagata（山形要人）（公財）東京都医学総合研究所　シナプス可塑性プロジェクト Mutations in the Tsc1 gene cause tuberous sclerosis complex (TSC). TSC patients harbor hamartomas in the brain and other organs. The neuropsychiatric symptoms of TSC patients include refractory epilepsy, autism spectral disorder and mental retardation. To investigate if the brain astrocytes contribute to the neuropsychiatric symptoms of TSC patients, we developed astrocyte-specific Tsc1 knockout mice (GFAP-Tsc1 cKO mice) and examined their phenotypes. Sociability was evaluated by 3 chamber test. GFAP-Tsc1 cKO mice showed impaired social memory. Immunohistochemical analyses revealed that GFAP-positive cells were increased in the brain, especially in the hippocampus, piriform cortex and amygdala. Moreover, the aberrant dendritic spines ware observed in dentate gyrus of the hippocampus, where one of the most severe gliosis was observed. These results suggest that the astrogliosis is involved in the development of neuropsychiatric symptoms of TSC. Because Tsc1 protein makes a complex with Tsc2 and inactivates Rheb protein together, Rheb could be activated in the astrocytes in GFAP-Tsc1 cKO mice. We tested whether activated Rheb is involved in the pathogenesis in GFAP-Tsc1 cKO mice. Treatment with Rheb inhibitors improved impaired social memory of the cKO mice. Gliosis and spine malformation in the dentate gyrus were also normalized by Rheb inhibitor administration. In addition, the cultured astrocytes from cKO mice induced abnormal spine shape in the cultured wild type neuron, and Rheb inhibitor recovered astrocyte-dependent abnormal spine morphogenesis in vitro. Thus, activation of the Tsc/Rheb signaling in the astrocytes could induce astrogliosis, affecting the spine morphology. The spine deformity may cause deterioration of neural function and would develop abnormal social behaviors observed in a mouse model of TSC.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-127 アセチルコリン神経系によるマウスアストロサイトのKir4.1および神経栄養因子BDNFの発現調節 Saki Shimizu（清水佐紀）¹，Masato Kinboshi（金星匡人）^1,2，Yukako Nakamura（中村優茄子）¹，Nishiki Oda（織田にしき）¹，Akio Ikeda（池田昭夫）²，Yukihiro Ohno（大野行弘）¹ ¹大阪薬大・薬品作用解析学 ²京都大院医・てんかん・運動異常生理学 Astrocytes are the most abundant glia cells and regulate neuronal excitability by maintaining ion homeostasis, metabolizing neurotransmitters and secreting neuroactive substances. Recent evidence illustrates that astrocytic brain-derived neurotrophic factor (BDNF) expression is specifically modulated by inwardly rectifying potassium (Kir) channel subunit Kir4.1 channel, which mediates the spatial potassium buffering function of astrocytes. Our previous study showed that blockade or expressional knockdown of Kir4.1 elevate BDNF expression in the primary culture of mouse astrocytes (Front. Mol. Neurosci., 10, 408, 2017). In order to explore the neural factors influencing on the Kir4.1-BDNF system, we investigated the effects of acetylcholinergic agents on mRNA expression of Kir4.1 and BDNF in primary cultured astrocytes of mice. Treatment of astrocytes with acetylcholine (3-30 μM) inhibited Kir4.1 expression and increased BDNF expression in a concentration-related manner. Both inhibition of Kir4.1 and enhancement of BDNF expression by acetylcholine (10 μM) were antagonized by mACh antagonist atropine (3 μM), but not by nACh antagonist mecamylamine (30 μM). In addition, inhibition of Kir4.1 expression by acetylcholine was significantly antagonized by the mACh M₁ selective antagonist pirenzepine (10 μM), which also inhibited acetylcholine-induced BDNF expression. The present results strongly suggest that acetylcholine inhibits Kir4.1 channel expression and increases BDNF expression via the activation of mACh M₁ receptor in astrocytes.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-128 シャルコー・マリー・トゥース病モデル L-MPZ マウスにおけるミエリン形成過程の解析 Yoshihide Yamaguchi（山口宜秀），Yoshinori Otani（大谷嘉典），Masayuki Takehara（竹原雅之），Tetsuhiro Nakajima（中島鉄博），Takuro Narazaki（楢崎琢朗），Jingjing Cui（崔晶晶），Hiroko Baba（馬場広子）東薬大薬機能形態学 Large myelin protein zero (L-MPZ) is a translational readthrough isoform of myelin protein zero (P0, MPZ). Recently, translational stop codon readthrough has become a key mechanism to modulate canonical gene function in mammals. Previously, we reported that abnormal increase of L-MPZ caused Charcot-Marie-Tooth (CMT) disease-like neuropathy including demyelination and axonal damage, suggesting that imbalance of translational readthrough may cause disease. However, the onset and progression of this neuropathy are still unclear. In order to clarify these disease processes, immunohistological analysis of L-MPZ in normal development and morphological analysis of demyelination models were performed. In sciatic nerves prepared from postnatal day (P) 0-21 and adult ICR mice, L-MPZ-positive signals were colocalized with myelin basic protein (MBP) in compact myelin at any age. These signals were dramatically increased during P0-10 and after that became constant but decreased in lysolecithin-injected demyelinated fibers. Additionally, L-MPZ signals were enriched in Schmidt-Lanterman incisures (SLI) and paranodal regions in the adult myelinated fibers, while P0 was mainly in compact myelin. These results suggest that L-MPZ is involved in the formation and maintenance of the PNS myelin. Morphological analyses of P7-28 of a mouse line (L-MPZ mouse) synthesized only L-MPZ demonstrated that thinner myelinated fibers and immature Schwann cells were started to increase from P14. Thus, excessive increase of L-MPZ prevents maturation of myelinating Schwann cells in developmental process.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-129 ラット三叉神経中脳路核におけるアストロサイト Akira Kawata（河田亮），Shingo Maeda（前田信吾），Tomonori Inoue（井上知則），Takao Tsuboi（坪井孝雄），Kouta Watanabe（渡部浩太），Tomohiro Kato（加藤智弘），Naomi Miyagi（宮城直美），Kazuyoshi Higashi（東一善），Osamu Takahashi（高橋理）神奈川歯科大学　口腔科学講座　神経組織発生学分野 The mesencephalic trigeminal nucleus receives proprioceptive sensory afferent of the trigeminal nerve from the jaw-closing muscle spindles and the periodontal ligaments, and also send a part of their axons to the motor trigeminal nucleus. Also in recent years, elucidation of glial cell-neuron network advances, the fact that the glia cells in the hippocampus and the cerebellum have responded variously in neural network has been shown. But, detailed research concerning the distribution and role of glia cells in mesencephalic trigeminal nucleus and around has not been shown. The aim of this study was to investigate the distribution of astrocyte in mesencephalic trigeminal nucleus. In this study, we used two type of astrocyte marker, glial fibrillary acidic protein (GFAP: an intermediate filament protein that is expressed by numerous cell types of the central nervous system including astrocyte and ependymal cells during development.) and glutamate aspartate transporter (GLAST: predominantly expressed in the plasma membrane, allowing it to remove glutamate from the extracellular space. It has also been localized in the inner mitochondrial membrane as part of the malate-aspartate shuttle.). As a result, GFAP and GLAST immunoreactivity was observed in cells of many regions of the brain stem including Me5. Their processes made contacts with mesencephalic trigeminal nucleus neuronal cell body on the same focal plane. Thus, these results suggest possibility the GFAP and GLAST positive cells in mesencephalic trigeminal nucleus participate in jaw-jerk reflex and periodontal-masseteric reflex.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-130 下垂体アデニル酸シクラーゼ活性化ポリペプチド (PACAP) はアストロサイトにおける乳酸輸送を制御する Yuki Kambe（神戸悠輝）¹，Yu Nakashima（中島優）¹，Nguyen Thanh Trung（Trung Thanh Nguyen）¹，Norihito Shintani（新谷紀人）²，Hitoshi Hashimoto（橋本均）^2,3，Takashi Yoshitake（吉武尚）⁴，Jan Kehr（Kehr Jan）⁴，Takashi Kurihara（栗原崇）¹，Atsuro Miyata（宮田篤郎）¹ ¹鹿児島大院・医歯学総合・生体情報薬理 ²大阪大院・薬・神経薬理 ³大阪大院・連合小児発達 ⁴カロリンスカ研究所・薬理 Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide, and PACAP signaling could play an important role in fear learning and memory. On the other hand, it is previously suggested that astrocyte-neuron lactate shuttle (ANLS) contributes to many forms of neuronal plasticity in the central nervous system (CNS), which includes fear learning and memory. However, it is still unclear what kinds of neurotransmitters evoke ANLS activation. In this study, we aimed to examine possible involvement of PACAP signaling in ANLS in the brain region related to fear memory consolidation/retrieval such as medial prefrontal cortex (mPFC), amygdala and hippocampus in both in vivo and in vitro. In cultured forebrain astrocytes, PACAP dose-dependently decreased glycogen contained in the astrocytes, but increased lactate in the culture supernatants. In the passive avoidance test, a rodent model of fear memory, ablation of PACAP and injection of the PACAP6-38 (PACAP antagonist) significantly inhibited long-term fear memory formation. In addition, memory acquisition significantly decreased glycogen, but increased lactate in PACAP (+/+) hippocampus. In contrast, memory acquisition failed to increase the lactate in (-/-) mice. Interestingly, the present report firstly suggested that memory retrieval also evoked ANLS activation in (+/+) amygdala and hippocampus, but significant increment of lactate were not detected in the same regions of (-/-) mice. In vivo microdialysis study further revealed continuous increment of hippocampal extracellular lactate concentration after single i.c.v. PACAP injection. These results suggested that brain ANLS activation by PACAP signaling, and PACAP-evoked ANLS could contribute to fear memory formation.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-131 ミクログリアー神経幹細胞連関とそのメディエーターに関する検討 Yukari Mogami（最上由香里）^1,2，Kaede Baba（馬場楓）³，Takeshi Suzuki（鈴木岳之）³，Kazue Hoshikawa（干川和枝）¹，Yasunari Kanda（諫田泰成）⁴，Kaoru Sato（佐藤薫）¹ ¹国衛研神経薬理 ²山梨大院医工薬理 ³慶応大薬 ⁴国衛研薬理 Microglia are important for the brain homeostasis and immune responses. In our previous report, we identified new developmental roles of microglia, i.e., activated microglia in the early postnatal subventricular zone (SVZ) enhance neurogenesis and oligodendrogenesis via cytokines. We are speculating that there exist temporal- and environmental- specific factors that modulate developmental functions of microglia. We found that the neural stem cells (NSCs) transiently expressed fractalkine in the rat SVZ from day 4-10 and microglia accumulated to these NSCs. Furthermore, microglia are the only cells that express fractalkine receptors (CX3CR1) in the CNS. We therefore investigated the relationships among microglia, NSCs, and fractalkine in this study. When neurospheres comprised of NSCs were co-cultured with mildly-activated microglia, NSC apoptosis was induced and the expression level of fractalkine was increased, while the morphology of microglia changed to more like resting type. Compound 18a, a CX3CR1 inhibitor, changed the morphology of microglia to the ameboid type. Furthermore, both of compound 18a and anti-fractalkine antibodies increased the number of apoptotic NSCs. These results suggest that mildly-activated microglia induce NSC apoptosis, and NSCs interacted with mildly-activated microglia then produced fractalkine, therby calming microglial activation and suppressing the NSC apoptosis.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-132 オリゴデンドロサイトおよびオリゴデンドロサイト前駆細胞における２光子in vivo カルシウムイメージング Shouta Sugio（杉尾翔太），Riho Ono（尾野里穂），Hiroaki Wake（和氣弘明）神戸大学　大学院医学研究科　システム生理学分野 Neurons were sending axons and connecting various regions to carry the sensory/motor information. A part of axons was wrapped by processes of oligodendrocytes (OCs) and formed myelinated axons. Once axons were myelinated, its conduction velocities were dramatically increased, approximately 50-fold higher than those of un-myelinated axons. Since, myelin tightly holds the axons by their complex lamina structure under the electron microscopy, it had been believed that myelin does rarely turn over under healthy condition. However, recent studies have been unveiled that the oligodendrocytes could be a plastic cells that are dynamically adapted their cell fate, morphology and myelination to the changes in neuronal activities, which could plastically modulate neuronal conduction velocities to alter the activity pattern of neuronal populations. Although, the physiological significances of activity-dependent myelination have been known in recent years, while the bidirectional signaling between neurons and OCs (including oligodendrocyte precursors: OPCs) that regulates activity-dependent myelination (for instance, Ca²⁺ transients via neurotransmitter or gliotransmitter) has not yet studied. To examine the spatiotemporal patterns of Ca²⁺ transients that regulate activity-dependent myelination, here, we performed two photon in vivo Ca²⁺ imaging using a transgenic mouse that expressed a fluorescent Ca²⁺ indicator (GCaMP6f) in OC and OPCs, and then compared the changes of Ca^{2+^{frequency and amplitude at three different situations; awake, anesthesia and after a motor learning acquisition. We found that the Ca²⁺ responses in processes of OCs/OPCs were varied; approximately 50% of processes in an OCs/OPCs were positively correlated with the changes in neuronal activities, the others were not. Our finding suggested that the Ca2+ transients regulating activity-dependent myelination should be lie in the various responses.}}		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-133 Ultrastructural analysis of microglia-neuron interaction following social defeat stress using serial electron microscopy Hirotaka Nagai（Nagai Hirotaka）¹，Junichi Ueyama（Ueyama Junichi）¹，Midori Nagai（Nagai Midori）¹，Chisato Numa（Numa Chisato）¹，Nobuhiko Ohno（Ohno Nobuhiko）²，Mikio Furuse（Furuse Mikio）³，Tomoyuki Furuyashiki（Furuyashiki Tomoyuki）¹ ¹Div Pharmacol, Kobe Univ Grad Sch Med ²Div Histol and Cell Biol, Dept Anat, Jichi Medical Univ ³Div Cell Struct, Natl Inst Physiol Sci Animal studies using various stress models have shown that excessive environmental stress induces depression-like behaviors with concomitant atrophic changes of neurons especially in the medial prefrontal cortex (mPFC) and the hippocampus. Recent progresses have revealed that inflammatory responses associated with microglial activation in the brain are critical inducers of dendritic atrophy of the neurons and depression-like behaviors, but the mechanism how the stress-induced brain inflammation leads to the altered morphology of the neurons and possibly neuronal dysfunction remains unknown. In the present study, we aimed to examine the interaction between the brain immune cells, namely microglia, and the neurons in the mPFC by using three dimensional electron microscopy. We subjected male C57BL/6 mice to either single or repeated social defeat stress and analyzed the brains from those stressed mice or from control mice which did not receive the stress by serial block face-scanning electron microscopy (SBEM). The pixel size of the serial electron microscopy images was as low as 6 nm and the thickness of each image slice was 60 nm, yielding the voxel size of 6 x 6 x 60 nm³ in each stack of images with the size of 30 x 30 x 30 μm³. With these high-resolution images, we successfully identified tiny processes of microglia and neurons. By segmenting and reconstructing microglia, axons and dendrites, we found that many neuronal components were surrounded and sometimes completely phagocytosed by microglia after the stress. This finding illustrates the usefulness of serial electron microscopy by SBEM in analyzing cell-to-cell interaction at the ultrastructural level. Using this technique, we are currently investigating the identity of neuronal compartments phagocytosed by microglia after the stress as well as functional significance of this microglia-neuron interaction.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-134 プリナージックおよびトル様受容体4活性化ミクログリアにおけるP2Y₂・P2Y₁₃受容体の発現と機能 Izumi Hide（秀和泉）¹，Yoichiro Morita（守田洋一郎）¹，Hiroko Shiraki（白榊紘子）¹，Yuhki Yanase（柳瀬雄輝）²，Kana Harada（原田佳奈）¹，Shigeru Tanaka（田中茂）¹，Norio Sakai（酒井規雄）¹ ¹広島大院医歯薬保神経薬理 ²広島大院医歯薬保皮膚科学 Microglia, the resident immune cells of the CNS, are potently activated by extracellular nucleotides and pathogen-associated molecular patterns (PAMPs) through purinergic and Toll-like receptors (TLRs), respectively. We previously reported that lipopolysaccharide (LPS) induced upregulation of P2Y₂ receptor via TLR4 in rat primary microglia. However, it remains unknown whether P2 receptors are regulated through purinergic activation as well as TLR4 and how the upregulated P2 receptors modulate microglial function. In this study, we investigated the effects of LPS and ATPγS on the expression of P2 receptors and the involvement of upregulated P2Y₂ and P2Y₁₃ receptors in cytokine expression. Stimulation of microglia by LPS and ATPγS caused a rapid and transient increase in P2Y₁₃ receptor mRNA by 3 hours and induced a delayed increase in P2Y₂ receptor mRNA up to 9 hours after stimulation. ATPγS itself did not elicit anti-inflammatory IL-10 mRNA expression, but markedly enhanced LPS-induced IL-10 expression which was significantly inhibited by both selective P2Y₂ and P2Y₁₃ antagonists, AR-C118925 and MRS2211, respectively. Furthermore, the expression of another anti-inflammatory cytokine activin A, a member of TGF-β superfamily, was also induced by LPS and ATPγS, and that was suppressed by AR-C118925 and more potently by MRS2211. In contrast, LPS and ATPγS-induced inflammatory TNF expression was significantly inhibited by AR-C118925, but not by MRS2211. Taken together, TLR4 and purinergic activation upregulated P2Y₂ and P2Y₁₃ receptors and these receptors are differently involved in the regulation of cytokine expression in activated microglia; anti-inflammatory IL-10 and activin A expression is mediated via P2Y₂ and more preferably P2Y₁₃ receptor, whereas inflammatory TNF expression is regulated through mainly P2Y₂, but not P2Y₁₃ receptor. These results indicate the possible participation of P2Y₂ receptor in microglia-mediated inflammation and a novel role of P2Y₁₃ receptor in anti-inflammatory response of microglia.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-135 注意欠如・多動性障害における活性化ミクログリア：[¹¹]PK11195 PETによる検討 Masamichi Yokokura（横倉正倫）¹，Kiyokazu Takebayashi（竹林淳和）¹，Akiyo Takao（高尾晃世）²，Tatsuhiro Terada（寺田達弘）³，Tomoyasu Bunai（武内智康）³，Yasuhiko Kato（加藤康彦）¹，Etsuji Yoshikawa（吉川悦次）⁴，Masami Futatsubashi（二ツ橋昌実）⁵，Hidenori Yamasue（山末英典）¹，Yasuomi Ouchi（尾内康臣）³ ¹浜松医大医精神科 ²山口大医 ³浜松医大医生体機能イメージング教室 ⁴浜松ホトニクス ⁵浜松光医学財団 Attention-deficit/hyperactivity disorder (ADHD) is one of major neurodevelopmental disorders characterized by inattentiveness and hyperactivity/impulsivity. Although a lot of genetic, clinical, and neuroimaging studies in ADHD have been reported, the precise neurobiological mechanisms underlying this disorder remain poorly understood. Biochemical studies on peripheral cytokines suggested possibility of altered immunological process in ADHD. Recently, neuroinflammation in central nervous system (CNS) has been reported to be involved in the pathophysiology of many psychiatric disorders. Especially, a number of studies have focused on a role of activated microglia in the neuroinflammatory response in a living human brain. Previous in vivo positron emission tomography (PET) studies have revealed elevated microglial activation in other psychiatric and developmental disorders (e.g. Alzheimer's disease, Autism spectrum disorder, Major depressive disorder, and Obsessive-compulsive disorder). To test a contribution of activated microglia to the pathophysiology of ADHD, by employing PET measurement. We investigated the microglial activation in individuals with ADHD compared with healthy subjects and any associations between the microglial activation and severity of ADHD symptoms. We recruited 24 drug-naive individuals with ADHD (mean age±SD, 32.3±8.0 years old) and 24 age- and sex- matched healthy subjects (32.1±8.6years old). All participants underwent PET measurement with radioligand [¹¹C]PK11195. Binding potential (BP_ND) of [¹¹C]PK11195 was estimated based on the simplified reference tissue model. We assessed symptom severities by using Conner's adult ADHD rating scale (CARRS), Wechsler adult intelligence scale 3rd edition (WAIS-III), and cognitive tasks for sustained attention, spatial working memory, and response inhibition from the Cambridge neuropsychological test automated battery (CANTAB). We tested a potential difference between microglial activation in the adults with ADHD and those in the healthy subjects and investigated any associations between the microglial activation and severity measures of ADHD. Our study might propose the involvement of activated microglia in the pathophysiology of ADHD and specific role of activated microglia on severity of ADHD symptoms.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-136 マウス全脳血管可視化手法の開発 Takeyuki Miyawaki（宮脇健行）^1,2,4，Etsuo Susaki（洲崎悦生）^2,3,4，Shun Yamaguchi（山口瞬）^5,6，Hiroki Ueda（上田泰己）^2,4,7，Yuji Ikegaya（池谷裕二）^1,8 ¹東京大学大学院薬学系研究科薬品作用学教室 ²東京大学大学院医学系研究科システムズ薬理学教室 ³JST さきがけ ⁴理化学研究所生命機能科学研究センター合成生物学研究チーム ⁵岐阜大学大学院医学系研究科高次神経形態学分野 ⁶岐阜大学研究推進・社会連携機構生命の鎖統合研究センター ⁷東京大学ニューロインテリジェンス国際研究機構 ⁸脳情報通信融合研究センター Cerebral vasculature is an anatomically and histologically inhomogeneous three-dimensional traffic network for blood cells, nutrients, oxygen, metabolic wastes, and signaling molecules, etc. Although structural and molecular analyses of cerebral vasculature are promising for providing fundamental insights into cerebral circulation and cerebrovascular diseases, which are major causes of death and disability, few studies have captured detailed structural information of molecularly identified vascular networks, due to technical limitations. Here, we introduce SeeNet, a method for near-complete three-dimensional visualization of intact cerebral vascular networks with high signal-to-noise ratios, compatible with molecular phenotyping. SeeNet employs perfusion of a novel multifunctional crosslinker, vascular casting by temperature-controlled polymerization of hybrid hydrogels, and a tissue clearing technique optimized for observation of vascular connectivity. SeeNet was capable of whole-brain visualization of molecularly characterized cerebral vasculatures at the single microvessel level. Moreover, SeeNet revealed a hitherto unidentified vascular pathway. SeeNet is quick, cost-effective, and reproducible. Thus, SeeNet will enable standardization of fine large-scale 3D tracing for molecularly identifying cerebral vasculatures, and advance our understanding of the structural and functional organizations of cerebrovascular networks.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-137 SRF転写コアクチベーターMRTFBの神経活動依存的核移行制御 Akiko Tabuchi（田渕明子）¹，Daisuke Ihara（伊原大輔）¹，Hiroki Tanabe（田邉広樹）¹，Shinnosuke Kato（加藤真之佑）¹，Shiori Imanishi（今西詩織）¹，Hiroyuki Sakagami（阪上洋行）² ¹富山大薬分子神経生物 ²北里大医解剖 Neuronal activity-dependent gene expression plays fundamental roles in brain development and function. In the last decades, intracellular signaling pathways including Ca²⁺ signaling and kinase cascade (synapse-to-nucleus signaling) triggered by neuronal activity have been identified and characterized to elucidate the molecular mechanism of activity-dependent gene expression. Accumulating evidence suggests that CREB and its cofactor, CREB-regulated transcriptional coactivator 1 (CRTC1) play central roles in regulating the expression of immediate early genes encoding synaptic proteins (Arc etc) and neurotrophins (BDNF etc). In this study, we demonstrate that, as well as CRTC1, a serum response factor (SRF) transcriptional cofactor, megakaryoblastic leukemia 2/myocardin-related transcription factor B (MKL2/MRTFB) translocates from the synapses to the nucleus in a neuronal activity-dependent manner. Our previous studies have shown that MKL/MRTF is highly expressed in the brain, is localized at synapses, and regulates dendritic morphology, synaptic maturation and Arcgene enhancer activity by using new antibodies that distinctly and specifically recognize endogenously expressed MKL1/MRTFA and MKL2/MRTFB proteins and RNAi-mediated knockdown of MRTF proteins. Here, by using these antibodies, we tested whether MKL2/MRTFB translocate from synapses to nucleus in cortical neurons. We found that membrane depolarization transiently induced nuclear translocation of MKL2/MRTFB. The nuclear translocation was also observed by stimulation with bicuculline/4AP, suggesting that synaptic activity mediates this process. Inhibitors for NMDA receptor and L-type voltage dependent Ca²⁺ channels completely blocked the nuclear translocation. Therefore, Ca²⁺ signaling via both Ca²⁺ channels is critical for the nuclear translocation. We further performed inhibitor experiments to identify which Ca²⁺ signaling pathway is required for the nuclear translocation. As a result, calcineurin phosphatase is involved. Furthermore, an inhibitor for actin polymerization, Latrunculin B, completely blocked activity-dependent nuclear translocation of MKL2/MRTFB. Taken together, these findings suggest that MKL2/MRTFB is another key player of the synapses-to-nucleus signaling not only in neuronal activity- and actin polymerization-dependent manner.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-138 TDP-43イントロンにおけるプロモーター領域の同定と機能解析 Minami Hasegawa（長谷川実奈美），Hirotaka James Okano（岡野 James 洋尚）慈恵医大総合医科学セ再生 TAR DNA binding protein 43kDa (TDP-43; encoded by TARDBP) is a major component of ubiquitin-positive neuronal inclusions in both amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). In recent years, it was revealed that more than 97% of ALS patients including familial and sporadic exhibit the TDP-43 pathology in motor neurons. While TDP-43 is well known to regulate RNA metabolism such as pre-mRNA splicing, mRNA stability and transport, TDP-43 is originally identified as a transcriptional regulator that binds to a regulatory element in the human immunodeficiency virus type1 (HIV-1) long terminal repeat known as TAR and suppresses HIV-1 gene expression. In previous study, TDP-43 transgenic mice showed progressive motor neuron death although TDP-43 knock out mice were embryonic lethal, indicating that TDP-43 expression must be strictly regulated in appropriate level and it is extremely important to examine the regulative systems. Although TDP-43 expression is regulated by negative feedback mechanism via alternative poly A site in 3' untranslated region, here we focused on transcriptional regulation of TDP-43. TDP-43 promoter region was shown by using promoter prediction database in previous study, however, no experimental study was demonstrated, thus it is unknown whether TDP-43 promoter is involved in the self-regulative systems. In this study, we identified TDP-43 promoter region and found its regulator candidate. More interestingly, we also found that TDP-43 intronic region possesses minor promoter activity. We hope that understanding transcriptional regulation of TDP-43 would be one of important clues to elucidate TDP-43 function.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-139 Vangl2との結合はPrickle2のユビキチン化とプロテアソーム依存性の分解を引き起こす。 Masashi Kishi（岸将史）¹，Tadahiro Nagaoka（永岡唯宏）²，Kinihiro Tsuchida（土田邦博）² ¹野崎徳洲会病院附属研究所精神・神経疾患研究部門 ²藤田医科大学　総合医科学研究所　難病治療学研究部門 The PET and LIM domain-containing protein, Prickle, plays a key role in planar cell polarity (PCP) in Drosophila. It has been reported that mutations in the PRICKLE2 gene, which encodes one of the human orthologues of Prickle, are associated with human autism spectrum disorder (ASD). To develop preventive and therapeutic strategies for the intractable disease, we studied the regulation of Prickle2 protein levels. Prickle2 levels were negatively regulated by a physical interaction with another PCP protein, Van Gogh-like 2 (Vangl2). The Vangl2-mediated reduction in Prickle2 levels was, at least in part, relieved by proteasome inhibitors or by functional inhibition of the Cullin-1 E3 ubiquitin ligase. Furthermore, the expression of Vangl2 enhanced the polyubiquitination of Prickle2. This ubiquitination was partially blocked by co-expression of a ubiquitin mutant, which cannot be polymerized through their Lys48 residue to induce target proteins toward proteasomal degradation. Together, these results suggest that Prickle2 is polyubiquitinated by the Vangl2 interaction in a Cullin-1-dependent manner to limit its expression levels. This regulation may play a role in the local and temporal fine-tuning of Prickle protein levels during PCP signal-dependent cellular behaviors such as scrapping and building of the mammalian synapses. The molecular interaction between Vangl2 and Prickle2 is a reasonable target for drug discovery for ASD.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-140 神経細胞における低温誘導性チロシンリン酸化シグナルの解析 Hiroshi Ohnishi（大西浩史），Mika Iino（飯野美香），Joji Kawasaki（川﨑穣二），Daiki Jingu（神宮大輝），Ayaka Motai（茂田井彩香），Kaori Imai（今井香織）群馬大院保生体情報 Mild hypothermia is known to be the therapeutic means for protecting the brain against ischemic, excitotoxic, or traumatic injury. However, cellular responses of neurons under hypothermic condition remains to be elucidated. SIRPα (Signal regulatory protein α) is a member of immunoglobulin superfamily membrane proteins. SIRPα contains tyrosine phosphorylation sites in its cytoplasmic region. Tyrosine phosphorylated SIRPα interacts with tyrosine phosphatase Shp1 or Shp2 and thereby activates its phosphatase activity. In the central nervous system, SIRPα is highly expressed in matured neurons. SIRPα undergoes tyrosine phosphorylation in the mouse brain in response to forced swim stress in cold water, and this signaling mechanism is implicated in regulation of the behavior (immobility) of mice during the forced swim. Hypothermia in cold water is the major cause of the induction of SIRPα phosphorylation during the forced swim. Furthermore, cold stress directly induces tyrosine phosphorylation of SIRPα in cultured neurons. Thus, tyrosine phosphorylation of SIRPα is a cold stress-responsive neuronal signaling. Here we further examined the effects of cold stress and another stress, extracellular sodium and chloride depletion, on tyrosine phosphorylation of SIRPα in cultured neurons. Our data suggest that intracellular Cl^-dynamics is a critical factor for the control of stress-induced SIRPα phosphorylation in cultured neurons. Cl^- channel-mediated cellular function, such as cell volume regulation, may be included in neuronal response to low temperature.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-141 ピロカルピン投与てんかんモデルマウス海馬におけるBRINPファミリー遺伝子の発現変化 Miwako Kobayashi（小林三和子），Daisuke Ochi（越智大輔），Eriko Uno（宇野恵梨子），Ichiro Matsuoka（松岡一郎）松山大学薬学部 BRINPs (BMP/RA-inducible Neural Specific Protein-1, 2, and 3) are family genes expressed in both the central and peripheral nervous system. BRINP1 is abundantly expressed in certain adult brain regions including cerebral cortex and hippocampus. Mice with disrupted BRINP1 gene exhibit abnormal behaviors such as increased locomotive activity and poor social activity which are analogous to symptoms of human psychiatric disorders such as schizophrenia (SCZ), autism spectrum disorder (ASD) and attention deficit hyperactivity disorder (ADHD). Among the three family members, BRINP1 expression in adult hippocampus is regulated in an activity-dependent manner by the intraperitoneal (i.p.) administration of kainic acid (KA), a glutamatergic agonist. In the present study, to elucidate physiological roles of BRINPs in hippocampus during epileptic conditions, we examined BRINP expression pattern in pilocarpine-induced seizure model mouse. Pilocarpine (380 mg/kg, i.p.) was administered to male C57BL/6J mice after pretreatment with scopolamine (0.9 mg/kg, i.p.) to minimize peripheral effect. After observation of continuous tonic/chronic convulsion in mice, diazepam (10 mg/kg, i.p.) was administered to terminate seizure. Mice administered scopolamine was used as control. Time-dependent mRNA expressions of BRINP family were determined by qRT-PCR using total RNA extracted from hippocampus of pilocarpine-treated mice. Both BRINP1- and BRINP2-mRNAs were increased by pilocarpine injection and their peak levels were sustained between 6 to 12 hrs after injection. On the other hand, BRINP3-mRNA levels were not changed by pilocarpine injection. We further investigated BRINP mRNAs expressions in hippocampus by in situ hybridization (ISH). Signals for BRINP1-mRNA were observed in both granule cell layer of dentate gyrus and pyramidal cell layer through CA1 to CA3 in control mouse hippocampus. In the pilocarpine-treated mouse hippocampus, signals of BRINP1-mRNA were increased in both granule and pyramidal cell layers and the increased levels in granule cells were prominent. Our previous study showed that BRINP1 was only member which was increased in hippocampus by KA-injection. In present study we found that BRINP2 was increased by pilocarpine-injection as well as BRINP1. Induction of robust BRINP1-mRNA expression in dentate granule cells in pilocarpine-treated mouse was confirmed by ISH.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-142 マウス視覚野における局所神経回路の違いによるPV陽性抑制性神経細胞の分類とクラスター型プロトカドヘリンガンマ欠損による影響 Nanami Kawamura（河村菜々実）¹，Etsuko Tarusawa（足澤悦子）¹，Yumiko Yoshimura（吉村由美子）²，Takeshi Yagi（八木健）¹ ¹大阪大院生命機能 ²生理研 The specificity of synaptic connections in the sensory cortex is fundamental for the proper processing of sensory information. It has been shown that excitatory and fast-spiking inhibitory neurons form neural networks on the basis of the specific synaptic connections in the rat visual cortex. However, the molecular mechanisms of the formation of specific neural connections between excitatory and inhibitory cortical neurons have not been elucidated yet. Our previous study has shown that reciprocal connections formed preferentially between clonally-related excitatory neurons are significantly decreased by the deletion of clustered protocadherins (cPcdhs) in the mouse barrel cortex, indicating that cPcdhs are involved in the formation of specific neuronal connections in the cerebral cortex. In mice, 58 cPcdh genes, which encode the cell-adhesion membrane protein cPcdhs, are organized into three gene clusters, α, β and γ. Individual neurons express distinct combinations of these isoforms. In this study, we investigated whether cPcdhγ are involved in the formation of the neural connections between excitatory and parvalbumin (PV) positive inhibitory cells. We used the conditional cPcdhγ deletion (cKO) mutants by crossing PV-cre mice and cPcdhγ flox mice. To test whether the deletion of cPcdhγ in PV cells affects the connection probability between pyramidal and PV cells, we performed triple-whole cell patch clamp recordings from layer 2/3 cells in the visual cortical slices obtained from 3-week-old mice. We found that the connection probability between pyramidal and PV cells in cKO mice was similar to that in the control mice. The amplitudes of unitary EPSCs and IPSCs were also similar between control and cKO mice. Surprisingly, we found the two types of PV cells according to the connectivity with pyramidal cells in control mice. One type of PV cells formed exclusively reciprocal connections with pyramidal cells, and the other received less excitatory inputs resulting in the low-reciprocity. Around 80% of PV cells can be categorized in the high-reciprocity. The effect of cPcdhγ-deletion on the degree of the reciprocity in PV cells would be discussed.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-143 マウス嗅神経細胞の匂い応答における逆作動薬効果 Shigenori Inagaki（稲垣成矩），Ryo Iwata（岩田遼），Takeshi Imai（今井猛）九州大院疾患情報 Each olfactory sensory neuron (OSN) expresses a single type of odorant receptor (OR) out of ~1,000 types in mice. OSNs expressing the same type of OR converge their axons onto the same glomeruli in the olfactory bulb (OB). In the OB, odor information is spatiotemporally represented in the glomerular map reflecting the affinity of odors to each OR. It is generally believed that odorants ""activate"" ORs and odor information is represented by the ""activation"" patterns of glomeruli in the OB. In this study, we performed in vivo two-photon Ca2+ imaging of OSN axon terminals in the OB, and found that ~5% of glomeruli show robust suppressive responses to odors, while ~25% demonstrated activation. As OSNs are known to show spontaneous activity without odors, this is most likely due to the suppression of spontaneous activity in OSNs. To examine a possible role for interglomerular presynaptic inhibition by GABAergic/dopaminergic short axon cells, we generated a mouse line with an OSN-specific knockout of GABAB receptors and dopamine D2 receptors. However, this knockout did not abolish the suppressive responses seen at OSN axon terminals. To examine whether unknown forms of pre-synaptic inhibition are involved, we also generated OSN-specific tetanus toxin light chain (TeNT) transgenic animals, in which synaptic transmission from all OSNs was blocked; however, we still observed robust suppressive responses at OSN axon terminals, suggesting that the suppression occurs non-synaptically. We therefore performed two-photon Ca2+ imaging of OSN somata in the olfactory epithelium in vivo, and found that the suppressive responses are already present at the OSN somata. Our results suggest that OSNs have spontaneous firing and that odorants act not only as agonists, but also potentially as inverse agonists in mammalian OSNs in vivo. The combination of excitatory and suppressive responses in OSNs may contribute to the rich spatiotemporal representation of odor information in the olfactory bulb.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-144 マウス嗅球における内側／外側に存在する対になる糸球体の匂い応答特性の解析 Tokiharu Sato（佐藤時春）^1,2，Ryota Homma（本間良太）¹，Shin Nagayama（永山晋）¹ ¹テキサス大学ヒューストン校　医学部　神経生物学解剖学、ヒューストン、USA ²新潟大学　脳研究所　システム脳病態学、新潟、日本 Axons of olfactory sensory neurons (OSNs) that express the same type of odorant receptor converge onto two glomeruli in the olfactory bulb (OB). One is located in the medial side and the other is in the lateral side of OB. Because these glomeruli are arranged symmetrically, it has been considered that there is a medial/lateral glomerular mirror maps in the OB. In contrast to the clear anatomical evidences, there is little functional study of the two maps because the majority of the medial glomeruli is located at the medial wall of the OB and thus are difficult to access in intact brain. Recently, symmetrical glomeruli that expressed trace amine-associated receptors (TAARs) are found both in the dorsal OB (Pacifico et al., Cell Reports, 2012). Taking advantage of these remarkable finding, we simultaneously recorded odorant-evoked responses to compare their functional properties directly. In this study, the activity of these symmetrical glomeruli was measured using in vivo wide-field calcium imaging technique with 125Hz temporal resolution. Two odorants, phenylethylamine and isopentylamine, were used to stimulate the TAARs glomeruli. For measuring neuronal activity in various types of neurons in the OB, we used Cre-dependent GCaMP3 expressing mice that were crossed with several types of Cre-driver mice (OSNs; OMP-Cre, GABAergic neurons; Gad2-Cre, dopaminergic neurons; DAT-Cre and mitral/tufted cells; Pcdh21-Cre). Contrary to our expectations, we could not find any differences in onset latency, rise time, decay time and peak amplitude of odor evoked activities between the medical and lateral glomeruli in all examined mice. What we found was that medial glomerulus has significantly larger respiration-locked fluctuation in the postsynaptic neurons of Gad2-, DAT- and Pcdh21-GCaMP3 mice. But the differences were not observed in the presynaptic neurons of OMP-GCaMP3 mouse. Interestingly, this trend was also observed in resting condition in pre-stimulation period. However, it is more prominent during odor stimulation. These results were consistently observed in the all experiments using different odorant concentrations. These results suggest that medial map in the OB promotes the respiration-locked rhythmical activity and the outputs may associate with the activity in higher brain centers where the phase-locked rhythm is used for the information process such as somatosensory coordination (Ito et al., 2014) and fear memory (Moberly et al., 2018).		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-145 風味形成によるマウス大脳皮質での味およびニオイ情報処理領域の変化 Yusuke Komine（小峰雄介）^1,2，Naoko Mizoguchi（溝口尚子）^1,3，Hideaki Sakashita（坂下英明）²，Kazuyo Muramoto（村本和世）¹ ¹明海大学歯学部・形態機能成育学講座生理学分野 ²病態診断治療学講座口腔顎顔面外科学Ⅱ分野 ³病態診断治療学講座薬理学分野 The flavor of food is formed in the brain by integrating olfactory and various oral sensory information with gustatory one. Of them, the integration of the odor and taste information is important in flavor formation. Generally, neural pathways for odor and taste are thought to converge in the orbitofrontal cortex as the secondary sensory cortex. However, some morphological and fMRI data have indicated that olfactory information should also be transmitted to the insular cortex (IC) including the gustatory cortex (GC). To address this issue, we investigated mouse cortical responses using in vivo optical imaging technique with flavoprotein autofluorescence when taste and odor stimulation was applied to the animal. We applied amyl acetate (AA) as an odor, and quinine (bitter taste) and sucrose (sweet taste) as tastants to the anesthetized mouse, and cortical responses to these stimulations were recorded with an in vivo imaging equipment. The stimulation by AA evoked cortical responses only within the piriform cortex (PC), while taste stimulation using quinine or sucrose resulted in showing the extensive responses in the IC. Simultaneous stimulation by both of odor and taste significantly expanded responsive area into the agranular part of the IC (AI), suggesting that the AI is a key center for flavor formation. Moreover, in order to investigate how the response to the formed flavor is represented in the cortices, we make mouse to form associative learning of taste and odor and compared the cortical responses to each chemosignal before and after learning. We trained mouse to learn AA as an aversive odor by associating it with quinine. In mice with aversive learning, AA-stimulation activated not only the PC but also the IC including the GC. Conversely, quinine-stimulation evoked significant response in the PC as much as the response in the GC in the same learned mice. Therefore, the signal processing for the single sensory information (taste or odor), which is originally processed only in a corresponding sensory cortex, changes plastically into that the same single sensory signal is simultaneously processed also in the cortical area for the another sensory modality, by forming the associative learning. Such changes in cortical responses may be helpful for the representation of flavor in the brain.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-146 腹側テニアテクタのシーンセル：環境や行動状況において特定の発火 Kazuki Shiotani（塩谷和基）^1,2，Yuta Tanisumi（谷隅勇太）^1,2，Koshi Murata（村田航志）³，Junya Hirokawa（廣川純也）¹，Kensaku Mori（森憲作）⁴，Yoshio Sakurai（櫻井芳雄）¹，Hiroyuki Manabe（眞部寛之）¹ ¹同志社大学大学院　脳科学研究科 ²日本学術振興会特別研究員DC ³福井大学　医学部 ⁴東京大学 The ventral tenia tecta (vTT) is a part of the olfactory cortex that receives both olfactory sensory signals from the olfactory bulb and top-down signals from the prefrontal cortex. To address the question whether and how the neuronal activity of the vTT is modulated by prefrontal cognitive processes such as attention, expectation and working memory that occurs during goal-directed behaviors, we recorded individual neuronal responses in the vTT of freely moving awake mice that performed learned odor-guided behaviors. Mice were trained to associate an odor (eugenol) in the odor port with the appearance of water reward in the reward port that is located at the left of the odor port. These mice were trained to associate a different odor (amyl-acetate) in the odor port with no-reward in the reward port. We found that the firing pattern of vTT neurons during the drinking behaviors showed clear tuning of individual neurons to distinct scenes (i.e., distinct environmental and behavioral contexts) of learned behaviors. Furthermore, spiking activity of these scene cells was modulated not only by the present scene but also by the future scene that the mouse predicted. These results indicate that different groups of vTT cells are activated at different scenes and suggest that processing of olfactory sensory information is handled by different scene cells during distinct scenes of learned behaviors. The scene effect on olfactory sensory processing in the vTT has implications for the neuronal circuit mechanisms of top-down attention and scene-dependent encoding and recall of olfactory memory.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-147 年齢に伴い増えるマウス外側嗅索での軸索シャフトの隆起構造の解析 Shin Nagayama（永山晋）¹，Frederik M Seibt（Seibt M Frederik）¹，Ryota Homma（本間良太）¹，Tokiharu Sato（佐藤時春）^1,2 ¹テキサス大学医学部ヒューストン校 ²新潟大学　脳研究所　システム脳病態学 Axons have unique compartments of presynaptic vesicle releasing sites called axonal varicosities and boutons. These swelling compartments are frequently seen in unmyelinated axons in a healthy brain. Interestingly, we discovered an axonal swelling structure that resembles varicosities in the lateral olfactory tract (LOT) that contains the bundles of myelinated axons from olfactory bulb mitral/tufted cells. This structure was consistently observed using multiple techniques, such as adeno-associated virus and electroporation labeling methods. Using Protocadherin21(Pcdh21)-Brainbow mice, which are generated by crossing Pcdh21-Cre (Cre expressing in mitral/tufted cells) mice with cre-inducible Brainbow mice, we analyzed this structure in detail. This mouse allowed us to observe a large population of axons as well as to identify each axonal structure. Interestingly, this structure was not detected in young (1-month old) mice but appeared in 2-month and older mice. The number and size of the structure monotonically increased in an age-dependent manner. Studies of immunohistochemistry revealed that the majority of swelling structures in LOT were Tom20 or p62 (mitochondria and autophagy marker protein, respectively) positive but SV2 (synaptic vesicle marker) signals were not detected in swelling sites of LOT. Furthermore, we could not observe calcium transient caused by the electrical stimulation of LOT in these structures using an acute slice tissue in which all LOT structures were preserved. These data suggest that the varicosity-like swelling structures observed in the LOT are not the ordinary vesicle releasing sites and may be associated with animal aging.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-148 匂い刺激によってナメクジ嗅覚中枢神経系に引き起こされるNO放出の蛍光イメージング（Ⅱ） Momo Murata（村田桃）¹，Kohei Ishida（石田康平）¹，Satoshi Watanabe（渡辺恵）²，Minoru Saito（斎藤稔）¹ ¹日大院総合基礎科学 ²国立精神・神経セ神経研 The land slug Limax valentianus has superior olfactory which can discriminate odor and have the ability of odor learning. The procerebrum (PC) of the slug is the olfactory nervous center for odor processing and learning. The PC shows an oscillatory local field potential (LFP) at about 0.7 Hz and a propagation of the neural activity from the distal to proximal regions. Previous studies reveal that odors modulate the central network dynamics. LFP frequency was increased by aversive odor stimulation while not by attractive odor stimulation. After odor aversive conditioning, LFP frequency was increased by innately attractive odor stimulation in isolated preparation. On the other hand, a gaseous neurotransmitter, nitric oxide (NO), functions specifically in the PC, which modulates the dynamics of central olfactory network and has been implicated in olfactory processing including learning. In present study, we attempted to observe NO spatiotemporal dynamics simultaneously LFP oscillation change when innately aversive or aversively conditioned odor was applied to the slug. Brain containing superior tentacle was isolated for preparation and the odor stimulation was applied to olfactory epithelium on superior tentacle by air flow using electric valves. For the conditioning, lip nerve was electrically simulated using glass suction electrode. The preparation was stained with an NO-sensitive dye, DAR-4M AM. The stained preparation was illuminated by LED and the fluorescence images were acquired through sCMOS camera. The LFP was obtained by using extracellular recording. The fluorescence intensity in the PC always continued to increase, indicating that NO is continuously released in the PC. The slope of intensity was increased by applying innately aversive and aversively conditioned odors whereas not by attractive odor. The LFP frequency was also increased by applying innately aversive and aversively conditioned odors. Additionally, these effects were blocked by NO synthase inhibitor, L-NAME. These results suggest that the increase in the NO release speed and LFP frequency in the PC are related to aversive response. Furthermore, the NO-released region in the PC will be also examined using the NO imaging.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-149 マウス味蕾における酸感受性イオンチャネルの発現 Natsuko Kumamoto（熊本奈都子），Yasuhiro Shibata（柴田泰宏），Takashi Ueda（植田高史），Shinya Ugawa（鵜川眞也）名古屋市大医機能組織学 In mammals, sour taste is detected by taste bud type III cells that respond to acidic pH and weak organic acids with electrical activity. Members of the acid-sensing ion channel (ASIC) family are proton-gated cation channels and candidates for sour-taste receptors or signaling components. Previous studies have shown that ASICs are expressed in human and rat taste cells and that ASICs are absent in taste buds at the anterior tongue of patients with an acquired sour ageusia. Expression of ASIC proteins in mouse taste cells remains unknown, partly because there are few subtype-specific antibodies. To investigate protein expression of ASICs in mouse gustatory papillae, we generated three kinds of knock-in mice expressing epitope- and fluorescence-tagged ASICs (V5-RFP-V5-ASIC1a, HA-BFP-HA-ASIC1b, and ASIC3-FLAG-EGFP-FLAG). Immunohistochemical analysis revealed that V5-immunoreactivity and HA-immunoreactivity were detected in the nerve fibers innervating the taste buds, while no FLAG-immunoreactivity was apparent in the gustatory papillae. To examine whether ASIC1a is expressed in type III cells of the ASIC1a knock-in mice, we performed double label immunostaining with an anti-V5 antibody and a type III cell marker SNAP25 or 5-HT, and found no significant difference in V5-immunoreactivity in type III cells between the knock-in and wild-type mice, suggesting that ASIC1a is hardly or not expressed in mouse type III cells. Further (electrophysiological) study will be needed to determine whether ASICs are directly involved in mouse sour-taste detection.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-150 マウスにおけるオルニチンによるうま味嗜好性の増強について Haruno Mizuta（水田晴野），Takashi Yamamoto（山本隆）畿央大学大学院健康科学研究科 Food contains various chemical substances. Among them, amino acids play a variety of important roles in taste as well as in nutrition, but few studies have examined the taste characteristics of L-amino acids. It is well known that miso soup with corbiculae is very delicious. Since corbiculae are rich in L-ornithine, L-ornithine, a non proteinogenic L-amino acid, may work as a palatability enhancer. To examine this possibility, by using adult C57BL/6 male mice, we employed the conventional 48-h two bottle preference test for water or miso soup with and without ornithine. The taste of L-ornithine itself had no particular taste. However, preferences were significantly increased when ornithine was added to miso soup at concentrations ranging from 1 to 30 mM. In the next step, additive effects of L-ornithine to single or combinations of basic taste solutions were examined. Addition of L-ornithine to a trinary mixture of NaCl, sucrose and monosodium glutamate (MSG), single solution of MSG and binary mixtures of MSG and sucrose or MSG and NaCl significantly increased the preferences. Addition of ornithine to 0.05M MSG increases the palatability equivalent to that of 0.2M MSG. The GPCR, class C, group 6, subtype A (GPRC6A) antagonist, calindol, abolished the preference-increasing action of ornithine. Chorda tympani responses also showed that MSG response was enhanced by L-ornithine and this effect was abolished by calindol. The present study shows that ornithine makes miso soup delicious by enhancing umami and the L-amino acid sensing receptor, GPRC6A, may be involved in the action of L-ornithine.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-151 RNA結合タンパク質hnRNP A/Bのマウス嗅覚神経細胞における機能の解明 Nanaho Fukuda（福田七穂）^1,2，Piergiorgio Percipalle（Percipalle Piergiorgio）³，Kevin Czaplinski（Czaplinski Kevin）⁴，Tomoyuki Fukuda（福田智行）⁵，Toshikuni Sasaoka（笹岡俊邦）¹ ¹新潟大脳研 ²奈良先端大バイオサイエンス ³New York Univ, Abu Dhabi ⁴Stony Brook Univ, NY, USA ⁵新潟大院医歯学 Heterogeneous nuclear ribonucleoprotein A/B (hnRNP A/B, Hnrnpab ) is an RNA binding protein with two RRM and glycine rich domains. We have previously shown that, in the mouse spermatogenic cells, hnRNP A/B directly binds to the 3'UTR of protamine2 (prm2) mRNA, and regulate its translational timing and efficiency during spermatogenesis (Fukuda et. al., PLOS Genetics, 2013). Hnrnpab is also expressed in the brain, where mRNA translation is strictly regulated in a spatio-temporal manner. We therefore set out a study to examine the role of hnRNP A/B in the mouse nervous system. In the expression analysis of hnRNP A/B in various parts of brain and sensory tissues, a high level of hnRNP A/B was detected in the olfactory sensory tissues. The immunohistological analysis of the olfactory neuroepithelium using neuronal markers revealed that the immature olfactory sensory neurons (OSNs) express high level of hnRNP A/B. We next performed the histological analysis of the nasal tissue of Hnrnpab KO mice and found that Hnrnpab KO mice have thinner olfactory neuroepithelium with a higher number of apoptotic bodies. In the olfactory bulb, the structure of the glomerular layer of Hnrnpab KO mice was found to be abnormal. These results suggest that hnRNP A/B regulates mRNAs essential for the formation and/or functions of OSNs. Further study on hnRNP A/B and their target mRNAs in the OSNs will elucidate the mechanisms and the role of mRNA regulation in neurons.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-152 ゼブラフィッシュ嗅覚忌避行動を司る神経回路メカニズムの解析 Miwa Masuda（増田美和）^1,2，Sayoko Ihara（伊原さよ子）^2,3，Naoki Mori（森直紀）³，Tetsuya Koide（小出哲也）¹，Nobuhiko Miyasaka（宮坂信彦）¹，Noriko Wakisaka（脇阪紀子）¹，Keiichi Yoshikawa（吉川敬一）³，Hidenori Watanabe（渡邉秀典）³，Kazushige Touhara（東原和成）^2,3，Yoshihiro Yoshihara（吉原良浩）^1,2 ¹理化学研究所脳神経科学研究センターシステム分子行動学研究チーム ²ERATO 東原化学感覚シグナルプロジェクト ³東京大学大学院農学生命科学研究科 In 1938, an Austrian ethologist, Karl von Frisch, reported the existence of Schreckstoff (scary stuff) in minnows, which is released from the skin of injured fish to notify its shoaling company about the presence of danger. Most fresh-water fish species including zebrafish display this olfactory alarm response which is important for fishes in risk avoidance even without visible presence of predators. Despite enormous efforts by many researchers, the identity of alarm substances still remains unknown. In this study, we first observed that upon application of conspecific skin extract, most zebrafish showed burst swimming followed by freezing at the bottom of the tank. This alarm reaction was not observed in olfactory epithelium-removed zebrafish, indicating the crucial role of the olfactory system. Next, we biochemically purified two small compounds from zebrafish skin extract, which individually activate distinct glomeruli in the olfactory bulb and synergistically evoke robust alarm reaction. Furthermore, we found the telencephalic nuclei that were activated only when fish were concomitantly stimulated by the two substances. These results demonstrate that the coincidence detection mechanism of two glomeruli activated by the two pheromone components underlies the emergence of olfactory alarm reaction in zebrafish.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-153 LATS1欠損マウスは蝸牛感覚上皮形成不全および細胞変性を呈する Takanori Nishiyama（西山崇経）^1,4，Masato Fujioka（藤岡正人）¹，Makoto Hosoya（細谷誠）¹，Ayano Mitsui（三井綾乃）¹，Chika Saegusa（三枝智香）¹，Naoki Oishi（大石直樹）¹，Tatsuhiko Harada（原田竜彦）³，Hideyuki Saya（佐谷秀行）²，Kaoru Ogawa（小川郁）¹ ¹慶應大医耳鼻咽喉科 ²慶應大医先端研 ³国際医療福祉大熱海耳鼻咽喉科 ⁴川崎市立川崎病院耳鼻咽喉科 Objective: LATS1 belongs to the family of AGC kinases and participates in the Hippo signaling pathway, which regulates organ size, self-renewal of stem cells, and cell differentiation. LATS1 phosphorylates and inhibits the activity of YAP and TAZ that are transcriptional coactivators of the Hippo pathway. Likewise, Cyclin dependent kinases inhibitors (CKIs) are mediator of cell cycle. It is known several CKIs, for example Cip1 and Ink4d, are necessary to maintain structure of inner ear and those deficiency cause hearing loss. We here report expression pattern and roles of Hippo pathway molecule, LATS1 in inner ear by investigating LATS1 deficient mice. Methods: Lats1KO mice (C57BL/6 background) were generated by conventional method. Hearing function was measured by Auditory Brainstem Response (ABR) and Distortion Product Otoacoustic Emission (DPOAE) under anesthesia. Histological analysis of organ of Corti was carried out by immunohistochemical staining using surface preparations of newborn and adult mouse cochlea. Superstructure of the organ of Corti was observed using Scanning Electron Microscope (SEM). Results of Lats1 KO mice were compared with their littermate controls. Results: Immunohistochemistry revealed expression of LATS1 in both inner and outer hair cells of the organ of Corti. Lats1 KO mice displayed disorganized polarity of inner and outer hair cells, degeneration and irregularity of stereocilia, revealed by either immunohisotochemistry or SEM. These phenotypes were observed not only in adult but also in newborn mice. In adult KO mice, higher threshold was observed than those of littermate controls in ABR and DPOAE. Staining for acetylated tubulin revealed misoriented kinocilium in Lats1 KO newborn mice, suggesting the abnormality in planar cell polarity (PCP) pathways. We therefore hypothesize that LATS1 is indispensable for kinocilium migration in the hair cells and thereby essential for hearing. We are now further investigating molecular mechanisms underlying control of PCP by LATS1.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-154 サリチル酸に誘発された耳鳴りはマウス聴覚皮質におけるギャップのオンセットに対する神経応答に影響を与える Motoki Yanada（簗田元揮）北海道大学大学院情報科学研究科 Tinnitus is a hearing disorder experienced by 10-15% of the population with a phenomenon in which subjective sound perception occurs without external sound input. A gap detection test for tinnitus assessment utilizes the acoustic startle reflex elicited by loud noise pulse and its inhibition by gap in noise preceding loud noise pulse. Tinnitus masks the gap in noise and results in gap detection deficits. However, the neural basis that induces gap detection deficits in the tinnitus state is unknown. Because the auditory cortex (AC) is necessary for the detection of brief sound gap, evaluating gap-evoked neural responses in the AC is an effective way to clarify the gap detection deficit mechanism. In this study, we analyzed auditory neural responses to the gap in the AC of pharmacologically tinnitus-induced mice combined with the behavioral gap detection test. First, tinnitus was behaviorally evaluated by the gap-prepulse inhibition of the acoustic startle reflex (GPIAS) after sodium salicylate (SS) injection, which is known to induce tinnitus. C57BL/6J mice were given intraperitoneal injections of SS (300 mg/kg B.W.), and the effect on gap detection was measured using narrowband background noise with four types of centered frequencies: 8, 12, 16, and 20 kHz. Tinnitus state was evaluated by comparing the result of GPIAS tests before and two hours after SS injection. As a result, significant gap detection deficits were found only for the tested noise at 16 kHz after SS injection, whereas those were not found for the noise at other frequencies. This indicated that SS-induced tinnitus occurred with a pitch around 16 kHz. Second, we recorded the gap evoked neural responses in the AC of urethane-anesthetized mice by a 16-channel silicon probe in order to examine the physiological effect of SS on gap detection in the AC. As a result, the phasic responses to the gap onset before SS injection significantly disappeared or weakened while those to the gap offset did not change. In addition, most of response durations to the gap before SS injection seemed to be shortened in the time axis. These results suggested that the change in neural responses evoked by gap onset after SS injection is one of the factors at the neural level of gap detection deficits. Analyzing the SS effects in the neural circuit of the AC enables us to elucidate the neural mechanism of tinnitus, and it will provide an insight for developing new treatment strategies of tinnitus in the future.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-155 大脳基底核は聴覚システムを多重階層的に制御している Ryohei Tomioka（冨岡良平）¹，Bo Zhou（周博）^2,3，Kenji Sakimura（崎村建司）⁴，Yuchio Yanagawa（柳川右千夫）⁵，Nobuaki Tamamaki（玉巻伸章）¹，Wen-Jie Song（宋文杰）^2,3 ¹熊本大院生命科学・医脳回路構造 ²熊本大院生命科学・医知覚生理 ³HIGOプログラム ⁴新潟大脳研基礎神経科学細胞神経生物 ⁵群馬大院医遺伝発達行動 The basal ganglia (BG) is known for its role in motor selection and motor control (Mink, 1996; Nelson & Kreitzer, 2014). While it receives input from virtually all areas of cerebral cortex (Selemon, Goldman-Rakic, 1985; Hunnicutt et al., 2016), its output is believed to target motor-related areas in frontal cortex via the thalamus (Mink, 1996). Recently, evidence is accumulating that the BG has cognitive functions in addition to motor functions (Helie et al., 2013), suggesting that the BG may have output targets other than motor-related cortical areas. To explore this possibility, we examined projections from the BG to the auditory pathway. Injection of the retrograde tracer Fast Blue into the auditory cortex of GAD67-GFP knock-in mice, resulted in abundant retrograde labelling of GABAergic neurons in the BG. Using the combination projection-genetic targeting strategy (Fenno et al, 2014), we found that GABAergic neurons in the BG innervated multiple structures in the auditory system, including the medial geniculate body, the inferior colliculus, and the auditory cortex. To examine whether GABAergic neurons in the BG have functional impacts on neuronal activities in the auditory cortex, we carried out neurophysiological experiments with optogenetic modulation. We found that activation of GABAergic neurons in the BG significantly suppressed spontaneous cortical activities. These results suggest a role of the BG in modulation of sensory functions. The BG-auditory circuits identified here might be part of a substrate for the cognitive function of the BG.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-156 スナネズミ聴覚野におけるコミュニケーション音声に対する選択的反応 Takafumi Furuyama（古山貴文）¹，Taisei Minei（嶺井大聖）²，Rina Hayase（早瀬莉奈）²，Kohta I Kobayasi（小林耕太）² ¹同志社大学　研究開発推進機構 ²同志社大学　脳神経行動工学研究室 Vocal communication is an important for many animals, including humans, to survive and to form a society. However, little is known about the brain region processing communication sounds. Primary auditory cortex is the first cortical stage to process the acoustic properties of spectro-temporally complex sound (e.g., communication sound). The purpose of this study was to investigate the role of the primary auditory cortex in vocal communication, and we assessed how selectively the region respond to communication sounds. The flavoprotein fluorescence imaging was employed for measuring the cortical activities in primary auditory cortex of Mongolian gerbils (Meriones unguiculatus). Subjects were deeply anesthetized by urethane (1.5 g/kg), and the skull of left temporal region was exposed. Auditory stimuli were 4 types of tone bursts (1, 4, 20, and 32 kHz) to confirm the tonotopic organization of recorded sites, and 7 types of communication sounds. All auditory stimuli were 80 dB sound pressure level (SPL) and were presented 20 times. Our data successfully showed the response of flavoprotein fluorescence of primary auditory cortex. The tonotopy was confirmed in A1 by using tone bursts. The cortical activities to communication sounds were observed at high frequency area of A1. The response of the area is highly selective to communication sounds, and suggests that the region of high frequency area in A1 is critically involved in processing communication sounds in Mongolian gerbils.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-157 ヒトiPS細胞由来内耳蝸牛有毛細胞の分化誘導 Tsubasa Saeki（佐伯翼）¹，Makoto Hosoya（細谷誠）²，Masato Fujioka（藤岡正人）²，Kaoru Ogawa（小川郁）²，Hideyuki Okano（岡野栄之）¹ ¹慶應大医生理 ²慶応医耳鼻科 Loss of hair cells in mammalian cochlea is irreversible once injured and results in permanent sensorineural hearing loss. Induction of hair cells from human pluripotent stem cells (hiPSCs) may provide platforms for disease modeling, eventually to therapeutics by drug screenings. Although reliable and robust induction method is indispensable for the biomedical application, previously reported methods was unfortunately poor especially in their induction efficiency and maturity of the induced hair cells. Moreover, the marker study for characterization of cochlear hair cells from hiPSCs was not clearly described before. In this study, we developed a novel strategy for differentiation of hiPSCs into cochlear hair cell linage. Firstly, we differentiated hiPSCs to otic progenitor cells with high efficiency using a previously reported method (Hosoya et al., 2017). Immunocytochemistry and qRT-PCR showed that otic progenitor cells expressed early otic progenitor markers, including PAX2, PAX8 and SOX2. Following treatment of induced otic progenitor cells with GSK-3βinhibitor CHIR99021, we observed upregulation of otic progenitor markers, PAX2, PAX8 and prosensory domain marker, LGR5. Then treatment of the cells with WNT signaling modulators promoted upregulation of various hair cell markers including ATOH1, MYO7A, MYO15A, BRN3C, PRESTIN by qRT-PCR analysis. Immunocytochemistry revealed that the induced hair cell-like cells expressed hair cell markers, MYO7A and BRN3C. We found some novel chemicals that promote differentiation of hiPSCs derived otic progenitor cells into cochlear hair cell linage. We will further refine induction protocol in its efficacy in hair cell differentiation and examine culture conditions to differentiate into mature cochlear hair cells.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-158 カルシウムイメージングによるマーモセット聴覚野からのミスマッチ陰性電位記録 Keitaro Obara（小原慶太郎）¹，Teppei Ebina（蝦名鉄平）¹，Yoshito Masamizu（正水芳人）¹，Shin-Ichiro Terada（寺田晋一郎）¹，Takanori Uka（宇賀貴紀）²，Misako Komatsu（小松三佐子）³，Noritaka Ichinohe（一戸紀孝）^3,4，Akiya Watakabe（渡我部昭哉）⁵，Hiroaki Mizukami（水上浩明）⁶，Tetsuo Yamamori（山森哲雄）⁵，Kiyoto Kasai（笠井清登）⁷，Masanori Matsuzaki（松崎政紀）¹ ¹東京大院医細胞分子生理 ²山梨大院統合生理学 ³理研CBS 高次脳機能分子解析一戸グループ ⁴国立精神・神経セ神経研微細構造 ⁵理研CBS 高次脳機能分子解析 ⁶自治医大分子病態治療研究センター ⁷東京大院医精神神経 The auditory mismatch negativity (aMMN) is an event-related potential that reflects preattentive detection of changes in repetitive and predictive tone sequence. Because the amplitude of the aMMN is attenuated in psychotic disorders including schizophrenia, the aMMN can be used as a biomarker for these disorders. Previous studies have shown that the aMMN consists of two mechanisms: (1) stimulus-specific adaptation of neural responses to a repetitive stimulus pattern, and (2) prediction error for the changes in repetitive stimuli (deviant detection). Among of the two mechanisms, the deviant detection is thought to be related to the biomarker of the psychotic disorders. However, it is unknown how the deviance detection is represented at a single neuron level in the auditory areas. To investigate the mechanisms of the deviance detection, we apply one-photon and two-photon calcium imaging of neuronal activities in the auditory areas of common marmosets. Marmosets are known as a highly vocal primate species and have three hierarchically connected auditory areas (core, belt, and parabelt), similar to humans. Moreover, because the cortical surface of the marmoset auditory areas is exposed to the outside of the lateral sulcus, these areas are suitable for calcium imaging. In this meeting, we will show the functional map of the core auditory areas obtained by using one and two-photon imaging techniques. We also show the deviant detection like responses recorded from the core auditory areas.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-159 非対称性振幅変化音への前聴覚野ニューロンの反応パターン Sohei Chimoto（地本宗平）山梨大医神経生理 Time envelope of natural sounds such as speech and musical sound are asymmetric under acoustic environments. Plucking the acoustic medium produces a sound with a quick attack in amplitude (damped sounds), while bowing the medium produces a sound with a slow increase (ramped sounds). Psychophysical experiments have shown that this temporal asymmetric features influence sound perception and identification. In the previous studies we showed that the higher auditory cortices as well as the primary auditory cortex (A1) include velocity-sensitive cells for the damped- and ramped-sounds and some neurons in the secondary auditory cortex (A2) and posterior auditory fields (PAF) had the sensitivities to direction of the amplitude change of the asymmetric sounds. It is known that the anterior auditory fields (AAF) just rostral to A1 has a tonotopic map and includes the neurons responsive to various kinds of sounds such as noise bursts, amplitude modulated sounds, and natural sounds as A1 does. However, little is known about the response patterns of AAF neurons during the asymmetrical stimuli. In the present study we recorded single unit activities from AAF of awake animals and examined response patterns during ramped and damped sounds. In the damped sound, the amplitude of sound wave exponentially decreases with a time constant set to 1/5th of the stimulus duration (2.5, 5, 10, 20, 40, 80, 160, and 320ms) and the peak amplitude of stimuli was 30, 50, and 70-dB SPL. The ramped stimulus is just the time-reversed version of the damped stimulus. The carrier frequency is set at the cell's best frequency or band passed white noise which produce the maximum response amplitude. Spike trains responding to each stimuli were used to construct peristimulus time histograms and the height of PSTH was transformed into the driven rate by subtracting the mean of background firing rates. We found that most of AAF neurons showed sensitivities to the abrupt change of stimulus envelope (edge cells) and a few neurons showed sensitivities to the slow amplitude changes during both damped- and ramped-sounds (slope cells). We also found most neurons in AAF show weak selectivity for damped- or ramped-sounds by comparing between the peak response amplitude during those asymmetric sounds. These results indicate that the majority of AAF neurons are tuned to the velocity (slow or quick) but not to the direction (attack or decay) of the amplitude change.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-160 刺激の予測と予測誤差に関わる脳波に与える視覚的曖昧さの影響 Kentaro Ono（小野健太郎）^1,2，Junya Hashimoto（橋本淳也）³，Ryosuke Hiramoto（平本亮介）³，Takafumi Sasaoka（笹岡貴史）^1,2，Shigeto Yamawaki（山脇成人）^1,2 ¹広島大学感性イノベーション拠点 ²広島大学脳・こころ・感性科学研究センター ³広島大学大学院教育学研究科 According to the predictive coding theory, the brain continuously generates predictions about upcoming sensory stimuli and updates it based on mismatch between a prediction and an incoming stimulus, called a prediction error. While neural correlates of prediction error have been reported as mismatch negativity (MMN), P300, late positive potential (LPP), neural correlates of a prediction have still been controversial. Some studies have reported a negative shift of brain potentials, which is called stimulus preceding negativity (SPN), during a prediction of a target stimulus. However, it is unknown whether the accuracy of prediction affect the SPN. Furthermore, it is still unclear how the accuracy of prediction is associated with the brain activity after presentation of a stimulus (P300 and LPP). Thus, the aim of the present study is to clarify how the accuracy of prediction affects the SPN, P300 and LPP. In the experiment, we used a S1-S2 paradigm and participants were asked to predict original pictures (S2) from different levels of mosaic (pixelated) pictures (S1). The brain potential was measured by electroencephalography (EEG) during prediction and after presentation of S2. The results showed that the subjective score of the accuracy of prediction was lower when the quality of S1 was low. Also, the subjective score of consistency between a predicted and a real S2 was lower when the quality of S1 was low. These results indicated that a rough mosaic S1 decreased the accuracy of prediction. Event-related potentials showed that the amplitude of SPN was increased when the quality of S1 was high. Furthermore, the amplitude of SPN at the right frontal electrode was correlated with the subjective score of the accuracy of prediction. After presentation of S2, the brain potentials like P300 and LPP were also associated with the quality of S1. Further, the subjective score of consistency between a predicted and a real S2 was correlated with the amplitude of LPP. These results suggest that the SPN not only reflects prediction as reported in the previous studies, but also reflects the accuracy of prediction. In addition, the accuracy of prediction was also associated with P300 and LPP, owing to the different level of prediction error caused by the different level of the quality of prediction.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-161 色恒常性の統一的な説明：ランドの２色法および#The Dress Masahiro Yamamoto（山本雅裕）東芝研究開発セ We have proposed that the color appearance of#The Dress can be explained in the DKL color space(1). In the DKL color space, the color distribution of natural objects shows a distinctive shape. Therefore, when the illumination color changes, it is expected that the color distribution perceived will be calculated from clear clues by their shape. We showed that the color appearance of #The dress could be calculated by Wasserstein metric with the illumination color as the constraint condition. Based on this idea, We attempted to consider the Land's two-color projection. To compare with the conventional calculation method by different spatial frequency filter of previous research(2), we made and tested the dissolve (apply the color layer as colored noise) version of Land's projection. The result shows that the original color is perceived by merely including 10% dissolve/noise. (It is a level to a calculation result be completely out of order in Deep Learning.) The average color by the calculation using the lowpass filter shows the redshift when the filter color is red. Because of the dissolve filter, the amount of redshift is not enough to show real filter color. It will show only a rough color direction. Therefore, it is necessary to change the application method of the spatial frequency filter. We suggest that two-step estimation, 1st searching the rough direction of color shift as luminance color 2nd mapping the contrast of objects in B/W image onto the color contrast in the color space. This constraint is similar to the color interpretation of # The Dress. The color distribution of natural object in the DKL color space creates constraint when converting from the contrast of B/W image to color. And the constraint enables mapping by KL divergence. We explain that Land's two-color projection can be interpreted by using natural color statistics in the DKL color space. For the original color to be confirmed by a dissolve filter, it is necessary to estimate from the contrast distribution to the color distribution, resulting in a transport problem with estimations of the color of the illumination light and filter like #The Dress. Our conclusion shows that both, Land's projection and #The Dress, are due to the function of the same color constancy and provide a model that works in common. (1) M.Yamamoto Neuro2017, Neuro2018 (2) A. Shapiro et al. i-Perception 9(1)(2018)

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-162 人工網膜の刺激効率のネコ外側膝状体単一ニューロン反応による評価 Tomomitsu Miyoshi（三好智満）¹，Hiroyuki Kanda（神田寛行）²，Takeshi Morimoto（森本壮）²，Takashi Fujikado（不二門尚）² ¹大阪大院医統合生理 ²大阪大院医感覚機能形成 We have been developed a novel retinal prosthesis, Suprachoroidal Transretinal Stimulation (STS), for photoreceptor degenerating diseases. The stimulating electrode array of STS is implanted into sclera and do not directly contact retinal tissue in order to avoid physical damage. To investigate the stimulation properties, we have recorded the single-unit responses from cat lateral geniculate nucleus (LGN) relay neurons. Previously we reported that the burstic discharges, which occured alternately on ON and OFF cells, were elicited by single stimulation of STS (Neuro2017). We also reported that this burstic response was due to the prolonged change of excitability by the interaction in the stimulated retinal circuits (Neuro2018). Here we investigated how stimulus frequency of continuous STS influenced the spike occurrence. The number of spikes of LGN relay neuron was investigated with continuous STS of various frequencies. The size of each electrode head in the implanted array was 0.5 mm in diameter and 0.3 mm in height, which was the same as the clinical device. The single-unit activities were recorded with metal microelectrode, and the number of spikes per second was analyzed. The single stimulation was biphasic, 1000 uA amplitude, and 0.5 ms/phase duration. The frequency of stimulation was changed from 1 Hz (1000 ms interval) to 100 Hz (10 ms). The number of the spikes per second elicited by continuous stimulation did not show simple profile such as single peak or trough against stimulation frequency. The maximum number of spike discharges was achieved by the stimulation not greater than 50 Hz. These results suggested that high frequency stimulation may less effective.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-163 網膜におけるバースト内発火パターンによる情報表現 Toshiyuki Ishii（石井俊行）^1,2,3，Toshihiko Hosoya（細谷俊彦）¹ ¹理研CBS 局所神経回路 ²東邦大院理生物分子科学 ³日本医科大医生理（システム生理） Understanding the rules by which neuronal spike patterns encode information is essential for elucidating the complex functioning of the nervous system. Neurons in various brain areas generate spike bursts, i.e., clusters of high-frequency action potentials separated by longer periods of silence. While the number of spikes within a burst is known to carry information, whether the interspike intervals (ISIs) within a burst also convey information is poorly understood. In particular, a burst of k spikes has k-1 intraburst ISIs, which can theoretically encode k-1 values representing information, but this possibility has never been tested. In this study, we demonstrate that such combinatorial coding occurs for retinal bursts. By recording ganglion cell spikes from isolated retinae of salamanders and mice, we found that intraburst ISIs encode oscillatory light sequences, which are much faster than the light intensity change encoded by the number of spikes. When a burst has three spikes, the two ISIs combinatorially encode the amplitude and phase of the oscillatory sequences. Analysis of the trial-to-trial variability suggested that intraburst ISIs are regulated by two independent mechanisms responding to orthogonal oscillatory components, one of which is common to bursts with different number of spikes. Therefore, the retina encodes multiple stimulus features by exploiting all degrees of freedom of burst spike patterns, i.e., the spike number and multiple intraburst ISIs.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-164 デジタルマイクロミラー技術による個々の細胞の信頼性の高いin vivo光遺伝学的刺激法 Ryo Aoki（青木亮），Andrea Benucci（Benucci Andrea）理研CBS 視覚意思決定 Circuit optogenetics with spatial-temporal patterned illumination is gaining much attention as a tool to dissect the neuronal functional connectivity within and across brain regions. Current methodologies are often cost ineffective, require high, possibly tissue-damaging laser power, and are not easily integrated with optical systems targeting deep brain regions (Yang and Yuste, 2018). Here, we present an approach based on a digital micro-mirror device (DMD) for a cost-effective, low laser power manipulation of neural activity with single-cell precision, millisecond temporal resolution, and easy integration with optical systems for patterned optogenetic stimulations in deep brain regions. First, using cortical slices of different thicknesses, we quantified the XYZ volumetric power density profile associated to a small excitation disk. At the focal plane, 120 um deep from the surface, a 25 um diameter disk (λ = 640 nm) was confined to a very small region (sigma = 11.6 ± 0.6 um, peak 0.956 ± 0.126 mW/mm^2). With depth, the disk diameter increased, as measured by a calibrated CCD camera focused on the bottom side of the sample and the light power rapidly and significantly decreased (sigma : 14.8 ± 0.8 um, peak 0.494 ± 0.040 mW/mm^2 at z=170 um, sigma = 21.2 ± 0.8 um, peak 0.131 ± 0.010 mW/mm^2 at z=220 um). Above the focal plane, we estimated the power increase from standard measures of light absorption and scattering in brain tissue (Helmchen and Denk, 2005). We then used the measured volumetric intensity profile, to estimate in vivo the probability of single-cell excitation in neurons expressing ChrimsonR and GCaMP8 in mouse V1. We adjusted the excitation disk diameter and the light intensity, to simulate light intensity profiles at different depths and distances from the center of the stimulation. The maximum average excitation probability was very small (Prob. = 0.31) and confined to a small volume (volume of Prob. > 0.25 was confined in a ~17 um lateral diameter and ~35 um axial diameter). In conclusion, our data supports the notion that single-cell excitation can be reliably achieved in vivo with single-photon DMD technology. We are currently applying this method to examine the plasticity properties of sensory representations in the mouse V1. We are also extending this methodology to GRIN lenses for patterned optogenetic stimulation in deep brain regions.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-165 視細胞リボンシナプスは光刺激に対する順応を調整する働きを持つ Yuko Sugita（杉田祐子）¹，Kenichiro Miura（三浦健一郎）²，Takahisa Furukawa（古川貴久）¹ ¹大阪大・蛋白質研・分子発生学 ²京都大院・医・認知行動脳科学 The ribbon synapse is a specialized structure which connect photoreceptor synapse terminal with bipolar cell and horizontal cell. The Pikachurin is a dystroglycan ligand in the retina, and is essential for the proper formation of ribbon synapse structure. Several lines of evidence suggest that the Pikachurin deletion specifically influences the ON pathway; the localization of mGluR6 is restricted to the postsynaptic site of ON bipolar cells in the ribbon synapses of the outer plexiform layer. Our previous study showed that the photoreceptor ribbon synapse contributes to the spatiotemporal frequency tuning of the retinal visual processing by which the late phase of OKR is mediated. However, it is still unclear how ribbon synapse structure contributes to visual processing underlying the initial phase of OKR. In the present study, we examined the dependence on ISIs (0-1707ms) in the OKRs of Pikachurin^-/-mice and compared with dependences on ISIs in the wild-type 129Sv/Ev (Taconic) mice and the TRPM1^-/-mice with dysfunction of the ON pathway. In general, two-frame animations elicited OKRs in the veridical direction without an ISI. As the ISI became longer, the OKRs reduced and eventually reversed their directions. We simulated the dependences on ISIs of individual mice by using a computational model of visual motion detection (the elaborated Reichardt detector) to estimate the characteristics of the temporal filters embedded in the visual system of mice. The frequency characteristics of temporal filters in Pikachurin^-/- mice showed no difference in the optimal temporal frequency, but showed stronger adaptation to an exposure to a sustained light signal than the wild type mice. This result suggests that the ribbon synapse is a factor regulating light adaptation of visual information processing. Further, these influences of Pikachurin deletion were different from TRPM1^-/- mice that showed significant difference in optimal temporal frequencies from the wild-type mice, whereas no significant difference in adaptation strength, suggesting that this result is not simply due to dysfunction of the ON pathway.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-166 Spatial distribution of hue selectivity in DKL color space in macaque early visual cortex Xiao Du（Du Xiao）¹，Ichiro Kuriki（Kuriki Ichiro）²，Xinrui Jiang（Jiang Xinrui）¹，Tao Zhou（Zhou Tao）¹，Hisashi Tanigawa（Tanigawa Hisashi）¹ ¹Qiushi Academy for Advanced Studies, Zhejiang University ²Research Institute of Electrical Communication, Tohoku University, Sendai, Japan. It is a fundamental question about color vision how cone signals are transformed into perceptual colors in the cortex. Using optical imaging, previous studies revealed functional structures for color processing in the early visual cortex, such as CO blobs in V1, CO thin stripes in V2, and color-sensitive domains in V4, and these structures are thought to play an important role in the transformation of cone signals. However, it is not known how hue selectivities based on cone opponency is organized as spatial arrangement in early visual cortex. In this study, we examined distribution of domains selective to individual hues in, so called, Derrington-Krauskopf-Lennie (DKL) color space that is based on the cone-opponent axes of the lateral geniculate nucleus (LGN). We performed intrinsic signal optical imaging on one monkey under anesthesia. Under 630 nm wavelength illumination, light reflectance was captured over a portion of the exposed cortex including V1, V2, and V4 by a CCD camera. Visual stimuli were presented on a carefully calibrated LCD monitor in front of the animal and to each eye separately using electromechanical shutters. The stimuli consisted of full screen drifting square-wave gratings of either chromatic or achromatic modulations in two spatially orthogonal orientations. The chromatic gratings were isoluminant color/gray or color/color stripes: the hue of color stripes was chosen from eight evenly spaced directions in an isoluminant plane of the DKL color space, in which four of them were along the L-M and S-(L-M) cardinal axes and the others were along two intermediate directions. The achromatic gratings had 10% luminance contrast. During the visual stimulation, we recorded reflectance changes associated with neural activity in the imaged region and created differential maps between stimulus conditions to visualize the spatial distribution of hue selectivities. Differential maps of isoluminant color/gray gratings minus luminance-contrast achromatic gratings revealed that different hue-selectivities were orderly localized within V2 and V4, regardless of whether the hue was chosen along the cardinal axes or the intermediate directions. In V1, on the other hand, the representation of hue selectivities was biased in favor of particular hues. We are now seeking the rules of hue representation in these visual cortices using DKL color space.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-167 Distinct projections from the lateral pulvinar to V2 thick/thin stripes in macaques. Toru Takahata（Takahata Toru），Ye Liu（Liu Ye），Jiaming Hu（Hu Jiaming），Songping Yao（Yao Songping） Zhejiang University It has been known that primate V2 is subdivided into at least three sub-compartments: thick stripes, thin stripes, and pale stripes, according to their reactivity to cytochrome oxidase histochemistry. Later, it has been revealed that these histochemical sub-compartments are associated with functional reactivity to distinct types of visual stimuli, such that thick stripes are more responsive to directional movement and depth coding, thin stripes are more responsive to color stimuli, and pale stripes are more responsive to form and orientation of the visual stimuli. Furthermore, these physiological properties are reasonably associated with connectivity from V1, as neurons in color-coding blobs preferentially project to thin stripes and neurons in interblobs preferentially project to thick and pale stripes. On the other hand, it was previously revealed that thick and thin stripes receive direct projections from the pulvinar complex of the thalamus, but pale stripes do not. However, it remains to be elucidated which parts of the pulvinar exactly send afferents to V2 stripes, and whether different subpopulation or sub-compartments of the pulvinar send parallel projection into different types of V2 stripe. Here, to address these questions, we injected different kinds tracers, BDA, CTB-Alexa-488 and CTB-Alexa-555, into three consecutive thick/thin stripes in V2, and examined retrograde labeling in the pulvinar of macaques. As a result, we found that there are a few patchy distinct labeling for each retrograde tracer, and that thick stripe-projecting compartments and thin stripe-projecting compartments are segregated, although they are located next to each other within the lateral pulvinar. Our study indicates a possibility that there are several parallel pathways within the pulvinar-V2 projection, similar to the manner of geniculo-striate projections.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-168 マウス頭頂連合野から一次視覚野への抑制投射は視覚野のコントラスト感受性制御に機能する Ryuichi Hishida（菱田竜一），Katsuei Shibuki（澁木克栄）新潟大脳研システム脳生理 Feedback projections from higher association areas to primary sensory cortex are thought to have an important role to control the cortical processing of sensory information. They are known to work for higher cognitive functions such as multimodal integration and attentional control, but it remains unclear about the underlying neural mechanisms. To better understand the mechanisms, we have investigated the inhibitory controls from posterior parietal cortex (PPC) to the primary visual cortex (V1) using transcranial flavoprotein fluorescence imaging in mice. We have revealed that the PPC negatively regulates V1 activity. Artificial activations of the PPC resulted in the reduction of V1 visual response. The PPC activations with two different intensities of electrical stimulation showed that the inhibition of V1 was stronger as the activity of the PPC increased. On the contrary, the inactivation of the PPC, the cutting of the cortical pathway from the PPC to V1 or muscimol injection to the area, resulted in the abolishment of the V1 suppression and the enhancement of on-response in V1 to LED light stimuli. These results indicate that the PPC has a function to inhibit V1 activity persistently. Interestingly, the PPC inactivation by the cortical cutting showed the enhancement of V1 response to the grating stimuli with high contrast and the reduction of the response to the ones with low contrast. These results may be explained by the contribution of the PPC to maintain the excitatory-to-inhibitory (EI) balance in V1. The sudden cutting of the cortical pathway could cause not only a partial disinhibition in V1 but also an increase in noise or spontaneous activities in V1 which is known to be related to a decrease in contrast sensitivity. Thus, the PPC may have function of adjusting the noise level of the visual cortex.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-169 ラットにおけるコントラスト感度および視覚刺激検出課題成績に対応したV1ニューロンの視覚応答とセロトニンによる調節効果 Akinori Sato（佐藤彰典）^1,4，Keisuke Tsunoda（角田圭輔）¹，Ryo Mizuyama（水山遼）^1,5，Satoshi Shimegi（七五三木聡）^1,2,3 ¹大阪大院生命機能スポーツ脳情報科学 ²大阪大全学教育推進機構スポーツ脳情報科学 ³大阪大院医スポーツ脳情報科学 ⁴大阪大生体統御ネットワーク ⁵大阪大ヒューマンウェアイノベーション Serotonin (5-HT) is one of the neuromodulators and released from the serotonergic neurons in the dorsal raphe nucleus to the whole brain in a behavioral context-dependent manner. It implies that 5-HT optimizes various brain functions including vision to suit the behavior of animal. To know the contribution of endogenous 5-HT to visual function of rats conducting visual detection, we previously examined the effects of fluoxetine (FLX), a selective serotonin reuptake inhibitor, on a behavioral contrast sensitivity (CS) measured using a two-alternative forced-choice visual detection task (2AFC-VDT). Intraperitoneal administration of FLX significantly increased CS comparing with no drug control condition, suggesting that endogenous serotonin improved behavioral CS. In this study, to clarify the neural mechanism underlying the CS improvement of FLX, we first constructed a new CS measurement system using the same visual detection task under head fixed condition. And then, we recorded the neural activity from the rat primary visual cortex (V1) using electrodes with multiple recording points during behavioral CS measurement in the presence and absence of FLX. Comparing to no drug condition, FLX administration did not significantly improve behaviorally-measured CS, but tended to facilitate visual responses of V1 neurons. To confirm whether and how the neural activities of V1 correlated with the task performance, that is, the success/failure (Hit/Miss) of visual detection, we compared visual responses between Hit and Miss trials for each neuron. Among 465 neurons recorded under no drug condition, 173 (37%), 75 (16%), and 217 (46%) neurons showed Hit-preference (larger response in Hit trials than Miss trials), Miss-preference (larger response in Miss trials than Hit trials), and no difference, respectively. Moreover, layer analysis demonstrated that the proportion of Hit-preference neurons for each layer were higher in layers 2/3 and 5/6 than layer 4, suggesting the possibility that success/failure of task is determined by the magnitude of visual signals from V1 to higher order visual areas, and that facilitation of the visual responses of V1 neurons by 5-HT improves behavioral CS. We are currently conducting the same experiments under higher dose conditions of FLX to obtain significant effects on CS and to examine how the responses of Hit preference neurons are modulated by 5-HT.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-170 網膜におけるON型／OFF型スターバーストアマクリン細胞の非対称性 Mie Gangi（雁木美衣），Toshiyuki Ishii（石井俊行），Makoto Kaneda（金田誠）日本医科大医生理（システム生理） Starburst amacrine cells (SACs) play an important role in the formation of direction selective responses in the retina. There are classified into ON SACs and OFF SACs based on their morphological properties. The ON SACs have cell bodies in the ganglion cell layer and arborize their dendrites in sublamina b of the inner plexiform layer (IPL). The OFF SACs have cell bodies in the inner nuclear layer and arborize their dendrites in sublamina a of the IPL. Because of the easy accessibility to the soma in the whole-mount preparation, the electrophysiological properties of ON SACs have been rigorously studied. The electrophysiological properties of OFF SACs were assumed to be same as ON SACs. However, in a previous set of experiments, we identified three ways in which the electrophysiological properties of OFF SACs were different from those of ON SACs. First, the P2X2-purinergic signaling pathway predominates in OFF SACs. Second, the glycinergic signaling pathway is predominantly used by ON SACs. Third, acetylcholine (ACh) synthesis might differ between ON and OFF SACs. In the present study, we examined whether a possible difference in ACh synthesis can produce further differences in cholinergic signaling pathways. We found that the application of ACh or carbachol (CCh) induced GABAergic IPSCs in both ON and OFF SACs, suggesting the formation of reciprocal dendritic contacts in the IPL. We also found that the actions of ACh/CCh differentially changed during development between ON and OFF SACs. These results indicate that ON- and OFF-pathways are not mirror-symmetric systems in the IPL and might differentially contribute to the direction selectivity in the retina.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-171 マウス網膜におけるAAVの感染指向性 Masashi Fukutome（福留雅史）¹，Tesshu Hori（堀哲崇）²，Chisato Maejima（前嶋千瀬都）²，Kenta Kobayashi（小林憲太）³，Satoru Morito（森藤暁）^2,4，Chieko Koike（小池千恵子）^1,2,4 ¹立命館大院生命 ²立命館大薬 ³生理学研究所 ⁴総合科学技術研究機構・システム視覚科学研究センター With an increasing number of causative genes identified, the widespread use of gene therapy becomes feasible. The number of retinal disease patients increases year by year, so the discovery of new gene therapy for retinal diseases has a high contribution. For gene therapy, it is desirable to have a delivery method with both high cell type specificity and high efficiency. The need for cell type specificity and high efficiency is particularly important for treating retinal degenerations, which may result from defects in the pigmented epithelium, in rods, or in cones. Viruses are potent gene delivery vehicles for the nervous system, but they suffer from non-specific infection. To address specificity and efficiency in targeting retinal photoreceptors, we have screened various adeno-associated virus (AAV) serotypes for infection patterns in the mouse retina following subretinal injection. AAV is currently the main viral vector that researchers use and further develop for gene therapy because it is considered to be non-pathogenic to humans and because it has been successfully altered to prevent its integration into the genome, thus eliminating DNA damage and unpredictable consequences. As the result, we have identified that AAV2/6 specifically and efficiently infected cone photoreceptors, compared to AAV2/5. It is possible to introduce a target gene to pyramidal photoreceptor cells with high probability, suggesting the possibility of new gene therapy for retinal diseases caused by pyramidal photoreceptor cells.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-172 視覚パタンの乱雑さ判断の神経基盤：経頭蓋磁気刺激による研究 Koki Kannaga（金長幸希）¹，Maki Hatanaka（畑中麻希）²，Daiki Yoshioka（吉岡大貴）¹，Takuya Miyagi（宮城拓弥）¹，Shigeki Takeuchi（竹内成生）^3,4，Yuki Yamada（山田祐樹）⁵，Hiroyuki Yamamoto（山本裕之）⁶，Yoshihiro Itaguchi（板口典弘）^1,2，Hirofumi Sekiguchi（関口浩文）⁴，Makoto Miyazaki（宮崎真）^1,2,3 ¹静岡大学大学院総合科学技術研究科情報学専攻 ²静岡大学情報学部 ³静岡大学創造科学技術大学院 ⁴上武大学ビジネス情報学部 ⁵九州大学基幹教育院 ⁶静岡大学保健センター Negative aftereffect occurs in the perception of random visual patterns (Yamada et al. 2013). The pattern randomness aftereffect is non-selective to the contrast polarity of visual patterns, though it is selective to pattern orientation. Based on these psychophysical properties of the pattern randomness aftereffect and previous neurophysiological findings, Yamada et al. predicted that the lateral occipital complex (LOC) is involved in the perceptual processing of visual pattern randomness. Using functional magnetic resonance imaging (fMRI), we previously found neural activity related to judgment of visual pattern randomness in the right LOC (Yamada et al. 2014). In the present study, we used transcranial magnetic stimulation (TMS) to investigate the brain-behavior relationship in judging visual pattern randomness. Eight participants judged whether the randomness of the target visual pattern (randomness index, ω = 1, 2, 3, 4, 5, 6 or 7 pixels; detailed in Yamada et al., 2013) was lesser or greater than a reference visual pattern (ω = 4 pixels). During the judgment task, TMS was applied to PO8 (using the 10-10 system) located over the right LOC. We used paired pulse TMS with an inter-stimulus interval of 10 ms. The TMS intensity was 40% of the maximum intensity of the stimulator (Magstim 200²), as used in a previous study applying TMS over the LOC (Bona et al. 2014). Based on reanalyzed results of our previous event-related potential (ERP) study (Miyagi et al. 2017), we set the TMS pulses to occur at 180 ms, 210 ms, and 250 ms after the onset of the target visual pattern. These intervals were selected to disrupt the ERP response to random visual patterns (latency: 210 ms on average), taking into account individual differences in the ERP response. A no-TMS condition was also included. The three TMS-interval and no-TMS conditions were randomly presented with a probability of 25%. As a result, TMS slowed the reaction time for judgment of visual pattern randomness, compared to the no-TMS condition. Moreover, TMS increased the just-noticeable difference for visual pattern randomness, that is, TMS decreased the perceptual resolution for visual pattern randomness. Thus, TMS at PO8 functionally disrupted judgment of visual pattern randomness, which provides evidence to support the causal role of the right LOC in the perceptual processing of visual pattern randomness.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-173 マウス一次視覚野におけるFast spiking抑制性細胞－錐体細胞間結合の発達 Mariko Yamamoto（山本真理子）^1,2，Takuji Iwasato（岩里琢治）^3,4，Yumiko Yoshimura（吉村由美子）^1,2 ¹生理研視覚情報処理 ²総研大院生命科学生理科学 ³国立遺伝研神経回路構築 ⁴総研大院生命科学遺伝学 The balance of excitation and inhibition is important for visual cortical function. We previously showed that fast-spiking (FS) neurons, a major subgroup of inhibitory interneurons, form strong inhibitory connections to pyramidal (Pyr) neurons, when they are reciprocally connected. In this study, we investigated the developmental process of synaptic connections between FS and Pyr neurons using mice at three developmental stages, P10-13 (before eye-opening), P14-16 (soon after eye-opening) and P21-26. We conducted dual whole-cell recordings from FS and Pyr neurons in layer 2/3 of visual cortical slices. In FS-Pyr neuron pairs, the proportion of reciprocally connected pairs was considerably higher than that of one-way excitatory or inhibitory connected pairs at P10-13, demonstrating that reciprocal connections were preferentially established before eye opening. The proportion of synaptic connections did not significantly change during development. At P10-13, the amplitude of IPSCs was not significantly different between reciprocally and one-way inhibitory connected pairs. The IPSC amplitude in one-way inhibitory connected pairs remained almost unchanged from P10-13 to P21-26, whereas that in reciprocally connected pairs significantly increased after eye-opening, resulting in the establishment of strong inhibitory connections in reciprocally connected pairs at P21-26. Next, we examined the effect of postnatal deletion of visual cortical GluN1, the essential subunit of NMDA receptor, on the development of synaptic connections using GluN1 conditional knock-out mice. The knock-out of GluN1 did not affect the proportion of synaptic connections analyzed at P21-26. In P21-26 KO mice, the IPSC amplitude in reciprocally connected pairs and one-way connected pairs slightly decreased and increased, respectively, compared with age-matched normal mice, leading no significant differences in the IPSC amplitude between these pairs. Thus, GluN1 may be required for regulation of inhibitory strength depending on synaptic connectivity. We also analyzed excitatory connections in FS-Pyr neuron pairs. We found no significant changes in the EPSC amplitude among normal mice at three stages. In addition, the amplitude was not significantly different between GluN1 KO and normal mice at P21-26. These results suggest that the strength of inhibitory connections in FS-Pyr pairs is modified after eye-opening depending on the synaptic connectivity using NMDA receptor-dependent mechanisms.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-174 時間順序課題によって計測した知覚潜時に及ぼす刺激呈示位置と視空間的注意の影響 Sayaka Fujimoto（藤本紗也加）¹，Hiroshi Tamura（田村弘）^1,2 ¹大阪大院生命機能 ²脳情報通信融合研究センター In this study, we aimed to clarify the relationship between retinal position (eccentricity and nasal-temporal visual fields) and perceived temporal order of visual stimuli. In the retina, stimulus presented in a peripheral visual field (> 24 degrees) is transmitted to the optic disc more quickly by 30 ± 7 milliseconds (mean ± standard deviation) than that presented at the fovea (0-4 degrees) (Sinha et al., 2017). Contrary, stimulus presented at the fovea are perceived 42.8 ± 18.9 ms faster than those presented in a peripheral visual field (30 degrees) (Rutschmann, 1966), meaning that information received at the fovea is quickly transmitted to the central nervous system. To resolve this conflict, we performed a series of psychophysical experiments, i.e., the temporal order judgement task. Stimuli were presented monocularly to the right eye. In the first experiments, we tried to replicate the previous psychological experiment (Rutschmann, 1966), and found that the perceptual latency to the stimulus at the fovea was 22 ms shorter than that in the peripheral visual field (p < 0.001, Wilcoxon signed bilateral rank test). In the second experiments, we dispersed the spatial attention by the periphery cue and found that the perceptual latency was comparable between the fovea and the peripheral visual field (p = 0.38). From these results, shorter perceptual latency observed at the fovea in the previous and in the first experiment is at least partially explained with spatial attention directed to the fovea. However, still there remains the contradiction between the retinal and perceptual temporal order, suggesting the presence of neural mechanisms that transmit information from the central visual field quickly. Because the optic disc is in the nasal side, stimulus presented to the temporal visual field is likely to be perceived faster. Therefore, in the third experiments, we compared perceptual latency between nasal and temporal visual filed and found that perceptual latency did not differ between them suggesting the distance to the optic disc did not contribute to the perceived latency. During the course of the third experiment, we found that, perceptual temporal order was reversed when stimulus onset asynchrony was 8 ms, suggesting that mechanisms other than neural transmission contribute to temporal order judgment.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-175 マウス側頭皮質における新規高次視覚野は、小領域ごとに異なる神経投射を示す Nana Nishio（西尾奈々）^1,2,3，Katsuei Shibuki（澁木克栄）³，Kenichi Ohki（大木研一）^1,2，饗場篤，崎村健司，澁木克栄 ¹東京大院医統合生理 ²東京大ニューロインテリジェンス国際研究機構 ³新潟大脳研基礎神経科学システム脳生理 The visual cortex of mice is a useful model for investigating the mammalian visual system. In primates, higher visual areas are classified into two parts, the dorsal stream and ventral stream. The ventral stream is known to be important for object recognition and called ""what"" pathway. In rodents, the lateral parts of the occipital cortex around the V1 are suggested to belong to the ventral stream, although the whole picture of the mouse ventral stream has not been elucidated. We recently demonstrated that the ectorhinal cortex (ECT, or TEa/36a), innervated by a ventral visual stream area POR, shows responses to moving object visual stimuli, and that the responsive regions in ECT were shifted depending on object size (Nishio et al., 2018). We further analyzed neuronal responses by using two-photon Ca2+ imaging and found that ECT neurons show selectivity to object size; neurons in the anterior part of the ECT show response preferences to large size object stimuli (˜20deg) and neurons in the posterior part show preferences to small size (˜2.5deg). These results suggest that neurons in the ECT represent object size. We analyzed neuronal projections to the ECT by using a retrograde tracer, and observed labelled cell bodies in visual cortices and several other areas including lateral amygdala. We also analyzed neuronal projections from the ECT by using AAV, and observed labelled axonal projections mainly in the tail of striatum and lateral amygdala. Interestingly, axonal projection patterns from the anterior and posterior parts of ECT were modestly different. Projections from anterior ECT localized in anterior parts of both the tail of striatum and lateral amygdala, compared to projections from posterior ECT. It is well known that the amygdala has broad functions including reward and fear leaning, and it has been reported that the tail of striatum is involved in threat prediction and retreat from novel objects. Thus, we hypothesize that the differences in neural functions and projections of the anterior and posterior ECT are involved in behavioral responses to real objects. We preliminarily conducted novel-object interaction tests with a large size novel object (H8cm) or a small novel object (H2cm), and mice showed tendency to approach to a small object more frequently than to a large object. Further research on the anterior and posterior ECT will provide insight into the understanding the what pathway in mouse temporal cortex.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-176 麻酔による上丘の方位選択性構造の違い Masatoshi Kasai（笠井昌俊），Tadashi Isa（伊佐正）京都大院医神経生物 The superior colliculus (SC) is a brainstem center which plays key roles in generating spatial attention and mediating the signal for sensory-motor translation. It is hypothesized that integration of spatial information is a critical process for selecting the target and shifting attention to it. However, it was difficult to examine neural responses across visual fields of the SC, particularly by recording activities of large neuronal population at a time. To overcome this limitation, we have developed a method of in vivo imaging to simultaneously capture visual responses from hundreds of neurons from the SC of anesthetized mice using the 2-photon laser microscope. In this study, we improved our previous method and will present a new technique for chronic imaging neuronal activities from the SC of awake head-fixed mice. GCaMP6f, Ca²⁺ sensitive fluorescent indicator, is delivered to the SC neurons by injection Adeno-associated virus (AAV) vector (AAV-hSyn-GCaMP6f) after removing small part of the cortex overlaid on the SC. To achieve long-term optical access to the SC and make a stable imaging window, a small glass cube is placed on the SC surface. After several weeks, we started Ca²⁺ imaging using the 2-photon laser microscope. Here, we found that the light-isoflurane anesthesia extensively alters the orientation selective response properties in the sSC. Specifically, neurons in light-isoflurane anesthetized condition showed stronger and sharper orientation selectivity. On the other hand, anesthesia with Ketamine-Xylazine combination showed similar orientation selective response to the awake condition. This result indicates that the orientation selectivity and generation of the orientation column-like structure might be regulated by wakefulness of the animal. This research has been approved by the ethics committee in Kyoto University for the care and use of experimental animals. There is no actual and potential conflict of interest in relation to this presentation.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-177 下側頭葉のリカレント結合が階層的カテゴリー分類に重要である Narihisa Matsumoto（松本有央）¹，Yusuke Taguchi（田口優介）^1,2，Masato Okada（岡田真人）³，Yasuko Sugase-Miyamoto（菅生（宮本）康子）¹ ¹産総研人間情報 ²筑波大院システム情報工学 ³東京大院複雑理工 We previously reported that face-responsive neurons in the anterior inferior temporal (AIT) cortex represent information about a global category, i.e. human vs. monkey vs. simple shapes, earlier than information about fine categories about the faces, i.e. facial expression and/or identity (Sugase et al., 1999; Matsumoto et al., 2005; Sugase-Miyamoto et al., 2014). The neuronal mechanisms of a hierarchical categorization in AIT remains unknown. In our previous study, we consisted a feed-forward deep neural network (FDNN) to compare information representation in each layer and the one encoded by a neural population in AIT with a visual stimulus set including human and monkey faces (Sugase-Miyamoto et al., 2017). We found the global categorization occurred in the lower layers of the FDNN and the fine categorization occurred in the higher layers. However, the time course of the hierarchical categorization could not be replicated by FDNN. In this study, we consisted a combined model of FDNN, i.e., Xception net (Chollet, 2017), and a recurrent neural network, i.e. Hopfield model (Hopfield, 1982), to investigate whether the combined model replicated the time course of the hierarchical categorization. The weights of the Xception net were pretrained by images of the imagenet database (Russakovsky et al., 2015). The weights of the Hopfield model were learned by covariance rule with 15 images taken from the database. When one of the learned images with noise was input to the combined model, the model output the different levels of categories, e.g. Poodle, Dog, and Human, in the time course. This result implies that recurrent connections in the Hopfiled model is important for the hierarchical categorization, suggesting that recurrent connections in AIT would be important for the hierarchical categorization.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-178 訓練なしで校正可能な視線計測システム Keiji Matsuda（松田圭司）¹，Takaaki Kaneko（兼子峰明）^2,3，Kenji Kawano（河野憲二）¹，Hideyuki Okano（岡野栄之）^2,3 ¹（国研）産業技術総合研究所人間情報研究部門 ²(国研）理化学研究所脳神経科学研究センター ³慶應義塾大学医学部 We have developed a non-invasive and inexpensive eye tracking system, which consists of infrared light illuminations, visible-ray cut filter, lens, digital high frame-rate infrared camera, computer, and software. The system has been used for monkey and human subjects. While the infrared light illuminates the eye, the black image of the pupil and the reflected image of the infrared illumination on the cornea are captured by the camera. The center of the pupil and the barycenter of the reflected image are calculated and tracked. The gaze vector towards the camera (gaze vector in camera coordinates) can be calculated from the pupil center and the barycenter of the reflected image. In our previous method, to translate the gaze vector from camera coordinates to object coordinates, we calculated coordination-transformation-parameters by instructing the subject to fixate small targets. But some subjects, for example, young children or marmosets, would not understand/follow the instruction to fixate the small targets for calibration. When the subject is untrained to fixate the stimulus, there is ambiguity to determine whether the subject really looks at the stimulus or not. Here we present a new system which solves the problem. It utilize the characteristics of slow tracking gaze movement which can only be elicited when a target moves slowly and smoothly. We displayed a moving visual stimulus in front of the subject to guide their gaze movements. We used `PsychoPy (https://www.psychopy.org/)' as the visual stimulus presentation platform and created a library for conveniently creating visual stimuli, sending information of the visual stimulus to our eye tracking software, iRecHS2. While the eye tracking system calculates the gaze vector in camera coordinates, it simultaneously receives the timing and position of the visual stimulus from moment to moment. By using the difference between the moving visual stimulus and the moving gaze vector in camera coordinates and ocular tracking delay, coordination transformation parameters from camera coordinates to object coordinates are calculated. By using the new calibration method, we successfully measured eye movements of marmosets without training. The program is available at https://staff.aist.go.jp/k.matsuda/iRecHS2/		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-179 化学遺伝学による扁桃体賦活化によりマカクザル腹側視覚皮質の社会的・情動的視覚情報表現が選択的に変容する Naohisa Miyakawa（宮川尚久）¹，Yuji Nagai（永井裕司）¹，Yukiko Hori（堀由紀子）¹，Takeshi Matsuo（松尾健）²，Takafumi Suzuki（鈴木隆文）³，Ken-ichi Inoue（井上謙一）⁴，Masahiko Takada（高田昌彦）⁴，Tetsuya Suhara（須原哲也）¹，Keisuke Kawasaki（川嵜佳祐）⁵，Takafumi Minamimoto（南本敬史）¹ ¹量研機構放射線医学総合研究所脳機能イメージング研究部 ²東京都立神経病院 ³情報通信研究機構, 脳情報通信融合研究センター ⁴京都大学霊長類研究所統合脳システム分野 ⁵新潟大学大学院医歯学総合研究科統合生理 A classical view of object vision emphasizes the hierarchical feed-forward connections from the lower to higher visual cortical areas, however, the contribution of the feedback projections from subcortical areas, such as amygdala-the brain region responsible for emotion and social processing-has been less attended. Here we investigate the role of amygdala in socio-emotional representation in the ventral visual cortex of macaque monkeys. Two monkeys received injection(s) of an adeno-associated viral vector expressing hM3Dq, an excitatory DREADD (Designer Receptor Exclusively Activated by Designer Drug), into unilateral amygdala. Several weeks after the injection, the monkeys underwent a positron emission tomography (PET) scan with a DREADD-selective ligand, [¹¹C]deschloroclozapine (DCZ), confirming in vivo hM3Dq expression in the amygdala. Neuronal recording studyElectrophysiological recording revealed that a low dose of DCZ administration significantly increased neuronal activity in the hM3Dq-expressing region. Further PET scans with [¹⁸F]fluorodeoxyglucose revealed that DCZ administration significantly increased metabolic activity not only in the hM3Dq-expressing site of the amygdala but also in severalsome patchy regions in the ipsilateral temporal visual cortex, including the anterior inferior temporal cortex and the superior temporal sulcus. This suggests that the amygdala enhances excitability of specific visual cortical sites. To examined the functional effect of amygdala excitation on the neural representation in the ventral visual cortex, one of the monkeys was received an open surgery to implant with an electrocorticogram electrode covering the ipsilateral temporal cortex. Early evoked visual response was recorded while the monkey was requiredperformed a simple fixation during a sequential presentation of visual stimuli, which had two properties: social (face or non-face) and emotional (neutral or negative). We applied support vector machine algorithm to decoded social and emotional information. We successfully decoded face and facial expression, but not emotion in general. Chemogenetic activation of the amygdala decreased decoding performance of facial expression (p< 0.05, chi-squared test) to chance level. These results suggest that the primate amygdala contribute to representation of socio-emotional information in the ventral visual cortex.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-180 視覚誘発性逃避行動における先行刺激による影響 Hiroshi Ishikane（石金浩史）^1,2,3 ¹専修大学心理 ²専修大学大学院心理学専攻 ³専修大学心理科学研究センター Looming objects elicit protective behaviors in both humans and animals. For example, frogs exhibit escape behavior (which is a protective behavior) in response to encountering looming objects. Previous studies have demonstrated that synchronized oscillatory spikes among dimming detectors (class-4 neurons) encode essential information for escape behavior in frogs. Thus, oscillatory activities related to escape behavior in frogs are useful model system to investigate the functional role of synchronous oscillations in the nervous system. It has been demonstrated that the frog retina transmits highly abstracted visual information to the brain than other animals. In addition, retinal ganglion cells are known to be affected by the visual stimuli presented before. In this study, two stimuli were presented successively to the frogs, and their escape behavior was monitored. We investigated how the first stimulus, which cannot trigger the escape behavior by itself, affected the escape behavior triggered by the second stimulus. Dark adapted bullfrog (Rana catesbeiana) was placed on a turntable which was surrounded by a transparent acrylic dome. Visual stimuli were presented by LCD. First, we confirmed that bullfrogs showed escape behavior in response to a large expanding dark spot (maximum diameter: 35 degrees). Next, we primarily presented a small expanding dark spot (maximum diameter: 8.75 degrees) which cannot elicit escape behavior by itself, and then presented a large expanding dark spot. The escape rate was significantly higher than that in the control condition (no preceding small spot). In the frog's retina, when a large stimulus which can elicit escape behavior is presented, class-4 neurons show long-range synchronous oscillations. If we pharmacologically inhibit the synchronous oscillation, the escape rate decreases, and if we strengthen the synchronous oscillation, the escape rate increases. Therefore, these long-range synchronous oscillations might play an important role in the visually guided escape behavior. The prime stimulus we used in this experiment might be too small to elicit the long-range synchronous oscillations, but might strengthen them elicited by the second stimulus.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-181 ショウジョウバエの色覚に関する高次ニューロンの機能解析 Tomoyuki Misawa（三沢智行），Junji Yamauchi（山内淳司），Takako Morimoto（森本高子），Yoichi Seki（関洋一）東薬大生命科学分子生命科分子神経科学 Vision is indispensable for most living things and color vision is one of the important elements that form visual information. Color vision is the ability to discriminate differences in wavelengths of light regardless of their intensity, and to do this, the presence of color opponent cells that compare information from photoreceptor cells with different spectral sensitivities is important. However, the mechanisms that form color opponent responses and the subsequent integration mechanisms of the opponent responses at higher brain centers are not well understood. In this study, in order to identify neurons underlying color vision and to reveal their functions in the neural circuits, we used Drosophila melanogaster as a model organism, which has a visual circuit structure similar to humans and is amenable to genetic manipulations. The visual information received by photoreceptors is processed through the lamina and medulla and is sent to the lobula. Then lobula columnar (LC) neurons receive the information in the lobula and project their axons into the central brain but the details of their functions are not well understood. First, to identify LC neurons that contribute to color vision, we performed color discrimination behavioral experiments using heat aversive learning. A few LC neuron types that were necessary for the color discrimination were identified. Furthermore, to ascertain whether those LC neurons respond specifically to color information and not to light or other factors, we evaluated spectral tuning properties of the LC neurons using calcium imaging. We also performed calcium imaging in second order neurons innervating the medulla and compared their responses to those of the LC neurons. Taken together, our results will help to elucidate color information processing pathways by identifying neurons related to color vision by behavioral and physiological approaches.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-182 ２フレーム仮現運動刺激はサル大脳皮質MST野の活動と追従眼球運動を逆転させた Aya Takemura（竹村文）¹，Kenji Kawano（河野憲二）^1,2，Kenichiro Miura（三浦健一郎）^1,3 ¹産総研システム脳科学 ²獨協医大医生理（生体情報） ³京都大院医認知行動脳科学 Two-frame movies presented with an inter-stimulus interval (ISI) between the first and second frames often induce sensation of motion in the opposite direction to the actual image shift. The motion information is extracted from dynamic visual images by the visual system and induces perception and movements of body and eyes. Ocular following response (OFR) is a tracking eye movement evoked by a sudden movement of a large-field visual stimulus with short latency, and a good example of behaviors that use the extracted visual motion information. Recently, it was reported that two-frame movies with ISIs also induce OFRs in the opposite direction to the actual image shift in primates. This phenomenon seems due to temporal filters with biphasic impulse response functions embedded in the visual system that underlie the OFR-generation. It is known OFRs were mediated by the neuronal pathway including the medial superior temporal (MST) area of the visual cortex. In the present study, we recorded single unit activities of direction selective neurons in MST of a monkey (Macaca fuscata), while presenting the two-frame movie stimuli which elicit OFRs, to understand their role in the reverse-OFRs. Each trial started by presenting a small fixation spot on a gray background. After the eye was positioned within ±1.5° of the fixation spot for a brief, randomized period, a sinusoidal grating (spatial frequency, 0.25 cpd, Mickelson contrast, 32%), whose orientation was orthogonal to the axis of preferred direction of each neuron, was displayed behind the fixation spot as the first frame. As the second frame, the grating was displayed with a 90° phase shift in neuron's preferred or anti-preferred direction with various ISIs (0-320 ms). To measure ocular and neuronal responses, we calculated changes in eye position and magnitudes of neuronal activities during the 60-ms starting from 60 ms and 40 ms after the second frame onset, respectively. We found that without ISI, the neuronal responses were larger when the stimulus stepped in the preferred than anti-preferred direction. However, this relationship was inverted when there were ISIs (> 10ms). The modulation of the neuronal activities was, in general, consistent with that of the OFRs regarding the dependence on ISIs. Since the neuronal responses preceded the ocular responses, the present results suggest a causal link between neuronal responses in MST and reverse-OFRs to two-frame movies presented with ISIs.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-183 スパースSTCによる、一次視覚野複雑型細胞の機能的サブユニットの同定 Kota S. Sasaki（佐々木耕太）^1,2，Takashi Tsukada（塚田尭）¹，Yunosuke Azuma（東侑之介）¹，Hirotaka Sakamoto（坂本浩隆）³，Yoshihiro Nagano（長野祥大）³，Yonghao Yue（楽詠灝）³，Masato Okada（岡田真人）^3,4，Izumi Ohzawa（大澤五住）^1,2 ¹大阪大院生命機能 ²脳情報通信融合研究センター ³東京大院新領域創成科学 ⁴脳神経科学研究センター One goal of sensory neuroscience is to understand how stimulus is transformed into neuronal responses so that they can be predicted for any given stimulus. Toward this goal, all functional elements should be discovered for the responses of single neurons. These elements can be recovered via spike-triggered covariance (STC) analysis by investigating pairwise relationship between stimulus pixels that potentially cause neuronal responses in visual system. STC analysis generally requires a tremendous number of spikes because of the curse of dimensionality. To alleviate this problem, we have devised a novel method (sparse STC) by using a sparse modeling (least absolute shrinkage and selection operator; LASSO) technique in the Fourier domain to help STC analysis. More specifically, superposing a minimal set of 4D waves, we modeled a variance-covariance matrix from which functional elements are extracted in STC analysis. First, the canonical model of a V1 complex cell (i.e., energy model) was used to characterize our method quantitatively. We found that Fourier LASSO estimated a variance-covariance matrix more accurately than the conventional STC method. As a result, functional elements were recovered with less noise in the sparse STC. To obtain results with the same S/N ratio, the number of stimuli was reduced by a factor of 5 to 10 in the sparse STC. Namely, approximately 10,000 stimuli were used in the sparse STC to recover the two functional elements of the model complex cell completely. Then, this analysis was performed to discover functional elements in actual complex cells recorded in monkey and cat primary visual cortex. For the first two elements, we obtained results qualitatively similar to the model cell. We wish to apply sparse STC to unveil functional elements that are yet to be discovered by previous techniques.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-184 サルV1野・V4野における、神経細胞の同期発火を元にした、視覚の集団表現の検討 Kouhei Motoya（本谷康平）¹，Kouki Shigekawa（重川幸輝）¹，Kota S Sasaki（佐々木 S 耕太）^1,2，Hiroshi Tamura（田村弘）^2,3，Izumi Ohzawa（大澤五住）^1,2 ¹大阪大院生命機能視覚神経科学 ²大阪府吹田市山田丘1-4 脳情報融合研究センター ³大阪大院生命機能認知脳科学 Sensory information is represented in the coordinated activity of a huge number of neurons in the cerebral cortex. Synchronous firing among them may underlie efficient transmission and representation of sensory information. The degree of neuronal synchrony in the visual cortex was examined in relation to orientation selectivity and/or physical distance in previous studies. Although orientation selectivity is one of the most remarkable characteristic of single neurons in visual cortex, multiple stimulus parameters usually affect their responses. Here, we investigate whether neuronal synchrony is related to selectivity to other stimulus parameters (e.g. spatial frequency) as well as to orientation. Penetrating the cortex with a 32-ch Michigan probe in presumed vertical direction, we recorded multiple single unit activities simultaneously in anesthetized and paralyzed macaque monkeys. These experiments were performed in V1 and V4 separately. To investigate synchronous activity of single units, cross-correlation analysis was performed for spike trains between all pairs of recorded neurons. The peak height of the cross-correlograms (CCGs) was used as a measure of neuronal synchrony. This measure was analyzed as a function of vertical distance between the neurons and similarity of their stimulus selectivity. Stimulus selectivity was evaluated for each neuron via a reverse correlation technique by presenting a rapid succession of flashed gratings defined by orientation, spatial frequency and spatial phase. The following analysis was performed for pairs of neurons whose activity was synchronous and whose stimulus selectivity was determined reliably (20 pairs in V1 and 106 pairs in V4). The degree of synchrony was strongly related to distance between neurons and similarity of orientation selectivity in V4 (3-way ANOVA, p < 0.001). In our small sample of V1 neuronal pairs, neuronal synchrony appeared to be related to similarity of their optimal spatial frequency. On the other hand, they did not seem to be related in V4 neuronal pairs.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-185 投射ターゲットおよび層特異的なV1神経細胞のサブネットワーク構造の解明 Ryosuke F Takeuchi（竹内 F 遼介）¹，Ami Tsuboi（坪井愛実）¹，Toshiaki Suzuki（鈴木俊章）¹，Masanari Onda（恩田将成）¹，Nao Morimoto（森本菜央）^1,2，Keisuke Isobe（磯部圭佑）³，Fumitaka Osakada（小坂田文隆）^1,2,4 ¹名古屋大院創薬 ²名古屋大高等研究院 ³理研光量子工学研究センター ⁴名古屋大未来機構ナノライフ研 Visual information processing is achieved by the hierarchical organization of the visual cortex. Mouse visual cortex consists of a primary visual area (V1) and at least nine higher-order visual areas (HVAs) which are functionally tuned to specific visual features. AL, one of the HVAs, specializes in processing fast moving, low spatial frequency visual stimuli, while PM, another HVA, specializes in processing slow moving, high spatial frequency stimuli. The functional specialization of HVAs can arise from the interaction between V1 and HVAs. Thus, revealing how information flows between V1 and HVAs is pivotal in understanding the mechanisms of vision. However, how HVA-projecting V1 neurons (HVAPs) integrate information from their presynaptic neurons remains unknown. Here we characterized the presynaptic networks of AL-projecting or PM-projecting V1 neurons (ALPs or PMPs, respectively). Monosynaptic inputs to ALPs or PMPs were mapped across the whole brain by a trans-synaptic viral tracing strategy. We injected rAAV2-retro-TVA-mCherry and rAAV2-retro-H2B-BFP-P2A-oG into either AL or PM, and then injected EnvA-pseudotyped, glycoprotein-deleted rabies virus expressing GFP into V1, based on both the retinotopic organization and the Allen Mouse Brain Atlas. We found anatomical differences in their presynaptic neurons between ALPs and PMPs. More PMPs and their presynaptic cells were distributed in the deeper layers of V1 than ALPs. Moreover, PMPs received many inputs from cortical areas such as retrosplenial cortex that are associated with executional functions, whereas most presynaptic cells of ALPs were in the visual areas. We next performed in vivo calcium imaging of ALPs or PMPs by 2-photon dual-plane microscopy to reveal how ALPs and PMPs represent information. Functional connectivity analysis revealed that correlation was higher for pairs of HVAPs in layer 2/3 than for pairs of other layer 2/3 neurons, suggesting the organization of projection-target-specific subnetworks responsible for processing specific attributes of sensory inputs. By contrast, pairs of HVAPs in layer 5 showed segregated correlation structures, suggesting that different subsets of HVAPs in layer 5 represent distinct information from other V1 neurons. From these results, we conclude that the projection-target-specific information flows from V1 to HVAs are achieved by integration of inputs from topographically and functionally segregated neurons through cortical layer-dependent pooling.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-186 霊長類における皮質脳波記録による自然視線行動下の皮質情報ダイナミクス Takaaki Kaneko（兼子峰明）^1,2，Misako Komatsu（小松三佐子）^3,4，Noritaka Ichinohe（一戸紀考）^3,4，Hideyuki Okano（岡野栄之）^1,2 ¹理研CBS マーモセット神経構造 ²慶應大医生理 ³理研CBS 高次脳機能分子解析 ⁴国立精神・神経セ神経研微細構造 Advances in neuroscience has accumulated snapshot understanding on how eye-movement is generated or how object identity is analyzed in several neural structures. However, it remains unknown how spatial and temporal interactions of multiple cortical areas generate sequences of active visual behavior under naturalistic condition. In the present study, we report the cortical information dynamics of visual behavior by recording electrocorticographic (ECoG) signal covering the entire hemisphere of the marmoset while the animal freely viewed naturalistic movie stimuli. We found that, under natural visual behavior, visual computation did not start from primary visual cortex rather MT complex and posterior parietal cortices which showed distinct pattern of efferent and re-afferent activity just after saccade onset. Furthermore, we showed that these rapid post saccade activities transmitted to, but not from, the early visual areas. We also found that information is not continuous flow even though the retina can sample visual information continuously, but rather a packet of information which travels from dorsal to ventral visual stream. Furthermore, we found active vision was intrinsically recurrent process where brain and behavior coordinated to maintain certain amount of neural activity. We observed that saccade triggered a transient neural activity and the following saccade occurred to refresh neural activity just before sum of a whole brain activity became a silent. These results provide several novel views of macro-level information dynamics which might be crucial for efficient and stable perception of natural active visual behavior. Understanding brain dynamics under natural vision may contribute not only to understanding how visual system is designed to work on real-life problems, but also to development of a new AI architecture which requires time-efficient on-time computation such as for robotics.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-187 同時多平面二光子イメージングによる神経回路の三次元解析 Masanari Onda（恩田将成）¹，Ryosuke F Takeuchi（竹内 F 遼介）¹，Toshiaki Suzuki（鈴木俊章）¹，Nao Morimoto（森本菜央）^1,2，Keisuke Isobe（磯部圭佑）^3,5，Fumitaka Osakada（小坂田文隆）^1,2,4,5 ¹名古屋大院創薬 ²名古屋大高等研究院 ³理研光量子工学研究センター ⁴名古屋大未来機構ナノライフ研 ⁵戦略的創造研究推進事業（CREST） One of the main goals of neuroscience is to understand how neurons organize neural circuits to perform computations that generate perception and behavior. Two-photon excited fluorescence microscopy, combined with in vivo labeling with genetically encoded indicators via viral vectors and transgenic animals, has allowed us to analyze the neural activity with high spatiotemporal resolution in living animals. Although two-photon imaging has been powerful in neuroscience research, conventional two-photon microscopes have been restricted to imaging from a single two-dimensional plane. However, the cortex is composed of six layers and each layer has distinct roles for cognitive and behavioral functions. Thus, three-dimensional, functional imaging is essential for understanding how cell types, neural circuits, and neural computations are implemented in the brain. Here we show simultaneous multi-plane two-photon calcium imaging to measure neural activity in the mouse cerebral cortex in vivo. We constructed a dual-plane two-photon imaging system using a spatial light modulator (SLM). We split a laser beam into two pathways; one laser beam directly excites one plane, and the other through SLM excites the other focal plane. The system can image two independent focal planes at 15 Hz each without mechanical movements of objective lens. We performed a proof-of-concept experiment showing simultaneous recording of the neural activity of hundreds of neurons at two different focal depths. Neurons expressing the calcium indicator GCaMP6 in the layers 2/3 and 5 of the mouse primary visual cortex (V1) showed spontaneous activity and visual responses to drifting gratings and natural images. Correlated activity between pairs of neurons in intra- and inter-layers decreased with the increasing distance of neurons. To reveal area- and layer-specific roles of projection neurons in the visual information processing, we next combined our three-dimensional two-photon imaging system with the rabies viral circuit mapping. We retrogradely labeled V1 neurons projecting to LM, a higher visual area, and recorded GCaMP6 signals from the layers 2/3 and 5. We analyzed the relationships between LM-projecting V1 neurons and other V1 neurons in the layers 2/3 and 5 to relate three-dimensional neuronal connectivity to circuit function. Our combined approach of three-dimensional two-photon imaging and viral circuit mapping will allow linking circuit mechanisms to complex computations and behaviors.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-188 神経障害性疼痛における脊髄Shati/Nat8lの役割とmGluR3の関与 Keisuke Miyamoto（宮本啓補）¹，Kazuyuki Sumi（鷲見和之）²，Kyosuke Uno（宇野恭介）²，Kazuhiko Kume（粂和彦）¹，Atsumi Nitta（新田淳美）²，Masahiro Ohsawa（大澤匡弘）¹ ¹名古屋市大院薬神経薬理 ²富山大院医薬薬物治療 Shati/Nat8l is characterized as an N-acetyltransferase-8-like protein (Nat8l) that catalyzes N-acetylaspartate (NAA) synthesis from aspartate and acetyl-coenzyme A. NAA is combined with glutamate and converted into N-acetylaspartylglutamate (NAAG), selective neurotransmitter for group II metabotropic glutamate receptor 3 (mGluR3), by NAAG synthetase. Neuropathic pain shows hyperalgesia and allodynia, but the most of the mechanisms remain unknown. Recently, we found that Shati/Nat8l knockout (Shati^-/-) mice represent the mechanical hypersensitivity. Thus we hypothesized that the neuropathic pain-induced mechanical hypersensitivity is caused by the decrease of Shati/Nat8l expression. In this study, we investigated the role of Shati/Nat8l to neuropathic pain in the spinal cord. Male Shati^-/- mice and wild type (Shati^+/+) mice were subjected to each experiment. The mechanical threshold was evaluated by von Frey filament test. Partial sciatic nerve injury (PSNL) model was used as neuropathic pain model. The mRNA level was quantified by real-time and semiquantitative reverse transcription-polymerase chain reaction or in situ hybridization. The expression of Shati/Nat8l mRNA in L4 to L6 spinal dorsal horn was attenuated in PSNL model mice as compared with sham operated mice. Lowered mechanical threshold by PSNL or Shati^-/- mice was diminished by rescue of Shati/Nat8l to spinal dorsal horn using adeno associated virus which codes Shati/Nat8l under the CMV promoter. Next, we investigated the detail mechanisms of the influence of Shati/Nat8l on mechanical nociceptive threshold. Intrathecal (I.t.) administration of NAAG (10 μg/animal) attenuated the mechanical hypersensitivity in Shati^-/- mice, which was inhibited by mGluR3 inhibitor LY341495 (0.3 mg/kg, intraperitoneal). However, i.t. treatment with NAA (10 μg/animal) did not affect the mechanical hypersensitivity in Shati^-/- mice. These results suggest that the decreased expression of Shati/Nat8l is involved in the hypersensitivity through the inhibition of NAAG-mGluR3 pathway in neuropathic pain.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-189 痛覚鈍麻マウスの順遺伝学スクリーニングによる新規疼痛候補遺伝子の同定 Tatsuhide Tanaka（田中達英）¹，Hiroaki Okuda（奥田洋明）²，Yuki Terada（寺田雄紀）¹，Takeaki Shinjo（新城武明）¹，Kazuya Nishimura（西村和也）¹，Ayami Isonishi（石西綾美）¹，Shoko Takemura（竹村晶子）¹，Kouko Tatsumi（辰巳晃子）¹，Akio Wanaka（和中明生）¹ ¹奈良医大医第2解剖 ²金沢大医機能解剖学 Pain significantly reduces quality of life (QOL) in various diseases. Although pain research has revealed pain-modulating cells, molecules and neural pathways, the cellular and molecular mechanisms of pain still remain largely unknown. We serendipitously noticed that pain responses to mechanical and chemical stimuli were extremely reduced in a bacterial artificial chromosome-transgenic (BAC-TG, Mlc1-tTA #Rhn) mice. Swelling and redness hardly occurred when chemical substances were injected into the hind paw of the mice. Since this TG mice harbors a BAC transgene (clone RP23-114I6), we first speculated that exogenous genes in the BAC may modulate pain behaviors. The exogenous gene expressions, however, were indistinguishable from those in naive mice. Based on a next hypothesis that the large transgene might affect endogenous gene expression and thereby influence pain behaviors, we performed forward genetic screening using next generation sequencing and cDNA microarray analysis. The BAC transgene was inserted into chromosome 8 and 20 gene expressions in the TG mice were decreased to less than 1/16 of those in wild type mice. Notably, three genes in the vicinity of the insertion site were almost knocked out. None of the three genes have been so far identified as pain-related factors. Whether these genes are involved in pain-sensing mechanisms is an open question. To elucidate the roles of the candidates in pain behaviors, we are investigating knock out (KO) and conditional KO mice.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-190 Loganin protects against chronic constriction injury-induced neuropathic pain by stimulating autophagic functions Bin-Nan Wu（Wu Bin-Nan）¹，Su-Ling Hsieh（Hsieh Su-Ling）²，Yu-Chin Chang（Chang Yu-Chin）¹，Yu-Chi Cheng（Cheng Yu-Chi）¹ ¹Department of Pharmacology, Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan ²Department of Pharmacy, Kaohsiung Medical University Hospital, Taiwan Since autophagy plays a key role in cell injury, thus, we aimed to investigate the regulation of autophagy function by loganin, a major iridoid glycoside, in rat sciatic nerve chronic constriction injury (CCI) model. Whether loganin affected autophagic functions in CCI-induced neuropathic pain remains unknown. To investigate the neuroprotective effect of loganin, we examined its activity on autophagic-lysosomal pathway in the rat CCI model. Sprague-Dawley rats were randomly divided into four groups: sham, sham+loganin, CCI and CCI+loganin. Loganin (5 mg/kg/day) was administered intraperitoneally starting at day 1 after surgery. Compared with the CCI group, thermal hyperalgesia and mechanical allodynia in loganin-treated group were significantly reduced. The membrane-bound of autophagic marker LC3B was increased in the spinal cord at day 7 after CCI. The ubiquitin-binding protein p62 binding to LC3 incorporates into autophagosomes which are degraded by autophagy. We observed that CCI led to the accumulation of p62, indicating a block of autophagosome turnover. The p62 expression was attenuated together with allodynia and hyperalgesia reduced at day 7 in the CCI+loganin group. Taken together, we suggest that loganin decreased CCI-induced neuropathic pain may be partly attributed to the promoting neuronal autophagy.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-191 オレキシン性下降性疼痛抑制系は脊髄のオレキシン１受容体を介してリナロール誘発性鎮痛に寄与する Yurina Higa（比嘉憂理奈）¹，Mitutaka Sugimura（杉村光隆）¹，Tomoyuki Kuwaki（桑木共之）²，Hideki Kashiwadani（柏谷英樹）² ¹鹿児島大学院医歯学総合研究科顎顔面再建学講座歯科麻酔全身管理学分野 ²鹿児島大学院医歯学総合研究科生体機能制御学講座統合分子生理学分野 Recently we reported that linalool odor exposure induced significant analgesic effects in mice. The effects were not observed in anosmic mice, indicating that olfactory input evoked by linalool odor exposure triggered the analgesic effects. In addition, hypothalamic orexin-containing neurons played a pivotal role for the analgesia because the effects were not observed in orexin-deficient mice. In this study, we further examined the contribution of descending orexinergic pain inhibitory pathway to the analgesic effects. Linalool was vaporized in an odor chamber at room temperature and the odorized air was ventilated into an observation chamber at constant rate (1 L / minute). A mouse was exposed to odorized air for 5 minutes in the observation chamber and then was performed a tail pincher test to measure the threshold for noxious mechanical stimuli. The contribution of orexinergic transmission in spinal cord to the linalool odor-induced analgesia was assessed by intrathecal injection of selective orexin receptor antagonists (SB334867 for orexin-1 receptor, and compound 29 for orexin-2 receptor) 10 minutes before the tail pincher test. The intrathecal injection of SB334867 completely suppressed the linalool odor-induced analgesia, whereas compound 29 did not show any significant effects, indicating the contribution of orexin-1 receptors for the analgesia in the spinal cord. Because the perikarya of orexinergic neurons were localized in the hypothalamus but not in the spinal cord, our results indicated that olfactory input evoked by linalool odor drive the orexinergic neurons in hypothalamus, and then the direct axonal input of the orexinergic neurons suppresses the nociceptive information via orexin-1 receptor in spinal cord. We will also discuss the suppression of immediately early gene expression induced by nociceptive stimulation in the spinal cord after linalool odor exposure. There is no state of conflict of requiring disclosure.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-192 マウス一次体性感覚野における痛覚応答細胞の領域・層特異的分布様式 Hironobu Osaki（尾崎弘展），Yoshifumi Ueta（植田禎史），Mariko Miyata（宮田麻理子）東京女子医大医神経生理 The primary somatosensory cortex (S1) processes not only tactile information but also nociceptive information. The tactile information from somatosensory thalamus is carried to the granular area in S1. Recently, we found the dysgranular area (Dys), adjacent to the granular area in S1, especially responded to noxious stimulation. Because it has been reported that a certain number of nociceptive neurons also exist in the granular area in S1, we compared the distribution and feature of nociceptive neurons between these two areas. We simultaneously recorded extracellular neural activates from supra- and infragranular layers in these two areas of mice and studied response properties to tactile and noxious stimuli at the single-neuron level. According to the response of a single neuron to tactile and noxious stimulation, we classified the neurons into three types; neurons preferred tactile input (T type), neuron preferred noxious input (N type) and neurons responding to both tactile and noxious inputs (T/N type). In layer II/III of the granular area, the majority (63%) of neurons were T type. On the other hand, in layer II/III of Dys, the majority (43%) of neurons were N type. The ratio of T/N type neurons was small (23%) in both areas. These data suggest that the layer II/III neurons process tactile and nociceptive information separately, and the neurons in the granular area mainly process tactile information and those in Dys mainly process nociceptive information. In layer V, the ratio of T/N type neurons increased to approximately 40% in both areas. This data suggests tactile and nociceptive information converged on a neuron at the cortical output layer in both areas. The area- and layer-specificities of nociceptive neurons distribution suggested that the hierarchical processing of nociception in S1.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-193 非同時性の陽性残効：触覚刺激の同時性判断における事前経験の影響 Kazuya Saito（齋藤和哉）¹，Koki Kannaga（金長幸希）²，Widjaja Kevin（Kevin Widjaja）²，Daiki Yoshioka（吉岡大貴）²，Yoshihiro Itaguchi（板口典弘）^1,2，Makoto Miyazaki（宮崎真）^1,2 ¹静岡大学情報学部情報科学科 ²静岡大学大学院総合科学技術研究科情報学専攻 Time perception is influenced by prior experience. For example, after exposure to audiovisual stimuli with a temporal lag (e.g., 235 ms), participants eventually perceived the temporal lag to be shorter than the actual lag (Fujisaki et al. 2004). This negative aftereffect is called "lag adaptation." In the present study, we investigated the aftereffect of synchrony (i.e., lag = 0) on synchrony judgment for unimodal tactile stimuli. For each trial, the participants (n = 12) received two successive pairs of tactile stimuli across the hands. The first stimulus pair was presented as the adaptor stimuli (i.e., prior), and the second pair was presented as the test stimuli. The inter-stimulus interval (ISI) between the adaptor and test stimuli was 500, 1000, or 2000 ms. For the adaptor stimuli, the stimulus-onset asynchrony (SOA) between two stimuli was -100, 0, or +100 ms, for which the appearance ratios were 25%, 50%, and 25%, respectively (i.e., synchrony-to-asynchrony ratios were evened). For the test stimuli, the SOA was -80 ms, -30 ms, -10 ms, 0 ms, +10 ms, +30 ms, or +80 ms. The participants judged whether the onsets of both test stimuli were synchronous or asynchronous. As a result, the proportion of the trials in which the test stimuli were judged as "synchronous" was significantly higher when the adaptor stimuli were synchronous (SOA = 0 ms) than when these were asynchronous (SOA = ±100 ms) under the 500- and 1000-ms ISI conditions. This suggests that a positive aftereffect occurred in the present task. The aftereffect disappeared under the 2000-ms ISI condition. Moreover, the proportion of the trials judged as "synchronous" was significantly lower under the 500- and 1000-ms ISI conditions than under the 2000-ms ISI condition, when the adaptor stimuli were asynchronous (SOA = ±100 ms). However, no significant difference in the judgement proportion was found among the ISI conditions when the adaptor stimuli were synchronous (SOA = 0 ms). These results suggest that the aftereffect was effective only when the adaptor stimuli were "asynchronous." Thus, after exposure to asynchronous tactile stimuli, the participants more frequently judged the test stimuli as "asynchronous." The asynchrony aftereffect on tactile synchrony judgment was not a lag adaptation but was consistent with the predictions based on the optimal Bayesian estimation model (Kording & Wolpert, 2004; Miyazaki et al., 2006).

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-194 分界条床核外側部CRHニューロンの痛み反応の性差における役割 Hiroko Hagiwara（萩原裕子），Toshiya Funabashi（舩橋利也）聖マリアンナ医大医生理 The bed nucleus of the stria terminalis (BST) plays an important role in controlling neuroendocrine and behavioral responses. We hypothesized the sex-specific involvement of CRH neurons in the dorsolateral subdivision of the BST in association with pain response. In the present study, we first determined whether CRH neurons in the dorsolateral BST specifically expressed Cre in CRH-ires-Cre mice (Jackson 012704). We crossed this strain with R26-CAG-LF-mTFP1 reporter mice (RBRC 05147). In situ hybridization with DIG-labeled cRNA probe for CRH mRNA revealed that mTFP1 fluorescence-expressed cells in the dorsolateral BST were also positive for CRH mRNA: 98% in males and 96.1% in females of mTFP1-expressed cells were CRH mRNA positive. This indicated that Cre expression drived by CRH promoter was functionally restricted in CRH neurons of the dorsolateral BST. Next, we injected into the dorsolateral BST with 0.6 μl of AAV9-hSyn-DIO-hM3D-mCherry (Addgen 44361) or AAV9-hSyn-DIO-hM4D-mCherry (Addgene 44362) in CRH-ires-Cre mice. We confirmed robust expression of mCherry positive cells in the dorsolateral BST. Three weeks after AAV9 injection, formalin test was carried out 30 min after Clozapine-N-Oxide (CNO, 0.5 mg/kg) treatment. CNO treatment in male CRH-ires-Cre mice expressed hM4D induced pain response compared to mice expressed hM3D. CNO treatment per see in wild male mice did not affect formalin test. We suggest that male mice are not sensitive for pain compared to female mice because CRH neurons in the dorsolateral BST are active.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-195 慢性疼痛患者のオフセット鎮痛における後帯状皮質の抑制:fMRI研究 Sushuang Yang（楊しゅくそう），Tianjiao LI（李天嬌），Takeshi Kamma（菅間剛），Eri Shinto（神藤慧玲），Atsushi Ito（伊藤篤史），Takashi Ota（太田隆嗣），Jiro Kurata（倉田二郎）東京医歯大心肺統御麻酔 [Introduction] Offset analgesia (OA), a disproportionately large decrease in perceived pain intensity following a slight, temporary increment of noxious thermal stimulus, is presumably mediated by endogenous pain modulation. Although patients with neuropathic and chronic pain showed attenuated offset analgesia, its central mechanisms remain unclear. The aim of this study was to investigate OA-relatd cerebral activity by functional magnetic resonance imaging (fMRI) and to analyze its difference between patients with chronic pain and healthy controls. [Method] We recruited 18 patients with various chronic pain disorders and 17 age-, gender-matched healthy controls. We applied noxious thermal stimulation on the left volar forearm while obtaining continuous pain ratings and whole-brain fMRI. We gave 3 kinds of paradimgs, 3 times each, in a pseudorandom order including OA: T1(46ºC, 5s)-T2(47ºC, 5s)-T3(46ºC, 20s); Constant: T1-T3 (46ºC, 30s); and Short: T1(46ºC, 5s)-T2(47ºC, 5s)-T3(32ºC, 20s). We analyzed OA-related brain activiy during T3 and its subdivisions, including T3-a (6s), T3-b (6s) and T3-c (8s), using ANCOVA analysis in Brain Voyager QX, as well as real-time pain perception changes (ΔOA) using repeated measures ANOVA by SPSS. We also recorded individual psychophysics including pain duration, current pain intensity, painDETECT Questionnaire, Short-Form McGill Pain Questionnaire, Pain Catastrophizing Scale, and Beck's Depression Inventory. Behavioral correlations were sought between cerebral activation and ΔOA using Pearson's correlation analysis. [Results] Compared to controls, patients showed a larger deactivation at the left posterior cingulate cortex [PCC, p(FDR)<0.05 and below] during OA-T3, especially during OA-T3-a and -b. On the other hand, patients showed decreased activation at the right dorsolateral prefrontal cortex, superior parietal lobule and precuneus during Constant-T3. ΔOA showed a negative correlation (P=0.04) with pain threshold and positive correlation (P=0.03) with pain duration in patients. [Conclusion] Enhnaced deactivation of the PCC, a core region of the default mode network, might be associated with impaired endogenous pain modulation in patients with chronic pain. Suppression of the prefrontal and parietal sensory assosiation areas during continuous pain in patients might implicate disturbed top-down regulation of pain processing in chronic pain.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-196 骨格筋における非活動性侵害受容器と交感神経の電気生理学的解析 Hiroki Ota（太田大樹）¹，Takanori Matsubara（松原崇紀）²，Harumi Hotta（堀田晴美）³，Kazue Mizumura（水村和枝）⁴，Toru Taguchi（田口徹）⁵ ¹帝京大学医療技術学部柔道整復学科 ²名古屋大学大学院医学系研究科神経性調節学分野 ³東京都健康長寿医療センター研究所自律神経機能研究室 ⁴日本大学歯学部生理学講座 ⁵新潟医療福祉大学リハビリテーション学部理学療法学科 Afferent and efferent fibers in the skeletal muscle play pivotal physiological functions such as sensory perception and hemodynamics, but the peripheral neural mechanisms are not fully understood. In particular, mechanically-insensitive afferents (MIAs) and sympathetic efferents have not nearly been investigated in terms of electrophysiology. Here we characterized these fibers in skeletal muscle using the single-fiber recording in vivo, with an index of axonal conductive properties, namely, activity-dependent slowing of conduction velocity (ADSCV) (Taguchi et al., 2010). Male SD rats were deeply anesthetized with sodium pentobarbital, and blood pressure, heart rate, rectal temperature and respiration rate were kept in a physiological range. Centrifugal and centripetal experimental setups were used for the recordings of efferent and afferent activities, respectively. In the centrifugal setup, the abdomen and the skin of lower leg were opened to make a pool. A bipolar stimulating electrode was placed under the lumbar sympathetic trunk. A recording electrode was placed under the gastrocnemius-soleus (GS) nerve and used to investigate ADSCV of sympathetic efferent fibers innervating the GS muscle. Twenty-one of 22 sympathetic efferent fibers showed small ADSCV (< 5%). In the centripetal setup with the similar, but opposite direction for recordings to the centrifugal setup, a stimulating electrode was placed under the GS nerve. ADSCV and responses of afferents to mechanical stimulus were recorded from teased fibers in the sciatic nerve. ADSCV of 118 mechanically-insensitive C-fibers showed a similar ADSCV pattern to sympathetic efferents recorded in the centrifugal setup (< 5%). Percentage of the presumed sympathetic efferents among all C-fibers (n = 197) in the GS was 59.9%, which was consistent with the previous anatomical study (Baron et al., 1988). Seventeen fibers exhibited larger ADSCV (> 10%), and were presumed to be MIAs. Three of the 17 MIAs acquired the responsiveness to mechanical stimulation 3-10 min after the intramuscular injection of a cocktail of inflammatory soup, consisting of bradykinin, histamine, serotonin and prostaglandin E₂, into the GS muscle. These results indicate that MIAs do exist in the skeletal muscle, and that MIAs and sympathetic efferents have different axonal conductive properties. The modal shift of MIAs from silent to active may contribute to peripheral mechanisms of muscular mechanical hyperalgesia.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-197 ショウジョウバエ幼虫の逃避行動における時間的制御の解明 Mami Nakamizo（中溝真未）¹，Jiro Yoshino（吉野次郎）¹，Shu Kondo（近藤周）²，Kazuo Emoto（榎本和生）¹ ¹東京大院理生物科学 ²国立遺伝研無脊椎動物遺伝研究室 Rapid and efficient escape behavior in response to noxious stimuli is essential for survival yet how noxious stimuli are transformed to coordinated escape behavior is still poorly understood. In Drosophila larvae, strong mechanical stimulation and high thermal stimulation induce corkscrew-like lateral turning (rolling behavior), followed by rapid forward locomotion (escape crawl). We (Yoshino et al, 2017) and other groups (Ohyama et al, 2015; C Hu et al, 2017) have successfully mapped the neural circuits that evoke escape behavior. Nevertheless, the mechanisms that determine the order and duration of each behavior giving rise to escape behavior remain elusive. In this study, we identify a pair of descending neurons, designated as the Callisto (Call) neurons, that control the duration of rolling behavior. Ablating Call neurons prolonged the duration of rolling behavior during noxious stimuli. Anatomical analysis of Call neurons shows that Call neurons directly innervate axon terminals of the nociceptive sensory neurons. Furthermore, genetic and functional data suggest that Call neurons express inhibitory neurotransmitter GABA and the duration of rolling behavior in response to noxious stimuli was remarkably longer in metabotropic GABA-B receptor null mutant. These data indicate that inhibitory circuit between Call and the nociceptive sensory neurons control the duration of rolling behavior via GABA-B receptor. Taken together, these findings demonstrate how the time-window of the escape behavior in response to noxious stimuli is regulated at the circuit, molecule as well as behavior levels.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-198 脊髄抑制性介在ニューロンに発現するmGluR4および8は疼痛伝達を修飾する Masamichi Okubo（大久保正道），Hiroki Yamanaka（山中博樹），Kimiko Kobayashi（小林希実子），Koichi Noguchi（野口光一）兵庫医科大学　解剖学講座神経科学部門 Glutamate is released as a neurotransmitter by most excitatory synapses in the nervous system. It also plays a key role in the physiological excitatory circuit in the spinal cord and is involved in pathological neurotransmissions such as inflammatory and neuropathic pain conditions. The actions of glutamate are mediated by different types of receptors that are ionotropic (iGluRs, AMPA, NMDA and kainate) and metabotropic (mGluRs) ones. Although it is well-studied expression of iGluRs, that of mGluRs is not fully elucidated in spinal cord. In this study, we examined the expression of mGluRs (mGluR1-8) and which subtype of mGluR is important for pain transmission in dorsal horn of spinal cord using inflammatory pain model in rats. RT-PCR revealed that mGluRs mRNAs, except for mGluR2 and 6, were detected in the spinal cord. We employed double labeling analysis, in situ hybridization histochemistry with immunohistochemistry, to examine precise distribution of each mGluR in neurons or glial cells in lamina I-II of spinal dorsal horn. It showed that mGluR1, 5 and 7 generally, and 4 and 8 frequently were expressed in neurons (mGluR1,5,7; > 90%, mGluR4, 8; approx. 60%). mGluR3 was expressed not only in neurons but also in oligodendrocytes. We next examined whether mGluR4 and 8 were expressed in excitatory or inhibitory neurons. It revealed that both mGluR4 and 8 were often expressed in inhibitory neurons (approx. 60%) more than in excitatory neurons (approx. 25%). These results led us to behavior test using formalin model, a typical inflammatory pain model. Intrathecal delivery of CPPG, antagonist for group III mGluR (mGluR 4, 7 and 8), suppressed nocifensive behaviors induced by intraplantar injection of formalin (1.5% in 50 μl). Furthermore, CPPG reduced induction of fos expression in excitatory neurons of dorsal horn induced by formalin injection. These findings suggest that mGluR4 and 8, which are especially expressed in inhibitory neurons, may have principal roles in modulation of pain signaling using mGluRs in spinal dorsal horn.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-199 昆虫の機械感覚系における異なる刺激情報の符号化をになう細胞集団 Hiroto Ogawa（小川宏人）¹，Kazuki Tanaka（田中寿希）²，Makoto Someya（染谷真琴）³，Hisashi Shidara（設樂久志）¹ ¹北海道大院理生物科学 ²北海道大院生命 ³理研CBS 知覚神経回路機構 In general, neurons in the sensory systems show distinct selectivity for the physical characteristics of stimulus such as direction and frequency. The selectivity to the stimulation parameters is represented as the tuning property. Many neurophysiological studies on the sensory systems have reported that the sensory tuning property is modulated depending on the other stimulus properties such as intensity and the changes in the tuning property have a great influence on the encoding of sensory information. However, the relationships between the stimulus intensity and tuning property or encoding accuracy for other stimulus characteristics of individual cells remain unknown. Moreover, the sensory information is represented as population activity of multiple neurons rather than single neuron activity. It is unclear how the sensory information on the multiple stimulus parameters including intensity and direction are encoded by neuron population. In order to address these questions, the neural system in which several cells are identified and their response properties are diverse at the same processing level is required. In this study, we used cricket cercal sensory system which was mechanosensory system to detect airflow dynamics. The directional information of airflow is processed by local circuit within the terminal abdominal ganglion and conveyed to the brain by ascending projection neurons including the several identified giant interneurons (GIs). GIs have selectivity to the direction of airflow and their directional selectivity differs according to the type of GIs. We measured the activities of individual GIs in response to airflow stimulus of various velocities from different angles using intracellular recording method. The present results revealed that the impacts of the stimulus intensity on the directional tuning varied by the type of GIs. In addition, the accuracy of direction or velocity encoding for individual and population of cells was analyzed. The stimulus direction was represented by the population activity of three pairs of GIs, which are LGI, GI 10-2, and GI 10-3, whereas the airflow speed was encoded by the cell group consisting of MGI, GI 9-2, and GI 9-3. This result suggests that the stimulus direction and intensity are separately conveyed by different groups of GIs, which may be related to their behavioral functions.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-200 脳卒中後疼痛ラットモデルの体性感覚野における神経活動の時空間パターン：膜電位感受性色素を用いたイメージング Kazuaki Nagasaka（長坂和明）¹，Ichiro Takashima（高島一郎）^1,2，Daigo Bando（坂東大豪）^1,2，Noriyuki Higo（肥後範行）¹ ¹産業技術総合研究所人間情報研究部門 ²筑波大学人間総合科学研究科 Central post-stroke pain (CPSP) is an intractable chronic pain that can occur several weeks after stroke at multiple sites in the somatosensory pathways. Several lines of studies demonstrated that stroke lesion of the ventral posterolateral nucleus (VPL) of the thalamus resulted in thermal hyperalgesia and tactile allodynia, typical symptoms of CPSP, in the extremities in human patients as well as in animal models using macaques and rodents. Currently, the neural mechanisms are not fully understood. An optical imaging using voltage sensitive dye (VSD) is powerful tools to visualize stimulus-dependent membrane potential and to reveal the spatiotemporal patterns of neural activity. Here, we performed optical imaging using VSD RH-795 to investigate sensory activity changes evoked by forelimb-stimulation that may underlie CPSP, using the rat model. To generate an artificial stroke, collagenase type IV was injected into the left VPL under ketamine and xylazine anesthesia. After 3 weeks following the injection, occurrence of both thermal hyperalgesia and tactile allodynia in the contralesional forelimb was confirmed by measuring its withdrawal threshold to thermal and mechanical stimuli. VSD imaging performed at 4 weeks after the injection under anesthesia showed a different activity pattern compared with those in the intact animals. In the intact rats, forelimb electrical stimulation evoked a brief depolarization in the forelimb area of the somatosensory cortex, and the signal amplitude increased with increasing stimulus intensity. In the CPSP rats, by contrast, the forelimb-evoked depolarization was propagated to the adjacent hindlimb areas of the somatosensory cortex and to the primary motor cortex. Moreover, the stimulus intensity-dependence was partly diminished. Our findings suggested that plasticity of sensory cortex after VPL lesion contributes to development of CPSP.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-201 触覚刺激によって喚起された感情状態の神経デコーディング Noriaki Kanayama（金山範明）^1,2，Masayuki Hara（原正之）³，Shigeto Yamawaki（山脇成人）² ¹産業技術総合研究所 ²広島大 ³埼玉大 Human emotion could be induced by multisensory integration of five senses and interoception. Of these sensory modalities, tactile stimulus should be an important sensory modality to understand the neural mechanism of our emotional response as a result of multisensory integration. Tactile inputs received at the somatosensory cortex could be integrated with visual and auditory inputs at the parietal and temporoparietal cortex and also would be integrated with olfactory, gustatory, and body internal information at the insular cortex, which suggests that the tactile information processing take a role to bridge the information about external object (mainly visual and auditory inputs) and the information about internal body state (mainly gustatory, olfactory, and interoception). However the neural correlates of the emotional state induced by the tactile stimulus was not fully clarified. In our study, we have scan the BOLD response by 3T fMRI scanner during positive and negative emotional state induced using tactile stimulator controlled by MRI compatible actuator. In this experiment, participants passively touched various materials inducing typically positive (e.g. fur) and negative (e.g. bristle) emotional response. We measured BOLD response during 4 seconds touching on the materials. We have conducted Multivariate pattern analysis to find the voxel differentiated by the emotional state (positive and negative) controlled by physical properties of touch. As a result, we found that the parietal cortex, the temporoparietal junction, and the insular cortex are the area which could decode the emotional state, positive and negative, with 60% or more accuracy rates. This finding suggest that the neural correlates of emotional state induced by tactile stimulation includes various sensory association area.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-202 視聴覚同時変化時のモダリティ間のカテゴリーの一致・不一致による脳磁場活動への影響 Emi Tanaka（田中絵美）¹，Tetsuo Kida（木田哲夫）^2,3，Ryusuke Kakigi（柿木隆介）^2,3，Minoru Hoshiyama（寳珠山稔）¹ ¹名古屋大　脳とこころの研究センター ²生理研統合生理 ³総研大院生命科学生理 Outer world is continuously changing, and our multisensory system has to follow and detect such continuous changes of environment. Processes for multisensory integration in the brain have been previously studied by providing multisensory inputs, but not by multisensory changes. In the present study, we assessed neuro-magnetic brain responses induced by simultaneous changes of multisensory stimuli in healthy participants. We manipulated inter-modal congruency of stimulus category (e.g., dog, sheep, cicada and cricket) before and after the changes during continuous presentation of visual and auditory stimuli thus resulting in pre- and post-change inter-modal categorical congruencies as two experimental factors. Subjects pressed either one of different two buttons depending on congruency of stimulus categories, e.g., they asked to press one button when voice and picture were congruent for any category, and another button when incongruent. A root sum square (RSS) for 204 sensors of a whole-head magnetoencephalographic (MEG) system was calculated, and peak latency and amplitude of each deflection of RSS was measured. First, pre-change inter-modal categorical congruency had a suppressive effect on neuromagnetic activity at temporal regions in the 200-ms post-change period, indicating the inter-modal congruency effect on the neural process in the primarily-modality specific region. Second, pre-change inter-modal congruency had a suppressive effect on activity at frontal sensors in the 600-ms post-change period. Thus, the history of inter-modal congruency of categories affects post-change cortical processes in different cortical regions. In addition, post-change inter-modal congruency had a suppressive effect on activities at occipito-temporal regions in the 250-ms post-change period. An interactive effect of pre- and post-change congruency was observed only for the 100-ms post-change response, suggesting the continuous history of inter-modal congruency affects post-change inter-modal categorical congruency at the relatively-early process. These results suggest that pre- and post-change inter-modal congruency differently affects spatially- and temporally-different cortical processes.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-203 Quantifying the contribution of body movements to the dynamics of the mouse parietal cortex during decision-making using videography and deep convolutional neural networks Federico Bolanos（Bolanos Federico），Ryo Aoki（Aoki Ryo），Andrea Benucci（Benucci Andrea） RIKEN Center for Brain Science In recent years, studies have shown that complex behaviors are associated with highly dynamic and variable neural activity, such as those observed when mice are engaging in psychophysical decision-making tasks. In particular it has been shown that a large portion of the variability in cortical neural activity can be explained by body moments. More so than the variability explained by decision-related task variables, such as trial difficulty, animal's confidence, degree of task engagement, etc. These highly variable and often overseen behaviors limit our ability to isolate the computational dimensions in the neural dynamics that are associated with the cognitive processes required to make a decision. This limitation is particularly important in areas known to represent sensory, motor, and decision-related signals, such as the posterior parietal cortex (PPC). We therefore imaged the activity of a population of neurons across days from the PPC of Camk2-tTA;TRE-GCaMP6s mice using 2-photon microscopy. We performed our imaging while the mice were engaged in a visual discrimination task, and also monitored in detail the body movements of the mice using two high-resolution video cameras. Utilizing an approach based on a convolutional neural network (DeepLabCut, Mathis et al., 2018) we extracted positional features from the videos that we then regressed against the 2-photon imaging data to estimate the contribution of each behavior on the neural activity. We extracted a total of 8 features: location and size of pupil, and location of left paw, right paw, snout, tail, whisking and licking. We observed that these movement related features significantly increase the dimensionality of the neural activity space, hence we expect to be able to identify non-overlapping dimensions along which cognitive variables may reside.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-204 運動出力の強度が自己生成音の知覚に影響する Nozomi Endo（遠藤希美）¹，Takayuki Ito（伊藤貴之）^2,3，Takemi Mochida（持田岳美）⁴，Tetsuya Ijiri（井尻哲也）¹，Kimitaka Nakazawa（中澤公孝）¹ ¹東京大学大学院総合文化研究科 ²Gipsa-lab, CNRS, Grenoble, France ³Haskins labs, CT, USA ⁴NTTコミュニケーション科学基礎研 Given that a stronger stroke produces a louder sound in the piano playing, the amplitude of the produced force in motor execution may affect our loudness perception of the sound through the auditory-motor interaction. Although auditory discrimination ability in tone perception can be improved through the training with self-generated tone, the mechanism of auditory-motor interaction in loudness perception has not been well known. In particular, the relationship between loudness perception and motor intensity is not fully investigated. The aim of this study is to examine whether the perceptual bias and accuracy in loudness perception are modulated depending on the amplitude of the produced force in finger movement. Thirty participants discriminated a difference in loudness between two stimulus sounds presented sequentially. The first sound was produced in synchronization with the force generation task by vertical movement of the finger. We set three force target (1N, 2N, and 4N) and this can be achieved with visual feedback. The second sound followed 1000 ms after without any movement production. Loudness of the first sound was always the same, and loudness of the second was varied. We also tested the condition that the loudness discrimination task without finger movement (non-motor condition) as control. Point of Subjective Equality (PSE) and Just Noticeable Difference (JND) were compared by two-way ANOVA with motor factor (Motor / Non-Motor) and force as within-subject factors. The main effect of both motor and force factors were significant in JND. As a result of multiple comparisons, JND was significantly increased when finger force strength was large. In addition, JND in motor condition was significantly greater than the one in non-motor condition. On the other hand, we do not find any significant differences in PSE. The results indicate that the loudness discrimination ability was reduced depending on finger force, although the shift of perceptual biases does not occur. The intensity control of motor execution may contribute to the tuning of loudness perception acuity.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-205 人為的な知覚神経の活性化により発現増加する成長因子は腫瘍増殖を促進させる Takashige Kondo（近藤貴茂）¹，Yusuke Hamada（濱田祐輔）³，Kazuki Tanabe（田邉一貴）¹，Kiwamu Sakurai（櫻井究）¹，Hitoshi Makebe（眞壁一志）¹，Michiko Narita（成田道子）¹，Naoko Kuzumaki（葛巻直子）^1,3，Akihiro Yamanaka（山中章弘）²，Minoru Narita（成田年）^1,3 ¹星薬大・薬理 ²名古屋大・環医研・神経2 ³先端生命科学研究センター (L-StaR) A growing body of evidence suggests that intractable pain can be associated with a decreased survival rate associated with a reduced quality of life in cancer patients. In contrast, early palliative care has shown to dramatically improve the survival rate of cancer patients. This notion was based on the supposition that pain plays a key role in tumor growth. However, little is known about the mechanism of tumor growth associated with pain. In the present study, we investigated whether activation of the nociceptive neuron could affect tumor growth. To artificially activate the nociceptive neuron, AAV6-hSyn-hM3Dq-mCherry (AAV-hM3Dq) was injected the nociceptive neuron. Two weeks after injection, we implanted Lewis lung carcinoma (LLC) cells around the sciatic nerve. Subsequently, a hM3Dq-specific ligand, clozapine N-oxide (CNO) (3 mg/kg i.p.) was repeatedly administered to AAV-hM3Dq-injected mice in order to increase neuronal signaling. Two weeks after implantation, tumor growth was dramatically increased in the AAV-hM3Dq injected-mice treated with CNO. Under these conditions, the expression levels of growth factors in the dorsal root ganglion (DRG) were increased by activation of the nociceptive neuron of AAV6-hM3Dq injected-mice by treatment with CNO. Furthermore, we found that tumor growth was dramatically promoted in growth factor injected-mice. These findings suggest that pain associated with activation nociceptive neurons may facilitate tumor growth associated with an increase in the production of growth factors in the DRG.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-206 トレッドミルベルト上のニホンザルによる自発的な四足歩行から二足歩行への転換過程における運動学的解析 Takashi Suzuki（鈴木享）¹，Akira Murata（村田哲）²，Masahiko Inase（稲瀬正彦）²，Katsumi Nakajima（中陦克己）¹ ¹岩手医科大学　医学部　統合生理学分野 ²近畿大学　医学部　生理学講座 The postural control system is distributed throughout the CNS. Neurophysiological studies to date have mainly concerned automatic aspects of postural control, i.e., maintenance of postural orientation and balance in a static or dynamic conditions. However, studies on voluntary aspects of postural control, e.g., volitional transformation of postural orientation during movement have been quite few. To investigate neural mechanisms related to volitional postural control during movement, as a critical first step, we kinematically analyzed behavioral processes of Japanese monkeys transforming locomotor patterns from quadrupedal to bipedal without interruption of on-going stepping on a treadmill. The subjects were two male monkeys. Lateral and back views of the walking animal were videotaped using two high-speed cameras. Stick figures of the head, trunk and limbs were drawn and the kinematic parameters were measured using an image-analyzing software. In the sagittal plane, the body axis angle was maintained at about 5° from the horizontal plane during quadrupedal locomotion. The postural transformation was initiated by forward movement of either hindlimb during the swing phase as a trigger. We defined this hindlimb as a trigger limb. The body axis started to right up at around the subsequent touchdown and reached near vertical with its angle of about 70° just before the liftoff of the trigger limb. Thus, the transformation was accomplished within a single step cycle. In the frontal plane, the head and hip positions fluctuated mediolaterally, less correlated with the step cycle during quadrupedal locomotion. Just before the transformation, the head and hip positions were shifted toward ipsilateral side of the trigger limb (postural adjustment for righting up). Next, these positions were conversely moved back to the contralateral side during the stance phase of the transformation (postural adjustment for walking bipedally) and again toward the ipsilateral side during the swing phase at the start of bipedal locomotion. Such left and right trunk shifts, each once in a step, led to cyclical trunk sway during stable bipedal locomotion. The results revealed two distinct postural adjustments: one before and the other during the postural transformation. Because the righting-up motion leads to successful performance of bipedal gait, our results suggest that voluntary postural transformation is a sort of postural adjustment preceding subsequent locomotor movements.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-207 視運動刺激中の脳卒中患者における重心動揺解析 Junya Komagata（駒形純也）¹，Atsushi Sugiura（杉浦篤志）¹，Daisuke Natori（名取大輔）³，Keita Asai（浅井敬太）³，Hiroshi Takamura（髙村浩司）²，Toshihiro Kitama（北間敏弘）¹ ¹山梨大総合分析実験セ ²健康科学大 ³石和共立病院 Stroke patients bear most of their body weight on the unaffected limb, resulting in distinct asymmetrical stance and weight bearing. Reduced weight bearing on the affected side has been closely related to increase in fall incidences; therefore, an important goal in stoke rehabilitation is to improve patients' abilities to bear weight on their affected side. The present study investigated whether broad visual scene motion (optokinetic stimulation, OKS) can help achieve this aim. Fifteen patients who could maintain a sitting posture without assistance were recruited. A stabilometric platform was used to determine the center of pressure (CoP). The patients were asked to sit quietly on the platform, which was fixed to a stool, in a dimly-lit room. For OKS, a pattern of random dots were projected onto a screen placed 100 cm in front of the stool. The image was moved continuously in either a horizontal (HOKS) or a torsional (TOKS) direction, with a velocity of 20 °/s. The static image was presented as a control condition. In each condition, four CoP parameters were examined: the total length of the sway path (SP), sway area (SA), sway mean, and sway vector. When HOKS and TOKS were directed toward the affected side, SP and SA were significantly greater than those in the control condition, indicating a decrease in postural balance stability. In addition, mean medio-lateral sway shifted significantly towards the affected side. The sway vector magnitude to the affected side (0° direction) was greater than that of the control condition, whereas no significant magnitude differences were observed for the forward (90°), unaffected side (180°), and backward (270°) directions. When OKS was moved toward the unaffected side, all CoP parameters were in a similar range to those in the control condition. Based on these results, we concluded that the OKS conditions used in the present study may be utilized to shift the CoP to the affected side in stroke patients. Therefore, this approach could be used as a balance training method for stroke patients.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-208 ネコの前肢リーチングに随伴する姿勢制御における頭頂皮質の役割 Mirai Takahashi（高橋未来），Toshi Nakajima（中島敏），Saori Miyagishi（宮岸沙織），Ryosuke Chiba（千葉龍介），Kazuhiro Obara（小原和宏），Kaoru Takakusaki（高草木薫）旭川医科大学　脳機能医工学研究センター The postural adjustment that precedes the onset of voluntary movement contributes to the appropriate performance of the movement. We have been studying mechanisms of postural control during forelimb reaching in cats. Cats were trained to maintain standing on force transducers, and to perform forelimb reaching forward to the target which was presented in front. The center of vertical pressure (CVP), which was calculated by ground reaction force, was continuously monitored. Forelimb reaching was performed under various target locations in space. To characterize postural change, we analyzed CVP at paw lift (the moment when the cat lifted the paw) and that at reach target (the moment when the cat reached the target). We found that both CVP positions were altered as a function of the target location. For each cat, we were able to identify the target location that minimized the distance between the two CVP positions. Our findings indicate that the cats estimated the CVP at reach target and provided the CVP which was close to the estimated one in advance of paw lift. Such a postural adjustment requires the mechanism that perceives the relation between the subject and the environment, so that it can predict how to achieve forelimb reaching depending on the relationship. Because the parietal cortex has been implicated in the integration of multimodal sensory information and the cognition of oneself and the external space, we postulated that this cortical area is one of the most important structures for the postural adjustment. In the present study, we microinjected muscimol, a GABA agonist, into parietal areas 5 and 7 of cats that were proficient in forelimb reaching, and examined how the microinjection affected the performance of the postural adjustment and reaching. Here we discuss the roles of these areas in the postural adjustment.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-209 脊髄損傷後の運動機能回復に伴う安静時皮質脳波の変化 Toshinari Kawasaki（川崎敏生）^1,2，Reona Yamaguchi（山口玲欧奈）³，Chao C Zenas（Zenas C Chao）¹，Masahiro Mitsuhashi（三橋賢大）^1,4，Satoko Ueno（上野里子）¹，Yukihiro Yamao（山尾幸広）²，Takayuki Kikuchi（菊池隆幸）²，Kazumichi Yoshida（吉田和道）²，Susumu Miyamoto（宮本享）²，Tadashi Isa（伊佐正）^1,3 ¹京都大院医神経生物 ²京都大院医脳病態生理脳神経外科 ³京都大ASHBi ⁴京都大院医脳病態生理臨床神経学 Spinal cord injury (SCI) inflicts severe damage on motor and somatosensory functions but accumulating evidence has shown that impaired movements recover considerably through rehabilitative training in some cases. In our previous studies showed that the rehabilitative training following the SCI induces neuronal plasticity in the central nervous system not only in the spinal cord but also in the cerebrum. In addition, other previous studies showed that resting-state brain activity reflects the state of brain network. However, the resting-state brain activity after the SCI has not been carefully examined. To address this question, we assessed the resting-state brain activity under both awake and general anesthesia conditions before and after the SCI in the macaque monkey. To record the brain activity, two 18-channel ECoG electrode arrays were implanted on both hemispheres covering the primary motor (M1), primary somatosensory (S1) and premotor (PM) cortices in the monkey. We longitudinally monitored brain activity of the monkey before and after the subhemisection of the spinal cord at C4/C5 for 5 months to investigate the change in the resting-state brain network structure associated with recovery of motor functions. Under the awake condition, the analysis of pairwise phase consistency (PPC) between contralesional M1 and ipsilesional PM revealed the transient increase in PPC at the beta-band range following the SCI. Furthermore, we calculated the Granger causality to investigate the direction of information flow between each brain area. As the result, the interhemispheric information flow from the ipsilesional PM to contralesional M1 in the awake state increased temporally after the SCI. On the other hand, interhemispheric information flow from ipsilesional PM to contralesional M1 increased immediately after the SCI and then that enhancement gradually decreased under the general anesthesia condition. These results suggested that the resting-state brain activity changed after the SCI and the change of brain network in the interhemispheric motor related areas after the SCI at the resting-state under the awake condition would be different from that under the general anesthesia condition. The role of interhemispheric interactions in the functional motor recovery will be discussed.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-210 視覚刺激はヒト間接的皮質-脊髄路興奮を増大させる Tsuyoshi Nakajima（中島剛）¹，Hiroyuki Ohtsuka（大塚裕之）¹，Shun Irie（入江駿）¹，Ryohei Ariyasu（有安諒平）¹，Tomoyoshi Komiyama（小宮山伴与志）²，Yukari Ohki（大木紫）¹ ¹杏林大学医学部統合生理学教室 ²千葉大学教育学部 Modulatory actions of inputs from visual system to cervical interneurons (IN) for arm muscle control are poorly understood in humans. In the present study, we examined whether photic stimulation (PS) modulates cervical IN systems that would mediate motor cortical excitation to motoneurons projecting to arm muscles. Healthy subjects, who all gave informed consents, were seated with recording of electromyograms from the right biceps brachii (BB) muscle. Flash stimulator for PS [white light, 50 μs duration, 2 J (relative energy)] was placed on 60 cm in front of the subject's eye. Transcranial magnetic stimulation (TMS) over the contralateral primary motor cortex and electrical stimulation of the ipsilateral ulnar nerve at wrist (NERVE) were delivered separately or in combination. A 10 ms inter-stimulus interval (ISI) for the combined stimulation (TMS behind) was used to give converging inputs on the upper cervical segments. PS was sometime delivered at 60 ms before TMS timing. Combination of TMS and NERVE gave rise to facilitation of motor evoked potentials (MEPs) in the BB. When the combined stimulation was delivered with PS, the ulnar nerve-induced facilitation of the MEP significantly increased in comparison to the control condition without PS. Furthermore, with recording of single motor units, a short-latency excitatory peak (i.e., di- or oligosynaptic effects) on post-stimulus time histograms following the combined stimulation was significantly facilitated by PS. Existence of spatial facilitation effects on the BB after combined stimulation at a short ISI (i.e., 10 ms) indicates that the facilitation takes place at premotoneuronal level in the cervical cord. The present findings therefore suggest that PS facilitates cervical IN systems that receive converging pyramidal tract and peripheral nerve inputs and excite arm motoneurons.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-211 齧歯類発達期における皮質脊髄路-運動ニューロン間の一過性シナプスの形成と退縮 Satoshi Fukuda（福田諭）¹，Takae Ohno（大野孝恵）¹，Naoyuki Murabe（村部直之）¹，Mizuho Niido（新戸瑞穂）¹，Hiroaki Mizukami（水上浩明）²，Keiya Ozawa（小澤敬也）^2,3，Toshihiro Hayashi（林俊宏）¹，Masaki Sakurai（桜井正樹）¹ ¹帝京大医生理 ²自治医大分子病態遺伝子治療研究 ³東大医科学研究所附属病院 Corticomotoneuronal (CM) direct connections linking motor cortex directly to spinal motoneurons (MNs) has long been believed to be seen only in higher primates that play an essential role in the cortical control of skilled movements. We recently reported a direct connection between the corticospinal axons and cervical MNs innervating forearm muscles in rodent (Maeda et al, 2016) and further demonstrated that those CM direct connections were shown to be eliminated until adulthood by the technique of retrograde trans-synaptic labeling using genetically-modified rabies virus (Murabe et al, 2016, 2018). It remains, however, unclear the accurate time window of transient direct connections because of the technical limitation to record excitatory postsynaptic currents (EPSCs) from spinal MNs in older ages. Because spinal MNs are known to be vulnerable in slices, developmental studies of functional connections in the spinal MNs, which require electrophysiological single cell recordings, were considerably limited especially in mature animals. In this study, we improved slice preparation technique to reduce the ischemic time period before immersing the removal spinal cord tissue into oxygen saturated cutting solution and succeeded in whole cell recordings from the spinal MNs of young adult mice (up to postnatal day 50). This enabled us to follow the direct CM innervation rate from infant to young adult and to analyze this regressive event during development.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-212 同名筋と異名筋運動ニューロンへの単シナプス性Ia促通に対する振動刺激の効果 Mitsuhiro Nito（仁藤充洋）¹，Takuya Yoshimoto（吉元拓也）¹，Wataru Hashizume（橋爪和足）¹，Masaomi Shindo（進藤政臣）²，Akira Naito（内藤輝）¹ ¹山形大医解剖一 ²松本大院健康科学 It is known that the amplitude of the Hoffmann(H)-wave of the soleus, mainly provoked by monosynaptic excitation of homonymous group Ia afferents, is suppressed by the tonic vibration stimulation (TVS) to its muscle belly or tendon in humans. The recovery time of the suppression is affected by changing the duration or frequency of TVS. In this study, effects of TVS on the monosynaptic Ia facilitation of homonymous and heteronymous motoneurons were evaluated using the H-waves of the flexor carpi radialis (FCR) and the tendon(T)-waves of the biceps brachii (BB), respectively. The H-waves of FCR were provoked by stimulating the median nerve trunk at the elbow in fourteen healthy human subjects. Details of monosynaptic Ia facilitation of heteronymous motoneurons were described in the previous report (Cavallari & Katz 1989). The T-waves of BB were induced by a mechanical quick tap applied to the distal tendon of BB (test T-wave) in twelve subjects. The amplitude of the T-wave was increased significantly by electrical stimulation to the median nerve at the elbow with the intensity below the motor threshold of FCR (conditioned T-wave) in all subject. When the TVS was applied to the FCR muscle belly at 100 Hz, the amplitudes of the H-wave of FCR and the conditioned T-wave of BB were suppressed in all subject. In both the H- and conditioned T-waves, the longer the duration (from 2 to 6 minutes) of TVS, the longer the recovery time of the suppression. The higher the frequency of TVS (57, 77 and 100 Hz for 6 minutes), the longer the recovery time. Since the amplitude of the test T-wave did not change throughout the experiment, changes in the duration or frequency of TVS should affect the transmission of group Ia afferent terminals mediating facilitation to the heteronymous motoneurons as well as to the homonymous one.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-213 随意運動のイメージはヒト頸髄介在ニューロンを介する間接的皮質-脊髄路興奮を促進させる Shun Irie（入江駿）¹，Tsuyoshi Nakajima（中島剛）¹，Shinya Suzuki（鈴木伸弥）^1,2，Ryohei Ariyasu（有安諒平）¹，Tomoyoshi Komiyama（小宮山伴与志）³，Yukari Ohki（大木紫）¹ ¹杏林大医統合生理 ²北海道医療大　リハビリテーション科学部　理学療法 ³千葉大学教育学部 Motor imagery is known to affect the reacquisition of motor functions after damages to the central nervous systems. However, it is unclear if motor imagery influences the cortico-motoneuronal (C-M) excitation mediated via cervical interneurons (INs), which may be important for functional motor recovery in animals. To investigate this, we examined the spatial facilitation of motor-evoked potentials (MEPs) induced by combined stimulation (CS) of the pyramidal tract and peripheral nerves. Healthy volunteers were included to record electromyograms (EMGs) from the right biceps brachii (BB). Transcranial magnetic stimulation (TMS) to the left motor cortex and electrical stimulation of the right ulnar nerve at wrist (NERVE) were delivered separately or in combination with an inter-stimulus interval of 10 ms (NERVE ahead). Sometimes, subjects were instructed to imagine elbow flexion with maximum effort during the stimulation without actual movements. CS facilitated MEPs in the surface EMG irrespective of motor imagery compared with algebraic sum of responses with separate stimuli. Motor imagery enhanced MEPs by separate TMS. Moreover, the CS-induced facilitation was significantly increased by motor imagery. Single motor unit recording also revealed increased facilitation during motor imagery, which was observed in peaks of the peri-stimulus time histogram 1-2 ms later than the onset latency. The present findings suggest that motor imagery facilitates indirect C-M excitations to arm motoneurons, which are mediated by the cervical IN systems (i.e., di- or oligosynaptic contribution). Thus, motor imagery could be a useful tool for functional motor recovery using the cervical IN circuitry.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-214 ヒト三角筋から上腕二頭筋への神経投射について：PSTHを用いた解析 Takuya Yoshimoto（吉元拓也）¹，Mitsuhiro Nito（仁藤充洋）¹，Wataru Hashizume（橋爪和足）¹，Manabu Jimenji（治面地学）¹，Takuji Miyasaka（宮坂卓治）²，Masaomi Shindo（進藤政臣）³，Akira Naito（内藤輝）¹ ¹山形大学医学部解剖学第一講座 ²帝京大学医療技術学部柔道整復学科 ³松本大学大学院健康科学研究科 Effects of low-threshold afferents from the anterior (DA), middle (DM) and posterior parts of the deltoid muscle (DP) to biceps brachii muscle (BB) motoneurons were studied in six healthy human subjects using a post-stimulus time-histogram method. The subjects sat in an armchair and put the supinated forearm on its armrest with the shoulder slightly flexed and the elbow semiflexed. As conditioning stimulation, electrical rectangular pulses (width 1.0 ms) with the intensity just below the motor threshold were delivered to the axillary nerve branch innervating DA, DM and DP with bipolar surface electrodes. BB motor unit firings provoked by very weak contraction (5% of maximal voluntary power) were recorded with a pair of needle electrodes. The stimulation to DA and DM induced facilitatory effects (facilitation) in 25/39 (64%) and 28/47 (59%) BB motor units, respectively. The latency and duration of the facilitation from DA and DM were 18.9±2.5 (mean±SD) and 2.3±0.9 ms, and 17.9±2.3 and 1.5±0.8 ms, respectively. Such facilitation was never provoked by pure cutaneous stimulation. The remaining 14 and 19 BB motor units received no effects by the stimulation. The central synaptic delay of the facilitation from DA and DM were 0.2±0.3 and 0.1±0.2 ms longer than that of the homonymous BB facilitation, respectively. The stimulation to DP produced no effects in 28/28 BB motor units. These findings suggest that BB motoneurons receive facilitation from DA and DM in humans. The facilitation should be mediated by group I afferents through a monosynaptic path in the spinal cord.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-215 小脳における感覚惹起性多経路信号の統合 Misa Shimuta（志牟田美佐）¹，Izumi Sugihara（杉原泉）²，Taro Ishikawa（石川太郎）¹ ¹東京慈恵医大薬理 ²東医歯大院医歯総合システム神経生理 The mutual connection between the cerebral cortex and the cerebellum has important roles in sensorimotor coordination and motor learning as well as in higher brain functions. The cerebellum receives various types of inputs via mossy fibers and climbing fibers. But it is not well-understood, from the functional aspect, what kinds of signals are conducted to the cerebellar cortex and how those multiple types of signals are integrated. To address these issues, we focused on somatosensory signals from the facial area of mice whose cortico-ponto-cerebellar pathway could be blocked by light illumination to the somatosensory cortex. When we gave tactile stimulation to the upper lip, excitatory synaptic currents in granule cells appeared in two distinct timings in the whole-cell recordings in vivo. The early response (~10 ms latency) was not affected but the late response (~30 ms latency) was suppressed by the light illumination, suggesting that they were trigeminal and cortico-pontine responses, respectively. Such two-phase responses were obtained in a wide area in crus II, from 5a- to 7+ aldolase C compartments, of the cerebellar cortex. Essentially the same result was obtained in the field potential recordings in awake mice. Similarly, in Purkinje cells, the early simple spikes were not affected but the late simple spikes were suppressed by cortical photoinhibition. Interestingly, the complex spikes were also suppressed in this manner. These results demonstrate that the cerebellar cortex integrates signals not only directly from the trigeminal nuclei but also from the somatosensory cortex via both mossy fibers and climbing fibers.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-216 細菌由来炎症因子LPSによる小脳発達異常 Haruko Otsuka（大塚暖子）¹，Noriyuki Asai（浅井紀之）²，Soon Tiong Kwong Thomas（Thomas Tiong Kwong Soon）¹，Yasunari Kanda（諫田泰成）³，Sachiko Yoshida（吉田祥子）¹ ¹豊橋技科大院工環境・生命工学 ²イチビキ株式会社 ³国立医薬品食品衛生研究所 Autism spectrum disorder (ASD) or mental disorder is increased during a recent decade, and multiple chemicals, for example, sodium valproate (VPA), have been associated with an increased risk of ASD. In early human ASD, cerebellar abnormality is one of the pathological focus points. On the other hand, fluctuation of internal immunity with infection or antibiotics could induce some mental disorders. Intestinal environment and inflammation plays a important role in the immune-homeostasis. Maintenance of intestinal flora and resistance of inflammation are required for healthy neuronal development. Lipopolysaccharides (LPS), characteristic components of the cell wall of Gram negative bacteria, are inducers of inflammation via TLR4 binding. Due to inflammation in pregnant bodies, LPS could induce autism, schizophrenia and other kinds of mental disorder of offspring. We have established the ASD-model rat with the administration of 600mg/kg VPA to embryonic day 16 p.o., and observed their cerebellar development. VPA-administrated rat showed the excess development of Purkinje cells and excess folds in the V to VI lobules of cerebellar vermis within two weeks after birth. This alteration is similar to human early ASD cerebellum. In this study, we investigated dose of LPS derived from E. Coli and administrated it to pregnancy rat at embryonic day 16 i.p. and observed the cerebellar development of offspring. LPS-administrated rat showed decrease of Purkinje cells and activation of microglia on the Purkinje layer at postnatal day 14. In addition, excess folding in cerebellar lobules were observed as same as cerebellum administrated VPA. We suggest that LPS would accelerate inflammation in developing cerebellar cortex and become irregular relationship between neuron and glia. Furthermore, the effects of some probiotic treatments against LPS-inflammation are reported.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-217 サル小脳核は同期サッカード中に時間予測および誤差の信号を伝達する Ryuji Takeya（竹谷隆司），Masaki Tanaka（田中真樹）北海道大院医神経生理 Neural control of synchronized movements must require temporal prediction and error signals. The former may directly regulate movement timing, while the latter may update temporal prediction for the subsequent movements. To explore neuronal correlates, we trained monkeys in the synchronized saccade task (Takeya et al., 2017-8) and conducted single neuron recording from the cerebellar dentate nucleus. In this task, the animals made sequential saccades to alternately presented visual stimuli. In the predictive condition, the target appeared periodically at a fixed interval of 400, 550 or 700 ms (selected randomly for each trial), and every synchronized saccade (within ± 20% of interval) was reinforced with liquid reward. In the reactive condition, the target interval was randomized in each trial, and every reactive saccade following the target onset (> 150 ms) was reinforced. These conditions were associated with different target color, and different trials were randomly presented in a block. So far, we have recorded from 54 dentate nuclear neurons that elevate activity before saccades. One third of these neurons exhibited greater firing modulation for predictive than reactive saccades. Approximately 74% neurons were clearly directional (> 20% modulation) while the others were not. These non-directional neurons (n = 14) showed predictive activity for target onset even during the initial few cycles, when monkeys generated reactive saccades. When the animals were presented with a block of longer stimulus intervals (700, 900 and 1100 ms), the predictive activity for the first few targets extended in time and peaked around the time of the mean of the stimulus intervals. These results suggest that neurons in the dentate nucleus may carry temporal prediction signals for target onset that could regulate the timing of synchronized saccades. Another group of dentate nuclear neurons (n = 14) exhibited postsaccadic activity. For a quarter of these neurons, the maximal firing rate and saccade latency (relative to target onset) showed a significant rank correlation, indicating that these neurons represented timing error of synchronized movements. Taken together, our results suggest that the lateral cerebellum might contribute to synchronized movements by generating both temporal prediction and error signals. A possible causal role of these signals is to be tested in the future study.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-218 視床下部外側野から小脳虫部へのオレキシン入力 Mitsuhiro Hashimoto（橋本光広），Hiroyuki Yaginuma（八木沼洋行）福島県立医大医神経解剖・発生 Cerebellar vermis malformation causes changes to the sleep-wake cycle, resulting in sleep disturbance. However, it is unclear how the cerebellum contributes to the sleep-wake cycle. To examine the neural connections between the cerebellum and the nuclei involved in the sleep-wake cycle, we investigated the axonal inputs in the mouse posterior vermis (lobule IX) by using an adeno-associated virus (AAV) vector (retrograde serotype, rAAV2-retro) as a retrograde tracer. When an AAV vector (rAAV2-retro) expressing enhanced green fluorescent protein (EGFP) was injected into the cerebellar lobule IX, the AAV vector (rAAV2-retro) retrogradely labeled neurons in the lateral hypothalamus (LH). A part of the EGFP-positive neurons in LH (approximately 30%) were expressing Orexin A that is involved in the transition from sleep to wake. To examine axonal projections from the LH, we unilaterally injected an AAV vector (serotype rh10) expressing humanized renilla green fluorescent protein (hrGFP) as an anterograde tracer into the LH. We observed hrGFP-positive axons in the cerebellar vermis and a part of the hrGFP-positive axons in the molecular layer were expressing Orexin A. These results indicated that the orexinergic neurons in LH directly project their axons to the molecular layer of cerebellar vermis. The direct connection between the LH and the cerebellar cortex suggests that the cerebellum is modulated by the orexinergic neurons in LH and participates in the neural network involved in the sleep-wake cycle.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-219 小脳へ投射する単一内側前庭核細胞の形態 Takahiro Ando（安藤貴弘），Izumi Sugihara（杉原泉）東医歯大院医歯総合システム神経生理 The interaction between the vestibular nuclei and the cerebellum is involved in the control of balance and equilibrium. The inputs to the cerebellum from the vestibular system are conveyed by mossy fibers that derive from the primary vestibular afferent nerve and from some vestibular nucleus neurons, and terminated mainly in the so-called vestibulocerebellum (flocculus, FL, nodulus, X, and the ventral uvula, IX). However, morphology of these fibers has not been fully clarified. To examine the anatomical characteristics of the projection to the cerebellum from the vestibular nucleus, first we located distribution of the cerebellum-projecting neurons in the vestibular nucleus with retrograde labeling with fluorescent tracer, True Blue (TB), injected into the flocculus or the nodulus. Labeled neurons were distributed bilaterally in the dorsal part of the medial vestibular nucleus (MVN), and the peripheral part of the superior vestibular nucleus (SVN), in both flocculus and nodulus injection. Then, we labeled these neurons anterogradely by injecting biotinylated dextran amine (BDA) into the dorsal part of the MVN. Anterogradely labeled mossy fiber terminals were distributed mainly in the bilateral nodulus, and ventral uvula, and moderately in the bilateral flocculus and also sparsely in the vermal part of the anterior lobules. Then, we reconstructed 32 cerebellum-projecting single axons nearly completely in other cases of BDA injection, from the identified cell body or from the point near the BDA injection site. These axons did not have ascending or descending branches that project to the oculomotor centers or the spinal cord. On the contrary, they showed various patterns of projections to the cerebellum, and some of them had branches that formed the commissural and the intrinsic connection within the vestibular nuclei. Based on their projection patterns, single axons were generally classified into three groups: (1) solely cerebellum type (n=14), (2) contralateral vestibular nucleus collateral type (n=11), and (3) ipsilateral vestibular nucleus collateral type (n=7). The first type of axons possessed 55 mossy fiber terminals on the average, suggesting that they send strong signals to the cerebellum. The second and third type of axons had innervation in the vestibular nuclei, showing an overlap between the commissural/intrinsic vestibular nucleus projections and the cerebellar mossy fiber projection.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-220 ゼブリン陽性、陰性プルキンエ細胞間での並行線維シナプス応答と興奮性の電気生理学的比較 Viet Tuan Nguyen-Minh（グエンミントゥアンヴィエト）¹，Khoa Tran-Anh（Tran-Anh Khoa）¹，Izumi Sugihara（杉原泉）^1,2 ¹東京医科歯科大学、システム神経生理学分野, ²東京医科歯科大学、脳機能統合センター Cerebellar Purkinje cells (PCs) that have a particular molecular expression profile are organized into separate longitudinal stripes, which have different topographic afferent and efferent axonal connections to be involved in variety of different functions. Expression levels of many molecules such as glycolysis enzyme aldolase C and glutamate transporter EAAT4 are co-expressed together among PC subsets, suggesting different physiological properties among them. In this study, we used whole-cell patch-clamp recording from PCs of different zebrin types in identified neighboring stripes in vermal lobule VIII in cerebellar slice preparations from Aldoc-Venus mice. Regarding basic cellular electrophysiological properties, no significant differences were observed in input resistance or in occurrence probability of types of firing patterns between Z+ and Z& mn; PCs. However, the firing frequency of the tonic firing type was higher in Z& mn; PCs than in Z+ PCs. In the case of parallel fiber (PF)-PC synaptic transmission, no significant differences were observed between Z+ and Z& mn; PCs in interval dependency of paired-pulse facilitation or in decay time course of synaptic current measured without or with the blocker of glutamate receptor desensitization. These results indicate that different expression levels of the molecules that are associated with the zebrin type may affect the intrinsic firing property of PCs but not directly affect the basic electrophysiological properties of PF-PC synaptic transmission significantly in lobule VIII. The results suggest that the zebrin types of PCs in lobule VIII is linked with some intrinsic electrophysiological neuronal characteristics which affect the firing frequency of PCs. However, the results also suggest that the molecular expression differences linked with zebrin types of PCs do not much affect basic electrophysiological properties of PF-PC synaptic transmission in a physiological condition in lobule VIII.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-221 ストレス応答に関わる細胞集団が淡蒼球と拡張扁桃体の境界領域に左右非対称に局在する Munenori Kanemoto（兼本宗則），Tomoya Nakamura（中村友也），Masahumi Kawaguchi（川口将史），Hiroyuki ichijo（一條裕之）富山大医解剖 The basal forebrain (BF) consists of structures including the medial septum, ventral pallidum, vertical and horizontal diagonal band nuclei, substantia innominata/extended amygdala and peripallidal regions. In mammals, the BF contains cholinergic, GABAergic, glutamatergic neurons and peptidergic interneurons. The BF is related to various aspects of emotional stress. However, circuit mechanisms for stress processing are still unclear. To explore stress-related neurons in the BF, patterns of neuronal activation were examined under stress by immunohistochemistry of immediate-early gene product (Zif268/Egr1) in C57BL/6J mice. Here, we found that novel neuronal clusters were situated in the boundary area of the ventral globus pallidus and the extended amygdala. By immunohistochemical analysis, the clusters were composed of neuronal cells (100%) exclusively with cell markers (NeuN, GFAP, calbindin-D28k), and included GABAergic neurons (84.8 ± 2.0%) with GAD65-mCherry mice. Although prototypic and arkypallidal GABAergic neurons are known to be in the GP, expression profile (parvalbumin, proenkephalin, FoxP2) suggested that the clusters were different from those GP neurons. In contrast, the clusters contained a small percentage of cholinergic neurons in the periphery of clusters (0.91 ± 0.6%), and were surrounded by cholinergic neurons, suggesting that the clusters were novel population in the extended amygdala. Next, Mice were immobilized in tubes for 60 min (immobilization stress, IMO). In the IMO group, the number of cells in the cluster were significantly larger than in the control group. In the intraperitoneal injection of diazepam before IMO (the IMO-diazepam group), the number of cells significantly smaller than in the IMO-vehicle group. The results indicated that the stress activated the clusters. Moreover, the clusters were asymmetrically placed in different positions along the anteroposterior axis for individual mice, indicating that the clusters were activated at particular positions in each mice. These results suggested that stress-related GABAergic neuronal clusters were asymmetrically arranged in the boundary area of the globus pallidus and the extended amygdala. The clusters participate in stress-related circuits, which are possibly activated as intersections in the BF.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-222 齧歯類の尾側線条体におけるドーパミン受容体D1及びD2発現ニューロンの特異的分布 Kumiko Ogata（緒方久実子），Fuyuki Karube（苅部冬紀），Yasuharu Hirai（平井康治），Fumino Fujiyama（藤山文乃）同志社大学　脳科学研究科 In the previous study, we first demonstrated the area in the caudal lateral striatum where the immunoreactivity for D1R was significantly poor than the other striatal area, so called D1R-poor zone. D1R-poor zone was located lateral to D2R-poor zone. D2R-poor zone has been already reported as the region where the immunoreactivities for D2R and pre-pro Enkephalin, markers for indirect pathway medium spiny neurons (iMSNs), were almost absent. Intriguingly we found the immunoreactivities for tyrosine hydroxylase and dopamine transporter were poor in both D1R-and D2R-poor zones. By using a retrograde tracer, we confirmed the density of labeled direct pathway MSNs (dMSNs) in D1R-poor zone was significantly lower, whereas that in D2R-poor zone was significantly higher than the dorsal striatum. However, no significant difference in the neuronal density was observed among these three areas. In the present study, we detected neurons expressing D1R and D2R by in situ hybridization to confirm the ratio of dMSNs and iMSNs to all neurons in D1R- and D2R-poor zones. As a result, D1R-poor zone comprises about 12% of dMSNs and about 79% of iMSNs, whereas D2R-poor zone comprises about 82% of dMSNs and about 4% of iMSNs. This result corresponded with that of previous tracer experiment we performed. In D1R- and D2R-poor zones, the immunoreactivity for calbindin was poor, but that for μ-opioid receptor was not so strong. Our findings showed the existence of unique area where various immunoreactivities were different from other striatal area and the distribution of dMSNs and iMSNs was biased in the sub-region of caudal lateral striatum. Our next step is to elucidate the neural circuitry of D1R- and D2R-poor zones and how this distinct distribution affects functional differences.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-223 新規に作成したノックインマウスMOR-CreERによるμオピオイド受容体陽性神経回路の可視化 Taro Okunomiya（奥宮太郎）^1,2，Itaru Imayoshi（今吉格）³，Hiroyuki Hioki（日置寛之）⁴，Chika Nishimura（西村知華）¹，Satoshi Yawata（矢和多智）¹，Ryoichiro Kageyama（影山龍一郎）⁵，Ryosuke Takahashi（髙橋良輔）²，Dai Watanabe（渡邉大）¹ ¹京都大院医生体情報 ²京都大院医神経内科 ³京都大院生命脳機能発達再生制御 ⁴順天大院医神経生物・形態 ⁵京都大ウイルス・再生研増殖制御システム Mu opioid receptor (MOR) is a mandatory component for opioid action. MOR-expressing neurons are widely distributed in the nervous systems. Many studies have reported the brain areas responsible for opioid analgesia, addiction or fear memory, using local administration of MOR agonists, antagonists or agonist-conjugated saporin. However, local administration of compounds affects not only postsynaptic MOR but also presynaptic MOR on many afferent fibers, which hampers precise understandings of cellular components responsible for opioid actions. Toward the understandings of circuit operation of MOR-expressing neurons, genetically modified mice in which Cre recombinase is selectively expressed in MOR-expressing cells should be useful. Here, we generated a novel inducible Cre knock-in mouse line MOR-CreER, in which the T2A sequence and the iCreERT2 gene followed by a stop codon replaced the stop codon in the exon 4 of the Oprm1 gene to gain genetic access to MOR-expressing cells. To visualize the distribution of recombination mediated by MOR-CreER allele, MOR-CreER mice were crossed with the Cre-dependent fluorescence reporter mice R26-CAG-LoxP-mTFP1. After tamoxifen treatment in adult mice, the brains and spinal cords were examined. We found that MOR-CreER allele mediated recombination in many areas of the nervous system including the olfactory bulb, caudate putamen (CPu), neocortex, hippocampus, midline and intralaminar thalamic nuclei, habenula, hypothalamus, interpeduncular nucleus, periaqueductal gray, superior and inferior colliculus, cochlear nucleus, raphe nuclei, medullary reticular formation, nucleus ambiguus, dorsal horn and dorsal root ganglion, in general agreement with previous studies. Interestingly, in the CPu, we found that mTFP1 positive cells were mainly located within strongly MOR immunoreactive regions, which is a cardinal feature of the striosome compartments. To achieve selective transgene expression in the striosome, next we infused a Cre-dependent adeno-associated virus vector encoding EGFP under neuron-specific promoter (AAV9-hSyn-DIO-EGFP) into the CPu of MOR-CreER mice. After tamoxifen treatment, we found that EGFP labeled neurons were selectively distributed in the striosome compartments. Thus, MOR-CreER mouse line is a powerful tool to visualize the MOR-expressing neurons in the nervous system and it would provide a new genetic tool to dissect the functions of MOR-expressing neural circuits such as the striosome compartments.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-224 質量分析による線条体投射神経領域のMBP (+)、MBP (-)膜画分の解析 Haruko Miyazaki（宮崎晴子），Nobuyuki Nukina（貫名信行）同志社大院脳科学研究科　認知記憶加齢部門 Myelin is an essential component to generate fast propagation of action potential in the nervous system. Myelin sheet has a high lipid content (70-75% of the dry weight) compared with other plasma membrane and forms a unique multilamellar structure to surround axon. Myelin basic protein (MBP) is a major component of myelin and can be used for specific marker of myelin. In the nervous system, not only "myelinated axons" but also "unmyelinated axons" exist. Recently, we found striatonigral fibers as a novel unmyelinated fiber tract in central nerve system (CNS). Unmyelinated fibers are not well investigated for a long time, because they are thought a minor population of axons in CNS. However, immunostaining with anti-Nav1.2, a specific marker of unmyelinated fibers, showed abundant distribution of unmyelinated fibers in CNS, and the results suggest that unmyelinated fibers could have some roles in CNS. To identify unmyelinated fiber specific proteins, we performed mass spectrometry analysis using MBP (+) and MBP (-) membrane fraction obtained from striatonigral fiber enriched region in mouse brain. To separate these two different fractions, we performed sucrose gradient centrifugation (0.32M/0.85M) using membrane fraction of striatonigral fibers. Western blot analysis showed that interface fraction was MBP (+) and pellet was MBP (-) fraction. After mass spectrometry analysis, we compared identified proteins between these two fractions. MBP (-) membrane fraction is a crude protein mixture but it could contain the unmyelinated fiber specific proteins.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-225 聴覚弁別課題獲得中の線条体領域の異なる機能 Susumu Setogawa（瀬戸川将）¹，Takashi Okauchi（岡内隆）²，Di Hu（胡迪）²，Nobuyuki Sakayori（酒寄信幸）¹，Mika Shigeta（重田美香）²，Keigo Hikishima（疋島啓吾）³，Hirotaka Onoe（尾上浩隆）⁴，Yilong Cui（崔翼龍）²，Kazuto Kobayashi（小林和人）¹ ¹福島医大生体情報伝達研究所 ²理研生命機能科学研究センター ³沖縄科学技術大院 ⁴京都大院医 The dorsal striatum is the key structure for associative learning linking sensory inputs to behavioral outputs through trial and error. The neural mechanisms underlying the acquisition of associative learning have been studied extensively using the operant conditioning task. Lesion and electrophysiological studies in rats have suggested that different striatal regions, including the dorsomedial and dorsolateral parts of striatum, associate with the sensory discrimination learning task. We previously conducted a small-animal neuroimaging with 2-deoxy-2-[18F] fluoro-D-glucose (FDG)-PET to assess dynamic changes of neural activation patterns in a two alternative auditory discrimination task with rats. Interestingly, the peak of FDG uptake in anterior (aDS) was detected during early learning phase, but that in the posterior striatum (pDS) was detected during late learning phase. To further investigate whether such different activation pattern drives different behavioral output in the action selection of auditory discrimination task, we performed transient striatal inactivation studies. We bilaterally injected the GABAa receptor agonist muscimol into the aDS or pDS, which was carried out 4 times at different time point during training. First injection was conducted on the second day from initiation of the task training. Second and third injections were conducted at one day after the success rate reached to 60% and 80%, respectively. Last injection was conducted at 1 week after the day the success rate reached to 80%. In early learning phase (60% criteria), aDS inactivation impaired the performance of the auditory discrimination task. In contrast, pDS inactivation decreased the success rate in late learning phase (80% criteria), but not early leaning phase. These results suggested that the different striatal areas are involved in performance of different learning phase.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-226 咀嚼の実行と想起にかかわる脳活動 Noriyuki Narita（成田紀之）¹，Sunao Iwaki（岩木直）²，Tomohiro Ishii（石井智浩）³，Taiga Fukuda（福田大河）⁴，Takashi Kaneda（金田隆）⁴，Takeshi Uchida（内田武）⁵，Ikuo Kantake（寒竹郁夫）⁵，Koh Shibutani（渋谷鑛）⁶ ¹日本大学松戸歯学部　口腔科学研究所 ²産業技術総合研究所自動車ヒューマンファクター研究センター ³日本大学松戸歯学部　有床義歯補綴学講座 ⁴日本大学松戸歯学部　放射線学講座 ⁵デンタルサポート ⁶日本大学松戸歯学部　歯科麻酔学講座 Studies of motor execution and related imagery have been conducted using physical functions, such as finger- or toe-tapping, swimming, or eating a meal. Those have shown that the prefrontal cortical area may be predominantly involved in motor imagery, while the predominant involvement of the primary sensorimotor cortex is considered to be motor executive function. Recently, motor imagery of swallowing, termed swallowing execution, was examined using fMRI, with the results indicating that motor imagery may enhance motor learning and performance in patients with post-neurologic injury. The present study was conducted to define cortical activities associated with chewing execution and imagery to establish neurorehabilitation strategies for patients with chewing deficits. Twelve healthy subjects were enrolled. Functional (T2* weighted) images were acquired using a 1.5-T Horizon MRI scanner (Intera Achieva 1.5 T, Philips Co.) and used to determine cortical activities. The subjects performed chewing imagery (CI) and gum chewing (GC), without odor or taste, as tasks, as well as rest (RE). Each task was performed for 30 seconds following by 30 seconds of RE, with each session repeated 5 times. Results for GC vs. Re showed primary sensorimotor cortex (M1/S1), primary visual cortex (V1), operculum (OP), lingual gyrus (LG), pallidus (PL), caudate (CD), amygdala (AMY), and para-hippocampus (PHC) activation. In contrast, results for CI vs. Re showed activation in the M1/S1, V1, OP, LG, insula (IS), temporal mid. (TM), precentral gyrus (PC), cingulate mid. (CM), and prefrontal cortex (PFC). Furthermore, results for CI vs. GC showed PFC, supplementary motor cortex (SMC), M1/S1, V1/V2, PC, IS, LG, TM, cingulate ant. (CA), CD, and putamen (PT) activities. We concluded that chewing execution specifically elicits activities in the limbic system, while chewing imagery specifically elicits top-down hierarchical cortex activities as part of the action planning of chewing.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-227 経路探索課題遂行中のサル前頭前野の軸符号化細胞の発見 Kazuhiro Sakamoto（坂本一寛）¹，Naohiro Saito（斎藤尚宏）²，Shun Yoshida（吉田隼）²，Hajime Mushiake（虫明元）² ¹東北医科薬科大医神経科学 ²東北大院医生体システム The prefrontal cortex (PFC) plays a crucial role in the cognitive processes known as executive functions that are necessary for goal-directed behavior in complex environments. To determine how the PFC employs its neuronal resources to adapt to ever-changing situations, the present study explored the tuning properties and time-development patterns of prefrontal neurons in male Japanese monkeys during a path-planning task. This behavioral task requires that a subject move a cursor to a final goal (FG) in a stepwise manner. We identified a neuronal group exhibiting activities associated with the first cursor direction (immediate-goal [IG] direction). The present analyses revealed that the neurons in this group displayed axis tuning along path directions and that this activity was transformed from vector tuning for FGs. The present study also attempted to distinguish putative interneurons (IN) from pyramidal neurons (PN) and found that both types of neuron shared similar tuning profiles. Thus, the present results from the macaque PFC demonstrated that axis-tuned cells and their dynamical transformation were involved in efficient spatial planning. These findings indicate that the brain is a complex system that manages its resources in a flexible and economical manner to adapt to uncertain environments.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-228 赤核はリーチング運動における運動誤差と標的誤差の適応に関与する Masato Inoue（井上雅仁）¹，Shigeru Kitazawa（北澤茂）^2,3,4 ¹大阪大学国際医工情報センター ²大阪大学大学院生命機能研究科 ³大阪大学大学院医学研究科 ⁴国立研究開発法人情報通信研究機構脳情報通信融合研究センター Errors in reaching could result from imperfect motor commands (motor error) or from target movements (target error). We have recently shown that motor error in the primary motor and the premotor cortices as well as in area 5 drives adaptation to compensate for the motor error, whereas target error in area 7 drives adaptation to compensate for the target error (Inoue et al., 2016; Inoue & Kitazawa 2018). The red nucleus (RN) receives inputs from the motor cortices and area 5 and sends output to the climbing fibers of the cerebellar cortex via inferior olivary nucleus, but not from area 7. We thus hypothesized that the RN provides signals for adaptation to the motor error but not for the target error. To test this hypothesis, we recorded neuronal activities of RN while monkeys made rapid reaching movement toward a visual target that appeared at a random location on a target screen. In one condition (prism condition) the motor error was augmented in a random direction by using a motor-driven prism device. In another (target-jump condition), the target jumped in a random direction to introduce target errors. We found 14 RN neurons that encoded motor error information during the prism condition, but unexpectedly found that seven of them encoded target error information during the target-jump condition as well. Further, three RN neurons encoded target error information in particular. Repetitive pairing of reaching movements with microstimulation to each or the 17 recording sites revealed that motor errors were compensated when the neuron of the stimulation site encoded motor error in particular (n = 7). By contrast, target errors were compensated when the neuron of the stimulation encoded target error (n = 10). These results suggest that the RN provides signals for adapting to the target errors in addition to those for adapting to the motor errors.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-229 難易度が高い運動実行中に側坐核ー運動野間の機能連関が出現する Michiaki Suzuki（鈴木迪諒），Yukio Nishimura（西村幸男）東京都医学総合研 Physical effort is a key issue for boosting motor performance. The nucleus accumbens (NAc) is suggested to be involved in effort-related behaviors from the aspect of decision-making. However, it is still unclear whether the NAc represents physical effort such as demanding motor execution. To clarify this question, we recorded local field potentials from the NAc and the primary motor cortex (M1) while a monkey performed demanding motor task. We, then, analyzed the neural activity in both areas, and investigated the NAc-M1 coupling related to different demanding motor execution. The task required the monkey to control the one-dimensional position of a cursor on a video monitor with wrist extension movement, and included the instruction phase and motor execution phase. One of the five targets reflecting different elastic force control conditions was presented on the screen during instruction phase. Then, the monkey was required to acquire the target after the go cue and to return the cursor to the start position after acquiring the target to receive juice reward. Five different force control conditions were classified into two situations, easy (free, weak) or demanding (accuracy, strong, strong-accuracy), based on difficulty in force control. The monkey received the same amount of reward/trial regardless force conditions. Results showed that the NAc activity during instruction phase was not different between different conditions. However, during motor execution the NAc activity at low frequency band (0-10Hz) was higher than that in easy conditions as well as the M1 activity. Furthermore, connectivity analysis showed that coherent activity between the NAc and M1 appeared at the low frequency band during demanding motor execution but not easy condition. Thus the NAc activity and the NAc-M1 coupling during motor execution was modulated and depends on demanding level of motor control. Those results suggest the functional contribution of the NAc to demanding motor control.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-230 運動学習と関連した白質における脂質の変化が一次運動野の神経活動に及ぼす影響 Kazuki Nishida（西田一貴）¹，Yoshihisa Tachibana（橘吉寿）¹，Shumpei Sato（佐藤駿平）²，Fumiyoshi Yamazaki（山崎文義）²，Mitsutoshi Setou（瀬藤光利）^2,3，Hiroaki Wake（和氣弘明）^1,3 ¹神戸大学大学院, 医学研究科, システム生理学分野, 神戸, 日本 ²浜松医科大学, 細胞分子解剖学講座, 浜松, 日本 ³CREST, JST There are abundant myelinated axons forming white matter in the brain. The myelinated axons can increase conduction velocity, thus contributing to the motor performance. Previously, human MRI studies have demonstrated that motor-learning related structural changes are detected in white matter. We have also demonstrated that oligodendrocytes facilitate axonal myelination in a neuronal-activity dependent manner in vitro. However, it remains unknown what kinds of molecules in the myelinated axons change during motor learning. In this study, we hypothesized that the profiles of lipids, the main components of the myelin, are changing during myelination in efferent axons associated with motor learning that affects on the alteration of neuronal activities in the primary motor cortex (M1). To test these issues, we used the liquid chromatography tandem mass spectrometry (LC-MS/MS) and matrix-assisted laser desorption/ionization Fourier-transform ion cyclotron resonance mass spectrometry imaging (MALDI FT-ICR MSI) to explore the lipids in the M1 of mice performed lever-pulling task with simultaneous monitoring of multiple M1 neurons using in vivo two-photon microscope. We found that sphingomyelin were increased unilaterally around white matter beneath M1 after motor training, and the number of task-related active neurons in M1 decreased in the process of motor learning. The Interhemispheric ratio of sphingomyelin distributed in the white matter was positively correlated with the number of success trials, while negatively correlated with the M1 activity in success trials. Here, we conclude that the change in lipid profiles in our brain is a neural basis for motor learning. In future, we will examine the motor-learning related changes in the metabolites of sphingomyelin (e.g., ceramide). We would also like to control animal's motor learning by manipulating the myelination by optogenetic stimulation of oligodendrocytes.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-231 優れてはいないが高感度のパーキンソン病患者に内在する潜在的運動適応能力: スマートデバイスを利用した研究 Ken Takiyama（瀧山健）¹，Takeshi Sakurada（櫻田武）^2,3，Masahiro Shinya（進矢正広）⁴，Takaaki Sato（佐藤剛章）⁵，Hirofumi Ogihara（荻原啓文）^5,6，Shin-ichi Muramatsu（村松慎一）^5,6,7，Taiki Komatsu（小松泰喜）^7,8 ¹東京農工大院工先端電気電子 ²立命館大理工 ³自治医大脳外科 ⁴広島大総科 ⁵鹿教湯病院 ⁶日本保健医療大理学療法 ⁷自治医大神経内科 ⁸日本大学スポ科 Parkinson disease (PD) is the second common neurodegenerative disease in the elderly. The decrease in dopamine in the striatum leads to cardinal motor symptoms including tremor, bradykinesia, and rigidity. Previous studies demonstrated the impaired implicit learning ability and the reduced sensorimotor adaptation ability in PD. These studies used visuomotor adaptation that was applied abruptly to evaluate motor adaptation. However, this method required explicit or cognitive ability to minimize the movement error. Here, we investigated the motor adaptation ability of the PD patients while decreasing the involvement of the cognitive ability and the burden to participate in the experiments. We utilize a smart-device-based experimental setting that is available to conduct motor adaptation experiments at any time and anywhere. Additionally, we relied on the gradually applied perturbation whose existence was aware of by 1 out of 54 healthy young participants in our previous study. A striking feature of the gradually applied perturbation is its difficulty to be noticed. The motor adaptation without awareness enables to investigate motor adaptation ability while avoiding the influence of task switch that involves the cognitive burden. We investigated the motor adaptation ability inherent in the PD patients (N=18, age=70.0±7.3), the age-matched elderlies (N=18, age=75.7±12.7), and the young healthy subjects (N=18, age=21.3±1.5). The PD patients performed the motor task during the on-state of their medication cycle, and the Hohen &Yahr stages of most patients were 3. There was a significant group effect on the magnitude of the motor adaptation (one-way ANOVA, F(2,51) = 6.75, p=0.0025); the PD patients showed the largest extent (Tukey's post-hoc test, p=0.0184 between the PD patients and the elderly, p=0.0032 between the patients and the young). In contrast, there was no significant group effect in the distance between the desired and actual adaptation pattern (one-way ANOVA, F(2,51)=1.5, p=0.232). In conclusion, the PD patients possessed larger but not better motor adaptation ability than the elderlies and the young. Motor adaptation can involve cerebellar function. Our finding supports the compensatory or paretic cerebellar functions inherent in the PD patients.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-232 機械刺激誘発嚥下における上皮型ナトリウムイオンチャネルの関与 Takanori Tsujimura（辻村恭憲），Makoto Inoue（井上誠）新潟大医歯総合研摂食嚥下リハ Food and/or liquid boluses are propelled from oral cavity to esophagus via pharynx during swallowing. A swallowing reflex evoked by pharyngolaryngeal stimulation plays a critical role not only in nutrition but also airway protection. Mechanical, taste and thermal stimuli are involved in swallowing initiation. Because impaired laryngeal mechanical sensation is associated with food bolus aspiration, it is important to know how the laryngeal sensory system regulates swallowing initiation. This study was performed to clarify the neuronal mechanism of mechanically evoked swallows. Urethane-anaesthetized Sprague Dawley male rats were used. A swallow was identified by activation of the suprahyoid and thyrohyoid muscles on electromyography. The swallowing threshold was measured by von Frey filament or electrical stimulation of the larynx. The number of swallows induced by upper airway distension or capsaicin application (0.03 nmol, 3 μl) to the vocal folds was counted. The effects of topical application of amiloride analogues (amiloride, benzamil, and dimethylamiloride), acid-sensing ion channel (ASIC) inhibitors (mambalgine-1 and diminazene), and gadolinium (0.3, 3 and 30 nmol, 3 μl) to the laryngeal mucosa on swallowing initiation were evaluated. A nerve transection study indicated that afferents carried by the superior laryngeal nerve play a primary role in the initiation of laryngeal mechanically evoked swallows. The mechanical threshold of swallowing was increased in a dose-dependent manner of amiloride analogues and gadolinium, but not ASIC inhibitors. The increased swallowing threshold by amiloride analogues or gadolinium was diminished following saline washout. The number of swallows by upper airway distension was significantly decreased by benzamil application. However, the initiation of swallows evoked by capsaicin or electrical stimulation was not affected by benzamil application. We speculate that the epithelial sodium channel is involved in the initiation of mechanically evoked swallows in the larynx.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-233 ゼブラフィッシュの逃避運動における左右のマウスナー細胞間の相反抑制の役割 Takashi Shimazaki（島崎宇史）^1,2，Masashi Tanimoto（谷本昌志）¹，Yoichi Oda（小田洋一）¹，Shin-ichi Higashijima（東島眞一）^2,3 ¹名古屋大学大学院　理学研究科　生命理学専攻 ²自然科学研究機構　生命創成探究センター ³総合研究大学院 During many behaviors in vertebrates, the central nervous system (CNS) generates asymmetric activities between the left and right sides to produce asymmetric body movements. For asymmetrical activations of the CNS, reciprocal inhibition between the left and right sides is believed to play a key role. However, the complexity of the CNS makes it difficult to identify the reciprocal inhibition circuits at the level of individual cells and the contribution of each neuron to the asymmetric activity. Using larval zebrafish, we examined this issue by investigating reciprocal inhibition circuits between a pair of Mauthner (M) cells, giant reticulospinal neurons that trigger fast escapes. Previous studies have shown that a class of excitatory neurons, called cranial relay neurons (CRNs), is involved in the reciprocal inhibition pathway between the M-cells. Using transgenic fish, in which two of the CRNs (Ta1 and Ta2) expressed GFP, we showed that Ta1 and Ta2 constitute major parts of the pathway. In larvae in which Ta1/Ta2 were laser-ablated, the amplitude of the reciprocal IPSPs dropped to less than one-third. Calcium imaging and electrical recording showed that the occurrence probability of bilateral M-cell activations upon sound/vibration stimuli was greatly increased in the Ta1/Ta2-ablated larvae. Behavioral experiments revealed that the Ta1/Ta2 ablation resulted in shallower body bends during sound/vibration-evoked escapes, which is consistent with the observation that increased occurrence of bilateral M-cell activations impaired escape performance. Our study revealed major components of the reciprocal inhibition circuits in the M-cell system and the behavioral importance of the circuits.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-234 鳴禽類の求愛行動における発声と身体運動の統合 Chihiro Mori（森千紘）^1,2，Ryosuke O Tachibana（橘亮輔）¹，Kazuo Okanoya（岡ノ谷一夫）^1,3 ¹東京大院総合文化研生命環境科学 ²日本学術振興会 ³理研CBS 認知行動連携 Many animal species communicate by multimodal signals, as in human speech with body gestures. In songbird species, communication through vocalization and motional signal is common in courtship displays (Ota et al. 2015, Ullrich et al. 2016, Iwama & Soma 2017). In the present study, we investigated how songbirds coordinate their body movement with learned song production. Male Java sparrow sings learned song to female during courtship with performing a stereotyped dance that includes hopping and bill-wiping. We recorded their courtship behavior especially focusing on head movement and song, and analyzed relationships between temporal patterns of them. The courtship tended to start with both hopping and bill-wiping, then the song followed later on. While the bill-wiping stopped during introductory notes of song, the hopping often continued until the end of song. We found that a hopping frequency was significantly higher during singing than during non-singing period, though hop timings were not associated with specific positions in the song. To reveal neural mechanisms for integration of multimodal communicative signals, we examined whether partial lesions on song-related brain area affect the courtship behavior. A preliminary result showed that partial pharmacological lesions of song premotor nucleus HVC changed songs, but didn't affect dance movements. These suggest that HVC is not involve in coordination of vocalization and motional signal. We are currently testing unilateral, bilateral, and reversible lesions to examine more detail.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-235 侵害受容刺激による呼吸制御に関わる腕傍核ニューロンの特性 Akiko Arata（荒田晶子）¹，Sotatsu Tonomura（外村宗達）²，Hirotaka Ooka（大岡裕隆）¹，Yumi Nakano（仲野由美）¹，Yoko Fujiwara-Tsukamoto（塚元葉子）³，Koichi Noguchi（野口光一）² ¹兵庫医大医生理（生体機能） ²兵庫医大医解剖（神経） ³羽衣国際大食物栄養 The sensation of nociceptive signals projects to the lateral parabrachial nucleus (LPB) of the pons via the dorsal horn; and LPB has also known as the system of inspiratory-expiratory (I-E) phase switching that contributes to the control of respiratory rate. The tight interaction between respiration and pain signals as nociception-respiration coordination were expected in LPB. In this study, we investigated the nociceptive-respiratory system using the pons-medulla-spinal cord preparation intact forelimb isolated from postnatal 0-2 days-old-rats. Respiratory activity was recorded from cervical fourth (C4) ventral nerve root. The C4 inspiratory rate increased significantly when a small amount of 2% capsaicin was injected into forelimb with pons, but the removal of pons had no effects. Moreover, C4 inspiratory rate also increased significantly with C8 dorsal root stimulation as a noxious stimulation. We examined the responded area of LPB stimulated by C8 dorsal root using optical imaging with voltage-sensitive dye, and the LPB neurons were recorded from the responded area using whole-cell patch-clamp. I-E neurons which were synchronized with the Inspiratory-Expiratory phase of C4 activity existed in the LPB and Kolliker-Fuse nucleus(KF). The spontaneous firing neurons which were not synchronized with the C4 activity were called non-respiratory neurons and existed in the LPB. In the external LPB, all I-E neurons and the half of recorded non-respiratory neurons were responded by C8 dorsal root stimulation. Non-respiratory neurons in the superficial layer of LPB received C8 dorsal root stimulation and showed a post-inhibitory rebound (PIR) caused by hyperpolarizing current pulse application. These results suggested that I-E neurons could directly receive noxious information, so I-E neurons were thought to be the core mechanism of nociceptive-respiratory coordination; the non-respiratory LPB neurons which expressed PIR might be participating in the onset mechanism of the nociceptive-respiratory relay network.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-236 Topographic representation of saccade vector in frontal eye field of common marmoset Chih-Yang Chen（Chen Chih-Yang）¹，Denis Matrov（Matrov Denis）¹，Kuan-Ting Ho（Ho Kuan-Ting）¹，Tadashi Isa（Isa Tadashi）^1,2 ¹Division of Physiology and Neurobiology, Department of Neuroscience, Graduate School of Medicine, Kyoto University ²Evolutionary Systems Neuroscience Group, Institute for the Advanced Study of Human Biology (ASHBi), Kyoto University Frontal eye field (FEF) is an important cortical area controlling saccadic eye movements. However, because this area is deep in the rostral bank of arcuate sulcus in macaques, which makes it difficult to access, whether it contains a continuous topographic representation of saccade vector remains undetermined. In the current study, we took advantage of the almost-flat cortical surface of common marmosets and systematically mapped the FEF with electrical microstimulations. We applied electrical microstimulations with tungsten electrodes in potential cortical areas identified by the atlas during the gap period, when marmosets performing gap saccade task to control the initial eye position and baseline state of eye fixation. The stimulations were applied with biphasic current at 300 Hz for 30 trains. We varied the stimulation amplitude in some cases but most of our stimulations were kept at 0.07 mA to ensure enough power to evoke saccades while minimizing the brain damage. We also systematically varied the initial fixation locations to identify whether the evoked saccades were vectors-based or toward a single endpoint. We successfully evoked saccades in area 8 and 45 according to the atlas. When the stimulation sites moved from medial to lateral, we observed a systematic decrease in amplitude and changing of direction from upper to lower visual field of the evoked saccades. The evoked saccades in these areas were vector-based. However, if the stimulation site was posterior to this region, the saccades became more toward a single endpoint. Taken together, we found a continuous topographic representation of saccade vectors in the marmoset FEF. Because several important cortical areas related to oculomotor systems are deep in the sulci of macaques, they are less understood. Using marmosets, we will be able to advance our knowledge on the organization of these areas and understand more about the primate oculomotor systems.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-237 新生ラット摘出脳幹ー脊髄標本にみられる呼吸ー体動連関における5-HT1A受容体の役割 Yoko Tsukamoto（塚元葉子）^1,2，Hirotaka Ooka（大岡裕隆）²，Toki Fujimoto（藤本乙希）¹，Chiaki Uchidda（内田千晶）²，Akiko Arata（荒田晶子）² ¹羽衣国際大・人間生活・食物栄養 ²兵庫医大医生理（生体機能） Respiratory rhythm is belonging to the medullary rhythm generator, and body movement is belonging to the spinal rhythm generator. Parabrachial Nucleus (PB) in the dorsolateral pons is a conjunctive system of medulla respiratory activity and spinal motor activity. It is known that serotonin strongly modulates the rhythm generator like respiratory or body movement. In the previous studies, we analyzed the relationship between respiratory rhythm and body movement using neonatal rat medulla-spinal cord preparation with/without pons; and we also examined the effect of serotonin on the relationship between respiratory rhythm and body movement in PB. We showed that the respiratory rhythm was synchronized to body movement when serotonin was applied in the pons-medulla-spinal cord preparation. On the other hand, even if serotonin was applied, the relationship between respiratory rhythm and body movement was not observed without pons. In this study, we examined the following things: the distribution of optical signals in the pons triggered by body movement; which of serotonin receptor participates in the relationship between respiration and body movement; and where is the serotonin receptor concerning with their relationship using immunohistochemical method. We found the optical signals induced by body movement in the dorsal lateral pons. The respiration-body movement coupling was abolished by 5-HT1A receptor blocker. A lot of 5-HT1A receptor immunoreactive cell bodies were found in lateral PB in the dorsolateral pons. These results suggest that pons is essential structure for the respiratory-body movement coupling, and 5-HT1A receptor in lateral PB may play a crucial role in the functional connection between medulla and spinal cord.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-238 一側一次運動野に対する経頭蓋的静磁場刺激は両半球間で異なる皮質興奮性の変化をもたらす Yasuyuki Takamatsu（高松泰行）^1,2，Satoko Koganemaru（小金丸聡子）³，Tatsunori Watanabe（渡邊龍憲）^4,5，Yoshihiro Yukawa（湯川喜裕）⁶，Masatoshi Minakuchi（水口雅俊）⁶，Ryota Shimomura（下村亮太）⁶，Tatsuya Mima（美馬達哉）¹ ¹北海道大院保健機能回復 ²立命館大院先端総合学術研究科 ³獨協医大医生理（生体情報） ⁴名古屋大院医リハビリ療法 ⁵日本学術振興会 ⁶村田病院リハビリテーション部 It has been reported that transcranial static magnetic stimulation (tSMS) over the primary motor cortex (M1) suppressed the cortical excitability by measuring motor evoked potentials (MEPs). Based on the theory of interhemispheric inhibition, we investigated whether tSMS over the left M1 had the facilitating effects of the cortical excitability on the right M1 and the changing of the excitability in each hemisphere showed inverse correlation. The design of this study was crossover trial, and right-handed 18 healthy subjects were included. The electromyogram was recorded from both first dorsal interosseous (FDI) muscles by single pulse transcranial magnetic stimulation. For tSMS, we used a cylindrical neodymium (NdFeB) magnet, which the surface magnetic flux density was 5340 G. A non-magnetic stainless-steel cylinder was used for sham stimulation. The duration of intervention was 30 min. We measured MEPs for both FDI muscles at before (Pre), immediately after (Post-0), 10 min after (Post-10), and 30 min after (Post-30) the intervention. Three-way repeated-measures ANOVA was used with Time as within-subject factor, and Hands and Conditions as between-subject factor. In case of significant interaction effects, we performed two-way repeated-measures ANOVA and post hoc analysis by using the Bonferroni correction for multiple comparisons. In addition, the correlation coefficient of the changing ratio between right and left hands at each time point with respect to the baseline was obtained. Time × Hands × Conditions interaction was significant. By two-way repeated-measures ANOVA, Time × Hands interactions was detected only for the real condition, and post hoc analysis showed that MEPs peak-to-peak amplitude was significantly decreased at Post-0, 10, and 30 compared to at Pre in the right hand. In the left hand, it was significantly increased at Post-0 compared to at Pre. Significant correlation of the changing ratio between right and left hands at each time point in both real and sham stimulation was not shown. This study might indicate that the suppression of the cortical excitability in the unilateral M1 applied tSMS induced the facilitation of the excitability in the contralateral M1. However, there was no linear correlation between the stimulation side and the opposite side, so it is necessary to further investigate the physiological mechanism by tSMS to M1 affects the other side.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-239 運動学習による皮質間連絡と視床-皮質投射の運動野神経回路変化 Jaerin Sohn（孫在隣）^1,2，Yoshiyuki Kubota（窪田芳之）^1,3，Yasuo Kawaguchi（川口泰雄）^1,3 ¹生理研基盤神経科学研究領域大脳神経回路論 ²日本学術振興会特別研究員PD ³総合研究大学院大学 For skillful movement, the neuronal circuitry in the motor cortex should be optimally organized. Repetitive training for a novel motor skill induces a reorganization of neuronal wiring in the motor cortex involving synaptogenesis, indicated by spine formation on pyramidal cell dendrites. However, the origin of inputs to the synapses on spines formed during motor learning has remained to be clarified. In order to depict the circuit remodeling in the motor cortex, here we characterized the presynaptic axon terminals innervating the newly-formed spines during motor learning. To observe spine dynamics in vivo under a two-photon microscope, we used Thy1-eGFP-M mouse line, in which green fluorescent protein is expressed in layer 5 pyramidal cells. To quantify motor learning, we applied success rate of single-seed reaching task. The spine formation rate at the dendritic tuft in layer 1 was significantly correlated with the success rate increase after the reaching training, indicating that the spine formation reflects the refining of the motor skill. To identify whether the presynaptic axon comes from the cortex or thalamus, we fixed the brains immediately after the two-photon microscopy, followed by immunohistochemistry for the excitatory presynaptic type markers (type I and II vesicular glutamate transporters), simultaneously with a excitatory postsynaptic marker. Confocal laser scanning microscopy at a synaptic resolution revealed that transiently-formed spines and persistently-formed spines received different excitatory inputs. The transiently-formed spines largely received the corticocortical inputs, whereas the persistently-formed spines were frequently innervated by thalamocortical inputs. Our finding provides a description of wiring dynamics during motor learning, indicating that corticocortical and thalamocortical networks play different roles in the cortical remodeling.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-240 脊髄後根神経節における好中球－神経の相互作用が実験的自己免疫性脳脊髄炎で生じる機械的アロディニアを引き起こす Yoshinori Hayashi（林良憲），Yuka Harada（原田ゆか），Jing Zhang（張競）九州大学大学院歯学研究院口腔機能分子科学 In multiple sclerosis (MS) patients, pain is a frequent and disabling symptom. However, the underlying mechanisms of pain in MS patients is poorly understood. In the present study, we have demonstrated that neutrophil accumulation in the dorsal root ganglion (DRG) is required for mechanical allodynia in experimental autoimmune encephalomyelitis, an animal model of MS, induced by myelin oligodendrocyte glycoprotein (MOG_35-55) immunization. Mechanical allodynia was observed in MOG_35-55-immunized mice, which preceded appearance of motor dysfunction. In addition, neutrophils were accumulated in the dorsal root ganglion (DRG). Neutrophil depletion abrogated mechanical allodynia caused by MOG_35-55 immunization. In contrast, adoptive transfer of MOG_35-55-stimulated neutrophils elicited mechanical allodynia in naive mice. Accumulated neutrophils in the DRG released neutrophil elastase in a cathepsin E (CatE)-dependent manner. Sivelestat, a selective neutrophil elastase inhibitor, suppressed mechanical allodynia. CatE-deficient (CatE^-/-) mice were highly resistant to MOG_35-55-induced mechanical allodynia. In addition, adoptive transfer of MOG_35-55-stimulated CatE^-/- neutrophils did not elicit mechanical allodynia in naive mice. Neutrophil-driven increased pain perception was mediated through the activation of protease-activated receptor 2 in DRG neurons. Activation of neutrophils by MOG_35-55 was mediated through toll-like receptor 4 (TLR4). MOG_35-55 also activated TLR4 in DRG neurons and induced chemokine (C-X-C motif) ligand 1 (CXCL1) which triggered neutrophil chemotaxis in the DRG. Finally, knockdown of CXCL1 or TLR4 in DRG neurons abrogated neutrophil accumulation in the DRG and mechanical allodynia after MOG_35-55 immunization. Thus, neutrophil-neuron crosstalk in the DRG is required for mechanical allodynia in experimental autoimmune encephalomyelitis. These findings suggest the mechanism of driving mechanical allodynia caused by MOG_35-55 and new strategy for preventing pain in MS patients.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-241 脳と腸の関係のモデルマウスの研究 Masami Kojima（小島正己）^1,3，Toshiyuki Tmizui（水井利幸）¹，Takamichi Tsukahara（塚原隆充）²，Konomi Matsui（松井このみ）^1,3 ¹産業技術総合研究所　バイオメディカル研究部門 ²株式会社栄養・病理学研究所、宇治田原町、京都、日本 ³大阪大学大学院生命機能研究科 Although the knowledge of brain is accumulated, how the function alters those of other organs and body. In the present study, we demonstrate that the actions of neuronal activity-induced brain-derived neurotrophic factor(BDNF) alters the function of tight junction (TJ) on intestinal epithelia. To show that, we compared the TJ function on the epithelia between wild-type and mutant mice with genetically-alteration in the BDNF gene. The mutant mice reduce the BDNF amount in brain by 64 % approximately and behavioral impairment of social interaction and anxiety. Interestingly, in the mutant animals, FITC-labeled dextran (MW 4000 kDa), which are administrated orally, penetrated into the blood through the intestinal epithelia TJ. However, the wild-type mice exhibited the penetration at significantly low levels compared to the littermate mutant animals. We will further show the impairment in social interaction test and other behavioral measurements. Anatomical analysis revealed that the population of thinner spines increased a and that of mushroom-head type ones. These results together suggest that the impairment of brain function leads to that of gut and TJ and germs invaded through the gut TJ render the body functions vulnerable.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-242 グルコース投与によるストレス誘導性交感神経系活性化の抑制 Naoko Yamaguchi（山口奈緒子）¹，Yoshihiko Kakinuma（柿沼由彦）²，Kaoru Mimura（三村郁）¹，Tomiko Yakura（矢倉富子）³，Munekazu Naito（内藤宗和）³，Shoshiro Okada（岡田尚志郎）¹ ¹愛知医科大学医学部薬理学講座 ²日本医科大学生体統御科学分野 ³愛知医科大学医学部解剖学講座 Exposure to stressor causes various systemic responses such as activation of the sympathetic nervous system and the hypothalamus-pituitary-adrenocortical (HPA) axis. Stress-induced sympathetic activation increases plasma levels of catecholamines (noradrenaline and adrenaline), resulting in elevations in blood pressure and heart rate. In addition, facilitated secretion of adrenaline from the adrenal medulla and pancreatic glucagon associated with sympathetic activation increase blood glucose level. We have previously reported that stress-related neuropeptides such as corticotropin-releasing factor and acute exposure to restraint stress increase the plasma levels of catecholamines, and that prostanoids such as prostaglandin E₂ and thromboxane A₂ in the brain, especially in the paraventricular hypothalamic nucleus (PVN), regulate the elevation of plasma catecholamine levels. Therefore, there is a high possibility that brain prostanoids are involved with stress-related changes in glucose dynamics. In the present study, we examined effects of intravenous glucose infusion on sympathetic activation, that is elevations of plasma catecholamine levels and changes in brain prostanoid levels, under restraint stress exposure in male rats. There were no significant differences in blood glucose levels between vehicle-treated stressed group and glucose-treated stressed group. However, intravenous infusion of 5% glucose solution effectively suppressed stress-induced elevation of plasma level of adrenaline, but not noradrenaline. Similarly, intracerebroventricular infusion of glucose solution also prevented the stress-induced elevation of plasma level of adrenaline, but not noradrenaline. Furthermore, we found that exposure to restraint stress increased thromboxane B₂, a stable metabolite of thromboxane A₂, in PVN dialysates, and the increase in thromboxane B₂ level was suppressed by intravenous glucose infusion. Our results suggest that glucose infusion can inhibit stress-induced sympathetic activation, and that prostanoids in the PVN are involved with this mechanism.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-243 ラットにおいてストレス関連性神経ペプチドのボンベシンは脳内5-HT₇受容体を介して中枢性に頻尿を誘発する Takahiro Shimizu（清水孝洋）^1,2，Shogo Shimizu（清水翔吾）¹，Naoki Wada（和田直樹）²，Shun Takai（高井峻）²，Nobutaka Shimizu（清水信貴）²，Youichirou Higashi（東洋一郎）¹，Katsumi Kadekawa（嘉手川豪心）²，Tsuyoshi Majima（馬嶋剛）²，Naoki Yoshimura（吉村直樹）²，Motoaki Saito（齊藤源顕）¹ ¹高知大医薬理 ²ピッツバーグ大医泌尿器 [Aims of study] Psychological stress exacerbates symptoms of bladder dysfunction including overactive bladder (OAB) and bladder pain syndrome (BPS), but the underlying brain mechanisms are unclear. Recently, we reported that bombesin (BB)-like peptides, which are stress-related neuropeptides, centrally induce frequent urination in rats. Under stress conditions, brain BB-like peptides can modulate activity of the serotoninergic nervous system, which has also been shown to modulate micturition. In this study, therefore, we examined the brain mechanisms for the BB-induced frequent urination in rats, focusing on 5-HT receptor subtypes, 5-HT_1A, 5-HT₂ and 5-HT₇, reported to control micturition in the CNS. [Methods] In urethane anesthetized (1.0 g/kg, ip) male SD rats (300-350 g), a catheter was inserted into the bladder to perform cystometry (12 ml/h saline infusion). Three hours after the surgery, BB or each 5-HT receptor antagonist was icv administered. (1) p-chlorophenylalanine (PCPA), a 5-HT synthesis inhibitor, was administered (200 mg/kg, ip) once a day for 2 days to induce acute depletion of brain 5-HT. After a day of the second administration, BB (0.03 nmol/rat) was icv administered. (2) WAY-100635 (WAY, a 5-HT_1A antagonist, 0.1 or 0.3 μg/rat), ritanserin (Ri, a 5-HT₂ antagonist, 0.3 or 1 μg/rat) or SB269970 (SB, a 5-HT₇ antagonist, 0.1 or 0.3 μg/rat) was icv pretreated 30 min before BB administration (0.01 or 0.03 nmol/rat, icv). [Results] (1) BB (0.03 nmol/rat, icv) significantly reduced intercontraction intervals (ICI) without affecting maximum voiding pressure. The BB-induced response was significantly suppressed by PCPA pretreatment. (2) BB at a lower dose (0.01 nmol/rat, icv) showed no significant effect on ICI, while it significantly reduced ICI in the presence of WAY, which can block the negative feedback control of 5-HT release. BB (0.03 nmol/rat, icv)-induced ICI reduction was significantly attenuated by pretreatment with SB, but not with Ri. [Conclusion] The brain BB system, which is implicated in psychological stress responses, is involved in frequent urination through the brain serotoninergic nervous system, and the frequent urination is mediated at least by brain 5-HT₇ receptors. Thus, brain BB and 5-HT₇ receptors might be useful targets for alleviation of stress-induced exacerbation of bladder dysfunction such as OAB and BPS.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-244 マウスの多重ストレスが帯状回皮質神経回路オシレーションへ与える影響 Rina Ito（伊藤吏那），Yasushi Hojo（北條泰嗣），Miki Hashizume（橋爪幹），Takayuki Murakoshi（村越隆之）埼玉医大医　生化学 The anterior cingulate cortex (ACC) is susceptible to stress. For therapeutic strategy of mental disorders, it is important to clarify the mechanism of emotional-cognitive deficits mediated by stress load. We analyzed the effect of multiple stress load on the network oscillation in ACC and behavioral performance (pre-pulse inhibition: PPI and forced swim test: FST) using male C57BL/6J mice (5-week old). Multiple stress load was composed of three types of the protocols, i. e., methalazoxyn ethanol acetate (MAM) injection during embryonic stage, maternal separation (MS) for 3 hours a day during postnatal 0-14 day, and chronic restraint stress (CRS) for 2 hours a day during postnatal 4-5 weeks. Mice were divided to 6 groups, control (Ctrl, no stress load), MAM (MAM stress alone), MS (MS stress alone), CRS (CRS stress alone) and 3S (multiple stress loaded) group. On the next day of the behavioral tests (PPI and FST), a coronal slice including ACC (450 μm) was prepared and network oscillation was evoked by kainic acid (KA, 3 μM, 1min). Network oscillation was recorded as field recording potentials in the superficial layers of cg1 sub region in the ACC. The power of network oscillation was calculated by the area under the curve of power spectrum density (PSD). The PPI levels was suppressed in MAM and 3S groups, compared with Ctrl group, and the immobility time tended to decrease in MS, CRS and 3S groups. On the other hand, power of network oscillation decreased in 3S group, but not in the single stress loaded groups. These results suggest that multiple stress load has more deleterious effects on the properties of network in ACC, than single stress load.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-245 プロスタグランジンE2は視床下部の室傍核に存在するコルチコトロピン放出ホルモン（CRH）ニューロンを活性化する Hiroyuki Igarashi（五十嵐敬幸）^1,2，Eric Stephen Kuebler（Kuebler Stephen Eric）¹，Julio Martinez-Trujillo（Martinez-Trujillo Julio）¹，Wataru Inoue（井上渉）¹ ¹Dept Physiol Pharmacol, Univ of Western Ontario, London, Canada ²Western BrainsCAN, London, Canada Inflammation activates the hypothalamic-pituitary-adrenal (HPA) axis, and the ensuing release of anti-inflammatory glucocorticoids provide critical negative feedback onto the immune system. An inflammatory mediator prostaglandin E₂ (PGE₂) is a key intermediate for this immune-to-brain signalling driving the HPA axis activation. We recently showed that PGE₂ potently inhibits the release of GABA onto the output neuroendocrine neurons of the HPA axis [corticotropin releasing hormone (CRH) neurons in the paraventricular nucleus of the hypothalamus (PVN)] via presynaptic EP3 receptors (Khazaeipool Z, Wiederman M, Inoue W., 2018). This suggests that PGE₂ permits the excitation of PVN-CRH neurons by a removal of GABA-mediated inhibition. However, it remains unclear how PGE₂ increases the activities of PVN-CRH neurons. Here, we examined PVN-CRH neurons' response to PGE₂ using Ca²⁺ imaging in acute brain slices. We expressed a genetic Ca²⁺ indicator GCaMP6s in PVN-CRH neurons by injecting cre-dependent AAV9 expression vector in CRH-ires-cre mice. Two photon real-time imaging revealed that bath application of PGE₂ (1 μM) triggered robust, oscillating Ca²⁺ elevations in a subpopulation of PVN-CRH neurons. These results support excitatory effects of PGE₂ on PVN-CRH neurons and also revealed heterogeneity of PVN-CRH neurons in their responsiveness to PGE₂.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-246 食事制限により誘導される胸腺内での軸索リモデリング Hiroshi Hasegawa（長谷川潤）¹，Razali Nurhanani（Nurhanani Razali）¹，Hirofumi Hohjoh（北條寛典）¹，Devi Bishnu Maharjan（Mah Bishnuarjan Devi）¹，Kimie Nakagawa（中川公恵）¹，Ken-ichi Mizutani（水谷健一）² ¹神戸薬科大　衛生化学 ²神戸学院大院薬　幹細胞生物学 Thymus is one of the primary lymphoid organs, which shows plasticity upon various biological stimuli. Previous studies have indicated that the decrease in size of the thymus, called thymic involution, is induced by a number of changes in physiological conditions, including ageing and pregnancy. Thymic involution is correlated with the decrease in immune activity, resulting in higher risks to infectious disease or cancer. Therefore, understanding of the mechanisms of thymic involution is important to maintain our immune activity and prevent diseases caused by its distortion. In order to reveal the molecular mechanisms of thymic involution, we have utilized mouse diet-restriction model. Diet-restriction for 48 hours in mice induced the reduced thymic weight over the level of weight reduction of the whole body. Tissue section analysis indicated the appearance of lipid droplets and condensed blood vessels in the diet-restricted thymus, without affecting basic structure of the thymus. Considering that the interaction between axon blood vessels and nerve axons regulate axon elongation during development, we hypothesized that the remodeling of blood vessels are accompanied by the remodeling of peripheral axons innervating to the thymus. Immunohistochemical analysis using anti-PGP9.5 antibody indicated the PGP9.5-positive axons were denser in the diet-restricted thymus, compared with control thymus. The expression of some neurotrophic factors were also affected by the diet-restriction. These results indicate that the diet-restriction caused the remodeling of blood vessels and peripheral axons. Analysis of the expression of T lymphocytes differentiation markers by quantitative RT-PCR indicated that the differentiation of T lymphocytes are also discomposed. Thus, diet-restriction modulate the peripheral axons in the thymus, possible contribute to the differentiation of T lymphocytes.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-247 AF4ファミリーに属する転写因子AFF4の視床下部摂食調節系に対する役割の検討 Tadasuke Komori（小森忠祐）¹，Tetsuya Nosaka（野阪哲哉）²，Toshio Kitamura（北村俊雄）³，Yoshihiro Morikawa（森川吉博）¹ ¹和歌山県立医大医解剖二 ²三重大院医　感染症制御医学分野 ³東京大医科学研究所　先端医療研究センター　細胞療法分野 Food intake is controlled by the hypothalamus that integrates a variety of peripheral signals, including hormones and nutrients. During fasting, food intake is stimulated by ghrelin, a stomach-derived hormone, via the activation of AMP-activated protein kinase (AMPK) and its downstream target, acetyl-CoA carboxylase (ACC), in the hypothalamus. Previously, we have reported that AFF4, a member of AF4 family of transcription factors, was induced by fasting in the hypothalamus of mice. In addition, AFF4 activates ACC in a hypothalamic neuronal cell line, GT1-7 cells (Komori et al., J Biol Chem, 2011). However, the in vivo roles of AFF4 in the hypothalamus during fasting remain unclear. In the present study, we analyzed AFF4-deficient mice to gain insights into the functional roles of AFF4 in the hypothalamus. We used heterozygous AFF4-deficient mice (AFF4^+/- mice), because more than 85% of homozygous AFF4-deficient mice die by postnatal day 1. There was no change in the amount of daily food intake between wild-type (WT) and AFF4^+/- mice. However, the amount of food intake after fasting for 48 hours was reduced in AFF4^+/- mice compared to that in WT mice. In addition, AFF4^+/- mice showed lower phosphorylation of AMPK and ACC in the hypothalamus during fasting. It has been reported that hypothalamic AMPK/ACC signaling induces the expression of orexigenic neuropeptides, neuropeptide Y (NPY) and agouti-related peptide (AgRP). Fasting-induced the expression of NPY and AgRP was reduced in the hypothalamus of AFF4^+/- mice compared to that in WT mice. These results suggest that AFF4 in the hypothalamus plays an important role in the regulation of food intake via the activation of AMPK/ACC signaling during fasting.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-248 ゼブラフィッシュ幼生の腸組織のカルシウムイメージングと光遺伝学的解析 Daiji Takamido（高御堂大慈）¹，Sayaka Nishida（西田さやか）²，Takuya Kojima（兒島卓也）²，Masataka Nikaido（二階堂昌孝）¹，Koichi Kawakami（川上浩一）³，Shin-ichi Okamoto（岡本晋一）¹，Kohei Hatta（八田公平）¹ ¹兵庫県立大院生命理 ²兵庫県立大理 ³国立遺伝研 The gut of zebrafish larva could be a good model for investigating the motor function of the digestive organs of vertebrates in vivo. We have reported properties of some of the circular smooth muscles and neurons involved in peristaltic reflex by using Ca ²⁺ imaging of smooth muscles and neurons as well as optogenetic techniques. In peristaltic reflex, a strong contraction occurs rhythmically on the oral side of the bolus, and in correspondence with it, a strong Ca²⁺ event is observed in the circular smooth muscles. We also observed peristalsis-like movement which occurs without bolus, and neuronal cell types which are active associated with this movement. Meanwhile, irrespective of the presence or absence of bolus, a contractile movement which is more regular and higher in frequency, corresponding likely to slow waves, has been observed. These two movements could exist simultaneously. In the circular smooth muscles, the short cycle Ca²⁺ events could be hardly or only weakly observed. In this research, in order to investigate the mechanism of formation of two types of contraction waves with different cycles, we express Ca²⁺ indicator GCaMP3 in various cell types and search cells which fire in association with each wave. Here, we demonstrate a distinctive non-neuronal cell group acts strongly in conjunction with slow waves, which may correspond to the interstitial cells of Cajal. In addition, we recently found Ca²⁺ events in the endodermal tissues, and some of the Ca²⁺ events were associated with local gut wall movement. When endodermal cells expressing ChR2 were stimulated by irradiation with blue light in the mid- or distal intestine, peristalsis like activity was induced. On the other hand, in the proximal intestine, amplification of the rhythmic activity was observed without changing its frequency. This discrepancy in the effect of activation of endoderm may indicate the difference between the neural circuitry in the proximal gut and mid- and distal intestines.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-249 LPSによるLHサージ状分泌抑制とミノサイクリン前投与による抑制解除へのkiss-1ニューロンの関与 Hitomi Fujioka（藤岡仁美），Atsushi Fukushima（福島篤），Toshiya Funabashi（舩橋利也），Tatsuo Akema（明間立雄）聖マリアンナ医大医生理 We previously reported that pretreatment with minocycline, a potent inhibitor of activation of microglia/macrophage, significantly prevented this suppression of ovarian steroid-induced LH surge by lipopolysaccharide (LPS) in ovariectomized (OVX) rats. In this study, we examined whether minocycline attenuated LPS-induced inhibition of activation of GnRH neurons in the preoptic area (POA) and kiss-1 neurons in the anteroventral periventricular nucleus (AVPV). LH surge was induced by estradiol and progesterone in OVX rats. Minocycline or saline was administered intraperitoneally once a day for 4 consecutive days including the day of the experiment. LPS or saline was injected intravenously at noon. The rats were fixed and brains were collected during 16:30-17:30. Blood were sampled at 11:00 h and just before sacrifice, and serum concentrations of LH were determined by radioimmunoassay. Serum LH levels at the time of LH surge was significantly reduced by LPS treatment. Double immunohistochemistry demonstrated that LPS treatment significantly reduced c-Fos immunoreactivity in GnRH immunoreactive cells without affecting GnRH cell number in the POA. Similarly, a combination with in situ hybridization of kiss-1 and immunohistochemistry for c-Fos illustrated that LPS treatment significantly reduced c-Fos expressing kiss-1 mRNA cells in the AVPV with no effect on the number of kiss-1 expressing cells as well. Minocycline pretreatment significantly attenuated the suppression of LH surge by the LPS treatment. Minocycline also significantly attenuated the reduction of c-Fos expression in GnRH immunoreactive cells in the POA and in kiss-1 expressing cells in the AVPV. These findings suggest that minocycline ameliorates LPS-induced suppression of the LH surge by improving the activition of kiss-1 neurons in the AVPV in turn GnRH neurons in the POA.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-250 いぐさがヒトの睡眠に与える影響 Takashi Kudo（工藤崇）¹，Etsuko Kato（加藤悦子）¹，Yuri Yoshimura（吉村友里）¹，Akari Tsunedomi（常冨愛香里）¹，Le Yang（楊楽）¹，Akiyoshi Honden（本傳晃義）¹，Taisuke Nakashima（中島大輔）¹，Toshinori Nakagawa（中川敏法）¹，Koichiro Ohnuki（大貫宏一郎）³，Makoto Kawaguchi（川口誠仁）²，Rinpei Sawada（澤田倫平）²，Shinsuke Nishida（西田伸介）²，Kuniyoshi Shimizu（清水邦義）¹ ¹九州大学農学研究院 ²熊本県農業研究センター、アグリシステム総合研究所、いぐさ研究室 ³近畿大、産業理工学部 Common rush (Juncus effusus) is the source of Tatami. In Japan, almost all common rush is produced in Kumamoto prefecture. But recently the import of tatami is markedly increasing and also Tatami made of paper is increasing. This cause the reduction of tatami farm family and it is possible it will disappear soon. One reason is that classical Japanese house which containing tatami room is decreasing and Western type house which do not have Tatami room is increasing. Generally people say that Tatami room have good effects on humans, but the scientific data is lacking. Previously our group showed that common rush has relaxation effects on human during daytime. In this project, we examined the effects of dried common rush on human sleep during night time. Ten young males and ten young females were attended as the subjects. We used two condition. One is that placebo Tatami (made of paper) was set in the sleeping room and the other is that Tatami was set. The subjects came to the sleeping room around 9:30 pm and some questionnaires were performed until 11:00 pm. The room right was turned off at 11:00 pm and EEG, ECG, and the movement during the sleep were measured. At 7:00 am the room right was turned on and some questionnaires were performed again. The data was analyzed depends on the group and compared in two groups. As for the psychological aspect, negative mood was significantly improved in Tatami group which is assessed by POMS2 test. In physiological aspects, sleep efficiency assessed by body movement was significantly increased in Tatami group. This is one-night short time experiments, therefore long-term experiments were needed to confirm the effects of Tatami.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-251 睡眠・覚醒制御および背側縫線核におけるグレリンの作用 Juhyon KIM（金主賢），Ryosuke Okumura（奥村亮典），Toshiki Tajima（但馬俊貴），Kazuki Nakajima（中島一樹）富山大学・工・生体情報 Ghrelin, an endogenous peptide produced in the stomach and some of the brain regions, plays several physiological functions including growth hormone secretion and feeding behavior. Ghrelin also affects regulation of sleep-wakefulness (Wake). Central administration of ghrelin increases amount of Wake, and decrease amount of both non-rapid eye sleep (NREMS) and REMS lasting for 60 min or longer. However, only total time spent in Wake, NREMS and REMS has been compared, and the detail of changes in each state has not been clarified. Also, brain regions of which ghrelin-induced wakefulness consists are remained unclear. Therefore, we examined effect of ghrelin not only on total time, but also on each duration and number of entry into Wake, NREMS and REMS sleep events on ghrelin- or saline-administrated rats (ICV) by sleep stage analysis using electroencephalogram and electromyogram. Then, the brains were performed immunohistochemical staining to detect c-fos and serotonergic neurons in dorsal raphe nucleus (DR), one of Wake-promoting nuclei. Total time spent in the Wake was increased, and in the both NREMS and REMS were decreased by ghrelin administration. Ghrelin induced no significant change in frequency of entry into the wake state, but increased in the duration. On the other hand, frequency of entry into the NREMS and REMS state were decreased, but no change was observed in the duration. In the DR region of control group, sparse c-fos expressions were observed. On the other hand, number of c-fos-expressing neurons was increased significantly in ghrelin group. Furthermore, number of c-fos-expressing serotonergic neurons in ghrelin group was also greater than that in control group. The present result indicates that ghrelin-induced Wake-promotion consists of duration extension of the wake event, and decreases of frequency of the NREMS and REMS event and is induced in part by activation of serotonergic neurons in the DR.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-252 ヒスタミンとCGRPによる脳血流循環の制御とinfra-slow oscillation リズムとの関係 Tomokazu Ohshiro（大城朝一），Hajime Mushiake（虫明元）東北大学　大学院医学系研究科　生体システム生理学教室 Distinct rhythmic oscillations of the brain activity are recognized at various frequency ranges (δ wave: 1 to 4 Hz, α: 8 to 12 Hz, β: 12 to 30 Hz and so on). Oscillations at much slower ranges (about 0.1Hz or less) are collectively known as infra-slow oscillations, whose origin and nature have not been well characterized. We previously showed that the slow electrical potential change at that slow frequency range in the anesthetized rat cortex reflected the cerebral vasomotion, a rhythmical oscillation of the cerebral arteries in their vessel wall diameter. To understand the mechanism underlying the rhythmical movement of the arteries, we have screened biologically active molecules which interfere the vasomotion. Here, we show that focall application of a biologically active amine, histamine and a short peptide, calcitonin gene-related peptide (CGRP) over the cerebral arteries disrupted the rhythmical oscillation of the arteries, suggesting that they may cooperatively control the cerebral vasomotion. Consistent with the observation that both molecules could act on the cerebral arteries, trigeminal nerves which are known to release CGRP, and distinct nerve fibers which expressed histidine decarboxylase, the major histamine producing enzyme, innervated the large cerebral arteries and they physically closely ran over the arterial smooth muscles. We also observed that tuberomammillary nucleus (TMN) in the hypothalamus, which is the sole source of the histamine in the central nervous system, showed a waxing and waning nerve activity synchronous to the infra-slow cerebral vasomotion. These findings suggest that a periodic release of histamine and/or CGRP from the artery-targeting nerve fibers triggers the constriction/dilation cycle of the cerebral arteries. We also found that chloride channel blockers (Fenamates, Etomidate, Pentobarbital and Picrotoxin), applied directly over the blood vessels, could also abolish the vasomotion. The finding suggests involvement of the chloride channels in generating the constriction/relaxation force of the blood vessel's smooth muscles during cerebral vasomotion.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-253 SIK1およびSIK2のPKAリン酸化部位変異マウスの睡眠覚醒行動 Minjeong Park（Park Minjeong）¹，Chika Miyoshi（三好千香）¹，Tomoyuki Fujiyama（藤山知之）¹，Miyo Kakizaki（柿﨑美代）¹，Aya Ikkyu（一久綾）¹，Jinhwan Choi（Choi Jinhwan）¹，Seiya Mizuno（水野聖哉）²，Satoru Takahashi（高橋智）²，Hiromasa Funato（船戸弘正）^1,3，Masashi Yanagisawa（柳沢正史）^1,4 ¹筑波大院　人間総合科学 ²筑波大院　人間総合科学 ³東邦大医 ⁴Univ. of Texas Southwestern Medical Center, Dallas, Texas, USA Through forward genetics research in which we screened randomly mutagenized mice for sleep abnormalities, we established a mutant pedigree which we termed Sleepy. Sleepy mutant pedigree shows short total wake time and increased sleep need with a dominant inheritance pattern. Moreover, we identified a splice mutation of Sik3 gene in sleepy mutant mice, resulting in the skipping of exon 13. SIK3 is a protein kinase which contains a well-conserved protein kinase A (PKA)-phosphorylation site, serine 551. The skipping of exon 13 results in a deletion of 52 amino acids including S551. Like sleepy mutant mice, Sik3 S551A knock-in mice also showed reduced total wake time. These results suggest that the phosphorylation of SIK3 S551 by PKA plays a crucial role in sleep behavior. In vertebrates, there are three SIK proteins, SIK1, SIK2, and SIK3. Importantly, SIK3 S551-equivalent serine residue is conserved at S577 in SIK1 and at S587 in SIK2. Hence, we hypothesize that SIK1 S577 and SIK2 S587 are involved in sleep/wake regulation, similar to SIK3 S551. To examine the role of the PKA phosphorylation site in SIK1 and SIK2 in sleep/wake behavior, we generated mutant mice in which SIK1 S577 and SIK2 S587 were substituted by alanine using CRISPR/Cas9 method. These mutant mice were subjected to EEG/EMG-based sleep analysis. Both Sik1 S577A mice and Sik2 S587A mice showed increased NREM sleep delta power. Furthermore, Sik1 S577A mice showed short time spent in wakefulness. These findings indicate the PKA recognition site of the SIK family commonly regulate sleep need.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-254 延髄によるレム睡眠中の血圧変動の調節 Yoshimasa Koyama（小山純正），Hayato Iwata（岩田逸人），Tatsusya Suzuki（鈴木達也），Kaname Mochizuki（望月要）福島大学共生システム理工学類 The medulla has a crucial role in the regulation of REM sleep and also contains a center of cardiovascular control, which regulate heart rate or blood pressure. During REM sleep, in addition to desynchronization of cortical activity or muscle atonia, large fluctuations of autonomic nervous systems occur frequently. Blood pressure fluctuation is one of the characteristic phenomenon of such autonomic signs. During REM sleep, blood pressure exhibits abrupt and large fluctuations, larger than those observed during waking, which is considered to reflect emotional changes during REM sleep. We have shown that the blood pressure fluctuation during REM sleep is under the control of REM sleep center in the mesopontine tegmental area and the amygdala. To clarify the role of the medullar neurons in the regulation of blood pressure during REM sleep, single neuronal activity in the medulla was recorded from unanesthetized, head-restrained rats to analyze the relation with blood pressure changes during sleep waking cycles. About half of the neurons recorded from the ventral medulla exhibited firing in close relation with blood pressure fluctuation during REM sleep. The onset of firing increase of these neurons preceded one to 3 seconds to the start of blood pressure increase, while the time preceding to the start of blood pressure increase during waking was less than one second. Some neurons discharged in close relation with blood pressure fluctuation during REM sleep but not with that during waking. The results suggest that the blood pressure fluctuation during REM sleep is regulated by the different mechanism from those regulating blood pressure changes during waking.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-255 Electroencephalographic characterization of the Sleepy mutant reveals a multi-spectral disorganization of NREM sleep dynamics accompanied by absence of active wakefulness markers Javier Diaz（Diaz Javier）¹，Hiromasa Funato（Funato Hiromasa）¹，Juan C Letelier（Letelier Juan C）²，Masashi Yanagisawa（Yanagisawa Masashi）¹，Kaspar E Vogt（Vogt Kaspar E）¹ ¹WPI-IIIS, University of Tsukuba ²University of Chile, Santiago, Chile. We recently reported mutant mice with significant sleep/wake abnormalities, obtained through a large scale EEG/EMG screening of a randomly mutagenized cohort (1). One of these mutants, named Sleepy, bearing a splicing mutation on the sik3 gene (with exon 13 skipping), exhibits increased daily amounts of NREM sleep with a constitutively elevated sleep need and EEG delta power during non-REM sleep. The molecular pathway for the regulation of sleep in which sik3 takes part is yet to be determined. To get a better insight into the Sleepy phenotype, we performed an extended EEG analysis whose interpretation relies on two main ideas. First, new studies suggest that EEG Gaussianity assessment (e.g. based on envelope analysis) reveal important EEG morphological aspects, invisible for classic spectral analysis (2). Second, new evidence supports the notion that in mice NREM sleep cycles through stages of graded sleep depth (3). Our main results show significant differences in phasic activity between Sleepy and wt mice during NREM sleep both at the sub-second and the infra-slow temporal levels, encompassing theta and sigma bands. In addition, we report an absence of theta rhythm during wakefulness in Sleepy, while this rhythm remains intact during REM sleep. Altogether these results reveal a disorganization in NREM sleep temporal dynamics in Sleepy, encompassing oscillations controlling sleep depth, while also lacking a marker of active wake (i.e. theta rhythm). As in several human sleep disorders, we can conceptualize the Sleepy phenotype in terms of a deteriorated sleep quality: somnolence-like wakefulness and increased NREM amount can be explained by NREM sleep that fails to reach stable deep-sleep. (1) doi:10.1038/nature20142. (2) doi:10.1016/j.neuroimage.2018.01.063. (3) doi:10.1126/sciadv.1602026

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-256 自由行動下のマーモセットにおける睡眠時脳波の測定 Iori Murai（村井伊織）¹，Teruaki Fujinaga（藤永輝明）¹，Maho Sugawa（須川真帆）¹，Yuka Kono（河野優華）¹，Teruo Kaneko（金子照生）¹，Keiko Tominaga（冨永恵子）²，Hirotaka Onoe（尾上浩隆）³，Kaoru Isa（伊佐かおる）³，Tadashi Isa（伊佐正）³，Hitoshi Okamura（岡村均）¹ ¹京都大院薬分子脳科学 ²大阪大院生命機能神経可塑性生理 ³京都大院医神経生物 Modern working style permits humans to adopt long working hours and shift works, which increases the danger to disrupt internal circadian rhythms. The rhythm disruption is closely linked to social stress leading to sleep disorders and life-style related diseases. However, this pathophysiology is difficult to reproduce in rodents, so the establishment of non-human primate model is needed for the detailed study. We recently have established an experimental system for examining the biological rhythms in a diurnal primate, common marmoset (Callithrix jacchus). In this presentation, we report the development of electroencephalogram (EEG) monitoring system in freely moving marmosets by using wireless communication. Three adult marmosets of both sexes (2-4 years old) were housed in a light-tight circadian behavior room (27-29 °C) with LED lighting. Animals were housed in light-dark or continuous light conditions. Illuminance was confined to 20-60 lux which enables animals to drink, eat and communicate freely. Animal activity was recorded by the attached moving sensors. The EEG electrodes were attached to the skull at the position +2 mm in front/+2 mm in the right side and -5 mm in front/-2 mm in the right side of the bregma. Electrodes for electrooculography (EOG) were attached to the upper and side part of the left orbit. Data was wirelessly transmitted to the receiver with a bandpass filter. The amplifier was mounted on the back of the marmoset. The activity levels of marmoset in free moving conditions were high in the light period and low in the dark period, which was synchronized with the environmental light/dark cycle. Similarly, we confirmed the clear diurnal variation of EEG, which α and β waves are dominant in the light period and θ and δ waves are dominant in the dark period. Even in the continuous light conditions, we could detect a clear change of circadian EEG rhythm maintaining REM and NREM sleep similar to light-dark cycle. Judged by the EEG and EOG, we also found the REM sleep periodically occurred in every 60-100 min during sleep in our illumination conditions in freely moving marmosets. The detection of awake, REM, and NREM in EEG in freely moving marmosets will contribute to clarifying the pathophysiology of the primate model of sleep-related diseases.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-257 Drosophila insulin-like peptide (dilp) 3による概日リズム制御 Sho Yamaguchi（山口翔），Jun Tomita（冨田淳），Kazuhiko Kume（粂和彦）名古屋市立大学大学院薬学研究科神経薬理学分野 Circadian rhythm relates to many physiologies such as sleep-wake cycle and metabolism. Clock mutant mice has larger consumption of food as well as longer circadian period. On the other hand, metabolism affect circadian rhythm. Leptin KO or High fat diet change the expression of circadian clock gene. Insulin, the secretion of which is regulated by circadian clock, adjust circadian clock in liver. Although, insulin receptor expressed in hypothalamus or hippocampus is known to regulate metabolism or cognitive function, the function of insulin in the brain is poorly understood. Drosophila melanogaster has 7 mammalian insulin homologs, Drosophila insulin-like peptides (dilps). Dilps regulate metabolism, development and activity. dilp2, dilp3 and dilp5 , expressed in pars intercerebralis (PI) of the brain, relate to fragmentation of sleep with age. In Drosophila as well as mammal, insulin signaling relates to memory function. dilp3 expressed in PI relates to age-related memory impairment. Recently, RNA-seq analysis has shown that dilp3 expresse in the lateral ventral neurons (LNvs). LNvs are the master clock neurons of Drosophila melanogaster. Additionally, Akt and target of rapamycin (tor), which are activated by insulin signaling, regulate circadian rhythm in Drosophila melanogaster. So, we hypothesized that dilp3 expressed in LNvs regulates circadian rhythm. Here, we found that knockout of dilp3 lengthened the circadian period. Using with GAL4-UAS system, we found that dilp3 knockdown in LNvs also lengthened circadian period. On the other hand, expression of dominant negative insulin receptor (InR) with timeless-GAL4 shortened circadian period. Interestingly, knockdown of secreted decoy of insulin receptor (SDR) in glial cells lengthened circadian period. Our results suggest that dilp3 expressed in LNvs regulates circadian rhythm through SDR and InR.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-258 視床下部MCH神経のレム睡眠時活動による海馬依存記憶の忘却 Shuntaro Izawa（伊澤俊太郎）^1,2，Ryo Inoue（井上崚）¹，Chowdhury Srikanta（Srikanta Chowdhury）¹，Toh Miyazaki（宮﨑杜夫）^1,2，Yasutaka Mukai（向井康敬）^1,2，Daisuke Ono（小野大輔）¹，Akihiro Yamanaka（山中章弘）^1,3 ¹名古屋大環境医神経系２ ²日本学術振興会特別研究員 ³CREST, JST Memory consolidation or erasure is occurred during sleep. However, the neural mechanism behind it has not been revealed. Injection of retrogradely tracing beads into the hippocampus, we found that melanin-concentrating hormone producing neurons (MCH neurons) were most population projecting from the hypothalamus to the hippocampus. MCH neurons are implicated in the regulation of rapid eye movement (REM) sleep. We confirmed that MCH neurons were active during REM sleep using fiber photometry. To reveal the role of MCH neurons in hippocampus-dependent memory, novel object recognition, Morris water maze and contextual fear memory tests were performed. Pharmacogenetical activation of MCH neurons impaired all these memory, whereas the inhibition or ablation of these neurons improved. These results suggested that MCH neurons activity impaired hippocampus dependent memory. We further performed optogenetic stimulation of MCH neurons and found that activation of MCH neurons during retention period impaired memory, but not in encoding or retrieval period. To reveal mechanism of memory impairment, slice patch clamp recordings from hippocampal pyramidal neurons was performed. Light stimulation of MCH nerve terminals in the hippocampus hyperpolarized and decreased firing frequency in hippocampal pyramidal neurons. We confirmed that hippocampus is targeted area of memory impairment by MCH neurons since optogenetic stimulation of MCH nerve terminals in the hippocampus impaired memory. Finally we tested whether activity of MCH neurons during REM sleep is involved in memory impairment. MCH neurons were sleep-wake stages dependently silenced (wakefulness, REM sleep, and Non-REM sleep) by closed loop system during memory retention period for 14 hr. The silencing during REM sleep significantly improved memory but not wakefulness or Non-REM sleep. These results suggested that MCH neurons impaired memory during REM sleep. Taken together, here we found that MCH neurons impaired memory by inhibiting hippocampal pyramidal neurons during REM sleep. This could be involved in effectively saving memory resources by erasing memories that are not important for survival.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-259 中枢性Erk5によるエネルギー代謝調節 Tetsuhiro Horie（堀江哲寛）¹，Gyujin Park（朴奎珍）¹，Yuka Inaba（稲葉有香）²，Takashi Iezaki（家崎高志）¹，Katsuyuki Kaneda（金田勝幸）¹，Hiroshi Inoue（井上啓）²，Eiichi Hinoi（檜井栄一）¹ ¹金沢大院薬薬理 ²金沢大新学術栄養代謝 Body weight homeostasis is maintained by balancing energy intake and energy expenditure. It is well-established that hypothalamic neurons in central nervous system (CNS) play a critical role in maintaining body weight and systemic energy homeostasis via sensing a variety of peripheral messengers including circulating hormones such as leptin and insulin. The extracellular signal-regulated kinase 5 (Erk5) is a member of the mitogen-activated protein kinase (MAPK) family and Erk5 is specifically phosphorylated and activated by MAPK/Erk kinase-5 (Mek5). Although Erk5 has been implicated in odor discrimination and long-term memory through its expression in CNS, little is known regarding the physiological importance of neuronal Erk5 in body weight and energy homeostasis so far. In this study, to reveal the in vivo role of Erk5 in hypothalamic neurons using a cell-specific knockout strategy, we used leptin receptor (LepR)-Cre mice, in which Cre recombinase is expressed specifically in LepR neurons. Erk5 deficiency in LepR-expressing neurons led to an obesity phenotype with an increase in white adipose tissue mass due to an increase in adipocyte size in female on normal chow diet, concomitant with an effective genomic deletion of Erk5 in the arcuate and ventromedial nuclei of the hypothalamus. Furthermore, Erk5 deficiency in LepR-expressing neurons showed impaired glucose tolerance and insulin sensitivity along with decreases in physical activity, food intake and energy expenditure. These results suggest that the Mek5/Erk5 axis controls body weight and systemic energy homeostasis possibly through its expression in hypothalamic neurons, thereby providing a novel target for metabolic diseases such as obesity and type 2 diabetes.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-260 ラット脳における[2,4-¹³C₂]β-ヒドロキシ酪酸と[1-¹³C]グルコース代謝の発生による変化 Koichi Sakogawa（迫川弘一），Tomoyuki Kanamatsu（金松知幸）創価大院工学研究科環境共生工 It is well known that glucose is the primary energy substrate for the adult brain. However, under the conditions of starvation, diabetes, and hypoglycemia, lactate and ketone bodies such as β-hydroxybutyrate (BHB) and acetoacetate support cerebral metabolism in the adult brain. It is also well known that monocarboxylic acid is cerebral fuel for the newborn and immature brain under normal physiological conditions. The purpose of this study was to investigate the developmental changes in metabolic pattern of BHB in the rat brain. The pups of Wistar rat in day 1, 4, 8, 12, 16, 26 (for the day of birth was day 0) were given a bolus intraperitoneal injection of [2,4-¹³C₂]-BHB or [1-¹³C]-glucose (0.5 g/kg-body weight, both 99% ¹³C enriched). After 30 min, the rats were decapitated and the blood was collected and the forebrain was dissected, weighed and frozen. The amino acid fraction extracted from each frozen tissue was employed for proton decoupled ¹³C-NMR spectroscopic measurement. Total ¹³C-signal intensity in Glu, Gln, Asp and GABA (total signal intensity of ¹³C) derived from [2,4-¹³C₂]-BHB was increased according to the development (from 1-day-old to 16-day-old, and then decreased at 26-day-old. However, total signal intensity of ¹³C derived from [1-¹³C]-glucose was weak from 1-day-old to 16-day-old, and then sharply increased at 26-day-old. These data may indicate that BHB is preferred substrate for energy source over glucose in the brain during early postnatal period and that metabolic switch between utilization of BHB and glucose exists at 16-day-old in the rat brain.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-261 扁桃体中心核におけるソマトスタチン陽性GABA作動性神経細胞の多様性 Shota Morikawa（森川勝太）¹，Kazuki Katori（香取和生）¹，Takuya Sasaki（佐々木拓哉）¹，Hiroyuki Hioki（日置寛之）²，Hideki Tamura（田村英紀）³，Yuji Ikegaya（池谷裕二）^1,4 ¹東京大院薬薬品作用 ²順天大院　医神経生物・形態 ³星薬大　先端生命科学研究センター ⁴脳情報通信融合研究センター The central nucleus amygdala (CeA) integrates adaptive behaviors, including defensive and appetitive responses. The CeA, a nucleus predominantly composed of somatostain (SOM) -expressing and protein kinase C delta (PKCδ) -expressing GABAergic neurons, is essential for Pavlovian fear conditioning and defensive behaviors. Cell type specific inactivation of SOM or PKCδ-expressing neurons results in impairment of specific behaviors. This indicate that genetically identified subpopulation of neurons in the CeA plays an important role in specific function. However, recent studies suggest this concept may not be sufficient to explain the complexity of neural circuit mechanism of the CeA. Here we show that SOM-expressing neuron in the CeA has different cell properties using histological analysis, viral tracing approach and electrophysiological analysis. To find new cell-type specific marker for SOM-expressing neuron, we performed double in situ hybridization in the mouse brain. Non-overlapping subpopulation of SOM-expressing neurons expressed VIP receptor 2 (Vipr2) or Dopamine receptor 3 (Drd3). Retrograde tracer experiments indicated that these neurons mainly project to the parabrachial nucleus (PBN). Furthermore, dopaminergic neurons co-expressing VIP in the ventral PAG innervated the CeA. These results suggest a new neural circuit mechanism of the CeA. Next, to reveal the electrophysiological property of Vipr2 and Drd3-expressing neurons and physiological function of VIP and Dopamine in the CeA, we conducted ex vivo patch clamp recording and Ca2+ imaging. Bath application of selective Vipr2 agonist showed a tendency to change the excitatory-inhibitory balance of PBN-projecting CeA neurons, and increased spontaneous activity of SOM-expressing neurons in the CeA. Finally, to observe neuronal activity of CeA neurons, we conducted unit recording during auditory fear conditioning and extinction. In this research, we propose that SOM-expressing neurons in the CeA has anatomical and functional heterogeneity.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-262 一夫一婦制のプレーリーハタネズミにおけるつがい形成依存的な恐怖学習減弱にはオキシトシンが介在する Yu Hirota（廣田湧），Shinichi Mitsui（三井真一）群馬大院保健 The presence of a conspecific mitigates stress and fear responses, and this phenomenon is called ""social buffering"". We reported in Neuroscience2018 at Kobe that in a monogamy rodent, prairie vole, male bonded with a female (partner) showed attenuated fear responses in fear-based learning tests than male cohabitated with a male conspecific, although social buffering effect requires the presence of a conspecific during stress presentation or recovery. In our case, the partner female is absent during conditioning or recovery. In other words, pair bonding with a female attenuates fear responses in male prairie voles despite the absence of a conspecific during or after conditioning. To find underlying mechanisms of the attenuation effect, we focused on roles of oxytocin which participates in pair bonding. We investigate the effects of an oxytocin antagonist (OXTA) on the pair bonding-dependent attenuation of fear learning and memory in male prairie voles. After a male prairie vole was cohabited with a naive female for 4 days, partner preference was assessed. Next day, a guide cannula was implanted aimed at the right cerebral ventricle. Afterwards, subjects were individually and undisturbedly housed for 5 days. Subjects were administrated 0.5 μg OXTA or vehicle, 30 minutes before they received foot shocks on a passive avoidance test apparatus. Latency to enter the dark chamber was measured at conditioning and memory test which performed 24 hours after the conditioning. Brains were prepared for cFos immunostaining 90 min after the memory test. The OXTA group, but not the vehicle group, showed significantly more prolonged latency at the memory test than conditioning. In addition, the latency at the memory test was significantly increased in the OXTA group in comparison with the vehicle group. These results suggest that oxytocin mediates the attenuation effect caused by pair bonding. Previously, we and other groups indicate that pair bonding increases oxytocin expression at the paraventricular nucleus of the hypothalamus. Hence, we consider that oxytocin mediates pair bonding-dependent attenuation of fear learning and memory.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-263 サル背側縫線核における情動と意思決定の協調表現 Masaharu Yasuda（安田正治），Yasumasa Ueda（上田康雅），Kae Nakamura（中村加枝）関西医生理 Our behaviors are influenced by emotional environments. For example, decision process may be achieved by different brain mechanisms under imminent threat and relaxed condition. Such use of environmental information to a range of adaptive choices is critical for survival in an ever-changing world. Recent studies have shown that the manipulation of serotonin, a critical neuromodulator of emotion and cognition, exhibits distinct behavioral effects depending on the emotional state. However, how serotonin contributes to adaptive behavior under distinct emotion is still unknown. To this end, we recorded single neurons' activity in dorsal raphe nucleus (DRN), a major source of serotonin, while monkeys performed a reversal choice task under various emotional contexts. In the task, after fixation on the central fixation point (FP, 1000-1500ms), two visual stimuli were presented in the left and right of the FP. The monkey chose one of them by making a saccade, followed by the receipt of a reward for one (correct), but no reward for the other one (wrong). During a block of trials, the reward-associated stimulus was identical, but it switched to the other one when the proportion of the correct choice reached the criterion. Before the choice task, we let two monkeys learn three emotional CS which were associated with reward (appetitive CS), tone (neutral CS), or air puff (aversive CS) in Pavlovian conditioning. During the choice task, one of the three pre-learned CSs was presented during the inter-trial-interval of consecutive three blocks. Thus, the animal performed the choice task under distinct emotional contexts. Indeed, pupil diameter was larger (monkey A and T) and heart rate was higher (monkey A) when aversive CS was presented, suggesting successful manipulation of the monkeys' emotion by CS presentation. Among 201 task-modulated DRN neurons, 133 (67/112 for monkey A, 66/89 for monkey T) changed those activities depending on the following choice before target presentation. Majority of them showed stronger activity in correct than in wrong choice trials (52 for monkey A and 52 for monkey T). Majority of such choice-predictive neurons showed stronger activities for appetitive (18) or aversive (22), rather than for neutral (7) CS condition, suggesting dual and cooperative representation of emotional arousal and optimal decision making in single DRN neurons. Such signal processing may enhance the animal's appropriate behavior under various emotions.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-264 前部帯状皮質は観察恐怖表出を制御する Kensaku Nomoto（野元謙作）¹，Madoka Nakamura（中村月香）¹，Hanako Murayama（村山花子）¹，Kazanari Nagai（永井一成）¹，Mayuko Miyakoda（都田真由子）¹，Kazutaka Mogi（茂木一孝）¹，Tsuyoshi Koide（小出剛）²，Takefumi Kikusui（菊水健史）¹ ¹麻布大獣医伴侶動物 ²国立遺伝研マウス開発研究室 Observational fear is the ability to exhibit fear-related behavior such as freezing and escaping when observing fear of a conspecific. Although previous studies have shown that the anterior cingulate cortex (ACC) plays an important role in observational fear, the underlying neural mechanisms are not well understood. To address this issue, we performed behavioral, neurophysiological, and chemogenetic experiments while animals performed the observational fear paradigm, in which one mouse (termed as an observer) observed fear-related behavior exhibited by a conspecific receiving footshock (termed as a demonstrator). First, we determined which sensory cue was important for observational fear by manipulating auditory, olfactory, and visual cues independently. We used wild-derived MSM mice as the observer, which exhibit greater fear-related responses than laboratory mice. Freezing behavior was facilitated by the presentation of stressed urine. The pixelated video clip was not as effective in evoking freezing behavior as the original video clip. The playback of the sound recorded when a mouse had received footshock did not affect freezing behavior. These results suggest that visual and olfactory cues are important for observational fear. Next, we recorded the neural activity of the ACC neurons during observational fear by using fiber photometry. We used a hybrid strain of MSM and C57BL/6 mice (MSMB6F1) as the observer, which exhibit greater freezing behavior than C57BL/6. We found that the ACC neurons of the observer were activated when the demonstrator received footshock. We did not see significant responses associated with freezing behavior of the observer. This suggests that ACC activities are involved in the perception of fear of a conspecific, but not in the execution of freezing behavior. Finally, we examined which pathway from the ACC is important for the expression of observational fear. We expressed DREADD receptors in the ACC neurons projecting to the periaqueductal gray (PAG). Our preliminary results showed that chemogenetic inhibition of the ACC-PAG pathway decreased freezing behavior of the observer, suggesting that this pathway plays an essential role in the expression of observational fear. In conclusion, we demonstrate that visual and olfactory cues have more impact on observational fear than auditory cues, and that the ACC is involved in the processing of sensory cues during observational fear, possibly through its projection to the PAG.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-265 慢性痛による拡張扁桃体神経回路の可塑的変化は不安を惹起する Naoki Yamauchi（山内直紀），Hiroshi Nomura（野村洋），Taiju Amano（天野大樹），Masabumi Minami（南雅文）北海道大院薬薬理 Chronic pain and psychiatric disorders often co-occur, suggesting common neuronal mechanisms that underlie these pathological states, yet the mechanisms are still unclear. The bed nucleus of the stria terminalis (BNST) is a limbic structure essential for regulating a variety of emotional states, such as anxiety, aversion, and reward seeking. This brain region is also involved in affective dimension of pain. Here, using spared nerve injury model mice of chronic pain, we show the neuroplastic changes in the synaptic transmission in I_h-current negative BNST neurons projecting to the lateral hypothalamus (LH) which can lead to maladaptive anxiety. First, consistent with previous reports, we found that chronic pain increased anxiety-like behaviors in elevated plus maze and light-dark box tests. Second, whole-cell patch-clamp recordings revealed that chronic pain increased spontaneous IPSCs and decreased spontaneous EPSCs onto I_h-current negative LH-projecting BNST neurons. This indicates that the E/I balance in these neurons were shifted toward inhibition. Interestingly, similar plasticity was detected in both contralateral and ipsilateral hemispheres to nerve injury. To test the causal role of this plasticity in behavioral anxiety, we bilaterally inhibited the LH-projecting BNST neurons using designer receptor exclusively activated by designer drugs (DREADDs). Inhibition of LH-projecting BNST neurons in naive mice induced behavioral anxiety without affecting their sensory components of pain. Together, these findings suggest that the maladaptive anxiety might be due to sustained suppression of the BNST-LH circuit that was induced by chronic pain.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-266 パブロフ型確率条件づけにおいて条件刺激に誘発された正または負の情動を反映する行動・生理指標 Shinya Nakamura（中村晋也），Toshiki Moritani（森谷叡生），Yasutaka Honda（本田保貴），Ken-Ichiro Tsutsui（筒井健一郎）東北大院生命科学脳神経システム In this study, we aimed to examine how positive and negative emotions are reflected in behavioral and autonomic responses under a well-controlled experimental condition in monkeys. We trained two Japanese monkeys in a probabilistic Pavlovian conditioning task with two distinct contexts. In each context, each abstract figure was associated with a specific reward/punishment probability (reward/punishment probability = 1/0, 0.5/0, 0/0, 0/0.5, 0/1 for one context; 1/0, 0.75/0.25, 0.5/0.5, 0.25/0.75, 0/1, 0/0 for the other) as a conditioned stimulus (CS). In both contexts, the monkeys held the touch key following the illumination of a red fixation spot to start a trial. One of the conditioned stimuli then appeared, and a delay period followed. After the delay period, the fixation spot turned from red to green, indicating the monkeys to release the key. A successful key release was followed by a reward (juice) or punishment (saline) delivery. Key release reaction time, spontaneous licking actions, and heart rate were recorded and analyzed as behavioral or autonomic measures. We found that these measures changed dependent on the reward/punishment probability in both contexts. The reaction time of the key release became shorter with higher reward probability and longer with higher punishment probability. The anticipatory licking actions during the CS, delay, and pre-release periods also increased with higher reward probability and decreased with higher punishment probability. Interestingly, the heart rate increased during the later delay period in trials in which the CS associated with higher punishment probability was presented, however such change was not observed when the CS associated with higher reward probability was presented. Thus, we found differential behavioral and autonomic measures reflecting positive and negative emotions elicited by conditioned stimuli in Pavlovian conditioning.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-267 δオピオイド受容体KNT-127はSNC80と異なり恐怖記憶形成後の消去学習を促進する Shoko Yanagisawa（栁澤祥子）¹，Daisuke Yamada（山田大輔）¹，Shinya Yanagita（柳田信也）²，Jun-Ichiro Oka（岡淳一郎）¹，Hiroshi Nagase（長瀬博）³，Akiyoshi Saitoh（斎藤顕宜）¹ ¹東京理科大院薬・薬理 ²東京理科大理工 ³筑波大学睡眠研究機構 Fear memory is an emotional function indispensable for living things to survive avoiding danger. δ-opioid receptors are present in many brain regions involved in emotion. We previously reported that selective agonists for δ-opioid receptors, KNT-127 and SNC80, produced potent anxiolytic-like effects in rodents. In this study, we evaluate whether KNT-127 and SNC80 influence contextual fear memory in the fear conditioning test in mice. Male C57BL/6J mice (8-9 weeks old) were used for behavioral experiments. On the conditioning day, the mice were trained with eight conditioning trials. Each trial consisted of a 1 s, 0.8 mA footshock (inter-trial interval 30 s). Twenty-four h following conditioning, the mice were injected subcutaneous administration (s.c) with drugs. After 30 min, the mice were reexposed to the same apparatus for 6 min (extinction training session). Twenty-four hours after extinction, each mice was placed back in the conditioning chamber and a 6-min test was performed (memory testing session). During each session, no footshock was given, and the mouse was observed every 1 min by a trained observer to assess its freezing behavior via a monitor connected to a video camera system. In extinction training session, KNT-127 and SNC80 significantly and dose-dependently reduced contextual conditioned freezing time in mice when compared to saline, respectively. In memory testing session, KNT-127 significantly reduced contextual conditioned freezing time when compared to saline, but SNC80 treated freezing time in mice were not significantly different among saline. These results suggested that KNT-127 produced the anxiolytic-like effects in extinction training session, and facilitated the fear extinction learning in memory testing session. On the other hand, it was suggested that SNC80 produced the anxiolytic-like effects in extinction training session, but produced no facilitation effects on fear extinction leaning in memory testing session. We proposed that although δ-opioid receptor agonists KNT-127 and SNC80 produced robust anxiolytic-like effects on contextual conditioned fear, these drugs have different mechanism on the extinction leaning in fear memory. We are currently studying molecular mechanisms on the extinction leaning after δ-opioid receptor agonist treatment.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-268 MAPKによるNpas4リン酸化は報酬関連遺伝子発現および行動を制御する Yasuhiro Funahashi（船橋靖広）¹，Anthony Ariza（アリザアンソニー）¹，Ryosuke Emi（恵美亮佑）¹，Yifan Xu（許伊凡）¹，Shan Wei（単偉）²，Ko Suzuki（鈴木航）¹，Sachi Kozawa（小澤祥）¹，Tesuya Takano（高野哲也）¹，Yoshimitsu Yura（由良義光）¹，Keisuke Kuroda（黒田啓介）¹，Taku Nagai（永井拓）²，Mutsuki Amano（天野睦紀）¹，Kiyofumi Yamada（山田清文）²，Kozo Kaibuchi（貝淵弘三）¹ ¹名古屋大院・医・神経情報薬理 ²名古屋大院・医・医療薬学・付属病院薬剤部 Dopamine is important for motor function, motivation, working memory and the reward system. Dopamine activates Mitogen-Activated Protein Kinase (MAPK) via Protein kinase A (PKA)/Rap1 in medium spiny neurons (MSNs) expressing the dopamine D1 receptor (D1R) in the nucleus accumbens (NAc), thereby regulating neuronal excitability and reward-related behavior. However, how MAPK regulates reward-related learning and memory through gene expression is poorly understood. Here, to identify the relevant transcriptional factors, we performed proteomic analysis using affinity beads coated with cAMP response element binding protein (CREB)-binding protein (CBP), a transcriptional coactivator involved in reward-related behavior. We identified more than 100 CBP-interacting proteins, including Neuronal Per-Arnt-Sim domain protein 4 (Npas4), which is a brain-specific basic helix-loop-helix transcription factor and regulates gene expression involved in synaptic plasticity. We found that Npas4 was phosphorylated by MAPK and the phosphorylation of Npas4 increased the interaction of Npas4 with CBP. The phosphorylation of Npas4 enhanced the exon I and IV-BDNF promoter activity. Furthermore, deletion of Npas4 in the D1R-expressing MSNs impaired cocaine-induced place preference, which was rescued by Npas4-WT but not by a phospho-deficient Npas4 mutant. These observations suggest that MAPK phosphorylates Npas4 in D1R-MSNs and increases its transcriptional activity to enhance reward-related learning and memory.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-269 幼少期ストレスが青年期の行動や海馬機能に及ぼす影響 Eri Segi-Nishida（瀬木（西田）恵里），Sara Nanno（南野沙良），Taiga Akuhara（悪原大雅），Akari Nishiyama（西山明香里）東京理科大生物工 Early life stress influences neuronal development and significantly increases the lifetime risk for the development of cognitive and affective pathology. It has been suggested that disruption in early life care impaired cognitive abilities in adulthood and truncated neuronal development through effects on diminishing cell proliferation and increasing cell death in the hippocampus in rodent models. However, the influences of early life stress in emotional behaviors and hippocampal development in adolescence remain understood. In this study, we examined the effects of early life stress by housing mouse dam with limited nesting and bedding material from postnatal days 2-9 and examined behaviors in 30 days of offspring; 1) open field test, 2) forced swim test, 3) novel object recognition test. We also investigated hippocampal differentiation including 1) cell proliferation, 2) cell survival, 3) expression of several differentiation markers. Adolescent mice stressed in early childhood showed an attenuation of anxiety-like behavior in open field test, an over-response of stress escape in forced swim test, and decreased recognition to a previously identified object. By immunohistochemical analysis, dentate gyrus of the hippocampus stressed in early childhood showed a significant increase in cell proliferation and survival, and dendritic elongation in the immature neurons. Collectively, our findings demonstrate that chronic early life stress induced by limited bedding greatly affects adolescent emotional behaviors and hippocampal development. It is necessary to clarify why anxiolytic-like behavior and hyperactivity by stress environment occur in adolescent mice stressed in early childhood and to investigate the relationship between these behaviors and hippocampal development in order to clarify the stress response in early childhood.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-270 運動効果に基づく新たなうつ病治療メカニズムの解明 Makoto Kondo（近藤誠），Yoshihisa Koyama（小山佳久），Yukiko Nakamura（中村雪子），Noriyoshi Usui（臼井紀好），Shoichi Shimada（島田昌一）大阪大院医神経細胞生物学 Major depression is a highly prevalent mental disorder affecting many people worldwide. Although selective serotonin reuptake inhibitors (SSRIs) are the most widely used antidepressants, a significant proportion of depressed patients do not achieve remission after initial treatment. It has been known that physical exercise provides neurogenic and antidepressant effects, and we recently demonstrated that the serotonin type 3 (5HT3) receptor is essential for exercise-induced hippocampal neurogenesis and antidepressant effects. In this study, we examined the 5HT3 receptor-mediated mechanism underlying hippocampal neurogenesis and antidepressant effects, and tried to establish novel prevention and treatment for depression, which is based on mechanisms of exercise-induced beneficial effects. Here, we showed that treatment with a 5HT3 receptor agonist induces antidepressant effects and increases hippocampal neurogenesis, in a fluoxetine-independent manner. In addition, histological analyses revealed that the 5HT3 receptor and insulin-like growth factor 1 (IGF1) are expressed in the same neurons in the hippocampal dentate gyrus. Furthermore, in vivo microdialysis and drug microinjection analyses showed that 5HT3 receptor agonist treatment increases extracellular IGF1 levels in the hippocampus, and that IGF1 signaling is required for the 5HT3 receptor-dependent hippocampal neurogenesis. Our findings suggest a novel mode of antidepressant action, distinct from fluoxetine-induced responses.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-271 神門への鍼治療は恐怖文脈条件付けしたマウスのすくみ反応を減弱させる Jungwon Hwang（黄晶媛），Masato Koike（小池正人），Hiroyuki Hioki（日置寛之）順天大院　医神経生物・形態 Acupuncture treatment on Shenmen (HT7) has been used for the treatment of mental disorders such as anxiety disorders and depression in oriental medicine. In the present study, to elucidate the influence of HT7 acupuncture on relaxation of fear memory, we examined freezing responses in contextual fear-conditioned (FC) mice with or without the acupuncture treatment on HT7. Thirty-six adult male C57BL/6J mice (8-12 weeks old) were moved from the context A (home cage) to the context B (foot-shock cage), and subjected to an electric foot-shock (0.5 mA, 2 seconds). The mice were immediately returned to the context A, and randomly divided into 2 groups, FC with HT7 acupuncture treatment (FC-HT7) group and FC with non-treatment (FC-none) group. In FC-HT7 group, 1 minute of the acupuncture treatment on HT7 was performed for 7 consecutive days, whereas sham-treatment was done in FC-none group for 7 days. We then placed the mice of 2 groups in the context A or B, and observed the behavior for 2 minutes. The mice of both groups in the context A showed almost no freezing response: FC-none and FC-HT7 groups displayed freezing responses for 1.5 and 1.3 seconds, respectively. The mice of FC-none in the context B showed freezing responses for 61.25 seconds, indicating that fear memory was strongly maintained even after 1 week. In contrast, freezing responses of the FC-HT7 group in the context B were significantly decreased to 30.86 seconds, as compared with FC-none group. These findings suggest that HT7 acupuncture treatment might be useful for reducing fear memory.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-272 気分変化による音楽嗜好性及び音楽嗜好性脳波の変化 Kiyohisa Natsume（夏目季代久），Hirokazu Sadahisa（定久裕和）九州工業大学大学院生命体工学研究科 Our previous study showed frontal gamma rhythm was related to music-preferences. Electroencephalogram (EEG) is affected with the change in the mood. We have studied how the mood changes the music preferences and the music-preferences-related EEG. Twenty-two male and one female participants (age: 23.7 ± 1.1; mean ± SD) took part in the experiment. They were divided into two groups the exclusion group and the control group. Cyberball task (CBT) was used to change the participant's mood. It is reported CBT induces social exclusion mood. In CBT the participants play catch with the other two players on the computer monitor, and the others throw the ball to you with the probability of 20 % (the exclusion group) and 33 % (the control group). Before and after the CBT, they answered the questionnaires of the state anxiety in the State-Trait Anxiety Inventory JYZ. After the questionnaires, the participants listened to music and evaluated the music preferences. The listening-music session consists of 4 periods, the 10-sec rest, the 40-sec music-listening, the 5-sec thinking-evaluation, and the 5-sec rating-preference. The participants were given 18 excerpts in the 1st listening session before CBT. In the second session after CBT, they listened to 22 excerpts including the first 18 ones. After listening to music, the participants had to rate its preferences in 6 linear scales. The higher scores indicated like excerpts. During the sessions, we recorded EEG. The time-averaged power spectrum of EEG using short time fast Fourier transformation was calculated. In results, the anxiety scores tended to increase after CBT in the exclusion group. The number of the excerpts which changed in the preference scores significantly increased more in the exclusion group than in the control group. The correlation coefficient between the preference scores and BPM became negative after CBT in the exclusion group. In addition, the EEG power had significantly smaller correlation with the preference scores after CBT. However, it kept the correlation with beat per minutes (BPM) of the excerpts. These results suggest before CBT, the brain processes the music preferences based on the related EEG, and after CBT, the participants become more anxiety, the related EEG is suppressed, and the brain processes the preferences using the BPM-related EEG.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-273 うつ病モデルマウスのミクログリアにおける前頭皮質の炎症関連遺伝子の発現変化 Mai Sakai（坂井舞）¹，Zhiqian Yu（兪志前）^1,2，Hiroshi Meguro（目黒博志）³，Hiroaki Tomita（富田博秋）^1,2 ¹東北大院医災害精神 ²東北大災害研災害精神 ³東北大医学部 It is well known that chronic stress-induced depression is a common hallmark of many psychiatric disorders including major depression disorder. Clinical studies report on elevated TNFα, IL-6 and decreased IL-10 in the patients with major depression. Elevated IL-6 has served as a consistent biomarker of depression and IL-10 is proposed to influence depressive behavior through its ability to counterbalance pro-inflammatory cytokine expression. Postmortem study showed that tmTNF was significantly increased in BA 46 from patients with major depression. Microglia are the resident macrophage-like populations in the central nervous system which play major role in the inflammatory process. Activated microglia are widely known to exert proinflammatory M1 and anti-inflammatory M2 functions. Although responses to chronic stress exposure have been widely characterized, few studies have examined the microglia activation and its response to chronic stress. However, microglial activation response to chronic stress such as chronic restraint stress had not been extensively explored. In this study, we subjected mice to 3 weeks restraint stress and allowed to recover for 3 weeks to investigate the stress and recovery impact on microglia activation. We found that restraint stress induced depression-like behaviors that were maintained 2 weeks after cessation of restraint stress. Following the restraint stress, microglia activation was significantly suppressed with decreased Tnf, Il1b, Il10 and Tgfb1 mRNA expression compared with controls. During the first week or two of recovery period, microglia activation and its M2 phenotype marker Cd86 mRNA expression displayed a significant up-regulation along with significantly increased Tnf, Il1b, Il10 and Tgfb1 mRNA expression. Furthermore, Il10 and Tgfb1 mRNA expression were significantly increased throughout the 3 weeks after cessation of chronic restraint stress. These findings demonstrated that a chronic stressful event could produce delayed immune response in the brain, may help us understand the microglia reversibility of the effects of chronic stress, which could also be used for further studies on anti-depression drug development.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-274 マウス逃避行動において上丘深層から扁桃体への神経回路が果たす役割 Kaoru Isa（伊佐かおる）¹，Thongchai Sooksawate（Sooksawate Thongchai）²，Sakura Ikeda（池田桜）¹，Kenta Kobayashi（小林憲太）³，Tadashi Isa（伊佐正）^1,4 ¹京都大院医神経生物 ²Dept of Pharm Physiol, Chulalongkorn Univ, Bangkok, Thailand ³生理研ウイルスベクター開発室 ⁴京大高等研究院 The superior colliculus (SC) is a brainstem center controlling the innate behavior, and especially, the output pathway from its deeper layer to the ipsilateral brainstem, particularly to the cuneiform nucleus (CuN), has been demonstrated to control the defense-like behavior. In our previous study, we succeeded in selectively expressing channelrhodopsin2 (ChR2) in the ipsilateral pathway by injecting FuGE-MSCV-Cre into the CuN and AAV-EF1a-DIO-ChR2-EYFP into the SC, both on the same side of mice. Then, blue laser stimulation (473 nm wavelength) of SC induced defense-like behavior in these mice. Interestingly, the defense-like responses were composed of 2 phases; firstly the short-latency upward head movements and secondly escape responses. The latter escape responses were context-dependent; backward retreat movements were induced when stimulated in an open field, while the same stimulus induced flight responses in a closed box, by which the animal flew to the corner. Therefore, it was considered that the latter response should be induced via the neural circuits under top-down regulation. In this regard, it was intriguing that immunohistochemical staining of axonal trajectories of the ipsilateral SC output pathways revealed that these axons have ascending collaterals targeting several subcortical centers, such as periaqueductal gray matter, lateral hypothalamus and mediodorsal thalamic nucleus lateral part, many of which are known to be connected to amygdala. In this study, we first studied whether the optogenetic activation of the ipsilateral SC output pathway induce excitatory responses in the central amygdala neurons under anesthesia, and we found that the amygdala neurons could be orthodromically activated with oligosynaptic linkage. On the other hand, we confirmed when the ipsilateral SC output pathway was stimulated under the passive avoidance test condition, the latency for the mouse to enter the dark room where it received the stimulation was significantly prolonged, which suggested that activation of the ipsilateral SC output pathway induced fearful emotion. For the further step, involvement of amygdala was tested by observing the effects of the bilateral amygadala lesions on the context-dependency of the defense-like responses induced by optogenetic activation in the open field vs closed box, and investigating their effects on behavior under the passive avoidance test condition. We will report the detail of the results in this presentation.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-275 マカクザルの内側前頭前野における化学遺伝学的神経抑制は誤信念帰属に基づく予期的視線偏向を消失させる。 Ryota Akikawa（秋川諒太）^1,2，Taketsugu Hayashi（林剛丞）^2,3，Keisuke Kawasaki（川嵜圭祐）²，Jun Egawa（江川純）³，Takafumi Minamimoto（南本敬史）⁴，Kazuto Kobayashi（小林和人）⁵，Shigeki Kato（加藤成樹）⁵，Yukiko Hori（堀由紀子）⁴，Yuji Nagai（永井裕司）⁴，Atsuhiko Iijima（飯島淳彦）^1,6,7，Toshiyuki Someya（染矢俊幸）³，Isao Hasegawa（長谷川功）² ¹新潟大院自然科学研究科 ²新潟大院医歯総合研究統合生理 ³新潟大学大学院医歯学総合研究科精神医学分野 ⁴量子科学技術研究開発機構放射線医学総合研究所脳機能イメージング研究部 ⁵福島県立医科大学付属生体情報伝達研究所生体機能研究部門 ⁶新潟大学医学部保健学科 ⁷新潟大学工学部工学科人間支援感性科学プログラム Theory of mind (ToM) is a fundamental social function to understand mental state of other individuals. Acquisition of ToM is evaluated by testing whether one predicts others' actions according to their beliefs, even when those beliefs are different from reality. Human neuroimaging studies using various false-belief (FB) attribution tasks have revealed the relationship between ToM and brain networks including the medial prefrontal cortex (mPFC). However, causal role of mPFC in ToM remains unsettled due to the lack of interventional studies to manipulate neuronal activity using appropriate animal models of FB attribution. In the previous year, we reported the macaque's ability to comprehend others' FBs in an anticipatory looking FB paradigm. In the present study we examined whether there is causation between the neuronal activity of the macaque mPFC and the spontaneous gaze bias to the FB targets. We injected a lentiviral vector incorporating hM4Di, an inhibitory DREADD (designer receptor exclusively activated by designer drugs), into the mPFC around area 9m. Six weeks following viral injection, we chemogenetically inactivated the mPFC by intramuscular injection of clozapine N-oxide (CNO), a specific ligand to hM4Di. Within 60 to 80 minutes after CNO injection, neuronal activity in the mPFC was significantly inhibited. We found that chemogenetic deactivation of the mPFC specifically altered the gaze bias to the FB target while leaving their abilities to track moving and hidden targets intact. Thus, our results indicate that neural activity in the macaques' mPFC plays a causal role in ToM. These findings suggest a novel view that macaques implicitly attribute mental states to others via operation of shared neural circuits with humans where the mPFC plays a pivotal role.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-276 Effects of cold treatments on behavioral modification in zebrafish Chun-Yung Chang（Chang Chun-Yung），Sok-Keng Tong（Tong Sok-Keng），Ming-Yi Chou（Chou Ming-Yi） Nation Taiwan University It is known that the physiological status of organisms changes according to environmental alterations, and changes from surroundings also induce different behaviors. Previous studies indicated that the changes in physiological status affect the nervous system directly and modulate behaviors in order to attain physiological homeostasis. However, the correlations of shifting physiological status and neural activity, as well as the underlying mechanisms of this process have not been fully understood. In the present study, we used zebrafish as the model to investigate how the environmental alterations modulate behavioral patterns and the corresponding neural activity. We treated adult zebrafish with cold environment, 18 degree Celsius, for 24 hours and then evaluated social and foraging behaviors of zebrafish. We found that the total amount of food intake of the zebrafish was significantly decreased after placed in cold water for 24 hours. On the other hand, there were no obvious changes in feeding motivation, aggressiveness, and social preference. To analyze the neural activity in the brain, we performed in situ hybridization of Egr-1, an immediate early gene. Our results indicated that the neural activity in the posterior part of parvocellular preoptic nucleus (PPp) was significantly increased after cold treatment for 1 hour and returned to normal level after cold treatment for 24 hours. These data suggested that the foraging behavior of zebrafish was suppressed when acclimating the cold. Within the acclimation process, there was a short-period of time that the neural activity of the hypothalamus was up-regulated. However, it is still not clear if there is a causality between neural activity in the brain and behavioral modification. Further investigation should be focused on the critical time periods and specific brain nuclei which cause the changes in physiological status and behaviors.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-277 ラットの援助行動への前部帯状皮質の関連 Nobuya Sato（佐藤暢哉）¹，Atsuhito Yamagishi（山岸厚仁）¹，Ayumu Inutsuka（犬束歩）²，Tatsushi Onaka（尾仲達史）² ¹関西学院大文総合心理科学 ²自治医大医生理 To examine the neural mechanism of prosocial behavior, we developed an experimental paradigm for occurring helping behavior in rats. Using the paradigm, we examined the involvement of the anterior cingulate cortex (ACC) in helping behavior. We used an experimental box that consists of pool and ground areas partitioned by a transparent plate in the helping task. The center of the transparent partition, there was a circular hole (90 mm in diameter) through which the rats could pass between the two areas. In front of the hole, there was a transparent circular door (95 mm in diameter) fixed with a fastener. When the helper rat opens the circular door, the soaked cagemate can escape from the pool area to the ground area. First, we found that c-fos expressions of neurons in the ACC increased, suggesting that the neurons in the ACC were activated by the helping behavior. We next examined the effect of stimulation of neurons in the ACC. The excitatory Gq-coupled designer receptors exclusively activated by designer drug (DREADD) hM3Dq or control GFP was expressed bilaterally in the ACC of the helper rats. Thirty minutes before each session of the helping task, the rats in both groups received intraperitoneal injection of clozapine-N-oxide (CNO), which selectively activates hM3Dq. As a result, the rats that neurons in the ACC were activated through DREADD showed almost no helping behavior whereas the rats in the control group, that were only administrated CNO, smoothly acquired the helping behavior. We also examined the effect of administration of oxytocin receptor (OXTr) antagonist to the ACC on the helping behavior. The acquisition of the helping behavior was impaired by bilateral microinjection of the OXTr antagonist into the ACC. These results suggest that the ACC is important for acquiring the helping behavior, and that oxytocin in the ACC has a role in the acquisition.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-278 マカクザルのミラーシステムおよびメンタライジングシステムにおける自他行動モニタリング中の神経活動およびその相互作用 Taihei Ninomiya（二宮太平）^1,2，Atsushi Noritake（則武厚）^1,2，Masaki Isoda（磯田昌岐）^1,2 ¹生理研認知行動発達 ²総研大院生命科学生理 Understanding others' actions and their consequences is of crucial importance for social life. Recent studies have revealed that the primate brain is equipped with two systems for processing such social information: the mirror system and the mentalizing system. However, it is still under debate as to how, if any, the two systems might interact during social information processing. To address this issue, we trained macaque monkeys to perform a role-reversal choice task with three kinds of partners, each of which had a different degree of similarity with the subjects: (1) a real monkey performing the task (real monkey condition, RM), (2) a filmed monkey performing the task that was replayed on a display (video monkey condition, VM), and (3) a filmed stick performing the task in the same way as the VM condition (video object condition, VO). Neural activities, both single units and local field potentials (LFPs), were recorded simultaneously in the ventral premotor cortex (PMv) of the mirror system and the medial frontal cortex (MFC) of the mentalizing system using linear-array multicontact electrodes. We found three sets of neurons in both cortical areas that were excited by the actions of its own, those of the partner, or both. The response magnitude of these neurons was modulated depending on the partner condition. The direction of information flow between the two areas was then investigated by applying a multivariate Granger causality analysis to the LFPs. We found that Granger information flow from PMv to MFC was increased in the order of RM > VM > VO during the observation of, as well as the execution of, the reaching movement. Conversely, Granger information flow from MFC to PMv was increased in the order of VO > VM > RM. These results suggest that the mirror system and the mentalizing system complement each other to extract action information from, and respond adaptively to, a wide variety of others. We propose that the mirror system primarily functions in interacting with others that are similar to oneself, whereas the mentalizing system does so with dissimilar others.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-279 シナプス形成抑制因子MDGAファミリー分子群の欠失が引き起こすE/Iバランス偏移がもたらす認知・行動異常 Tohru Yamamoto（山本融）¹，Daiki Ojima（尾嶋大喜）¹，MD Razib Hossain（Hossain MD Razib）¹，Atsushi Tada（多田篤史）^1,2，Naohiro Kurokawa（黒川直弘）²，Ken Nagasawa（長澤研）²，Kazuaki Takuma（琢磨和晃）²，Yuhki Nakai（中井雄規）³，Hiroo Takahashi（高橋弘雄）¹，Takashi Tominaga（冨永貴志）³，Yasushi Kishimoto（岸本泰司）² ¹香川大医分子神経生物 ²徳島文理大香川薬生物物理 ³徳島文理大香川薬神経科学研 Neurexin (NRXN)-Neuroligin (NLGN) pathway plays a pivotal role in synapse formation and maintenance. We isolated GPI-anchored IgSF molecule MDGA1/2 and found that they interfere NRXN-NLGN association though their direct interaction with NLGNs. We reported that haploinsufficiency of MDGA2 elevated excitatory/inhibitory balance (E/I balance); increased density of excitatory synapses, enhanced mEPSC frequency and amplitude, and altered LTP. We also recently showed that loss of MDGA1 reduced E/I balance; increased density of inhibitory synapses, enhanced mIPSC frequency, and also altered LTP. Our observations suggested that MDGAs deficient mice could be a useful animal model for investigating behavioral and cognitive impairments caused by E/I imbalance, and we further analyzed the behavioral characters of MDGAs deficient mice including cognition, social interaction, social dominance, fear, and psychiatric disorder-related behavior. MDGA2 deficient (E>I) mice showed ASD-like impaired social interaction with repetitive behavior; however, MDGA1 deficient (E

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-280 母性行動におけるOXTR発現神経はLSではGABAによって制御されMPOAではプロラクチン産生を制御することで貢献する Shizu Hidema（日出間志寿）¹，Keisuke Sato（佐藤佳亮）¹，Saori Yada（矢田紗織）¹，Yumi Takahashi（高橋友海）¹，Hiroaki Mizukami（水上浩明）²，Katsuhiko Nishimori（西森克彦）¹ ¹東北大院 ²自治医大遺伝子治療研究部 Studies with mice deficient in oxytocin or oxytocin receptor (Oxtr) genes have suggested that this system has essential functions in the overall regulation of social behaviors, including maternal behavior. Postpartum females lacking Oxtr gene (Oxtr -/-) showed impaired maternal behavior. Oxtr-Venus knockin (Oxtr Venus/+) mice were generated in order to identify OXTR-expressing neurons. They showed several nuclei with high densities of OXTR expressing neurons, including the lateral septal nucleus (LS) and medial preoptic area (MPOA), which have been implicated to have a critical role in the regulation of maternal behavior. The number of OXTR neurons expressing Fos was increased in the MPOA and LS of Oxtr Venus/+ mice when they engaged in maternal behavior. To determine whether OXTR in the LS and MPOA played a critical role in regulating maternal behavior, we ablated Oxtr expressing neuron in the LS or MPOA of Oxtr Cre/+ female mice using AAV-flex-dtA, and observed impaired maternal behavior after parturition. Elimination of OXTR in the LS of floxed OXTR mice did not result in a significant decrease in maternal behavior. Next, we observed the maternal behavior of Oxtr Cre/+: Gad67fx/fx with the reason that almost all OXTR expressing neurons in LS are GABAgic. Restricted eliminaton of Gad67 in OXTR expressing neurons showed serious impairment in maternal behavior. It is suggested to be important that the most of OXTR neurons in LS are GABAgic for maternal behavior. However, the elimination of OXTR in MPOA of floxed OXTR mice impaired maternal behavior and survival rate of their pups were serioiusly decreased. These data suggest that OXTR expressing neurons in MPOA are significant for maternal behavior and lactation. Thus, present study implies that OXTR signaling LS or MPOA region have different roles for maternal behavior with different mechanisms.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-281 脳内aromataseとヒトの性格ーPET研究 Kayo Takahashi（高橋佳代）¹，Takamitsu Hosoya（細谷孝充）^1,2，Kayo Onoe（尾上嘉代）¹，Hisashi Doi（土居久志）¹，Yasuhiro Wada（和田康弘）¹，Yasuyoshi Watanabe（渡辺恭良）¹ ¹理化学研究所生命機能科学研究センター ²東京医科歯科大学生体材料工学研究所 Aromatase is an estrogen synthetic enzyme and is distributed in various tissues, i.e., gonad, fat, liver, muscle, brain, and breast. It has been reported to be involved in the several brain functions such as cognition, behavior, emotion, and pathophysiology of Alzheimer's disease and autistic spectrum disorder. Aromatase has been reported to be involved in aggressive behaviors in genetically modified mice and in personality traits by genotyping studies on humans. However, no study has investigated the relationship between aromatase in living brains and personality traits including aggression. We performed a positron emission tomography (PET) study in 21 healthy human subjects using ¹¹C-cetrozole, which has high selectivity and affinity for aromatase. Before performing PET scans in humans, subjects answered the Buss-Perry Aggression Questionnaire and Temperament and Character Inventory to measure their aggression and personality traits, respectively. A strong accumulation of ¹¹C-cetrozole was detected in the thalamus, hypothalamus, amygdala, and medulla. Males showed a tendency to have higher aromatase level in the brain regions. Females showed associations between aromatase levels in subcortical regions, such as the amygdala and supraoptic nucleus of the hypothalamus, and personality traits such as aggression, novelty seeking, and self-transcendence. In contrast, males exhibited associations between aromatase levels in the cortices and harm avoidance, persistence, and self-transcendence. The association of aromatase levels in the thalamus with cooperativeness was common to both sexes. The present study suggests that there might exist associations between aromatase in the brain and personality traits. Some of these associations may differ between sexes, while others are likely common to both.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-282 集団飼育環境におけるマウスの個体識別能力を定量化する新規行動課題の開発 Masaya Fujiwara（藤原昌也）¹，Naoya Saito（斎藤直哉）²，Ayano Kameike（亀池彩乃）²，Yusuke Makino（牧野友祐）^1,2，Toshihiro Endo（遠藤俊裕）³，Masaki Kakeyama（掛山正心）^1,2 ¹早稲田大学重点領域研究機構環境医科学研究所 ²早稲田大学人間科学学術院 ³フェノバンス・リサーチ・アンド・テクノロジー合同会社 We report here a novel behavioral task measuring the ability of mice to recognize other mice using IntelliCage apparatus. In human, one can easily identify other individuals who they are and recognize what role they are, and this individual recognition ability is the basic function to build a relationship with other individuals. Mice are also known to be able to identify individuals, but it is unknown whether mice can recognize its role like human. IntelliCage is a computer-based fully automated behavioral analysis system for RFID-tagged mice in a group-housed environment (a large standard plastic cage (55×37.5×20.5 cm) equipped with four triangular operant learning chambers (15×15×21cm, corners, hereafter) that fit into each corner of the cage). In the individual recognition task, 16 male mice were group-housed in an IntelliCage (8 experimental and 8 target mice). Each experimental mouse (follower) was assigned one own target mouse, and the follower can access drinking water only at the corner in which its own target mouse lastly visited. Thus, each follower mouse had to identify the target mouse from cagemates and follow it to have drinking water as a reward. In the experiment, 16 male mice were divided into 2 groups of 8 mice each (4 follower and 4 target mice). In one group, the task was performed in the light period everyday (light phase group), the other group performed in the dark period everyday (dark phase group) for 20 days. Number of following behaviors to the target was gradually increased and decreased number of following behaviors to other 7 mice was gradually decreased throughout the test in both light and dark phase groups, showing that mice can acquire this task. We also found that the following score of dark phase group was lower than that of light phase group, indicating that the precision of individual identification is lower in the dark phase than in light phase. We next assessed the new target allocated task, in which the new target mouse was allocated to another target mouse within the same group. Mice in both groups acquired to follow the new target within a week. These results suggest that the mice can not only identify other individuals, but also they can recognize the role of it, in other words, mice can update individual information in the brain.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-283 ヨシノボリ属の生殖的隔離を司る行動および神経基盤の解析 Masafumi Kawaguchi（川口将史）¹，Koji Matsumoto（松本浩司）²，Naoyuki Yamamoto（山本直之）³，Kei Nakayama（仲山慶）⁴，Hanako Hagio（萩尾華子）³，Hironori Izumi（和泉宏謙）⁵，Fumikazu Suto（須藤文和）⁶，Yasunori Murakami（村上安則）⁷，Hiroyuki Ichijo（一條裕之）¹ ¹富山大医解剖 ²愛媛大附属校 ³名古屋大院生命農 ⁴愛媛大沿岸環境科学セ ⁵富山大生命科学先端セ ⁶国立精神・神経セ神経研 ⁷愛媛大院理工 Freshwater goby fishes (Rhinogobius spp.) comprise some dozen species. Although several species sympatrically inhabit a geographic region, the phylogenetic analysis revealed that each species can maintain its genetic uniqueness. Thus, the reproductive isolation among Rhinogobius species has been nearly established, though hybrid populations are also reported as rare cases. To address the behavioral mechanisms of reproductive isolation, we performed the combinatorial breeding experiments between R. flumineus and R. fluviatilis, sharing the upstream of a river. Males of R. flumineus exposed to conspecific females repeatedly exhibited components of the courtship behavior, such as crawling, head shaking and tail raising. In contrast, females of R. fluviatilis presented to R. flumineus males were intimidated by the heterogeneous males by means of dorsal fin spreading, gaping and attacking. Furthermore, R. flumineus males showed the same reaction on the females located in a neighbor tank, where males need to distinguish females using only the visual perception. These observations indicated that R. flumineus males discriminate characteristics of females between two species in association with the visual information, and select the appropriate behavior (courtship or aggression). Thus, the reproductive isolation between R. flumineus and R. fluviatilis is supported by the premating isolation that enables these two species to keep their genetic uniqueness. Next, to shed light on the neural mechanism underlying the behavioral selection in association with the discrimination, we examined the neurons expressing c-fos in whole brains of R. flumineus males by means of in situ hybridization, and compared the distribution patterns of c-fos positive neurons between courtship and aggressive behaviors. Both males with courtship and aggression expressed c-fos in the optic tectum and the lateral part of dorsal telencephalon, which are centers for visual processing. The c-fos positive neurons in the most rostral region of preoptic area, the lateral hypothalamus, and the periaqueductal gray were related to the courtship behavior, while the aggressing males exhibited c-fos expression in the caudalmost part of intermediate pituitary. These distinct patterns of neuronal activity might generate the distinguishable behaviors between courtship and aggression.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-284 処女雌と授乳期雌マウスの内側視索前野のエストロゲン受容体陽性細胞における仔の呈示時の神経活動性の比較 Tomoaki Murakawa（村川友哲），Aki Takahashi（高橋阿貴），Sonoko Ogawa（小川園子）筑波大学行動神経内分泌学研究室 Estradiol plays a major role in maternal behaviors in the postpartum period by acting on the medial preoptic area (MPOA). Although several studies have reported that exposure to pup stimuli activates more MPOA neurons in the postpartum, compared to virgin females, the profile of activated cells has not been specified. Since two types of estrogen receptors, ERα and ERβ, are localized in the MPOA, we investigated in both virgin and postpartum female mice whether and how ERα or ERβ expressing cells are activated during pup exposure. We used newly developed transgenic mice whose ERβ positive cells were labeled with red fluorescent protein (RFP). In the postpartum group, ERβ-RFP mice were exposed to one unfamiliar male pup (3-5days old) in their home cage for 15min on postpartum day (PND) 5, after they had been separated from their own pups for 2days, starting on PND 3. Mice in the virgin group were tested similarly after individually housed for 3days. All mice were sacrificed at 90min after pup exposure and brain tissues were collected for immunohistochemical detection of ERα, ERβ-RFP, and c-Fos, as a marker of neuronal activity. We found there were three distinctive cell populations: 1) single ERα expressing; 2) single ERβ expressing; and 3) ERα and ERβ co-expressing, similarly in both virgin and postpartum brains. Pup exposure activated single ERα expressing cells, which were localized in the ventro-medial area of the MPOA, more prominently in the postpartum group compared to the virgin group. Very few single ERβ expressing and ERα and ERβ co-expressing cells were activated. These results suggest that activation of ERα expressing cells may be critically involved in the induction of maternal behavior in postpartum female mice. Supported by KAKEN #15H05724 to SO.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-285 社会性の形成に関わる神経基盤の探索 Noriyoshi Usui（臼井紀好）^1,2，Hideo Matsuzaki（松﨑秀夫）^3,4，Yoshihisa Koyama（小山佳久）²，Yukiko Nakamura（中村雪子）²，Makoto Kondo（近藤誠）²，Shoichi Shimada（島田昌一）² ¹阪大院・医・共同研 ²阪大院・医・神経細胞生物 ³福井大・子どものこころ ⁴阪大院・連合小児発達 ASD is a neurodevelopmental disorder resulting in pervasive abnormalities in social communication, repetitive behaviors, restricted interests, and hypersensitive or hypoesthesia. The domestic prevalence rate is as high as 1-3%, and the effective treatments are not currently other than instruction on post-diagnosis education and parenting methods for families. ASD research is conducted all over the world and despite the identification of more than 1,000 associated genes to date, it has not yet reached the understanding the neural basis underlying symptoms and the establishment of an effective treatment method. Therefore, in this research, we focused on difficulties in social communication and decided to identify the neural basis of social nature. First, we created the model mice (SI mice) which display social deficit by manipulating the social environment during juvenile period, and confirmed the reductions in social behaviors. Furthermore, from other behavior analyses, we found increased anxiety-like behaviors in SI mice. Next, we conducted gene expression analysis in the prefrontal cortex, and identified novel candidates associated with sociality. These genes were involved in transcriptional regulations and synaptic functions. Since the plasticity of the neural circuit was suggested by changes in the social environment, we are carrying out histological analysis to screen histological phenotypes. In this meeting, we would like to discuss our latest findings on the neural basis underlying social communication.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-286 物体に対するエージェンシーの誤帰属とその神経科学的メカニズム: Human Connectome Projectパイプラインを用いた大規模サンプル研究 Hiroki Tanaka（田中大貴），Atsushi Miyazaki（宮崎淳），Takayuki Fujii（藤井貴之），Toru Ishihara（石原暢），Muneyoshi Takahashi（高橋宗良），Tetsuya Matsuda（松田哲也）玉川大学脳科学研究所 Attribution of intentionality or agency to objects is fundamental to empathy, perspective taking, and prosocial behavior. However, overuse of such attribution induces a delusion (perceiving intentionality in obviously inanimate objects), disturbing adaptive social relationships often seen in patients with schizophrenia (Ciaramidaro et al., 2015; Riemar, 2017). Previous social neuroscience studies revealed that STS was more activated while participants watched high-intention animations; however, whether this activation is also involved in over-attribution of agency is unclear. Previous studies did not examine relationships between brain activation and attribution of agency. Therefore, we directly investigated brain areas related to a ""false alarm of agency"" in participants who perceived intentionality in random-moving objects. Using the publicly available Human Connectome Project dataset (Glasser, 2013; Glasser et al., 2016), we analyzed resting-state fMRI (rsfMRI) and task fMRI (tfMRI) data obtained from 1048 healthy participants (563 women, age range: 22-37 years). Using the HCP protocol, we generated resting-state functional connectivity maps containing 15, 25, 50, 100, and 300 regions by independent component analysis. In tfMRI, participants engaged in the following social cognition task (Castelli et al. 2000; Wheatley et al. 2007): They were presented with 20-second video clips in which several shapes moved. After watching video clips, participants judged if the shapes had mental interaction or random movement. Judgment of random stimuli as exhibiting mental interaction was regarded as a false alarm. In total, 164 participants showed at least one false alarm. In all resting-state functional connectivity maps, contrasting brain activation during mental and random conditions in a common brain component significantly predicted a false alarm (FDR corrected p<.05). The component did not include STS, but included bilateral PCC and precuneus, both within the default mode network. Thus, atypical brain activation in the default mode network, in which patients with schizophrenia show abnormality (Garrity et al., 2007; Ongur et al., 2010), may be involved in over-attribution of agency to objects.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-287 Analyses for the role of ESR1 polymorphisms in mouse genome Lalithadevi Mallarapu（Mallarapu Lalithadevi），Akira Tanave（Tanave Akira），Yuji Imai（Imai Yuji），Tsuyoshi Koide（Koide Tsuyoshi） National Institute of Genetics Estrogen receptor 1, ESR1 (or ER-Alpha) is a predominant subtype of estrogen receptors (ERs) which plays a major role in estrogen-mediated proliferation, bone metabolism, sexual development and social behavior. The structure of ESR1 known to code for a multiple transcript variants resulting from alternative splicing. These splice variants are mostly produced by the combination of exon-skipping and insertion of short exon(s). Many of these transcript variants are translated into nuclear receptor proteins with altered structure and function. The Esr1 gene structure is different between human and laboratory mice. But, mouse is the extensively used model animal to study ESR1 regulation and function in vivo. The mouse Esr1 gene consists of 8 conventional coding exons and 10 transcript variants. Within the mice strains, the structure of Esr1 gene showed marked difference in wild derived mice strain MSM/Ms (MSM) when compared to laboratory mice strain C57BL/6J (B6). We identified two insertions in intronic (insertion A) and exonic (insertion B) regions of B6 genome when compared to MSM. Some of these insertions are absent in human, similar to MSM. In my study, I am trying to clarify the role of these insertions in the function of ESR1. In order to study the role of these insertions (A and B) we developed a Esr1ΔA and Esr1ΔB mice using B6 through CRISPR/Cas9-mediated gene editing. During breeding, we observed that Esr1ΔB heterozygous mice showed reduced reproductive potential and survival of pups. However, the breeding of Esr1ΔA mice was normal. Here we would like to study whether these changes in Esr1ΔB heterozygous mice are due to change in behavior of parents or hormonal changes which leads to low pup survival. Previous studies showed that ESR1 plays a major role in regulating maternal and aggression behaviors. Based on these studies, we are focusing on maternal and aggression behaviors in Esr1ΔB mice. Preliminary experiments on maternal behavior showed that Esr1ΔB heterozygous and homozygous mice were reluctant to retrieve pups (took more time) when compared to the wild type control. These results indeed provided a glimpse of change in maternal behavior in Esr1ΔB mice.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-288 自閉スペクトラム表現型の指標としてのマウスのハドリング行動 Masaki Kakeyama（掛山正心）^1,2，Masaya Fujiawara（藤原昌也）²，Seico Benner（ベナー聖子）³，Hidenori Yamasue（山末英典）³ ¹早稲田大学人間科学学術院 ²早稲田大学重点領域研究機構環境医科学研究所 ³浜松医科大学精神医学講座 Analyzing social behavior has strongly been needed Disruption of social behavior, one of Problems in social behavior is one of major symptoms of autism spectrum disorders (ASD), and there has been strong need for behavioral index of mice to address ASD pathogenesis. We have recently developed a video-analysis-based system, the Multiple-Animal Positioning System (MAPS), to automatically and separately analyze the social behavior of multiple individuals in group housing condition, and reported that group-housed male mice exhibited huddling behavior within several hours even with the unfamiliar stranger mice. In human, the distance during social interactions, that is social proximity, is known to depend on the relationship between the others, and thus, social proximity can be an index of social behavior. In this study, the huddling behavior of mice was investigated to estimate as a possible new behavioral index contributing ASD pathogenesis including re-analysis of our previous findings. First, social isolation of male mice during adolescence leads he mice reared in social isolation stayed as far away from each other as possible and took two days for all four of them to finally huddle together, showing that adolescent social isolation results in deficit of formation of social relationship in adulthood. Second, the phenotype of these socially isolated mice was partially rescued by cohabitation with group-housed mice. The fact that social proximity of mice can be affected by the social experience in early life and at the time of co-housing might indicate the social proximity of mice is similar to that of human in terms not only phenotype but also its development. In human, administration of oxytocin has reported to affect experimental measures associated with social behavior and ASD core symptoms, and thus, if administration of oxytocin altered social proximity in mice, it can be a good index for ASD pathogenesis. Effect of oxytocin in mice will also be discussed.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-289 コモンマーモセットにおける聴覚情報処理に関わる神経応答 Naho Konoike（鴻池菜保）¹，MIKI MIWA（三輪美樹）¹，Kosuke Itoh（伊藤浩介）²，Katsuki Nakamura（中村克樹）¹ ¹京都大霊長研高次脳 ²新潟大脳研 Common marmosets (Callithrix jacchus) are small primates and display a unique social behavior, such as exchange the species-specific calls. They have come to be used in many laboratory experiments because of genetic manipulability to produce primate disease models. In the current study, we aim to reveal the neural mechanisms underlying auditory processing of marmosets. The two adult female marmosets (Age: 8 and 7 years old / Body weights: 380g and 400g, respectively) were used. The subjects were implanted in the unilateral auditory cortex with a 16ch micro-electrocorticogram (ECoG) array (NeuroNexus). For one subject, the array was placed in the subdural space, whereas the array for another subject was placed on the dura. All experimental procedures were approved by the animal welfare and animal care committee of the Primate Research Institute of Kyoto University. We recorded ECoG from the marmosets sitting on primate chair and passively listening to vocalization of conspecifics. In addition, we attempted to investigate dynamic neural processes of naturally behaving and freely moving marmosets. The wireless device (TBSI) was also used and recorded neural activities from free moving marmosets in the same conditions. We acquired and analyzed the neural activity focused on the relationship with each species-specific call, and compared the neural data from subdural and epidural arrays. We also recorded the auditory steady state response, which was used to assess neural synchrony for specific frequencies in schizophrenia and in animal models. These recording methods would provide useful neural data for evaluation of the primate disease models and clinical research.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-290 法的判断の神経相関：法的専門知識と法秩序に対する信頼は差を生むか？ Takeshi Asamizuya（浅水屋剛），Ai Takahashi（高橋愛），Hiroharu Saito（斎藤宙治），Ryosuke Higuchi（樋口亮介），Go Naruse（成瀬剛），Shozo Ota（太田勝造），Junko Kato（加藤淳子）東京大院 The neural correlate of the legal judgment of ordinary people has been underexplored, whereas the lay-judge system is an important part of the court system in many countries, including Japan, Korea, USA, and UK. Inside the MRI machine, participants were asked to read vignettes about criminal procedure cases and make legal judgments. With the close cooperation with legal specialists, five murder cases were made to examine the effect of the repentance of defendant on the judgment of the appropriate punishment. We focused on the brain activity that is associated with the legal judgment when the defendant does or does not show repentance. The novelty of our approach is examining the difference in brain activation between legal professionals and lay people. On campus, we recruited two groups of participants, i.e., students on campus who are not specialized in law and law students who have just passed the national bar exams. We also recruited lawyers who have had experienced legal practice. We also examined a difference in rsfMRI between legal professionals and lay people. The results also indicate that the strength of support for the legal system dissociates brain activation when non-law students make a legal judgment. We examine whether and to what extent the legal expertise and the attitude toward legal system make difference in participants' neural correlates when they make legal judgments.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-291 サル扁桃体における社会的情報と情動情報の表現 Koji Kuraoka（倉岡康治），Kae Nakamura（中村加枝）関西医科大学医学部生理学講座 Macaque monkeys are expert at processing social and emotional information. Social and emotional information are independent in nature although these two are closely related. However, it is not clear whether they are computed separately or conjointly. To this end, we developed a behavioral task in which the monkey performed a saccade task under different social and emotional context. First, we confirmed that the monkey obtained social information from the gaze of other individuals. After fixation on a central fixation point (FP), a target dot was presented on left or right of the FP to which it made a saccade to obtain a liquid reward. An image of a monkey or cartoon face, looking left or right, was briefly presented before target presentation. Saccade reaction times were significantly shorter when the gaze direction of the monkey image was consistent with that of the target than when they were inconsistent. In contrast, the gaze direction of cartoons did not affect saccade reaction times, indicating that gaze-inducted saccade modulation was bound to the social images. Next, we examined the monkey's preference of images by a choice task. Among four facial stimuli, two (A&B) led to a large reward while the rest (C&D) led to a small reward. Further, A&C represented an honest gaze looking at the direction of forthcoming target while B&D represented a dishonest gaze opposite to the target. As expected, the monkey chose the stimuli leading to a large reward more often than that to a small reward. Moreover, rate of choosing the stimuli associated with honest gaze was significantly higher than those with dishonest gaze, indicating decision making based on social preference. The amount of reward and the direction of the gaze did not have an interacted impact on the monkey's choosing behavior, indicating independent processing of social and emotional information. Amygdala has been implicated in processing of social and emotional information. To investigate whether and how they are computed at a single neuron level, we recorded the activity of neurons in the monkey basolateral amygdala during the saccade task. Among neurons that responded to a stimulus, we found that number of neurons showing a significant effect of social or emotional factor are larger than number of neurons showing both effects. These results indicate that the monkey discriminates social and emotional information, and each of those is separately computed by the basolateral amygdala neurons.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-292 マウスの従順行動における扁桃体基底外側核および腹側海馬の役割 Hiromichi Nagayama（永山博通）¹，Yuji Imai（今井悠二）²，Yuki Matsumoto（松本悠貴）^1,2，Tsuyoshi Koide（小出剛）^1,2 ¹総研大生命科学遺伝学 ²国立遺伝研系統生物研究セマウス開発 Some animals approach and explore toward humans. These behaviors are known as active tameness and considered as a behavioral basis of affiliative relationship between animals and humans. It has been considered that domestication causes some changes in brain as well as behaviors in animals. However, precise neurobehavioral mechanism of active tameness is still unclear because lack of knowledge to study neural mechanism associated with domesticated animals. Thus, our study aimed to identify neurons causing the tameness by using a mouse model of domesticated animals. We established two types of mouse groups tameness and non-tameness. Tameness groups are offspring of a selective breeding using wild-derived heterogeneous stock (WHS). Individuals contacting towards a human hand for longer times were mated repeatedly through the breeding. On the other hands, a mating without any selection was conducted for control groups. Using these mouse groups, we investigated various behavioral characteristics of tame mice. Furthermore, differences of brain activity during a hand presenting stimulus was investigated to reveal what kind of emotions and which brain regions was important for the tame behavior. At first, we found that two groups of mice exhibited exploratory and anxiety-like behaviors at same levels in novel environment and object exposures. However, behavioral differences were observed in some social conditions. Tame groups explored other individuals more frequently while non-tame groups avoided in three types of sociability tests. Next, we found that basolateral amygdala (BLA) and ventral hippocampus were not activated by hand presentation stimuli in tame mice but these regions of non-tame mice were activated. It was reported that these regions associate with social anxiety. Thus, we hypothesized that the low-activity of these brain regions caused an insensitive to specific anxiety which was important as basis of tameness. Now, we were trying to improve the tameness by a suppression of the anxiety using anti-anxiety drug. We want to discuss about the relationships between tameness, sociability and anxiety based on recently results.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-293 雄型社会行動の調節に果たすエストロゲン受容体αとアンドロゲン受容体の役割 Shoko Sagoshi（佐越祥子）¹，Kazuhiro Sano（佐野一広）¹，Masahiro Morishita（森下雅大）²，Shinji Tsukahara（塚原伸治）²，Anders Agmo（Agmo Anders）^1,3，Sonoko Ogawa（小川園子）¹ ¹筑波大学　行動神経内分泌学研究室 ²埼玉大学　理工学研究科調節生理学研究室 ³University of Tromso, Tromso, Norway Testosterone (T) plays an essential role on masculinization of immature brains and facilitation of male-type social behavior by acting on the masculinized brain later in adulthood. T not only acts through androgen receptors (AR) but also acts on estrogen receptors (ER) after being aromatized to estradiol intracellularly in the brain. In our previous study, we generated ERα-cell specific AR knockout (KO) mice and examined the effects of AR deletion only in the ERα-positive cells on male-type social behavior. We found that lack of AR on ERα expressing cells almost completely abolished male sexual behavior but did not affect aggressive behavior. However, greatly elevated levels of serum T found in gonadally intact KO mice compared to wild-type (WT) mice may have a compensatory effect on the levels of aggression. In the present study, we examined the effects of testosterone propionate (TP) treatment on recovery of aggressive behavior in gonadectomized male mice. Male KO and their WT were tested once for sexual and aggressive behaviors at 3 weeks after gonadectomy to confirm suppression of the behavior. On the next day, they were implanted subcutaneously with either 10 or 20mm silastic capsule (i.d. 1.98mm, o.d. 3.18mm) containing crystalline TP. They were then tested for sexual and aggressive behavior repeatedly starting on 7 days after capsule implantation. We found that TP successfully restored aggressive behavior in both KO and WT mice. On the other hand, unlike WT mice which showed substantial levels of sexual behavior by 10 days after capsule implantation, KO mice failed to respond to TP treatment. Detailed analysis of AR and ERα distribution in a number of brain regions in the social behavioral network confirmed that AR expression was greatly reduced in the medial preoptic area, bed nucleus of the stria terminalis, medial amygdala, ventromedial nucleus of the hypothalamus and periaqueductal gray in KO mice compared to WT mice. These results suggest that activation of AR and ERα expressed on the same cells is necessary for the induction of sexual behavior, but not critical for aggressive behavior. Unlike sexual behavior, T may facilitate aggressive behavior by primarily acting on ERα after being aromatized to estradiol, and/or acting on AR expressed in ERα negative neurons. The latter possibility needs to be tested in future studies by treating mice with dihydrotestosterone, non-aromatizable androgen.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-294 マウス養育行動のメラニン凝集ホルモン欠損による影響 Chitose Orikasa（折笠千登世），Yoko Kato（加藤陽子），Harumi Katsumata（勝又晴美），Shiro Minami（南史朗）日本医大先端研 Melanin-concentrating hormone (MCH) is considered to have an integrative role in neuromodulating and physiological functions such as feeding, energy balance, sleep, stress, anxiety and cognition. There also have been several evidences of MCH system in maternal behavior. Here we examined the maternal behavior by the tet-off system to ablate MCH neurons specifically in the lateral hypothalamic area using MCH-tTA; TetO DTA bigenic mice. Pup survival rate after parturition in female mice which ablated MCH neurons congenitally was lower than that of control females even their litter size at birth was equal. We also examined the parental behavior and sexual behavior in virgin female and male MCH-tTA; TetO DTA bigenic mice at 11 weeks of age. Sexually naive female mice which ablated MCH neurons were less interested in pups and maternal care when compared to control mice during pup presentation test. In contrast, sexually naive male mice with ablations of MCH neurons showed more attack to pups than that of control. Since MCH play a role in regulatory systems including its food intake and energy homeostasis, their body weight was significantly lower in MCH ablated groups than in control groups, however, in their estrous cyclicity, no significant difference was observed. The effect of ablation of MCH neurons on mating behavior was not found. These results suggest that MCH neurons deficiency inhibit parental care to the pups in mother, and in both sexually naive female and male mice. Our data indicate the possibility that MCH neurons play a role in neuromodulating mechanism of parental behavior in rodents.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-295 内側前頭皮質および背外側前頭前野への反復経頭蓋磁気刺激がサルの競争的エサ取り行動に与える影響 Yoshiaki Ikeda（池田義晃）¹，Takayuki Hosokawa（細川貴之）^1,2，Hitoshi Nagano（長野人士）¹，Atsuhiro Saita（齋田淳裕）¹，Shinya Nakamura（中村普也）¹，Kenichiro Tsutsui（筒井健一郎）¹ ¹東北大院生命科学脳神経システム ²川崎医療福祉大医療技術学部 Macaque monkeys show various social behavior to the others depending on their hierarchical status in society. In this study, we aimed to investigate the neural substrate of social behavior by manipulating the activity of the medial frontal cortex (MFC) and dorsolateral prefrontal cortex (DLPFC), which are implicated in emotional control and social behavior, by using repetitive transcranial magnetic stimulation (rTMS). Two Japanese monkeys were engaged in a competitive food-picking task, in which they faced each other and competed to pick up pieces of sweet potatoes, each of which was put in the individual wells of a board placed in front of them. We bilaterally applied 1 or 10 Hz rTMS, which is known to have an inhibitory or facilitatory effect on the stimulated brain areas, targeting to the MFC or DLPFC of one of these monkeys and evaluated its task performance with or without the rTMS session. These monkeys were comparable in the social rank and usually did not hesitate to pick up the food pieces even from the wells near the counterpart. After 1 Hz rTMS to the MFC, the monkey became to pick up less food pieces from the wells near the counterpart, whereas, 10 Hz rTMS to the MFC did not induce such behavioral change. In contrast, after 10 Hz rTMS to the DLPFC, the monkey became to pick up more food pieces from the wells near the counterpart, whereas, the monkey tended to pick up less food pieces from the wells near the counterpart after 1 Hz rTMS to the DLPFC. Together, these results indicate that the MFC and DLPFC have opposite functions in emotional control and social behavior, and that these areas complementarily function for appropriate social behavior to the others.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-296 ビタミンB1及び食餌性マグネシウムの欠乏による脳内炎症と海馬依存性記憶障害 Ryuhei Tsuji（辻竜平），Mizuki Miyahara（宮原瑞希），Shohei Takahashi（高橋翔平），Tatsurou Serita（芹田龍郎），Satoshi Kida（喜田聡）東京農大院・農・バイオ Malnutrition including deficiencies of Vitamins, mineral and amino acids has been shown to impair the function of brain. Vitamin B1 (Thiamine) deficiency causes Wernicke-Korsakoff's syndrome in human that displays severe deficits in learning and memory. Our previous studies have shown that mice that recovered from pyrithiamine-induced thiamine deficiency (PTD mice) display chronic impairments of hippocampus-dependent memory. Furthermore, we observed that PTD mice show significant decreases in the number of neurons and spine density in hippocampus. Our findings suggest that PTD impairs hippocampus-dependent memory by degenerating hippocampus. On the other hand, we examined effects of dietary magnesium deficiency (MgD) on learning and memory and found that mice fed an MgD diet show deficits in hippocampus-dependent contextual fear, spatial and social recognition memories. However, in contrast to observations in PTD mice, MgD mice showed normal spine density and morphology of hippocampal neurons. To understand and compare molecular mechanisms of hippocampus-dependent memory impairments in PTD mice and MgD mice, we performed RNA-seq analyses of hippocampus using the next-generation sequencing. We analyzed transcriptome of hippocampus of PTD and MgD mice. Interestingly, PTD mice showed abnormal expressions of inflammation-related genes in the hippocampus; PTD mice showed significant and strong increases in mRNA levels of TNFα, IL1α, IL1β, IL6, TLR2, IL17RA, Ccl7, Spp1, MAPKAPK2 and M1 microglia markers. MgD mice also showed similar abnormal expressions of inflammation-related genes in the hippocampus. However, MgD mice showed significant but weak increases in mRNA levels of Ccl7, and normal mRNA levels of TNFα, IL1β, IL6, MAPKAPK2. Taken together with our previous observations, these results indicated that PTD induces strong neural inflammation with degenerating hippocampus, whereas MgD induces weak neural inflammation without affecting hippocampal morphology. In summary, our findings suggest that malnutrition such as PTD and MgD induces memory impairments that are associated with neural inflammation and that stronger and more prolonged inflammation by malnutrition may lead to brain degeneration.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-297 Modulation of dentate granule cell activity during fear memory extinction in freely moving mice Alvaro Carrier Ruiz（Carrier Ruiz Alvaro）^1,2，Yuki Sugaya（Sugaya Yuki）^1,2，Masanobu Kano（Kano Masanobu）^1,2 ¹Graduate School of Medicine, The University of Tokyo ²WPI-IRCN, UTIAS, The University of Tokyo Granule cells (GCs) in the hippocampal dentate gyrus mediate the extinction learning of episodic memories. Nevertheless, little is known about how this is achieved, especially in behaving animals. To address this issue, we expressed the calcium sensor GCaMP6f in GCs of adult C57BL/6 mice by means of stereotaxic adeno-associated viral vector injection. We recorded fluorescent signals representing Ca²⁺ transients in freely moving animals by using a miniature microscope during acquisition, retrieval and extinction of a cued fear memory. Control mice were exposed to cues without foot shocks during the conditioning period and all of the recorded data were analyzed by Inscopix Mosaic and GraphPad Prism software. After conditioning, cue exposure triggered freezing behavior, confirming the successful retrieval of cued fear memory. Along with extinction of the fear memory, cue-triggered freezing behavior decreased significantly in conditioned mice. Our imaging data revealed a significant increase in the number of active GCs during cued fear extinction, which was also confirmed by the immunolabeling of Fos positive cells in the granule cell layer. Furthermore, GCs of conditioned mice exhibited a transient increase in firing frequency and a higher cue selectivity during extinction sessions. Finally, we found that chemogenetic inhibition of GCs decreased cue-induced freezing, whereas generalized ablation of GCs increased cue-dependent freezing and disrupted cued extinction. Overall, our results suggest that GCs are necessary for the retrieval and extinction of cued fear memory. By transiently increasing their activity, dentate GCs may promote synaptic plasticity necessary for extinction learning of fear-conditioned episodes.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-298 結合ニューロフィードバックの作業記憶成績への長期効果 Masahiro Yamashita（山下真寛）¹，Tomohisa Asai（浅井智久）¹，Hiroshi Imamizu（今水寛）^1,2 ¹ATR認知機構研 ²東京大院人文社会行動文化・心理 Working memory refers to a short-term memory to guide a goal-directed behavior. The ability has been linked with left fronto-parietal network (LFPN) connectivity that is known as a language network. It remains elusive whether working memory performance can be improved by strengthening within-network connectivity of the LFPN. To address this question, we utilized a connectivity-based real-time fMRI neurofeedback (NF) approach. We hypothesized that increasing connectivity by NF training would strengthen connectivity even at resting state and improve working memory performance. To select participants for the NF sessions, we first examined behavioral performance in an adaptive N-back training. Fifty-nine healthy young participants (age 23.4 ± 2.2 years) underwent an adaptive letter N-back training in consecutive 5 days. We selected relatively under-performing 15 participants (age 23.1 ± 2.9 years) based on their average N-back level at the final training day. The participants spent four sessions of NF training, in which 10 participants received real-feedback score that represented connectivity-strength (NF condition) while the other 5 participants received sham-feedback that was obtained from other participants' feedback score (sham condition). Participants underwent test sessions within one week before and after, and also after more than a month (Pre/Post/Follow-Up-test). In the test sessions, fMRI activity was recorded during 3/4-back task and resting state. Additionally, working memory performances were measured using digit/fractal N-back tasks. Our preliminary results showed that 60%/60% of participants in NF/sham group increased connectivity after the training sessions. On average, we found increased connectivity at Post-test in both groups (33%/54% increase in NF/sham group), but only NF group maintained the increase at Follow-up-test (26%/13% increase in NF/sham group). NF group improved the digit/fractal N-back performance at Post- and Follow-up-test, whereas sham-group did not improve the performance. Both NF and sham group may increase connectivity during training sessions. Resting state connectivity increased both groups immediately after the NF training but only real-NF maintained the effect after a month. The three types of memory contents are maintained through verbal rehearsal strategy according to previous studies. Connectivity NF on LFPN may generally improve working memory that requires verbal rehearsal.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-299 海馬時間細胞における相対的時間情報表現 Akihiro Shimbo（新保彰大）^1,2，Ei-Ichi Izawa（伊澤栄一）²，Shigeyoshi Fujisawa（藤澤茂義）¹ ¹理研CBS 時空間認知神経生理学 ²慶應大文心理 How our brain represent time? Although time itself is hypothetical entity, using it is important for our daily-life. In particular, interval timing, the ability to measure temporal information in seconds to minutes range, is contributed to various behaviors, such as foraging, decision making, associative learning, sequential and motor learning. Several studies reported that hippocampal pyramidal neurons can represent seconds to minutes range duration. So, population of hippocampal neurons shows sequential firing which fills the gap between events (Pastalkova et al., 2009; MacDonald et al., 2011), and these neurons are termed `time cell' because of similarity of place cell. Although time cells may represent elapsed time, the characteristics of time cells in interval timing has not been understood yet. For example, what type of temporal information are represented by the time cells? Do they encode the absolute duration (absolute type), relative percentage of the duration (relative type), or context-dependent temporal information (remap type)? To sort the three types of of temporal information encoded by time cells in interval timing task, we changed the length of interval which subjects are required to measure within one session. In this task, rats were required to discriminate two different interval, running on a treadmill, for making a correct choice and experienced different sets of intervals in task conditions, i.e. they experienced 5 s or 10s interval in block 1, 10 s or 20 s interval in block 2, and then 5 s or 10 s interval again in block 3. We defined neurons which shows similar activities in bolck1 and 2 as absolute type, neurons which fired at similar percentage of duration in block1 and 2 as relative type, and neurons whose activities was totally different between block 1 and 2 as remap type. We recorded neural activities in dorsal CA1 during running on the treadmill, and found that a subset of CA1 neurons showed firing activities depending on time from a start of running. Moreover, we sorted neuron into three types. The majority of time cells are sorted into relative type. Using these relative types of time cells, we decoded the temporal information correctly. Rats were also required to choose short or long after intermediate duration (7.1 s or 14.2 s). Decoding temporal information from relative type are differentiated depending on the rats' choice. These results indicated that relative type of time cells related to subjective time of rats.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-300 文脈恐怖記憶における海馬の機能的構築の検討 Shoko Arai（荒井翔子）¹，Constantine Pavlides（Pavlides Constantine）^1,2 ¹筑波大院　人間総合科学行動神経科学 ²The Rockefeller University, New York, USA Using immediate early genes (IEG) as indicators of neuronal activity, previous studies from our laboratory observed that activated neurons formed clustered ensembles of a few active neighboring cells in the dorsal hippocampus both in spatial and sequential memory tasks (Pavlides et al., 2019; Nakamura et al., 2010; Cho et al., 2017). However, these studies were performed using a 2D analysis and involved a limited part of the dorsal hippocampus. In the present study, we examined whether a cluster-type organization also occurs in contextual memory task. We analyzed functional organization three-dimensionally in CA1 field of dorsal and ventral hippocampus. The dorsal hippocampus is known to play an important role in spatial information, while the ventral hippocampus is important for processing emotional information. We further separated CA1 into distal and proximal parts, which receive distinct projections from the entorhinal cortex, that involve different types of information. Twenty-four hours after rats were contextual fear conditioned, shock group animals were placed in the same context again and the duration of freezing response was measured. Ninety minutes after the memory test (peak time of Zif268 protein expression), the rats were perfused for IEG detection. Three control groups were also included: 1) shocked, immediately removed from context; 2) context, non-shocked; and 3) home cage controls. The brains were cut coronally in 60 μm thick sections and Zif268 protein expression was labeled by fluorescent immunohistochemistry. The sections were captured every 3 μm. Consecutive 3 sections were concatenated for each region. Signals above the thresholds of average + 2SD optical density were counted as Zif268 immuno-reactive cells. Topographic organization was performed by determining XYZ coordinates for each Zif268 IR cell, calculating average distances between a large number of cells and defining a cluster as cells falling within the average + 2SD distance. Cells further away were part of a different cluster or were single cells (i.e., not a cluster). Using a 3D analysis, we found a cluster-type organization in the CA1 field of dorsal hippocampus both in the transverse and septotemporal axis.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-301 ラットの条件刺激呈示順序に依存した条件性風味選好に関わるCCK媒介性シグナルの役割 Keisuke Shinohara（篠原恵介）¹，Kana Izumiya（泉屋佳奈）²，Yasunobu Yasoshima（八十島安伸）¹ ¹大阪大院人間科学行動生理 ²大阪大人間科学行動生理 Animals often learn to prefer neutral flavors associated with the intake of sugar. The sugar-elicited conditioned flavored preference (CFP) is based on both of flavor-taste (F-T) and flavor-nutrient (F-N) associations. Myers and Whitney (2011) reported that hunger state influences the expression of learned preference for flavors associated with nutrient. In this study, rats ware sequentially presented with two flavors as conditioned stimuli (CSs) paired with intra-gastric glucose infusion as an unconditioned stimulus (US). In the result, preference for the early-occurring flavor was expressed only when rats were hungry, whereas rats preferred the late-occurring flavor regardless of hunger state. In present study, we examined whether this phenomenon occurs in oral glucose-CFP which may be mediated by F-T and F-N associations. In experiment 1, female Wistar rats (n = 10) were trained with two distinct flavor CSs in succession (each for 8 minutes) per session under food restriction. They were exposed alternately between session with CSs (Early(+), Late(+)) paired with 12% glucose US and session with different CSs (Early(-), Late(-)) unpaired with US (each 8 sessions). After completion of the training, two-bottle choice tests were conducted after overnight deprivation (hungry test) and 90 minutes after giving of a daily amount of normal chow (fed test). During the series of these tests, rats were experienced all of four comparisons: Early(+) vs. Early(-), Late(+) vs. Late(-), Early(+) vs. Late(+), and Early(-) vs. Late(-). In the result, Late(+) was preferred over Late(-) only in fed test, whereas Early(+) was not preferred over Early(-) in both tests. Moreover, rats also preferred Late(+) over Early(+) only in fed test, which is a unique feature of oral glucose-CFP. In experiment 2, the other rats (n = 8) were conducted training and tests in a similar manner to experiment 1, except that they received an intraperitoneal injection of a Cholecystokinin-A (CCK-A) receptor antagonist devazepide (300 μg/kg) or vehicle before the fed test in a within-subject design. In the result, devazepide injection diminished rats' preference for Late(+) over Early(+), whereas they preferred Late(+) over Late(-) irrespective to drug injection. These results suggest that CCK-mediated signals partially modulates the expression of rats' learned preference for the late-occurring flavor over the early-occurring flavor in a glucose-containing meal.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-302 恐怖記憶消去を制御する脳領野同定による消去機構の組織学的解析 Ryouta Tachibana（立花亮太），Ryouta Tachibana（立花亮太），Kida Satoshi（喜田聡）東京農業大院 Extinction of fear memory is thought to be a therapeutic target for emotional disorders such as post-traumatic stress disorders (PTSD). Conditioned fear memory is thought to be a rodent model of PTSD. Memory consolidation is a process to stabilize short-term labile memory, generating long-term memory. Importantly, the reactivation of conditioned fear memory by re-exposure to the conditioned stimulus (CS) in the absence of the unconditioned stimulus (US) initiates extinction, a new inhibitory learning that weakens fear memory expression. We have shown that extinction memories are consolidated through a molecular mechanism similar to consolidation including the requirement of new gene expression and CREB-mediated transcription in the medial prefrontal cortex (mPFC) and amygdala (Mamiya et al, 2009). In this study, to further understand mechanisms for extinction of contextual fear memory at the systems level, we tried to identify brain regions that regulate contextual fear extinction. To do this, we measured the c-fos expression induced through CREB-mediated transcription when contextual fear memory is extinguished. Mice were trained with a single footshock (0.4 mA, 2 sec) and 24 hours later, re-exposed to the training context for 30 min. c-fos positive cells were counted 90 min after the re-exposure using immunohistochemistry. Consistent with our previous observations, significant increases in c-fos positive cells were observed in the mPFC and amygdala after extinction learning. Importantly, we found a significant increase in c-fos positive cells in lateral habenula. Our result suggests that the lateral habenula play critical roles in fear memory extinction.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-303 海馬神経新生エンハンサーによるニコチン依存性場所嗜好性記憶の忘却促進 Ayaka Minami（南朱夏），Satoshi Kida（喜田聡）東京農業大学大学院 It is important to develop biologically based effective treatments for addiction disorders of drug, alcohol and gambling. From this view, forgetting, erasure and extinction of memory for addiction are candidates of therapeutic targets of these disorders. Addiction memory displays some aspect of an associative memory between pleasure induced by drug (unconditioned stimulus; US) and environmental context (conditioned stimulus; CS). Our previous study showed that neurogenesis enhancers such as treatment with memantine (MEM), an uncompetitive antagonist of the N-methyl-D-aspartate glutamate receptor, promotes forgetting of hippocampus-dependent fear memory in mice (Ishikawa et al., 2016). In this study, we tried to develop a treatment for drug addiction by facilitating forgetting of addiction memory. To do this, we examined the effects of MEM treatment on memory for nicotine addiction using the conditioned place preference (CPP) task that enables to form a place preference memory associated with drug. Mice were exposed to the conditioned chamber consisting of white and black compartments for 20 min per day for 4 consecutive days and were assessed time spent in white and black compartments, respectively (day 1-4; Habituation). Then mice were received systemic injections of vehicle or nicotine (0.3 mg/kg) before a 20 min exposure to white or black compartment, respectively, and exposed to both compartments once a day for 5 consecutive days (day 5-9; Conditioning); white compartment (CS) is associated with nicotine treatment (US). Twenty-four hours after the conditioning at day 9 (day 10), mice were exposed to the conditioned chamber for 20 min and assessed time spent in white and black compartments, respectively (Test). Nicotine group exhibited significantly more time spent in white compartment compared to vehicle group, indicating that nicotine-treated mice formed a nicotine-associated CPP memory. To examine whether increasing in adult hippocampal neurogenesis promotes forgetting of this CPP memory, mice received systemic injections of MEM (50 mg/kg body weight) once a week for 4 weeks. Importantly, MEM-treated nicotine group exhibited significantly less time spent in white compartment compared to vehicle-treated nicotine group. These observations suggest that neurogenesis enhancer promotes forgetting of memory for nicotine addiction. Forgetting of memory for addiction may contribute to develop effective treatments for addiction disorders.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-304 ニワトリヒナの巣外套吻側部は刻印付けの想起に必要である Naoya Aoki（青木直哉），Toshiyuki Fujita（藤田俊之），Shinji Yamaguchi（山口真二），Koichi J Homma（本間光一）帝京大学薬学部 Filial imprinting of domestic chick is a typical example for early learning. In imprinting, newly-hatched chicks follow a moving object and then memorize it. The intermediate medial mesopallium (IMM) is assumed to be the most critical brain area because it is necessary for the acquisition of imprinting. However, when the bilateral IMM regions were lesioned several hours after the imprinting training, lesioned chicks can remember the imprinting object. Therefore, the IMM was thought to be necessary for the short-term retention of imprinting memory. These suggest that the imprinting memory in the IMM is transferred to other brain areas. Recently, we found that intermediate hyperpallium apicale (IMHA) has neural connections from the IMM and that it is one of the brain areas responsible for imprinting recall. Because the imprinting memory was partially remained after the IMHA lesion, other regions can also be responsible for the imprinting recall. In this study, we tried to find the brain region which is necessary for the imprinting recall besides the IMHA. Then, we injected the anterograde tracer (biotinylated dextran amine) into the IMM. As a result, we found massive neural connections from the IMM to the nidopallium dorsocaudale (Ndc), which is thought to be functionally analogous to the prefrontal cortex of mammals. In addition, to examine whether the Ndc is necessary for the imprinting recall, the bilateral Ndc regions were lesioned 24 hours after imprinting training. The preference scores of chicks significantly decreased after the Ndc lesion. We concluded that the Ndc receives the information of imprinting from the IMM, and retain it as a long-term memory.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-305 鳴禽における腹側被蓋野・黒質ニューロンの聴覚応答は社会的文脈に依存した修飾を受ける Shin Yanagihara（柳原真）¹，Maki Ikebuchi（池渕万季）²，Chihiro Mori（森千紘）¹，Ryosuke O Tachibana（橘亮輔）¹，Kazuo Okanoya（岡ノ谷一夫）^1,2 ¹東京大院総合文化研生命環境科学認知行動科学 ²理化学研究所脳神経科学研究センター As in human speech acquisition, social interactions are crucial for vocal learning in songbirds. During development, juvenile zebra finches listen to a song directly from a live tutor, and faithfully imitate the tutor song. However, passive exposures to a tutor song presented from a speaker in the absence of a tutor lead to a poor imitation of the tutor song. These behavioral studies highlight the importance of social interactions with a tutor during vocal learning, but neuronal mechanisms underlying the social facilitations in vocal learning are still not well understood. Here, we hypothesized that social interactions with a tutor is rewarding for juvenile, and hearing a tutor song directly from a live tutor enhances the activity of brain reward circuitry in juvenile leading to faithful imitation of the tutor song. To test this, we examined whether the social interactions with a tutor influence the activity of midbrain ventral tegmental area (VTA) and substantia nigra pars compacta (SNc) neurons in juvenile zebra finches. To this end, we chronically recorded multiple single neuron activity from VTA/SNc in freely-behaving birds. Consistent with the previous electrophysiological studies, a group of VTA/SNc neurons exhibited singing-related or movement-related activity. We also found that another group of VTA/SNc neurons exhibited auditory responses to a tutor song presented through a speaker, and these responses were considerably modulated by social context. Some of these song-responsive neurons exhibited enhanced auditory responses when a juvenile was in the presence of a tutor compared to alone. Furthermore, similar enhanced responses were observed when a juvenile listened to the tutor song directly from a live tutor. These results suggest that a group of VTA/SNc neurons may code social rewards and play a crucial role in social facilitation of vocal learning.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-306 哺乳類外側手綱核 - 正中縫線核神経回路は痕跡恐怖条件付け学習に必要 Taku Sugiyama（杉山拓）¹，Takehisa Handa（半田剛久）¹，Yoshie Ito（伊藤世志江）¹，Tanvir Islam（Islam Tanvir）¹，Akira Uematsu（植松朗）²，Joshua Johansen（Johansen Joshua）²，Hitoshi Okamoto（岡本仁）¹ ¹理化学研究所　脳神経科学研究センター　意思決定回路動態研究チーム ²理化学研究所　脳神経科学研究センター　学習・記憶神経回路研究チーム To survive in the world, it is important for animals to learn to associate external stimulus with consequent results (reward or punishment). Even if there is a considerable time-gap between events, animals can do this. To examine brain mechanisms for associating temporarily separated events, trace conditioning has been used. In trace conditioning, conditioned stimulus (CS) and unconditioned stimulus (US) are temporarily separated by non-stimulus period (trace period). To make CS-US association by trace conditioning, CS information should be maintained by the time US is given. However, it is still unclear how CS information is maintained in the brain. We considered that the lateral habenula (LHb) is a good candidate to maintain CS information, because of the following reasons. First, during aversive conditioning, LHb is activated by both aversive US and CS. Second, LHb neuron indirectly regulates hippocampal activity via median raphe (MnR) serotonergic (5HT) neuron. Hippocampus is a necessary brain region for trace conditioning. Then, we hypothesized that ""the LHb-MnR pathway contributes to CS information maintenance in aversive trace conditioning"". To verify our hypothesis, we used trace fear conditioning (TFC) with optogenetics and in vivo calcium imaging. Using projection specific optogenetics, we inhibited activity of MnR projecting LHb neurons during trace period, and found that CS-US association was impaired. In in vivo calcium imaging, MnR-projecting LHb neurons showed tonic increase of GCaMP6f signal after US exposure. Interestingly, increase of GCaMP6f signal was sustained over 2 min, and GCaMP6f signal level gradually increased as US exposure was repeated. We thought that the tonic increase of GCaMP6f dynamics reflects tonic firings of LHb neurons. Our behavioral and imaging results suggest that sustained activities of MnR projecting LHb neurons contribute to CS information maintenance. In Previous works, it was shown that LHb and hippocampus have inputs from MnR 5HT neurons, and 5HT can increase excitability of these neurons. Taken together with our results and previous reports, we hypothesized that ""Sustained activity of LHb neuron is supported by recurrent inputs from MnR 5HT neurons, and activity of MnR 5HT neurons are maintained by LHb activity. MnR provides 5HT inputs to hippocampus for maintaining CS information"".		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-307 皮質のカルシウム活動は海馬の脳波の状態に依存する Yoshiaki Shinohara（篠原良章）¹，Shinnosuke Koketsu（纐纈真之介）¹，Hajime Hirase（平瀬肇）²，Takatoshi Ueki（植木孝俊）¹ ¹名古屋市立大学医学研究科・統合解剖学講座 ²理化学研究所・神経グリア回路研究チーム Phase-locked synchronization between the hippocampus and cerebral cortex has been reported during working memory task and spatial navigation. However, the spatio-temporal dynamics of the hippocampus and wide cortical areas is largely unknown. To analyze synchronized activity between the hippocampus and various cortical areas, functional imaging methods such as PET and fMRI lack the temporal resolution of neural activity. Owing to the large volume conductance, surface EEG recording cannot precisely identify the location of activity source. Our transgenic mouse line (G7NG817), which expresses the calcium indicator G-CaMP7 in astrocytes and the majority of excitatory neurons in the cortex, enables us to observe the temporal dynamics of cortical calcium in 30Hz-100Hz transcranially. Utilizing this mouse line as an experimental tool, we performed simultaneous recording of hippocampal EEGs and imaging of cortical calcium activities. We found that the cortical calcium dynamics was closely correlated to hippocampal EEG status. Calcium activities observed during theta and non-theta states showed distinct spatial and temporal patterns. Cortical calcium level is higher in wide areas of the cortex during the theta state, and the temporal calcium fluctuation is relatively mild. During non-theta states, basal calcium level is lower, but larger cortical calcium elevations that spread over cortical areas co-occurred with hippocampal ripple oscillations. The cortical calcium transients were observed in various vision-related areas, and co-occurred with ripples showed distinct spatial patterns to each other event of the ripples. The finding implies a rapid and dynamical switching of synchronized activities between the cortex and hippocampus during non-theta states.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-308 Parallel hypothalamic inputs to the hippocampus selectively modulate contextual and social memory Mingxiao Gu（Gu Mingxiao）¹，Shuo Chen（Chen Shuo）¹，Linmeng He（He Linmeng）¹，Weiyi Zheng（Zheng Weiyi）¹，Arthur J.Y. Huang（Huang Arthur J.Y.）¹，Adam Z. Weitemier（Weitemier Adam Z.）¹，Denis Polygalov（Polygalov Denis）¹，Kana Namiki（Namiki Kana）²，Hiroshi Hama（Hama Hiroshi）²，Hiroyuki Hioki（Hioki Hiroyuki）³，Atsushi Miyawaki（Miyawaki Atsushi）²，Thomas J. McHugh（McHugh Thomas J.）¹ ¹Laboratory for Circuit and Behavioral Physiology RIKEN Center for Brain Science ²Laboratory for Cell Function Dynamics, RIKEN Center for Brain Science ³Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine Episodic memory requires converging streams of contextual, social and temporal information along the longitudinal axis of the hippocampus. Work in both rodents and humans suggests that specific types of information preferentially engage distinct hippocampal subregions. For example, the dentate gyrus (DG) is involved in contextual recognition, CA1 is crucial for temporal coding and CA2 is required for social, but not contextual memory. Some of this specialization can be related to local circuit architecture. However, how content-specific modulatory inputs shape this mnemonic process still remains unclear. Here we show in mice that parallel, anatomically distinct projections from the supramammillary nucleus (SuM) of the hypothalamus to the dentate gyrus (DG) and CA2 hippocampal regions modulate memory in a content-specific manner. At the population level, both contextual novelty and social novelty activate the supramammillary nucleus (SuM); however, circuit-based tracing and recordings in a newly developed SuM-Cre transgenic mouse reveal that DG-projecting SuM neurons respond to contextual novelty whereas those targeting CA2 are preferentially engaged by social novelty. Furthermore, inhibition of the SuM-DG pathway leads to deficits in recognizing contextual change while stimulation of the SuM-CA2 projections impairs the expression of social memory. This segregation of social and contextual signals in the hypothalamus suggests bottom-up novelty signaling can flexibly modulate hippocampal memory.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-309 アルツハイマーモデル動物海馬領域からの、測光電極法による電気活動とカルシウム動態の同時記録 Munenori Ono（小野宗範），Harunori Ohmori（大森治紀），Shinji Muramoto（村本進司），Sachiko Yamaki（山木幸子），Fong Xu（許峰），Lanlan Ma（馬蘭蘭），Zubo Wu（呉祖波），Nobuo Kato（加藤伸郎）金沢医大院医生理 It is well known that Alzheimer disease (AD) patients have severe learning disabilities. In the early stage of AD, several studies have suggested that soluble amyloid beta induced the disorder in the neural activity and the intracellular calcium dynamics, which are very likely to be relevant to the learning disability as well as the progression of cell death in AD. However, the neurophysiological disorder in early stage of AD has not been fully elucidated. In this study, to examine the disorder in the neural activity and calcium dynamics in vivo, we utilized a photometric patch electrode (PME, Hirai et al., 2015), with which simultaneous recording of fluorescence and electrical signals were available. Using a PME, we recorded the local field potential (LFP), spike activities, and the signals of fluorescent calcium indicator in the hippocampus, where is closely related to learning ability and spatial cognition. The recordings were performed in dorsal hippocampal CA1 region of model mice of AD (3xTg), with which spatial cognitive defects were confirmed in the early stage of AD. During the PME recording, the mouse was placed on a treadmill to monitor the movement, and head-fixed via a metal rod. The spatial learning ability of the individual animal was assessed by Morris Water Maze (MWM) test in advance of the PME recording. The PME recordings showed that the calcium signal increased when the theta oscillation dominated in LFP during locomotion. The transient rise of the calcium signal was also seen sporadically while the animal was stationary. At the rise of the calcium signal, the fast gamma oscillation in the LFP was observed. In the AD model animals, the rise of the calcium signal was slower than that in the wild type animals, which suggested that the synchronization of the neural activities in the AD model animals might be degraded.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-310 C型ナトリウム利尿ペプチドとその受容体の働きは視覚的刷込み成立に重要である Tomoharu Nakamori（中森智啓），Yurino Chiba（千葉ゆりの），Haruka Ueno（上野晴香），Ami Makita（牧田愛美），Tsugumi Okubo（大久保つぐみ），Kasumi Hirai（平井霞），Hiroko Ohki-Hamazaki（浜崎浩子）北里大一般教育 To understand the mechanism of neural plasticity in early childhood learning, we investigated possible roles of natriuretic peptide (NP) family and their receptors in chick visual imprinting. The NPs include structurally related peptides that have essential roles in body fluid balance and cardiovascular homeostasis. In avians, several structurally related molecules are comprised in the NPs: renal NP (RNP), B-type NP (BNP) and C-type NP (CNP1, CNP3 and CNPP), and three receptors for the NPs have been identified. CNP3 mRNA was restrictedly expressed in the visual Wulst (VW, considered as mammalian visual cortex), which is an essential region for the process of visual imprinting. And we had found that expression of CNP3 in the VW was higher during the critical period (post hatching day 1; P1) of imprinting than after the critical period (P7). Then, we focused on CNP3 as a candidate which is involved in the process of visual imprinting. In this study, we examined the expression and function of its receptors. Two NP receptors, NPR1 and NPR2, were expressed widely in the chick telencephalon, including the VW region in which CNP3 was expressed. Next, we examined whether CNP3 and NPR1, as well as NPR1 and NPR2, co-localize in the VW cells. CNP3-positive cells were negative for NPR1 and resided in proximity to the NPR1-positive cells. Most of the NPR2-positive cells expressed NPR1. And we tested the ability of these peptides to stimulate chicken NPR1 or NPR2 in HEK293 cells expressing either of the receptors. The activation of NPR1 was stronger with CNP3 than with other subtypes of CNP. In the VW, CNP3-expressing cells were negative for NPR1, but they resided in the vicinity of NPR1-expressing cells. Moreover, when we injected CNP3 into the VW or ventricle of P1 chicks brain before imprinting training, it became possible to establish imprinting behavior with a shorter training time than usual. In addition, when the change in expression levels of CNP3 with time passed from imprinting learning was examined, we found that the expression level of CNP3 transiently increased at three hours after training. These results suggest that CNP3 and its receptor NPR1 in the VW may have functional roles in the early learning.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-311 側坐核ドーパミンD2受容体を介する弁別学習 Takeshi Sawada（澤田健）^1,2，Yusuke Iino（飯野祐介）^1,2，Kenji Yamaguchi（山口健治）^1,2，Mio Tajiri（田尻美緒）^1,2，Sho Yagishita（柳下祥）^1,2，Haruo Kasai（河西春郎）^1,2 ¹東京大院医 ²東京大ニューロインテリジェンス国際研究機構 Dopamine 2 receptors (D2R) are densely expressed in the striatum, and related to the symptoms of neuropsychiatric disorders such as schizophrenia. High-affinity binding of dopamine to D2R has implicated that the D2R detect a transient reduction in the dopamine concentration (DA dip). However, the nature of D2R-dependent behaviours which can account for the therapeutic effects of antipsychotic drugs, D2 antagonists, has been elusive. Here, we found in mice that tone-induced reward learning in the nucleus accumbens (NAc) was markedly generalized dependently on D1R, while the over-generalized learning was restricted by discrimination learning. The DA dip was necessary for the discrimination learning, while, to our surprise, not for extinction. To elucidate how such narrow DA dip is detected, we investigated accumbal slice, and found that as short DA dip as 0.4 s induced enlargement of dendritic spines of D2R expressing spiny projection neurons (SPNs) in the presence of an agonist of adenosine A2 (A2A) receptor when they were preceded by two-photon uncaging of caged-glutamate. Inhibition of CaMKII and the A2A receptor reduced both the synaptic plasticity in slice and the discrimination learning in vivo. Finally, we found that discrimination learning was impaired in a psychosis mouse model and was rescued by a D2R antagonist. Thus, our data demonstrate that D1R and D2R cooperatively regulate the generality and specificity of associative learning, and suggest that psychotic symptoms such as delusion is caused by impaired discrimination of environmental cues.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-312 実行順序を考慮した、恐怖条件付け記憶に対する急性断眠ストレスの影響 Miki Hashizume（橋爪幹）¹，Rina Ito（伊藤吏那）¹，Rie Suge（菅理江）²，Yasushi Hojo（北條泰嗣）¹，Takayuki Murakoshi（村越隆之）¹ ¹埼玉医大医　生化学 ²埼玉医大医　生理 Basolateral amygdaloid complex (BLA) is essential for emotional learning. Especially, cued fear conditioning is deeply associated with the BLA while both BLA and hippocampus are essential for the contextual fear memory. Several researches report that enhanced slow oscillatory activity (˜1 Hz) during slow-wave sleep (or NREM sleep) was important for memory consolidation and synaptic plasticity. It is also reported that projection neurons of basolateral nucleus (BL, a ventral part of BLA) showed slow rhythmic activity and burst-fired at a low rate (˜1 Hz) preferentially during slow-wave sleep, suggesting that slow oscillatory activity in BL during REM/NREM sleep may be involved in memory consolidation. Therefore, we investigated whether acute sleep deprivation (SD) affects the maintenance of emotional memory associated with BLA, in relation to our previous studies in which slow inhibitory oscillation (0.1-3 Hz) observed in BL projection neuron was attenuated by acute SD. In the present study, we performed tone-dependent fear conditioning (FC) in juvenile rats (P23-30) in combination with acute SD, thereafter followed by electrophysiological recordings. Rats were conditioned by tone (3 kHz, 10 sec) co-terminated with electrical shock (1 mA, ˜1 sec duration) in the conditioning chamber. Five tone-shock pairings were repeated with 60-100 sec intervals before (FC-SD group) or after (SD-FC group) housing for 3 hours in the water-immersed cage, where the animal was unable to sleep in order to avoid drowning. About 24 hours after conditioning, tone-associated fear memory retrieval was examined by monitoring freezing behavior in a different chamber and contextual fear memory was tested 1 hour later. Then the oscillation activity of BL projection neuron was recorded 2 hours later. The total time of freezing induced by five tones, was significantly shorter in both the SD-FC group and the FC-SD group, than that in control group which was conditioned in the same way without SD. On the other hand, contextual fear memory was not significantly affected, although a slight decrease was observed in SD-FC group. The power of oscillation tended to be decreased in conditioned rats in all the three groups regardless of SD. These results suggest that SD disturb acquisition/consolidation of the emotional memory primarily associated with the amygdala, which could be related with the reduction of the oscillation in BL projection neuron.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-313 ショウジョウバエの嗅覚記憶における価値情報のデコーディング Daisuke Yamazaki（山崎大介），Makoto Hiroi（廣井誠），Takashi Abe（阿部崇志），Kazumichi Shimizu（清水一道），Tetsuya Tabata（多羽田哲也）東京大定量研神経生物 Associative memory is formed by the coupling of conditioned (CS) and unconditioned (US) stimuli. Therefore associative memory is based on the changes of the valences to CSs. In Drosophila odors (CS) and punishment (US) information are integrated in the mushroom bodies (MBs) consisted of Kenyon cells (KCs). KCs are constituted by three major subtypes, α/β, α'/β', and γ neurons in morphological and developmental aspects. We focus on the γ KCs innervated by the dopaminergic neurons (DANs) conveying the various valences. We have shown that they can be further classified by cAMP-responsive element (CRE) activity. CRE-positive (γCRE-p) and -negative (γCRE-n) γ neurons are essential for aversive and appetitive olfactory memory, respectively, and they have the reciprocal antagonistic activity. However, the dynamics of the aversive or appetitive information have not been well understood in KCs. In this study we observed the γ neuron activities by in vivo calcium imaging and found the complemental and cooperative activity between γCRE-p and γCRE-n neurons during memory formation. We show the essential components of their activity patterns required for memory formation through the optogenetic behavioral analysis. These results demonstrate that combining in vivo imaging and optogenetic behavioral analysis would be a good tool to understand the information dynamics involving the feedback network and it is expected to decode the valence information in the fly brain.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-314 Pre-configured Ensembles are Recruited into Upcoming Hippocampal Engram Khaled Ghandour（Ghandour Khaled）^1,2,3，Noriaki Ohkawa（Ohkawa Noriaki）^1,2,3，Chi Chung Alan Fung（Fung Chi Chung Alan）^2,4,12，Hirotaka Asai（Asai Hirotaka）^1,2，Yoshito Saitoh（Saitoh Yoshito）^1,2,3，Takashi Takekawa（Takekawa Takashi）^2,5，Hirofumi Nishizono（Nishizono Hirofumi）^2,6，Masaaki Sato（Sato Masaaki）^4,7,8，Masamichi Ohkura（Ohkura Masamichi）^7,8,11，Junichi Nakai（Nakai Junichi）^7,8，Yasunori Hayashi（Hayashi Yasunori）^8,9,10，Tomoki Fukai（Fukai Tomoki）^2,4,12，Kaoru Inokuchi（Inokuchi Kaoru）^1,2 ¹Graduate School of Innovate life sciences, University of Toyama, Japan ²Japan Science and Technology Agency (JST), CREST, Japan ³JST, PRESTO, University of Toyama, Toyama, Japan. ⁴Okinawa Institute of Science and Technology (OIST) ⁵Faculty of Informatics, Kogakuin University, Japan. ⁶Life Sciences Research Center, University of Toyama, Japan. ⁷Graduate School of Sciences and Engineering, Saitama University, Japan. ⁸Brain Body System Sciences Institute, Saitama University, Japan. ⁹Kyoto University Graduate School of Medicine, Japan. ¹⁰RIKEN Brain Science Institute, Japan. ¹¹Graduate School of Clinical Pharmacy, Kyushu University of Health and Welfare, Japan. ¹²RIKEN Center for Brain Science, Japan. The process of determining the allocation of information to particular neurons and synapses within a neural network is known as the theory of `Memory allocation`. These particular set of neurons are termed engram cells, in which activating these engram cells either by physiological or artificial input can drive the recall of that specific event. However, how memory is allocated to these specific subpopulations of neurons is still poorly understood. Here we show that spontaneously evoked "pre-configured" ensembles formed during pre-learning sleeping periods are highly correlated with those formed during novel experience only in engram cells. A compatible imaging system was established to observe the neuronal activity of CA1 neurons and the labelled engram cells; through a photoconvertible fluorescent protein Kikume Green Red (KikGR). The neuronal activity of hippocampal CA1 neurons was observed, through Ca²⁺ influx with G-CaMP7 in freely-moving animals by miniature head-mount fluorescent microscopy. Our advanced imaging system of engram cells and non-engram cells provides deeper insights into the dynamics of the neural activity across several memory processing stages. Using Non-negative Matrix Factorization (NMF) analysis, engram cells showed that sub-ensembles detected during learning were highly correlated with sub-ensembles found during pre- and post-learning sleep sessions as well as retrieval sessions, however, these correlations were abolished upon exposure to a different context. In contrast, these features were not observed in the non-engram cells. These findings suggest that the hippocampal network is pre-configured into cellular sub-ensembles that could rapidly be used to encode the near future novel experience.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-315 海馬CA3反回回路は学習後の固定化・想起プロセスに関わる Masanori Nomoto（野本真順）¹，Emi Murayama（村山絵美）^1,3，Tatsuya Haga（芳賀達也）^3,7，Noriaki Ohkawa（大川宜昭）^1,3,4，Shin-ichi Muramatsu（村松慎一）^5,6，Tomoki Fukai（深井朋樹）^2,3,7，Kaoru Inokuchi（井ノ口馨）^1,3 ¹富山大学大学院医学薬学研究部（医学）生化学講座 ²沖縄科学技術大学院大学（OIST） ³CREST, JST ⁴さきがけ, JST ⁵自治医科大学医学部, 神経内科 ⁶東京大学医科学研究所, 遺伝子・細胞治療センター ⁷理化学研究所, 脳神経科学研究センター Hippocampus is a major site for multimodal convergence, and critical for memory association between the events. Mathematical simulation models suggest that CA3 recurrent circuit, upstream of CA1, where input information reverberate, is involved in the associative memory processing. However, the role of CA3 in the associative memory processing is still obscure. To clarify this hypothesis, we used the mutant mice specifically lacking the N-methyl-D-aspartate (NMDA) receptor gene in CA3 (CA3-NR1 KO mice) and in vivo CA3 calcium imaging using the combination of a microendoscope nVista and an AAV vector expressing a calcium indicator, G-CaMP. The CA3-NR1 KO mice showed the impairments of long-term, but not short-term, cued-fear memory retrieval in the light fear conditioning (LFC) and auditory fear conditioning (AFC) tasks. Optogenetic silencing of CA3 during cue presentations in the test sessions inhibited long-term memory retrievals in both LFC and AFC tasks. These behavioral results suggest that CA3 is involved in the cued-associative memory processing. To understand the network mechanism that is important for the memory processing, we performed in vivo CA3 or CA1 calcium imaging through the learning-, consolidation-, and -retrieval phases in an LFC task using the CA3-NR1 KO mice. Correlation analysis of transient activity in CA3 showed that KO group showed less correlation activity through consolidation and retrieval phases compared with control group, suggesting that deficit of CA3 NMDA receptor that leads to a deficit of CA3 recurrent plasticity, affected the memory processing after the training (e.g. consolidation and long-term memory retrieval). These behavioral and calcium imaging results strongly suggest that involvement of the CA3 recurrent circuit in the memory consolidation and retrieval. We have also parallelly recorded and analyzed the hippocampal CA1 LFP and mouse behavior during resting state after the learning, to demonstrate the physiological mechanism that involves for the memory processing.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-316 空間記憶固定化における海馬と前帯状皮質の細胞活性 Ayaka Bota（棒田亜耶花）^1,3，Akihiro Goto（後藤明弘）³，Alessandro Luchetti（Luchetti Alessandro）²，Tanvir Islam（Islam Tanvir）²，Masamichi Ohkura（大倉正道）¹，Junichi Nakai（中井淳一）¹，Hajime Hirase（平瀬肇）²，Yasunori Hayashi（林康紀）^1,2,3 ¹埼玉大学理工学研究科 ²理化学研究所　脳神経科学研究センター ³京都大学医学研究科 The memory initially formed in the hippocampus (HPC) is subsequently transferred to other parts of the brain through memory consolidation process. The role of the anterior cingulate cortex (ACC) in this process has been proposed from studies detecting active neurons by in situ hybridization of immediate early gene. However, what information ACC neurons represent and whether it is different from other areas of the brain have not been clear. We thus imaged neuronal activities during a spatial memory task in HPC and ACC. To longitudinally image the Ca2+ dynamics of large populations of neurons in HPC and ACC in freely behaving mice, we combined the chronic miniature microscope system (nVista) and transgenic mice that express a fluorescence calcium indicator protein G-CaMP7. In our spatial memory task, mouse alternatively exposed with two distinct environments (""A(square track)"" → ""B(open field box)"" → ""A""). This task was repeated for 9 days with calcium imaging in HPC or ACC, respectively. We found a small proportion of excitatory neurons (named ""spatial context cell"") in ACC that respond to an entire open field or a large chunk of a running track, which is not seen in hippocampal place cells. These spatial context cell gradually increases on day 9. The delayed appearance of spatial context cells is consistent with their role in consolidation of memory on the context used in the experiment. In addition, we did not observe spatial context cell which are active in both context A and B. Therefore, activity of these cells may distinguish context A versus B. Then, we exposed the animal to a new environment after they went through 9 days of training to examine whether repeated exposure to the same context is required to shape these neurons rather than just running. We used three different square track context, A, B and C. The mice were first trained in context A-B-A-B for 9 days and then exposed to C on day 9 after the last session. The fraction of spatial context cell increased in day 9 for context A and B. In the unfamiliar context C, the fraction of spatial context cell is significantly decreased. This suggest the appearance of spatial context cells depends on formation of specific contextual memory rather than behavior itself. Overall, these results indicate that the formation of spatial context cells occurs context specific way in time course consistent with memory consolidation process.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-317 クラスター型プロトカドヘリン欠損の海馬細胞集団活動における影響 Hirotaka Asai（浅井裕貴）^1,2，Noriaki Ohkawa（大川宜昭）^1,2,3，Yoshito Saitoh（斎藤喜人）^1,2,3，Khaled Ghandour（Ghandour Khaled）^1,2,3，Hirofumi Nishizono（西園啓文）⁴，Mina Matsuo（松尾美奈）⁴，Teruyoshi Hirayama（平山晃斉）^7,8，Shin-ichi Muramatsu（村松慎一）^5,6，Ryosuke Kaneko（金子涼輔）⁹，Takeshi Yagi（八木健）⁷，Kaoru Inokuchi（井ノ口馨）^1,2 ¹富山大医生化学 ²CREST ³JSTさきがけ ⁴富山大　生命科学先端研究支援ユニット　動物実験施設 ⁵自治医科大　神経内科 ⁶東大　医科学　遺伝子・細胞治療センター ⁷大阪大院生命機能　時空生物学　心生物学 ⁸徳島大院医歯薬学　機能解剖 ⁹群馬大院医学　附属生物資源センター Clustered protocadherins (cPcdhs), a large subgroup of adhesion molecules, are important for neural morphology, such as axonal projection and dendrite spread. cPcdhs have two unique characteristics: diversity based on the complex combinations of expressing isoforms in each cells and highly specific interaction with each other. Therefore, cPcdhs are thought to have crucial roles in forming diverse neural circuits. However, little is known whether deletion of cPcdhs affects neural ensemble activity. We hypothesize that cPcdhs configure neural ensembles and then deletion of cPcdhs affect neural ensemble activity. In order to investigate the role of cPcdhs in neural activity, we conducted in vivo Ca²⁺ imaging and immunostaining using cPcdhs-deletion mice. Ca²⁺ imaging in the hippocampal CA1 showed that cPcdhβs-deletion 1) increased the size of cell ensembles synchronously activated, which were extracted with non-negative matrix factorization analysis and 2) reduced repetitive activity, revealed by correlation matrix analysis, during home cage session and novel context exposure session compared to wild type mouse. The large size ensembles were not frequently activated. The expression of cFos induced by novel context exploration was slightly reduced in the hippocampus of cPcdhβs-deletion mice. Thus, deletion of cPcdhβs affects the size of cell ensemble.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-318 嗅周囲皮質/嗅内皮質ネットワーク可塑性への遅延性不活性化カリウム電流の関与：マウス脳スライスの膜電位イメージング Riichi Kajiwara（梶原利一）¹，Yoko Tominaga（冨永洋子）²，Takashi Tominaga（冨永貴志）² ¹明治大理工・電気電子生命 ²徳島文理大香川薬神経科学研 The rhinal cortices, such as the perirhinal cortex (PC) and entorhinal cortex (EC), are located within the bidirectional pathway between the neocortex and the hippocampus. At odd with tracing studies on the pathway, physiological studies indicate that perirhinal transmission of neocortical inputs to the EC occurs with an extremely low probability. Our previous study in rat brain slices indicated that an increase in excitability in deep layers of the PC/EC border initiated the neural activity transfer from the PC to the EC. In the present study, we hypothesized that such change in network dynamics are not incidental observations but rather due to the plastic features of the perirhinal network which links together with the EC. To confirm this idea, we analyzed the network properties of the neural transmission throughout the rhinal cortices and the plastic behavior of the network by performing a single-photon wide-field optical recording technique with a voltage-sensitive dye in mouse brain slices that contained the PC, EC, and hippocampus. We first aimed to seek a condition that we can test the PC-EC neural transmission by partial suppression of γ-aminobutyric acid (GABA) A system. In such condition, the spread of neural activities across the border of the PC/EC was not observed by the electrical stimulation delivered to superficial layers of the PC. Next, we examined the effect of low concentration (40 μM) of 4-aminopyridine (4-AP) on PC-EC transmission, which known to specifically block the slowly decaying K current (D-current). The D-current expressing cells are abundant at deep layers of the PC. The 4-AP enhanced neural activity in the PC, which eventually propagated to the EC via the deep layers of the PC/EC border. Interestingly, washout of 4-AP was unable to reverse the entorhinal activation to the previous state. The change in the network property persisted for more than one hour. This is not limited to the observation caused by the application of 4-AP, because once PC-EC transmission occurred by repeated burst stimulation to the neurons in the perirhinal deep layers without treatment with 4-AP, even stimulation of the superficial layers, which was previously ineffective at initiating PC/EC transmission, allowed for effective neural transmission to the EC. These results indicate the long-lasting enhancement of neural transmission, suggesting the existence of plasticity in the perirhinal-entorhinal network.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-319 前頭前野における異なる神経回路が社会行動と社会的認知記憶を制御する Toshiya Miyabayashi（宮林敏也），Kae Sakurauchi（櫻内華恵），Toshiyuki Tanimizu（谷水俊之），Rie Ishikawa（石川理絵），Satoshi Kida（喜田聡）東京農大院農バイオ Social recognition (SR) memory contributes to discrimination of novelty and familiarity in mice. We have shown that mice form a SR memory when mice are exposed to a novel juvenile mouse for 3 min, but not 1 min. We also identified brain regions required for SR memory; gene expression in the medial prefrontal cortex (mPFC), anterior cingulate cortex (ACC), basolateral amygdala (BLA) and hippocampus (HP) is required for consolidation of SR memory. Furthermore, our in silico network analyses suggested that mPFC and HP are important to generate SR memory, whereas ACC and amygdala are important for coordinating brain activity when social interaction is initiated, by connecting with other brain regions. In this current study, we tried to identify functional neural circuits regulating SR memory and social behavior based on the result of in silico analyses. To do this, we first tried to identify functional neural connections in the mPFC by combining retrograde tracing (retrobeads; RB) with immunohistochemical staining of activity-dependent c-fos expression. Mice were exposed to a juvenile mouse for 3 or 1 min, respectively, whereas control group was not exposed to it. We measured the number of positive cells of RB and/or c-fos. ACC, paraventricular thalamic nucleus (PV), claustrum (Cla) and ventral CA1 showed significantly more c-fos⁺/RB⁺ double positive cells in 3 min group, whereas BLA showed significantly more these double positive cells in 1 and 3 min groups, compared to control group, respectively. We next investigated functional roles of amygdala (AMY)-mPFC and PV-mPFC pathways on social behaviors and recognition memory. To do this, we examined effects of inactivation of AMY-mPFC or PV-mPFC using optogenetics. Mice were expressed archaerhodopsin-T (ArchT) in the AMY or PV using adeno-associated virus. Interestingly, optogenetic inactivation of AMY-mPFC, but not PV-mPFC, during the exposure to juvenile mouse inhibited social behavior. In contrast, this inactivation of PV-mPFC, but not AMY-mPFC, immediately after the exposure blocked the formation of SR memory. These results suggest that AMY-mPFC pathway contributes to expression of social behavior, whereas PV-mPFC pathway are required for the formation of SR memory. Taken together, our findings suggest mPFC regulates both social behaviors and recognition memory formation by receiving functional projection from BLA and PV, respectively.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-320 新生ニューロンの神経回路への組み込み過程と空間記憶・学習における役割 Yusuke Suzuki（鈴木裕輔）¹，Ozgun Mavuk（Mavuk Ozgun）^1,2,3，Takuma Nakagawa（中河拓摩）^1,2，Mayumi Yamada（山田真弓）^1,2,3，Itaru Imayoshi（今吉格）^1,2,3 ¹京都大院生命高次生命 ²京都大生命科学研究科附属生命動態研究センター ³京都大ウイルス・再生研 Neurogenesis continues throughout adulthood in the mammalian brain. In the adult brain, neural stem cells (NSCs) exist only in two regions, the subventricular zone (SVZ) of the lateral ventricle and the subgranular zone (SGZ) of the hippocampal dentate gyrus (DG). It has been reported that NSCs continuously generate new neurons over time and newly born neurons are integrated into the functional networks of the olfactory bulb and the hippocampal dentate gyrus, respectively. To analyze the long-term contribution of new neurons generated by NSCs in the adult brain, we used Ascl1CreERT2 knock-in mouse strain, in which tamoxifen administration induced Cre recombination in NSCs and transient amplifying cells of the adult brain. In vivo lineage tracing with Ascl1CreERT2; R26R-stop-mTFP1 mice showed that the proportion of mTFP1 positive adult-born neurons increased with time and expanded in the granular cell layer of the olfactory bulb and hippocampal DG. Next, we focused on the functional roles of the newly born neurons. There are several evidences that newly born neurons are important for various brain functions. However, it remains unknown how they function to spatial memory formation and retention. In this study, we ablated newly born neurons in the adult brain by using the GFAP-TK transgenic mouse strain. This strain expresses herpes simplex virus thymidine kinase (TK) under the control of GFAP promoter, rendering mitotic NSCs sensitive to the antiviral drug valganciclovir (VGCV). We confirmed a near complete ablation of neurogenesis in the olfactory bulb and hippocampal DG. These VGCV treated GFAP-TK mice were subjected to Barnes maze (BM) test. The BM test is one of the main behavioural tasks used to validate spatial learning and memory, and the novel 3 meter diameter-BM test was launched in order to more precisely capture subtle changes of the behaviours, by slowing-down the learning processing. We found that significantly different spatial exploration strategy between the GFAP-TK and the littermate controls, especially being obvious in situations where slightly different from that in initial spatial memory acquisition. Our ablation study combined with a high sensitivity memory test provides evidence that adult neurogenesis influences the behaviours related to spatial memory formation and retention.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-321 リアルタイムCa²⁺イメージングによる恐怖記憶再固定化時および消去時の海馬神経活性動態 Hotaka Fukushima（福島穂高），Tatsurou Serita（芹田龍郎），Satoshi Kida（喜田聡）東京農大生命科学バイオ Memory retrieval induces two opposite memory processes; reconsolidation and extinction. Reconsolidation maintains or enhances fear responses, whereas extinction weakens them. We have investigated mechanisms of regulation of fear memory after retrieval using inhibitory avoidance (IA) task because this task is thought to allow discriminating reconsolidation and extinction phases at the time point when mice enter into dark from light compartment (LC) (Fukushima et al., 2014). Using this paradigm, we have shown that re-exposure to the LC induces enhancement of IA memory through reconsolidation, while 10 min re-exposure to the dark compartment (DC) induces extinction of IA memory. In this study, to understand mechanisms of transition from reconsolidation phase to extinction phase after retrieval, we examined neural activity dynamics in the hippocampus in reconsolidation (in the LC) and extinction (in the DC) phases of IA memory. To do this, we performed in vivo real-time Ca²⁺ imaging of hippocampal CA1 neurons expressing GCaMP6f in freely moving mice. Mice were trained and 24 hrs later, re-exposed to the LC and then stayed in the DC for 10 min to extinguish IA memory after their entry from the LC (re-exposure). Control group did not receive an electrical footshock during the training but were re-exposed to the light and dark compartment twice (re-exposure and test). We measured the number of Ca²⁺ event in the hippocampal CA1 neuron during the re-exposure and test, respectively. Hippocampal CA1 neurons exhibited comparable Ca²⁺ event frequency in the light and dark compartments during the re-exposure. In contrast, neuron populations showing increased (Up neurons) or decreased (Down neurons) Ca²⁺ event frequencies during the test session compared to the re-exposure session were observed in the light and dark compartments, respectively. Importantly, chi-squared test showed a significant difference in the proportion of Up neurons and Down neurons between extinction group and control group. Interestingly, extinction group showed significantly more Down neurons in the LC during the test compared to control group, suggesting that acquisition and maintenance of fear memory extinction suppressed the activity of some neural populations that is associated with "fear". These observations suggested that the activities of some neural populations are correlated with fear status.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-322 海馬における神経炎症性サイトカインTNFαは恐怖記憶想起を負に制御する Shohei Takahasi（高橋翔平），Hotaka Fukushima（福島穂高），Satoshi Kida（喜田聡）東農大・農・バイオ Pavlovian fear conditioning generates fear memory, reflecting an association between the conditioned stimulus (CS) and unconditioned stimulus (US). This CS-US association is stabilized through gene expression-dependent memory consolidation, generating a long-term fear memory (LTM). Interestingly, brain microglia responses to stress including fear conditioning and generates tumor necrosis factor α (TNFα), a key mediator of neuroinflammation. TNFα has been suggested to induce synapse/spine degeneration and inhibit long term potentiation (LTP). Interestingly, previous studies have suggested that hippocampal TNFα plays critical roles in regulation of contextual fear memory (Yu et al., 2016). However, roles of hippocampal TNFα in fear memory processes remain unclear. Here we examined effects of micro-infusion of TNFα into hippocampus on encoding, consolidation and retrieval of contextual fear memory. Mice were trained with a single footshock (0.4 mA, 2 sec, training) and 24 hours later, assessed freezing responses by the re-exposure to the training context for 3 min (test). Mice showed normal LTM during the test when mice received micro-infusion of TNFα (20 ng/0.5 μl/side) 6 hrs before the training, suggesting that mice micro-infused TNFα show normal encoding and consolidation of contextual fear memory. Interestingly, hippocampal TNFα micro-infusion 6 or 18 hrs, but not 2 or 24 hrs, before the test significantly reduced freezing responses during the test, compared with control group. These observations suggested that hippocampal TNFα micro-infusion impairs retrieval of contextual fear memory with critical time window affecting memory retrieval. To examine effects of TNFα-infusion on memory retrieval at the molecular level, we examined effects of this TNFα micro-infusion on c-fos expression that is regulated in the neural activity-dependent manner and induced in response to memory retrieval. We measured the number of c-fos positive cells at 90 min after the test. Consistent with behavioral observations, mice micro-infused TNFα 6 hrs before test shows significantly less c-fos positive cells in the CA1 area of the hippocampus compared with control group, confirming that micro-infusion of TNFα blocks memory retrieval. Collectively, our findings suggest that hippocampal TNFα negatively regulates fear memory retrieval.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-323 味覚嫌悪学習における島皮質-扁桃体基底外側核間の共同的な神経活動の役割 Konami Abe（阿部こなみ）¹，Marin Kuroda（黒田真鈴）¹，Yosuke Narumi（鳴海陽介）¹，Yiki Kobayashi（小林祐樹）²，Shigeyoshi Itoahra（糸原重美）²，Taiichi Furuichi（古市貞一）¹，Yoshitake Sano（佐野良威）¹ ¹東京理科大学 ²理化学研究所, 脳神経科学研究センター Memory is thought to be stored in a subset of neurons activated during learning. These neurons are reactivated during memory retrieval. In the amygdala and hippocampus, it has been shown that the selection of this subset of neurons is regulated by neural excitability and CREB (cAMP response element binding protein) activity during learning. However, it is not well known about the interaction of cell assembly between brain areas during recruiting and storing a given memory into these neurons. Conditioned taste aversion (CTA) is associative learning in which a taste (such as saccharine; conditioned stimulus [CS]) with the experience of malaise (in this experiment, it was induced by i.p. of LiCl; unconditioned stimulus [US]). It is believed that the insular cortex (IC) and basolateral amygdala (BLA) are required for formation and retrieval of CTA memory. We previously showed CREB levels determine which insular cortical neurons go on to encode a given conditioned taste memory and the functional connectivity between IC and BLA is increased in strong US conditioning group compared with weak US conditioning group. Here, we tested whether neurons with higher excitability levels in the IC and BLA are selectively involved into a CTA memory trace by manipulating neural excitability during conditioning in a subset of neurons and analyzing c-fos expression in those neurons. Consistent with previous findings in auditory fear conditioning, highly excitable neurons during conditioning were preferentially expressed c-fos following CTA memory retrieval in BLA. However, increased neural excitability during learning did not bias c-fos expression following retrieval in IC when neural activity was only manipulated in IC. Interestingly, IC neurons with higher excitability during learning were preferentially expressed c-fos following memory retrieval when both of IC and BLA was manipulated during learning. These findings suggest that coordinated activation of IC and BLA determines which subset of neurons recruit a given memory in the insular cortex.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-324 情動記憶を消去する前頭前野の神経回路動態 Akira Uematsu（植松朗），Tomoya Duenki（でゅんきともや），Joshua P Johansen（じょはんせんじょしゅあ）理化学研究所脳神経科学研究センター Organisms learn to respond emotionally to stimuli that predict unpleasant or rewarding outcomes in order to increase survival rates. When these predicted outcomes are omitted, these emotional memories and their associated behavioral responses are reduced through a process called extinction learning. Previous research indicates that the ventromedial subregion of the prefrontal cortex (vmPFC) is important for extinctions of both appetitive and aversive memories. Anatomically the vmPFC sends broad projections throughout the brain and each set of projecting neurons forms distinct population in vmPFC. However, little is known about whether specific projection-defined population or combinations of them encode aversive and appetitive extinction and whether, and if so, how the neural dynamics in vmPFC cell populations differentiate distinct forms of extinction. To understand these questions, we first performed a series of experiments in which we optogenetically manipulated projection-specific vmPFC cell populations in rats. Animals first learned to associate different auditory cues with either sucrose solution or foot shock and then the auditory cues were presented alone in extinction. During extinction, each projection-specific cell population was optogenetically inhibited and the effect on extinction learning and memory was assayed. We found that each population has unique functional roles in appetitive and/or aversive extinctions. Next, to examine vmPFC neural representations during appetitive and aversive extinctions, we recorded calcium dynamics of vmPFC excitatory neurons with miniature microscope and tracked same neurons during appetitive and aversive extinctions. Our preliminary data show that population activity dynamics in vmPFC differed between appetitive and aversive extinction. Further results will be discussed in the poster.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-325 ショウジョウバエ求愛条件付けに必要な脳神経回路の探索 Shoma Sato（佐藤翔馬），Takaomi Sakai（坂井貴臣）首都大院理工学生命科学 The fruit fly Drosophila melanogaster has been used as a model organism to elucidate the mechanisms of learning and memory. Various behavioral assays for measuring memory have been isolated such as courtship conditioning which is based on male courtship behavior. A male fly usually courts actively toward a female to success copulation. However, a mated female rejects the male courtship to block successful mating. Thus the male is exposed to the stressful condition by pairing with the non-receptive female. After this experience, the male courtship activity is reduced even toward a virgin female. This experience-dependent courtship suppression is considered to be based on learning and memory. Although the neural bases of innate male courtship behaviors have been studied, the neural circuits underlying courtship memory is poorly understood. In this study, to identify the neural bases of courtship memory, we measured neural activities in the brain related to courtship conditioning. In the male fly brain, mushroom bodies (MBs) considered to be the memory center play a crucial role in courtship memory, and the male-specific P1 neuron cluster (P1) is known as the courtship command neuron. Therefore, we hypothesized that the relationship between the MBs and P1 is involved in courtship memory. To monitor activated neurons after by courtship conditioning we employed transcriptional reporter of intracellular Ca²⁺ (TRIC) system. Since TRIC is a binary expression system which transcriptional activity is regulated by calmodulin and a calmodulin target peptide, TRIC induced transcription depends on intracellular Ca²⁺ concentration. We observed TRIC-induced GFP signaling to monitor activated neurons by courtship conditioning. We are going to explain the neural circuit underlying courtship conditioning. In addition, we will show real time neural activities in the identified neural circuit during courtship conditioning by in vivo Ca²⁺ imaging experiments.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-326 一過性中強度運動による海馬CA1を介した記憶固定化促進に関わる脳部位 Koshiro Inoue（井上恒志郎），Akihiko Yamaguchi（山口明彦），Isao Morita（森田勲）北海道医療大 Background and Purpose: Our previous studies in rats demonstrated that an acute moderate exercise (AME) after learning boosts memory consolidation via new protein synthesis in dorsal hippocampal (dHip) CA1. The dHip CA1 receives modulatory inputs from the basolateral amygdala (BLA), ventral tegmental area (VTA) and locus coeruleus (LC) regions, and the activation involves in the protein synthesis dependent memory consolidation. To identify any potential regions mediating the enhanced memory consolidation by AME, this study investigated the AME-specific activation of afferent input to the dHip CA1 using a neural activity marker c-fos and a retrograde tracer Cholera Toxin b subunit (CTb). Methods: Adult male Sprague-Dawley rats were unilateral injected with CTb into the dHip CA1, and were habituated to the treadmill apparatus through the 10-days of running program. On the test day, the rats explored two identical objects for 5 min in an open-field arena to learn the positions. Immediately after the learning, the rats performed 20 min of AME (20 m/min) or rest (no-AME) in treadmill. In the two groups, single or double immunostaining cells for c-fos and CTb were counted in the dHip CA1(no-injection side), BLA, VTA and LC. Results: There was no significant difference in the exploring distance (locomotor activity) and time of objects (learning level) between AME and no-AME groups. By contrast, AME increased the number of total c-fos⁺ cells in dHip CA1. Furthermore, in only LC, AME increased the number of total c-fos⁺ cells and the percentage of c-fos⁺/CTb⁺ cells in total CTb⁺ cells. However, these changes in LC had no significant correlation with the changes of total c-fos⁺ cells in dHip CA1. Conclusion: The results indicate that post-learning AME increases neural activity in LC, and leads to activation of dHip CA1-projecting neurons in LC. Although further detailed analysis is required, the LC might be a crucial region in modulating the AME-induced improvement of memory consolidation.		7月27日（土）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-327 認知の柔軟性における海馬新生ニューロンの役割 Shota Ando（安藤翔太）¹，Junya Koga（古賀淳也）¹，Sayumi Yokota（横田紗弓）¹，Kazumasa Tanaka（田中和正）¹，Thomas J McHugh（Thomas McHugh）²，Tatsuhiro Hisatsune（久恒辰博）¹ ¹東京大学新領域創成科学研究科 ²理化学研究所　脳神経科学研究センター Neurogenesis in the dentate gyrus (DG) of the hippocampus and lateral ventricle occurs throughout life and contributes to a variety of cognitive processes, including context discrimination, forgetting and cognitive flexibility. Interventional methods such as ablating dividing or neural stem cells in the DG with drugs or x-irradiation have been widely used to analyze the function of these hippocampal newborn neurons (NBN), but these approaches can also impair the proliferation of glia and damage surrounding tissue. Here we use transgenic mice in which the tetanus toxin light chain (TeTX) is specifically expressed in mature NBNs (NBN-TeTX mice) to address how the loss of transmission from this selected group of neurons impairs cognition. To evaluate the influence of NBN on cognitive flexibility, we first examined extinction learning in the contextual fear conditioning paradigm. NBN-TeTX and control mice both acquired robust contextual fear memory, however during extinction we observed significantly slower learning in the NBN-TeTX mice compared to controls. Additionally, to test reversal learning we used Morris water maze. Days of acquisition were divided into the initial and then the reversal sessions (the location of platform was changed). Although mice in both groups learned similarly in the initial, a significant deficit in NBN-TeTX mice was observed in the reversal. Furthermore, we performed operant learning to assess flexibility in voluntary behavior. In this test mice were first trained to lick in response to a light cue to receive a water reward (Go task). Following acquisition the mice were then retrained to restrain their lick response during the same light cue (No Go task). In the Go task we did not observe any significant difference between the genotypes, whereas we observed a significant acquisition deficit in NBN-TeTX mice in the No Go task. These results suggest that although initial learning was not affected, reversal learning was suppressed by impairing NBN synaptic transmission. Our behavioral data are consistent with a role of NBNs in cognitive flexibility. Our experiments will test a hypothesis that NBNs in the DG are crucial for hippocampal/OFC interactions underlying these behaviors. Furthermore, in this study, we also investigated the lateral ventricle that was not previously examined in the NBN-TeTX mice of this laboratory.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-329 時計遺伝子periodが発現するLNdsは長期記憶の維持に必要である Yuki Suzuki（鈴木悠希），Yuto Kurata（倉田裕斗），Takaomi Sakai（坂井貴臣）首都大院理工学生命科学 The period (per) gene was originally identified in Drosophila as a circadian clock gene. We have reported that per is also essential for the formation of long-term memory (LTM) (Sakai et al., 2004). There are about 150 per-expressing neurons in the adult brain. Although a neural cluster, ventral lateral neurons (LNvs), including per-expressing neurons acts as the circadian pacemaker neurons, per-expressing neurons involved in LTM are still unclarified. To identify critical per-expressing neurons in LTM, we examined whether electrical silencing and knockdown of per in a part of per-expressing neurons impairs LTM. To induce electrical silencing and per knockdown in a part of per-expressing neurons, the binary gene expression system (GAL4/UAS) was used. In this study, we used several GAL 4 lines with GAL4 expression in LNds. When these GAL4 lines were used, the electrical silencing and per knockdown in GAL4-expressing neurons induces LTM impairment, suggesting that LNds are involved in LTM. To achieve LNd-specific gene expression, two binary gene expression systems (GAL4/UAS and LexA/LexAop) combined with FLP (FLP/FRT) were used. Using this system, we confirmed that LNd-specific electrical silencing also induces LTM defect. Thus, we concluded that the neural activity of LNds is necessary for LTM. Next, we examined whether LNds are involved in consolidation, maintenance, and recall of LTM. Induction of shibire^ts1 in the target neurons can inhibit synaptic transmission in a temperature-dependent manner. F₁ flies between UAS-shibire^ts1 and LNd-specific split-GAL4 line was used in the experiments. We confirmed that the disruption of synaptic transmission in LNds during memory maintenance phase impires LTM. These results indicate that LNds are essential for LTM maintenance.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-330 ショウジョウバエにおける仮想求愛学習法の確立 Tomohito Sato（佐藤智士），Shoma Sato（佐藤翔馬），Takaomi Sakai（坂井貴臣）首都大院理工学生命科学 Animals can modulate their behavior through their learning in nature. The experience-dependent behavioral modification, also called "behavioral plasticity", should be based on the neural plasticity in the brain. Understanding molecular and cellar mechanisms underlying behavioral plasticity is one of the challenging goals in neuroscience. For direct analysis of the learning process, artificially control of the memory is a powerful strategy. If the fixed animals can form memories under microscopy, live imaging of memory process in the brain could be performed. In such view point, Drosophila melanogaster is a nice model organism to establish virtual learning method for memory formation because various genetic information and tools are available for regulating neural activities. Courtship conditioning is one of the traditional behavioral paradigms to measure Drosophila memory. Although virgin females are receptive to courting males, mated females are unreceptive. Conditioned males that have been repeatedly rejected by mated females gradually reduces their courtship activity. However, neural circuits or their operating principles to induce courtship suppression after courtship conditioning is still unclarified. We found that thermogenetical activation of DA1 projection neurons, which relay 11-cis-vaccenyl acetate (cVA) information, causes plastic courtship suppression. cVA is known as the inhibitory sex pheromone emitted from mated females. Thus, it is possible that the experience of continuous cVA inputs induce courtship suppression. To further examine that artificial activation of cVA/DA1 pathway induces courtship suppression, we will show whether the activation of cVA/DA1 pathway by optogenetic tools also induces courtship suppression.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-331 学習による小脳構造マクロ可塑性の分子細胞神経機構の解明研究 Chiaki Sugai（菅井智昭）¹，Yuki Hori（堀祐樹）²，Takasi Hanakawa（花川隆）¹ ¹国立精神・神経セ脳病態統合イメージングセ ²理化学研究所　生命機能科学研究センター Intelligence relies on an ability to encode learned experiences into the long-term memory storage and to retrieve necessary information from the storage on demand. The neurobiological mechanism supporting the encoding is called neuroplasticity. In human studies, it has become clearer that neural plasticity following experiences can be visualized using MRI, which measures macroscopic indices such as gray matter volume and fractional anisotropy. We may call these MRI-based observation as 'macroneuroplasticity'. However, the mechanisms of 'macroneuroplasticity' at the cellular and molecular levels remain completely unknown even though some hypotheses have been proposed such as synaptogenesis, neurogenesis, changes in glial cells, and angiogenesis. Here we investigated the effects of operant conditioning learning on changes in regional gray matter volume in rats using voxel-based morphometry (VBM) analysis of MRI. Long-Evans rats underwent an operant conditioning for three days, and MRIs were acquired before and after the learning. In the learning group (n = 20), the rats were attached with a custom-made head-fixed attachment, and they learned to pull a spout-lever in response to an auditory cue for a water reward. After a rat was fixed to the task system with the attachment, the rat started to learn how to use the spout-lever via an operant conditioning scheme. In the control group (n = 20), rats were also attached with the head-fixed device, but they did not undergo the operant conditioning task. Whole-brain three-dimensional T2-weighted MRIs were obtained before the learning and after three days of the learning for the learning group. The same inter-scan interval was applied to the control group. We performed VBM analysis of rat MRIs with SPM8 for a small animal. A statistical analysis was performed using two-way ANOVA (p < 0.001, uncorrected). The VBM analysis revealed significant volume changes of gray matter in the cerebellum, especially lobule V and VI, in the learning group in comparison with the control group. This cerebellar region was consistent with the part of the cerebellum, which showed increases in glucose metabolism during the same operant behavior as measured with positron emission tomography (Hori et al. Neuroimage 2019). In conclusion, we have established an experimental model to examine biological underpinnings of the macroneuroplasticity. Using this model, we will investigate histological changes after learning in the cerebellum.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-332 新規プロテインキナーゼA結合タンパクAkain1 欠損マウスは痛覚感受性と類似条件の分別に異常を示す Kazuki Fujii（藤井一希）^1,2，Yumie Koshidaka（腰高由美恵）²，Mayumi Adachi（安達真由美）²，Yuko Yanagibashi（柳橋裕子）²，Mina Matsuo（松尾美奈）²，Yasunori Aizawa（相澤康則）³，Keizo Takao（高雄啓三）^1,2 ¹富山大学大学院生命融合科学教育部認知・情動脳科学専攻行動生理学講座 ²富山大学研究推進機構研究推進総合支援センター生命科学先端研究支援ユニット ³東京工業大学生命理工学院 Cyclic AMP (cAMP) -dependent protein kinase (PKA) is a ubiquitous serine/threonine kinase. PKA plays a key role in the signaling of many G protein-coupled receptors through the consequent production of cAMP. The specificity of PKA actions is achieved by controlling its cellular localization through a family of A-kinase anchoring proteins (AKAPs). AKAPs localize PKA to specific intracellular sites and spatially restrict intracellular signaling events. AKAP-PKA interactions control various cellular processes, including the regulation of neuroplasticity. A-kinase anchor inhibitor 1 (Akain1) is a novel PKA-binding protein with a unique function in PKA signaling. Akain1 competes with other AKAPs (e.g., AKAP1 and MAP2) for PKA binding and seems to cancel intracellular localization of PKA. In particular, Akain1 is preferentially expressed in neural tissues. How Akain1 affects brain function and behavioral characteristics, however, is unclear. To elucidate the function of Akain1, we generated Akain1 knockout (KO) mice on the C57BL/6J background using the CRISPR/Cas9 genome editing system. Akain1 KO mice were subjected to a comprehensive battery of behavioral tests. In the hot plate test, the latency to the first hind paw response on the hot plate was significantly longer in Akain1 KO mice than in their wild-type (WT) litter mates, suggesting that Akain1 KO mice have decreased pain sensitivity compared with WT mice. In the pattern separation test, Akain1 KO mice exhibited impaired performance in distinguishing between two similar contexts. In the Barnes maze test, Akain1 KO mice and WT mice learned the fixed escape box position at the same rate. For reversal learning, in which the target was moved to the opposite side of the maze, the Akain1 KO mice and WT mice learned the new escape box position at similar rates. While the time spent investigating the new target was significantly longer than the original target, there was no significant difference in Akain1 KO mice, suggesting that Akain1 KO mice had deficits in behavioral flexibility. Although further investigation is necessary to clarify the underlying mechanisms of the behavioral effects of Akain1 deficiency, these findings suggest that Akain1 has a critical role in pain sensitivity and discrimination of similar contexts.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-333 A hypothalamic novelty hub in the mammalian brain Linmeng He（He Linmeng）^1,3，Shuo Chen（Chen Shuo）¹，Mingxiao Gu（Gu Mingxiao）¹，Weiyi Zheng（Zheng Weiyi）¹，Arthur J.Y. Huang（Huang Arthur J.Y.）¹，Adam Z. Weitemier（Weitemier Adam Z.）¹，Denis Polygalov（Polygalov Denis）¹，Kana Namiki（Namiki Kana）²，Hiroshi Hama（Hama Hiroshi）²，Hiroyuki Hioki（Hioki Hiroyuki）⁴，Atsushi Miyawaki（Miyawaki Atsushi）²，Thomas J. McHugh（McHugh Thomas J.）^1,3 ¹Laboratory for Circuit and Behavioral Physiology, RIKEN Center for Brain Science ²Laboratory for Cell Function Dynamics, RIKEN Center for Brain Science ³Department of Life Sciences, The University of Tokyo ⁴Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine The ability to recognize and encode information incongruous with previous experience is critical for survival in a changing world and novelty signals across the brain can enhance attention, perception and memory. However, novelty can be heterogeneous in type and salience depending on the information that it is driven by. Although the importance of subcortical regions such as the ventral tegmental area and locus coeruleus in broadly signaling novelty has been well established, these diffuse monoaminergic transmitters have yet to be shown to convey specific information regarding the type of environmental changes that drive them. Thus, whether novelty with distinct salience, such as contextual and social novelty, is differently processed and routed in the brain and, if so, via which neural circuits, remain unclear. Here we identify a previously unknown novelty hub in the hypothalamus of mammalian brain-the supramammillary nucleus (SuM). Unique about this surprisingly understudied region is that it not only responses broadly to novel stimuli, but segregates and selectively routes different types of novelty information to discrete subfields of the hippocampus. Taking advantage of precise genetic access enabled by a newly developed SuM-Cre transgenic mouse, we found that novelty channeling from the SuM significantly modifies hippocampal-based memory. This content-specific routing of novelty signals represents a previously unknown mechanism that enables the hypothalamus to flexibly modulate selective components of cognition.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-334 視空間作業記憶課題を遂行中のラットの大脳皮質における広域多点局所電場電位記録と機能的結合性解析 Masato Ohi（大井真人）¹，Yasutaka Honda（本田保貴）¹，Takayuki Hosokawa（細川貴之）^1,2，Shinya Nakamura（中村晋也）¹，Kei Oyama（小山佳）³，Bestmann Sven（Sven Bestmann）⁴，Ken-Ichiro Tsutsui（筒井健一郎）¹ ¹東北大学　大学院　生命科学研究科　脳神経システム分野　宮城県　日本 ²川崎医療福祉大学　医療技術学部　岡山県　日本 ³国立研究開発法人量子科学技術研究開発機構放射線医学総合研究所脳機能イメージング研究部　千葉県　日本 ⁴Department of Clinical and Movement neuroscience, UCL Institute of Neurology, London, UK To elucidate the dynamics of large-scale cortical networks for the working memory function, we performed simultaneous recording of local field potentials (LFPs) in various cortical regions in rats during the performance of a visuospatial working memory task. In a trial, rats were required to lick a spout of the same direction as a visual cue, while the disappearance of the visual cue and the presentation of spouts as response targets were intervened by a delay period lasting for 2 seconds. A correct response was rewarded with 20 microliters of sucrose solution coming out from the chosen spout. The target of LFP recordings were cortical regions such as dorsal and ventral medial prefrontal cortices (dmPFC, vmPFC), anterior and medial parieto-occipital cortices (aPOC, mPOC), primary and secondary visual cortices (V1, V2) and primary motor cortex (M1). Then we conducted Granger causality analysis between every possible combinations of recording points to evaluate changes of functional connectivities during the task performance. As a result, we found a contrasting functional connectivity pattern between the cue presentation period and the delay period. During the cue presentation period, the connectivities in the posterior-anterior direction in the occipital and parietal regions, i.e. V1/V2 to POCs, were prominent, possibly reflecting the visuo-spatial processing being performed in a bottom-up manner. On the contrary, during the delay period, the connectivities in the anterior-posterior direction linking the frontal regions and the parietal or occipital regions, i.e. mPFCs to POCs and V1/V2, were prominent, possibly reflecting the top-down signal to maintain visuospatial information in working memory.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-335 嗅周皮質におけるターゲット情報のダイナミックなコーディング Tomoya Ohnuki（大貫朋哉），Yuma Osako（大迫優真），Yoshio Sakurai（櫻井芳雄），Junya Hirokawa（廣川純也）同志社大院脳科学 Cortical neurons flexibly respond to multiple task-events characterized by distinct computational aspects, such as cue, action, and reward. There are two possible explanatory scales, dynamic population code realizing computational flexibility and single-neuron code associating relevant information across different task-events. The former often emphasizes highly distributed representations across neurons, suggesting that diverse activities of individual neurons are essentially uninterpretable at single-neuron level. On the other hand, the latter highlights subsets of neurons which encode relevant information across different task-events and consider them to be neural representations of higher-order information. These interpretations were independently developed, and it is unclear how neurons contribute to dynamically evolving population states with their individual ability to integrate relevant information across task-events. To address this issue, we analyzed neural activities in the perirhinal cortex. We trained five rats to perform a two-alternative forced-choice task where they chose a target port (left/right) associated with a presented cue to obtain water reward. We used randomly-interleaved visual and olfactory cues to investigate coherent choice-target representations. A time-resolved population analysis revealed that population patterns representing choice targets dynamically evolved through the task. The population encoding patterns became unstable in erroneous trials, indicating that the target code was relevant to successful task performance. Interestingly, the population patterns during the cue and reward periods were substantially reversed and therefore allowed us to efficiently decode the choice targets across the task periods. This decodability largely depended on a subset of neurons characterized by activation across those task-periods. Our results clarify the contribution of flexible but structured single-neuron responses across task events to population dynamics, suggesting their role in linking relevant information represented by different population states.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-336 脳内ネットワークの周波数特異性 Shogo Kajimura（梶村昇吾）^1,2，Jonathan Smallwood（Smallwood Jonathan）³，Kenji Matsumoto（松元健二）⁴ ¹京都工芸繊維大 ²ATR脳情報通信総合研 ³York University, York, UK ⁴玉川大脳研 Brain is organized by multiple networks that consist of distributed brain areas. Well-defined brain network derives from clustering algorithms with resting-state functional connectivity in low-frequency band (< 0.1 Hz). However, recent studies have shown that higher frequency bands (> 0.1 Hz) might also have network organization with frequency-specific properties. Thus, revealing the frequency-specific network structures will deepen understanding of the network-based information processing and provide a novel framework to approach to neural mechanisms of human cognition and pathological conditions. Approaching to the high frequency data need to remove the effect of physiological noises (cardiac/respiratory cycle) and collect data with shorter TR (< 1.0 s) by multiband sequence. In this research, we used the data of 140 participants obtained from an open resource data (NKI-RS) scanned with TR=645ms and have cardiac/respiratory data. In the preprocessing step, we removed the effect of cardiac/respiratory noise and decomposed into multiple frequencies (0.005-0.052Hz, 0.052-0.106Hz, 0.106-0.206Hz, 0.206-0.439Hz) by wavelet transformation algorithm. Then, we applied a clustering algorithm to each frequency band data and detected the most appropriate network structure. Finally, we applied the dynamic causal modeling algorithm for estimating the existence of inner- and inter-frequency effective connection across networks and assessed whether the communication across frequency support human cognition by applying a machine learning algorithm and classifying the phenotypic properties. Application of the clustering algorithm revealed that the appropriate clustering number reduced by increasing frequency, indicating detailed brain network in the lower frequency bands are integrated in the higher frequency bands. In addition, application of the dynamic causal modeling algorithm revealed that inter-frequency connection is more likely to exist in higher cognitive function networks (default mode, central executive) than primary function networks (visual, sensory) and such connections converge in 0.052-0.106Hz. Furthermore, pattern of inter-frequency network connections mainly related to 0.052-0.106Hz successfully classified high/low score of personality trait and temperament scales including IQ. These results show that networks are integrated in higher frequency and networks in 0.052-0.106 Hz are the hub of inter-frequency interaction and important for human cognition.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-337 音楽に合わせた運動により誘発されるポジティブな気分は前頭前野賦活と実行機能向上に関わる：機能的近赤外線分光法研究 Kazuya Suwabe（諏訪部和也）¹，Kazuki Hyodo（兵頭和樹）²，Takemune Fukuie（福家健宗）¹，Genta Ochi（越智元太）¹，Kyeongho Byun（Byun Kyeongho）¹，Kazuki Inagaki（稲垣和希）¹，Yousuke Sakairi（坂入洋右）¹，Hideaki Soya（征矢英昭）¹ ¹筑波大学　体育系　ヒューマン・ハイ・パフォーマンス先端研究センター ²明治安田厚生事業団　体力医学研究所 A great deal of attention has been focused on physical exercise benefits, in terms of improved mental health such as mood and cognitive function. Our recent studies have consistently shown that a single bout of exercise elicits increased task-related brain activation mainly in dorsolateral part of the prefrontal cortex (DLPFC), which results in improved executive function measured by Stroop task performance (Yanagisawa et al., 2010; Byun et al., 2014; Kujach et al., 2018). Since positive affect could modulate the PFC activation, it is tempting to hypothesize that exercise-induced positive mood may facilitate the beneficial effect of exercise on executive function. Here we thus examined the role of the music in the exercise benefits, since it would be a powerful tool to boost one's mood during exercise. Thirty-three young adults performed ten minutes of moderate intensity (50% VO_2peak) pedaling exercise with two experimental conditions; listing to one's favorite music and to beep at steady tempo. Ratings of perceived exertion (RPE) were assessed at the end of the exercises. Participants performed a color-word matching Stroop task (CWST) before and after the exercise sessions. Stroop interference time (the contrast of reaction time for neutral task and incongruent task) was analyzed as an index of executive performance. During the CWST, prefrontal activation was monitored using functional near-infrared spectroscopy (fNIRS). Music decreased RPE and increased positive mood (vitality) compared to metronome control. There was a trend, but not significant, for improvement in Stroop task performance in music condition. There was no significant difference between conditions on prefrontal activation. Correlation analyses revealed significant correlations among increased vitality, shortened Stroop interference time and increased task-related activation in the left-DLPFC and the right-frontopolar area. These results suggest that listening music during moderate exercise may have a facilitatory effect in enhancing prefrontal activation and executive performance, and the effects are influenced by exercise-induced mood changes.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-338 自己組織化マップによる多次元ニューロンデータの解析 Yoshiya Matsuzaka（松坂義哉）東北医科薬科大学神経科学教室 Recent innovation of research methods in neuroscience enabled simultaneous data acquisition from large population of neurons while simultaneously identifying their morphological and immunohistochemical properties as well as functional characteristics. The increasing scale of simultaneously recorded neuronal population and the number of features to analyze have made it impractical to categorize neurons by subjectively set criteria of humans. Application of pattern recognition by artificial neural network would be a viable solution to objectively categorize neurons with diverse anatomical and physiological features. In this study, I used Kohonen's self-organizing map (SOM) to analyze neuronal data recorded from monkeys while they were performing a two choice arm reaching task in which a color cue instructed the monkeys either to reach to or away from the spatial cue which subsequently appeared either on the left or the right of the working space. The neuronal data were recorded from the posterior medial prefrontal cortex (pmPFC), presupplementary and supplementary motor areas (preSMA and SMA, respectively). Each neuron was characterized by eleven features that were defined by their temporal profile of spiking activity and the areas where they were recorded. The multidimensional data of 492 neurons were categorized by the SOM. The SOM successfully identified several clusters among the recorded neurons. Further, the SOM revealed the clusters which were either common across or specific to respective cortical area that would have been unnoticed by conventional methods, clarifying the similarities as well as dissimilarities between the cortical areas examined. These findings indicate that SOM would be a potent tool in analyzing neuronal data having large number of data dimension.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-339 サル眼窩前頭皮質の行動決定における役割 Tsuyoshi Setogawa（瀬戸川剛）¹，Takashi Mizuhiki（水挽貴至）^1,2，Narihisa Matsumoto（松本有央）³，Fumika Akizawa（秋澤文香）²，Ryosuke Kuboki（久保木亮介）²，Barry J Richmond（リッチモンドジェイバリー）⁴，Munetaka Shidara（設楽宗孝）^1,2 ¹筑波大 ²筑波大院　人間総合科学 ³産総研　人間情報研究部門 ⁴Lab. Neuropsychol., NIMH/NIH, Bethesda, MD, USA When faced with having to choose one from some alternatives, animals, including humans, will normally choose more valuable options than less valuable ones. Previous studies have reported that neuronal activities in orbitofrontal cortex (OFC) are related to the subjective values of offered options. Here we studied whether 1) neurons in OFC encode the difference in value between offered options, and 2) there is a causal link between OFC neuronal activity and choice. We trained two monkeys to perform a decision-making task. In this task, two choice targets were presented sequentially at the center of the monitor. Each choice target represented a schedule of 1, 2 or 4 trials of a simple visual color discrimination to be rewarded with 1, 2 or 4 drops of liquid reward after schedule completion. The length and brightness of the choice targets were proportional to the required number of the visual-discrimination trials and the amount of liquid reward, respectively. After the sequential target presentation, the two choice targets simultaneously reappeared, one on each side of the fixation point. The monkey indicated its choice by touching the corresponding bar (left or right) in the primate chair. Following a choice of one target, the chosen reward schedule task was started. The subjective reward values of each choice target were estimated from monkeys' choice behavior by an exponential discounting model. We recorded 256 single neurons from OFC (Monkey P: 137, Monkey H: 119). For 56/256 (21.9%) of the neurons, the neuronal firing during the second target presentation period was significantly correlated with the subtraction between the first target value and the second target value, suggesting that these neuronal responses encode the difference in value between two offered alternatives. Later when the choice occurred, that value difference signal had mostly disappeared (only 12/256; 4.7%) and a signal indicating the chosen value emerged (53/256; 20.7%). To examine the causal relationship between these neural activities and choice, we injected muscimol into a small regions of OFC rich in neurons coding for choice-related values. Inactivating this tissue led the monkeys to choose slower and more likely to choose the less valuable alternative, when the difference in value was small. These results suggest that OFC neurons code for value information that could be used to guide choices, and these signals have a direct influence on the choice.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-340 内部モデルに基づいた意思決定とマウス前頭皮質の神経表現 Kosuke Hamaguchi（濱口航介），Dai Watanabe（渡邉大）京都大院医生体情報 The representation of the world in the brain is called "internal model" and is thought to play an important role in decision making. For a foraging animal, knowing that a resource (i.e. food) is limited would be crucial, because, after consumption, one could quickly choose the next action to find the other resources. To study the influence of internal model in decision making, we trained head-fixed mice to conduct a two-choice foraging task with limited resources. Within each trial block, the rewarding action (lick left/right) is fixed. Once a fixed amount of water reward was delivered, the block ends and rewarding action is reversed. An efficient strategy to solve this task is win-stay, lose-shift; stick to the rewarding side until failed. However, well trained mice developed intriguing behaviors that seem to predict the end of the block, such as predictive reversal near the end of the block, or reversal without failure. This suggests that the mice understood the structure of the task. To study the neural representation of the internal model guided decision making, we conducted two-photon calcium imaging from the frontal cortex of the mice which includes the areas involved in orofacial motor control. We found that a subset of neurons displayed choice and reward specific ramping activity that increased its activity toward the movement initiation. Furthermore, the amplitude of ramping activity changes as a function of reward count within the block. A large fraction of ramping neurons decreased its activity as the animal collected more rewards. This is consistent with the dynamics of action value computed with an internal model of limited resources. Our data indicates that ramping activity in the mouse frontal cortex encode the variables of internal-model guided decision making process.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-341 前頭前野に投射しているﾓﾉｱﾐﾝ神経細胞が急性ｽﾄﾚｽ直後の意思決定課題に関与する Natsuko Kubota（久保田夏子），Maina Ishida（石田舞奈），Seiichiro Amemiya（雨宮誠一朗），Takeshi Nishijima（西島壮），Ichiro Kita（北一郎）首都大院人間健康行動生理 Extensive evidences have indicated that decision-making is modulated by acute stress, but the strategy of decision-making to acute stress immediately before a learned task and its neural mechanisms are not well understood. In decision-making, the prefrontal cortex (PFC) plays a key role in optimizing decision-making strategies, including attention, motivation, flexibility, impulsivity, and exploration. Additionally, several studies have suggested that brain monoaminergic system that originates from the brainstem and projects to the PFC can affect the strategies in decision-making processing, and is known to be sensitive to various stressors. Thus, it is possible that acute stress immediately before a learned task, which is considered as familiar choice situation, may modulate the decision-making processing via activation of monoaminergic neurons projecting to the PFC. Here, we examined the involvement of monoaminergic system projecting to the PFC in a learned choice task immediately after acute stress in rats, using observation of behavior and immunohistochemistry for c-Fos and Fluoro-Gold (FG, a retrograde tracer). FG was injected into the PFC before the experiment. T-maze choice task (3 pellets vs. 1 pellet) was conducted on two consecutive days to make it the learned task, and the high-reward side was constant throughout two days and the test day (i.e., day 3). In the test day, half of the rats received mile restraint stress for 30 min (the other group left in the normal cage as the control) immediately before the test session of T-maze choice task. In the test session, we recorded the number of VTE (vicarious trial and error) and the choice time at the T-shaped choice area, and transit time from the choice area to reward area, as well as the number of choice of high-reward side (CHR). Although acute restraint stress did not affect the CHR, the number of VTE and choice time were increased after the acute stress. In addition, the expression of c-Fos in the FG-positive neurons in the dorsal raphe and the locus coeruleus, which are the major sources of serotonin and noradrenalin neurons respectively, was enhanced during the choice task after acute stress. These results indicate that the strategy of decision-making on the learned choice task immediately after acute stress may be affected via activation of monoaminergic system projecting to the PFC, especially serotonergic and noradrenergic systems, suggesting enhancement of deliberation by acute stress.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-342 DREADDによる、報酬を基にした柔軟な意思決定におけるサル前頭眼窩野の役割の解明 Kei Oyama（小山佳）¹，Yukiko Hori（堀由紀子）¹，Yuji Nagai（永井裕司）¹，Toshiyuki Hirabayashi（平林敏行）¹，Naohisa Miyakawa（宮川尚久）¹，Atsushi Fujimoto（藤本敦）¹，Koki Mimura（三村喬生）¹，Ken-ichi Inoue（井上謙一）²，Mark AG Eldridge（Eldridge AG Mark）³，Richard C Saunders（Saunders C Richard）³，Tetsuya Suhara（須原哲也）¹，Masahiko Takada（高田昌彦）²，Makoto Higuchi（樋口真人）¹，Barry J Richmond（Richmond J Barry）³，Takafumi Minamimoto（南本敬史）¹ ¹量研機構放医研　脳機能イメージング研究部 ²京都大　霊長研統合脳システム ³Lab Neuropsychol, NIMH, Bethesda, USA To survive in a dynamic world, animals must rapidly adapt to novel environments and optimize their behavior to obtain rewards. The primate orbitofrontal cortex (OFC) is thought to contribute to reward-based adaptive decision-making. However, a causal relationship has not yet been established, mainly because the reversible inactivation of large and non-continuous areas of the brain, like the bilateral OFC, is technically challenging in non-human primates. In this study, we applied the chemogenetic technique, Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) to reversibly inactivate OFC in macaque monkeys. Three monkeys underwent open surgery to receive multiple injections of an adeno-associated viral vectors expressing a silencing DREADD, hM4Di, into OFC bilaterally. Several weeks after the injections, we performed a positron emission tomography (PET) scan with a DREADD-selective ligand, [11C]deschloroclozapine (DCZ) and confirmed in vivo that hM4Di expression covered OFC bilaterally as intended. Reward-based adaptive decision-making was tested with a modified reversal learning task in which the monkeys were required to choose either of two visual stimuli presented on a computer screen. On each day, five new visual stimuli were paired with 1, 2, 3, 4, or 5 drops of juice. In each trial, two stimuli were randomly selected from the five. To maximize the rewards, the monkeys had to choose a better option by learning the session-specific stimulus-reward associations (learning phase). After the performance was stabilized (optimal choice >80%), the stimulus-reward associations were reversed (S1-R5, S2-R4...S5-R1). Now to maximize the reward, the monkeys had to select the previously inferior stimuli (reversal phase). After several months of training, the monkeys learned to choose a better option in both the learning and reversal phases within several tens of trials in each day's session. Intramuscular injection of the novel DREADD agonist, DCZ, did not impair initial learning but was followed by impaired performance during the reversal in all three monkeys. This impairment is attributed to the OFC-DREADD inactivation, because the injection of DCZ alone did not affect the performance in a control monkey without DREADD expression. These results suggest that the primate OFC plays a critical role in reward-based adaptive decision-making when a situation requires overriding of the previously established associations.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-343 交尾意思を決定するショウジョウバエ雌のフィードフォワード神経回路 Hiroshi Ishimoto（石元広志）¹，Azusa Kamikouchi（上川内あづさ）² ¹名古屋大, 大学院理学研究科, 栄養神経科学講座 ²名古屋大, 大学院理学研究科, 生命理学専攻 Females make decision whether to accept or reject the courting male during a series of courtship ritual. This process is widely appeared in many animal species, however, little is known about the neuronal and molecular mechanisms underlying it. Female mating behavior of fruit fly, Drosophila melanogaster, is one of the behavioral models in which to elucidate how the brain controls the mating process at the molecular, cellular, and neural circuit levels. At the early phase of courtship ritual, female flies presumably discriminate the encounter and then proceed to further valuation of the potential mating partner. As the first reaction of this process, female flies escape, fend, and kick away the courting male fly to acutely avoid unfavorable mating. These pre-mating processes of the female flies suggest the existence of the neural circuits in the female brain that regulate behavioral switching from courtship rejection to acceptance. Here, we explored the neuronal nature of the behavioral switching of the female pre-mating behavior, by applying advantages of molecular-genetics of Drosophila melanogaster. To identify responsible neurons for the pre-mating behavior of virgin female, we suppressed the activity of target neurons in female flies by using Gal4/UAS system and examined the behavior by quantitatively detecting fly copulation semi-automatically with a computer program. We found that suppression of a sub-cluster of dopaminergic (DA) neurons significantly increased copulation rate and hastened copulation latency. This DA neurons connect with a downstream neurons forming a feedforward circuit that was divided into at least two structural and functional classes. A class of cholinergic neurons positively modulates courtship rejection and the other class, composed by GABAergic neurons, negatively modulates the cholinergic neurons. This recurrent circuit forms a feed-forward motif. These findings provide perspectives on how the pre-mating response of virgin females controlled at the neural circuit level.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-344 リスク選択における報酬期待との統合処理に関与するサル腹外側前頭前野の役割 Ryo Sasaki（佐々木亮）¹，Norihiro Takakuwa（高桑徳宏）¹，Tadashi Isa（伊佐正）^1,2 ¹京都大学大学院医学研究科 ²京都大学ヒト生物学高等研究拠点 One would need to flexibly take risky option efficiently depending on the returns, although it is well known that we do not prefer taking too much risk. Recent studies showed that many brain areas represent positive and/or negative reward-related parameters like expected value, reward probability, size, timing and risk etc. However little is known about where and how those parameters are integrated to make a decision. To answer this question, we investigated how the animals (Macaque monkey) handle their risky choice in the situation whether they might get reward with high risk and high return or low risk and low return condition. We defined the risks as the reward probability and the returns as the reward size. We also compared this in the condition with different size of expected values to see how the expected value affects monkeys' risk seeking. Once the monkey fixated his gaze on the fixation point, two targets were presented simultaneously at horizontally symmetrical positions relative to the fixation point. The monkey was required to choose one of the two targets by a saccade to get reward in certain reward probability and expected value assigned to the color of the selected target. The monkey successfully learned the assignment of the color to the combination of expected value and the risk/return. We found that monkey preferred risky choice as default mode which is consistent with other previous studies. Interestingly, we also found that the monkey took more risk when the expected value was relatively small. These results suggest that the monkey makes a choice by integrating both risk and expected value. We next investigated the causal role of a variety of brain areas in the behavioral choices by reversible inactivation with microinjection of a GABAA receptor agonist, muscimol. We were careful to identify the target area and successfully localized it using MRI image injecting the gadolinium in advance. When muscimol was injected into bilateral ventrolateral prefrontal cortex (vlPFC) while the monkey was performing the task, the sensitivity to risky choice was gradually weakened over time, although the sensitivity to expected value was remained to be consistent. Interestingly, the interaction between risk and expected value was also weakened. Our results suggest that the integration of risk/return and expected value might be accomplished in vlPFC. We will also discuss about the role of the other brain areas such as OFC, NAc and ACC.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-346 自己開始運動に先行する自発的脳活動 Honami Sakata（酒多穂波）¹，Kosuke Itoh（伊藤浩介）¹，Yuji Suzuki（鈴木雄治）¹，Katsuki Nakamura（中村克樹）²，Masaki Watanabe（渡辺将樹）¹，Hironaka Igarashi（五十嵐博中）¹，Tsutomu Nakada（中田力）¹ ¹新潟大脳研 ²京都大霊長研 An action can be triggered by an external stimulus input, and can also be initiated endogenously in the brain without any external stimulus; the neural mechanisms responsible for the latter type of action remain poorly understood. In this study, we investigated the neural mechanisms underlying self-initiated movements as an example of endogenously generated actions. Self-initiated movement is a movement initiated at one's free timing, and it is preceded by readiness potential (RP) on electroencephalogram. RP is the gradual buildup of negative potential, starting from ≧1 s before a self-initiated movement. Recently, RP was described using an accumulator model, which proposes that stochastic neural activities that occur spontaneously in the absence of stimulus inputs accumulate over a period of time until reaching a threshold, at which point an action occurs. According to this model, the slope of RP should reflect the rate of increase of spontaneous neural activities. Experiment 1 confirmed this prediction by showing that the RP slope and time interval of each movement (waiting time, WT) are negatively correlated. Experiment 2 sought to identify the brain regions wherein this accumulation of increased neural activities occurs. We measured and compared the brain activities of subjects under 2 different conditions i.e., while performing self-initiated movements and during externally triggered movements using functional magnetic resonance imaging. Self-initiated movements were preceded by a gradual increase in the brain activities in the right fronto-parietal areas, precuneus, insula, visual cortex, and auditory cortex, as well as in the supplementary motor area. This finding suggests that spontaneous activities in multiple brain regions, including sensory cortices, such as visual cortex, are responsible for the occurrence of self-initiated movements. Finally, Experiment 3 examined if interferences in the increasing brain activities influences actions. We observed that the presentation of visual stimuli during the waiting time for self-initiated movements prolongs the WT. Thus, pre-movement accumulation of neural activities in distributed cortical areas, including visual cortex, was likely to be causally related to self-initiated movements. In conclusion, pre-movement increase in spontaneous neural activities in distributed brain regions, as described by the accumulator model, is responsible for the generation of self-initiated movements.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-347 ショウジョウバエにおいてセロトニン作動性ニューロンが聴覚応答行動時間を制御する Nao Morimoto（森本菜央）^1,2，Ryosuke F Takeuchi（竹内 F 遼介）¹，Ayano Ishida（石田彩乃）¹，Azusa Kamikouchi（上川内あづさ）³，Fumitaka Osakada（小坂田文隆）^1,2,4,5 ¹名古屋大院創薬 ²名古屋大高等研究院 ³名古屋大院理 ⁴名古屋大未来機構ナノライフ研 ⁵JSTさきがけ・CREST Animals control their behavioral output in response to external sensory cues depending on their values. Such behavioral response is controlled under multiple time scales but its underlying neural mechanisms remain unclear. We explored these mechanisms by using the fruit fly Drosophila melanogaster as a model. In flies, the courtship songs produced by males not only seduce female flies, but also increase the mating drive of males, leading to the typical chasing behavior. This behavior of males has long been known as auditory behavioral responses, in which song stimulation induces immediate chasing behavior and its gradual decrease over time. In our previous studies we found that combination of female and song stimulation suppressed the behavioral decrease, suggesting that the chasing behavior can be modulated in a value-dependent manner. Here we aim to reveal how the temporal dynamics of the chasing behavior were modulated over time. To characterize the molecular factor of the temporal dynamics change, we have performed an RNAi screen using pan-neuronal GAL4 driver and found that knocking down of 5-HT2BR disrupted the gradual decrease of the chasing behavior. This result suggests that serotonin is involved in controlling the temporal dynamics of the chasing behavior. The fly brain has about 90 serotonergic neurons, which are divided into 10 clusters. To determine which serotonin clusters are responsible to the temporal dynamics change, we ablated each serotonin cluster by the conditional expression of Kir2.1 or by the TrH RNAi tool. Ablation of one cluster accelerated the decrease of auditory behavior without changing the amount of the onset behavioral response. These results indicate that this serotonergic cluster is responsible for keeping the behavior, raising the possibility that this serotonergic cluster codes the values of the external cues.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-348 タスクスイッチ中のサル前頭前野情報表現の解読 Hironori Kumano（熊野弘紀）¹，Keisuke Kawasaki（川嵜圭祐）²，Takafumi Suzuki（鈴木隆文）³，Isao Hasegawa（長谷川功）²，Takanori Uka（宇賀貴紀）¹ ¹山梨大院医統合生理 ²新潟大院医神経生理 ³情報通信研脳情報通信融合研究センター Switching behavior based on multiple rules is a fundamental ability to behave properly according to context. We have previously shown that neurons in the lateral parietal area (LIP) integrate relevant information preferentially depending on context. We hypothesized that the prefrontal cortex (PFC) control the speed of integration depending on context. In this study, we examined neural activity from a wide range of PFC by recording electrocorticogram (ECoG) signals while a monkey performed the reaction time task switching paradigm. A Japanese macaque (Macaque fuscata) was trained to switch between a direction discrimination task and a depth discrimination task. On a given trial, the color of the fixation point (FP) indicated whether the monkey had to discriminate motion direction (upward or downward) or stereoscopic depth (far or near). The monkey was allowed to make a saccade as soon as a decision was made. Difficulty of the tasks was varied by changing the percentage of coherently moving and binocularly correlated dots in the visual stimulus. We first customized ECoG electrodes that can cover all surfaces of the PFC including the lateral PFC (64 channels), the medial PFC (32 channels), and the orbitofrontal cortex (OFC; 32 channels). During the task, visual response and saccade related response were observed in various areas within the PFC. We examined whether task rule can be decoded from the event related potentials (ERPs). We used a linear Support-Vector-Machine (SVM) to decode task rule by classifying the ERP in a 20-ms window that slid every 10ms from 100 ms before FP onset to 400 ms after FP onset. We found that ERP in all areas of the PFC encoded the task rule (significantly above the pre-FP period). Peak classification accuracy was observed as early as 80 ms after the FP onset in the OFC. Medial PFC exhibited peak accuracy at around 100 ms after the FP onset. The peak accuracy in the lateral PFC was delayed compared to other areas and was observed at 200 - 300 ms after the FP onset. We suggest that representation of task rule is first encoded within the OFC and propagates in wide areas of the PFC.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-349 Free finger tapping tempo reflects intrinsic brain rhythms (2): assessed by resting-state fMRI Muneyoshi Takahashi（高橋宗良）¹，Takayuki Fujii（藤井貴之）¹，Sai Sun（Sun Sai）²，Atsushi Miyazaki（宮崎淳）¹，Takuya Akashi（明石卓也）³，Morimichi Furudate（古舘守通）⁴，Chao Zhang（Zhang Chao）⁵，Toru Ishihara（石原暢）¹，Hiroki Tanaka（田中大貴）¹，Haruto Takagishi（高岸治人）¹，Shinsuke Shimojo（下條信輔）^1,2，Tetsuya Matsuda（松田哲也）¹ ¹玉川大脳研 ²Div Biol & Biol Eng/Comp & Neural Sys, Caltech, Pasadena, CA, USA ³岩手大工 ⁴岩手大技術部 ⁵福井大工 The mean rate at which a person repeats a simple movement is called a personal tempo. A number of studies have shown that its variability within individual is quite smaller than that across individuals. Thus, the personal tempo is often regarded to represent his/her intrinsic characteristics in relation to their personal traits and communication skills. In terms of neural correlates, a functionally connected structure known as the default mode network (DMN) allegedly provide a fundamental rhythm especially during the resting state. Its irregularity is often observed in psychiatric and/or communication disorders such as schizophrenia, depression, and autism. The aim of this study is to explore a link between our physical personal tempo and the intrinsic fluctuations in the brain activity. A total of 37 university students were participated. Resting-state functional MRI was conducted using Siemens 3T-MRI scanner. We then employed three types of the metronome-guided finger tapping task outside the scanner. Each task type consisted of (1)one-minute Forced phase, in which the participant required to tap a sensor device as accurately as possible following the metronome, followed by (2)two-minute Free phase, in which they needed to keep tapping but at their own pace without any guiding tone from the metronome. We gave 7.0 Hz in the fast condition and 0.25 Hz in the slow condition, in the phase (1). In the natural condition, we presented a tempo that each participant felt natural for tapping. This natural tempo was measured ahead of time, asking the participant to tap with a rhythm that they feel natural for 10 seconds. Using Group ICA on the resting-state fMRI data (Smith et al., 2014), we extracted DMN and calculated the fractional of amplitude of low frequency fluctuations (fALFF) as a measure of spontaneous activation level (Zou et al., 2008). As a result of cross correlation analyses between fALFF and the mean tapping frequencies among three conditions, we found that there was significant positive correlation exclusively in their natural condition. Spontaneous activation level of DMN during the rest is known to increase by undirected attentional fluctuations such as mind-wandering, including in the case of ADHD. The present findings, therefore, imply that the intrinsic physical rhythms may represent a level of attentional instability during rest, reflecting activation level of DMN, and that free tapping can provide a quick assessment as such.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-350 The relationship between implicit leadership in interpersonal rhythmic interaction and trust in a partner Takayuki Fujii（藤井貴之）¹，Muneyoshi Takahashi（高橋宗良）¹，Sai Sun（Sun Sai）²，Atsushi Miyazaki（宮崎淳）¹，Toru Ishihara（石原暢）¹，Hiroki Tanaka（田中大貴）¹，Haruto Takagishi（高岸治人）¹，Shinsuke Shimojo（下條信輔）²，Tetsuya Matsuda（松田哲也）¹ ¹玉川大脳研 ²Div Biol & Biol Eng/Comp & Neural Sys, Caltech, Pasadena, CA, USA Interpersonal rhythmic interaction creates a pleasant feeling and an asymmetric EEG synchronization between leader and follower (Yun et al., 2012), suggesting that these dynamic factors strengthen social bonds. Most studies of social interaction focus on explicit interpersonal relationship (e.g., leader and follower) defined by the experimenter. However, such relationships often emerge unconsciously in a self-organized manner in our daily life. The psychological effect of these implicit interpersonal relations on social decision making has not been made clear. This study aims to investigate the influence of implicit interpersonal relationship on decision making regarding trust in partners, which is the fundamental element of social bonds. A total of 44 pairs of university students participated. A cooperative finger-tapping task was carried out by two participants of the same sex. Participants were instructed to tap as synchronously as they could (cooperative finger-tapping task). The participants' finger movements during the cooperative finger-tapping task (60 sec) were recorded by markers placed on top of the index finger using motion-capture cameras (VICON). We measured the tapping lag of the participants from the movement of the index finger. A tap prior to that of the partner was labelled as ""lead"" while a tap after the partner was labelled as ""follow."" We found that such tapping leadership sometimes reversed but still showed a bias within a 60-sec session. We calculated the lead rate of the tapping within each session. Immediately after the tapping task, the participants played a trust game individually in a private booth. In the trust game, participants were asked to indicate the amount of money they would transfer to the partner. We used the offer amount in the game as an indicator of trust in the partners. The majority (>80 % approx.) of tapping lags between two participants were within 50 msec. This is mostly below the time-lag detection threshold, and participants did not pay attention, thus the leader-follower relationship was mostly implicit. Yet, the higher the lead rate was, the lower the offer amount was (i.e., a negative correlation). This trend could not reach statistical significance when participants played the trust game with an anonymous player. Our results suggest that implicit lead/follow relationships may have a critical link to explicit decision making regarding partners.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-351 Free finger tapping tempo reflects intrinsic brain rhythms (1): assessed by EEG Sai OF Sun（Sun Sai OF）¹，Sai Sun（Sun Sai）¹，Daw-An Wu（Wu Daw-An）¹，Takuya Akashi（Akashi Takuya）²，Morimichi Furudate（Furudate Morimichi）³，Chao Zhang（Zhang Chao）⁴，Tetsuya Matsuda（Matsuda Tetsuya）⁵，Muneyoshi Takahashi（Takahashi Muneyoshi）⁵，Shinsuke Shimojo（Shimojo Shinsuke）^1,5 ¹Division of Biology and Biological Engineering/Computation and Neural Systems, California Institute of Technology, Pasadena, CA, USA ²Faculty of Science and Engineering, Iwate University, Morioka, Japan ³Division of Technical Support, Iwate University, Morioka, Japan ⁴School of Engineering, University of Fukui, Fukui, Japan ⁵Tamagawa University Brain Science Institute, Tokyo, Japan Each individual has his/her own tempo in walking and other activities, preference for music, etc. Such preference could be linked with intrinsic/spontaneous brain rhythms, cognitive abilities and personality traits/life styles. Spontaneous brain rhythms may play a crucial role by providing endogenous constraints to sensory-, cognitive-, or motor-driven activity (Fries et al., 2001). Studies have documented intrinsic brain activity during resting wakefulness with several brain rhythms and a large-scale brain network involved (Mantini et al.,2007). We ask whether endogenous brain rhythms during resting wakefulness are correlated with (or perhaps drive) the spontaneous tempo of simple movements (such as finger tapping). Using a new design of spontaneous finger tapping task combined with EEG, we aimed: 1) to develop a new method of testing spontaneous movements, and evaluate its reliability and sensitivity; 2) to examine the neural correlates of spontaneous movements, 3) to evaluate the relationship between resting wakefulness and free tapping. 23 participants tapped using their fastest/slowest/natural tempo with their eyes closed. At the one-minute mark, a beep signaled that they could tap freely for the following two minutes. An Arduino tapping device (1000Hz sampling rate) was developed to record the time-points of tapping. The natural condition is always performed after counterbalanced fastest/slowest conditions to avoid priming effects. This paradigm was designed to obtain one's own intrinsic tempo, and to examine its robustness. Results can be summarized:1) After the beep, the natural free tapping did not change its tempo, but the fastest and the slowest tapping merged into the natural or nearby tempo. 2) Natural tapping had smallest variance relative to the other two conditions. 3) Naturally tapping yielded reduced motor evoked potentials (MEPs) and modulation, suggesting less efforts or conflicts. 4) Natural tapping yielded reduced theta peak frequency and peak amplitude, also indicating the specialty of intrinsic rhythms with less efforts, conflicts, or cognitive control. 5) An enhanced default mode network (DMN) connectivity was found only during natural tapping. Altogether, the natural tapping involves less mental efforts or cognitive control, thus possibly related to DMN activity. Our behavioral and EEG paradigm provide a simple and objective diagnostic tool for various mental status, including earlier Parkinson's Disease and ADHD.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-352 コモンマーモセットの視覚探索課題の開発 Takuro Ikeda（池田琢朗），Katsuki Nakamura（中村克樹）京都大学霊長類研究所高次脳機能分野 The common marmosets (Callithrix Jacchus) are becoming an important model for both basic and clinical neuroscience. Their brain structure, which is similar to human, and their relatively fast and high reproduction rate make them a good model for neuroscience research combined with genetic engineering techniques. However, unlike humans and macaque monkeys, there are very few experimental tasks for marmosets to examine their cognitive functions. We arranged a typical visuomotor task for human/macaque monkey and developed a new task for marmoset. This visual oddball search task requires marmosets to search an oddball target among other distractors. Two marmosets (male, 6yo 410g and 7yo 360g) were trained for the task. The marmosets were seated in a monkey chair with their head fixed and the eye position was recorded at 1000 Hz during experiment (Eyelink, SR Research). Each trial started with presentation of a central fixation spot. After 400 ms of fixation, a stimulus array consisted of six objects (one target and five distractors) was presented. The five distractors were same while a target was different from distractors in one or two visual features (color, orientation, or spatial frequency). The monkeys were required to select the target within 1000-1500 ms by eye movement, although it got more reward if it could select the target faster. The task is relatively simple and works well for marmosets. Overall performance was significantly better than chance (percent correct > 80%), and the marmosets could select the target with single saccade in most of the time. The average reaction time of the first saccade was ~210 ms, which is similar to those of humans and macaque monkeys. The current task condition focus on simple visual attention, possibly mediated by bottom-up mechanisms. However, this task allows us to test visual sensitivity and feature-based visual attention by changing the difference between the target and distractors. As the neuronal system underlying visual and oculomotor functions are highly preserved in marmosets, macaques, and humans, this task will enables trans-speices comparison of visual attention.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-353 言語関連領域への経頭蓋直流電気刺激による発話運動統御への影響 Kengo Matsuhashi（松橋憲吾），Ryuichiro Hashimoto（橋本龍一郎），Yasuto Yada（矢田康人）首都大院人文科学言語科学 When a speaker receives auditory feedback of his/her own voice with a 200-ms delay, the delay causes dysfluency in speech. This phenomenon is called the delayed auditory feedback (DAF) effect. Previous neuroimaging studies suggested that the DAF induces cortical activity changes in the frontal and temporal language-related regions. However, few studies have examined the DAF using brain stimulation. We investigated the speech rate during a reading aloud task under DAF and normal auditory feedback (NAF) conditions while applying a transcranial direct current stimulation (tDCS) to the left inferior frontal cortex (LIFC) and superior temporal cortex (LSTC) separately. Change in speech rate induced by tDCS was evaluated with reference to the rate under the sham stimulation condition both for DAF and NAF. In total, 14 healthy volunteers (6 females; age = 20.92 ± 1.43) participated in the study. Each volunteer participated in either (1) 2 mA (N = 8) or (2) 1 mA stimulation session. Analysis of the 14 subjects combining both sessions showed that the anodal stimulation to LIFC significantly decreased the speech rate compared with the sham and the cathodal stimulation on both DAF and NAF conditions (p < 0.05). The same result was confirmed even in the 2 mA session alone for both DAF and NAF conditions, although the effect of anodal stimulation was attenuated in the 1 mA session in which no significant change in speech rate was observed. For the LSTC, the cathodal stimulation significantly increased the speech rate on both DAF and NAF conditions only in the 1 mA session (p < 0.05). These results suggested that, although the effects of tDCS to language-related regions on speech rate varied depending on several parameters including polarity, intensity and target area of stimulation, the anodal stimulation to LIFC most consistently decreased the speech rate regardless of DAF and NAF conditions possibly by changing cortical excitability associated with speech motor control.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-354 心の理論に関わる脳領域間のネットワークと物語理解の関連 Hiroto Ito（伊藤潤人）¹，Koji Jimura（地村弘二）²，Asuka Terai（寺井あすか）¹ ¹公立はこだて未来大学 ²慶應大院理工基礎理工 Theory of Mind (ToM) is the function to make inferences with respect to others' mental states and is utilized to understand or predict others' behavior. Recognizing others' mental states is required to grasp others' thought including characters' thought within the context of the story. ToM is related to language processing, particularly in the aspect of making sense during communication and story comprehension. Some previous researches have suggested relationships between ToM and language processing (story comprehension, understanding other's utterances etc.). Therefore, our previous research examined the relationships between activation of the 31 brain regions related to ToM, elicited by the story comprehension task in contrast to the math task, and performance of the story comprehension task using the functional MRI data provided by the Human Connectome Project (HCP), and showed a gender difference of the relationships. The purpose of this research is to elucidate relationships between network connectivity in the 31 brain regions relating to ToM during the story comprehension task and performance of the story comprehension task. The network connectivity of each subject was estimated using correlation coefficients among the regions computed based on Blood Oxygenation Level Dependent (BOLD) time-course signal during the story comprehension task. Across all subjects, each edge between two regions in the network is related to the scores of the story comprehension task using a linear regression model and the important edges were extracted the significance of the predictive power using significance test. The results suggest that the network among cingulate, cerebellum, inferior parietal lobule and superior frontal gyrus has an influence on the score and the influence varies depending on gender.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-355 感覚刺激と意味の対応付けへのヒトミラーシステムの関与 ~脳波μ抑制による検討~ Hiroki Kataoka（片岡大貴），Jiro Okuda（奥田次郎）京都産業大学院先端情報学 In our daily use of language, we may express many different meanings by one same linguistic phrase depending on situations. This feature of context-dependent usage of language is thought of as one remarkable aspect of the human language system. To realize this multiple usage of the linguistic system, it is needed to dissociate one meaning attached to a phrase and then to associate another meaning. These processes have been studied in terms of de-grounding (dissociating a meaning from linguistic symbols) and grounding (associating another meaning with the symbols). It has been proposed that mirror neurons in the brain may contribute to the process of grounding, by providing connection between self-relevant behavioral meanings attached to the symbols and observation of the physically-presented symbols, since the mirror neuron shows discharge both during execution of specific motor acts and during observation of similar acts. On the other hand, relationship between de-grounding and the mirror neuron has been less investigated. In the present study, we investigated possible association between de-grounding and mirror system activity in the human brain by recording mu-band power suppression with electroencephalography (EEG) as an index of the mirror neuron discharge. In our experiment, we artificially associated color information with meaning of bodily act. First, we asked subjects to associate two pairs of a colored disk and an action word (task 1). Second, we presented the subjects with a disk with gradually changing color, and asked them to judge timing of changes of meaning of the disk (task 2). Additionally, we also asked them to judge timing of changes of perception of color itself (task 3). As a result, dissociation of the action meaning from the disk (de-grounding) occurred after the subjects judged perceptual change of the color. On the other hand, association of the alternative meaning with the disk (grounding) occurred before the perceptual change of the color. We confirmed mu-band power suppression at the parietal electrodes during presentation of the action words in the task 1 as the index of the mirror system activity. In the analysis of the EEG power in the task 2, the mu-band power at the parietal electrodes significantly decreased from 4 seconds before de-grounding, and then to at the latest 10 seconds after de-grounding. The present results indicate possibility of involvement of the human mirror system in processes of de-grounding.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-356 鳴禽による歌のリズムの文化的学習 Masashi Tanaka（田中雅史），Kentaro Abe（安部健太郎）東北大院生命科学 Acoustic communication is widely utilized by animals to convey their internal state and environmental information. Sound can be readily detectable with high temporal resolution, which allows efficient coding of complex information. Indeed, auditory signals in animal communication are often highly complex—which we appreciate in the birdsong, a sequence of skillful vocalizations produced by songbirds. Songbirds learn songs from others and transmit the tutor's song across generations by precisely imitating its temporal modulation of pitch, amplitude, and timbre. To analyze the complex birdsong, studies have mainly focused on the stereotyped component of the song called "syllables", and analyzed spectral and temporal properties of syllables. However, such analysis often fails to capture dynamic aspects of song information such as melody, dynamics, and rhythm. Here we illustrate how a songbird species, the zebra finch, dynamically controls the song rhythm by using our newly developed software. Our analysis demonstrates that estimated song tempo is generally stable at ~10 Hz throughout a song bout and across song bouts in normally-reared zebra finches. In contrast, song tempo of zebra finches that were raised without a tutor was much less stable, suggesting that the song rhythmicity is not innate, but is learned through cultural transmission. Moreover, we found that tempo fluctuation in female-directed courtship songs tends to be more stereotyped than undirected, solitary songs in normally-reared zebra finches. Therefore, temporal variability of songs can convey information about the quality and internal state of the signaler. Since recent studies on songbirds have begun to elucidate detailed neural circuitry for singing, the songbird is a useful model in which to explore the mechanism and function of the rhythmicity prevalent in animal communication, from the chirp of crickets to human music.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-357 概念融合による特徴創発に関連する神経基盤 Asuka Terai（寺井あすか）¹，Junichi Chikazoe（近添淳一）²，Takaaki Yoshimoto（吉本隆明）²，Norihiro Sadato（定藤規弘）²，Koji Jimura（地村弘二）³ ¹公立はこだて未来大学 ²自然科学研究機構生理学研究所 ³慶應大理工生命情報 Combining two concepts as conceptual blending creates novel features, which are atypical characteristics of the original concepts. Similarly, interpretations of metaphor "topic (A) is vehicle (B)" involve creative features that are not typical characteristics of the words (A and B). The present study examined neural dynamics during the generation of creative features in conceptual blending. During fMRI scanning, human participants were presented a pair of concepts ("A" and "B"). The concept pair has a novel metaphorical, a familiar metaphorical, or a literal meaning, in the form of "A is B". They were asked to freely point out features associated with the presented pair, and to make vocal responses in the scanner. After scanning, they rated semantic unrelatedness between their generated features and the concept words. The features were then classified into creative or non-creative feature based on the rating of semantic unrelatedness. The results were compared to the findings of the previous research regarding to metaphor comprehension to examine the influence of the metaphorical context to the emergence. Behavioral analysis revealed that the non-creative features were less generated from metaphorical pair and the creative features were more infrequently generated. The findings of the metaphor comprehension show a similar tendency. In imaging analysis, fMRI signal was first extracted for each of the time windows before the response, allowing the analysis of temporal change in brain activity that preceded feature generation. The interaction effect of response types (creative and non-creative features) and temporal change was observed in middle frontal gyrus and inferior frontal gyrus, which were not involved in feature emergence during metaphor comprehension process.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-358 社会的知識における行動知覚と人格特性認知の連合 Masahiro Okamoto（岡本正博），Satoshi Eifuku（永福智志）福島県立医大医システム神経科学 We make inferences about others' behaviors based on what we know about them. Inability to make such inferences may cause serious difficulties when interacting with other people. A recent study revealed that personality traits were correlated with the prediction of others' actions, and that brain activity during prediction was partially explained by the perceived traits of third-party targets by study participants. Conversely, how we "know" about others' traits remains unclear. When reading a novel, we make inferences about the characters based on their behaviors. Such an experience raises important questions about what kind of behaviors and speeches lead us to identify a specific trait not only when reading novels but in a real life. In the present study, participants were asked to rate people close to them on 20 personality traits, such as good-naturedness and impudence. In addition, the participants provided reasons for rating each personality trait, with concrete episodes. As a result, principle component analysis on the ratings revealed three components (approachableness, extraversion, gentleness). Correspondence analysis identified verbs characterizing each component: talking, listening, and seeing for the "approachableness" component; drinking, participating, and connecting for the "extraversion" component; and thinking, working, and acting for the third component. Document classification successfully classified each episode into one of the three components based on the frequency of the listed verbs. Taken together, these results suggest that specific verbs are associated with specific traits. In our next study, we are going to investigate how these verbs are represented in the human brain.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-359 脳活動符号化モデルを用いた物語読み聞き不変的な意味表現の解明 Hiroto Q Yamaguchi（山口裕人）^1,2，Tomoya Nakai（中井智也）^1,2，Shinji Nishimoto（西本伸志）^1,2,3 ¹大阪大院生命機能 ²情報通信研究機構脳情報通信融合研究センター ³大阪大院医 We perceive language in two forms through different sensory modalities; written texts through the visual modality and spoken words through the auditory modality. Despite such a difference, we understand the same semantic contents. Under unimodal conditions, previous studies have utilized an encoding modeling analysis to comprehensively understand how semantic contents are represented in the brain (Huth et al., 2012, 2016). However, it remained unclear if these models captured modality-invariant semantic representations. To address this issue, we conducted fMRI experiments to record whole-brain activity under two modality conditions. In one condition, human participants listened to naturalistic monologues for three hours. In the other condition, they read the transcribed version of the same stories. To estimate the similarity of the semantic representations under two conditions, we performed modality-wise encoding modeling analysis. We firstly extracted sensory, phonemic, and semantic features that corresponded to the lower-to-higher conversion of the stimulus in each modality condition. We extracted visual features using a motion energy model (Nishimoto et al., 2011) and auditory features using a modulation transfer function model (Chi et al., 2005). Phonemic features were quantified as one-hot vectors that correspond to phonemes. We transformed words into semantic vector representations using Word2vec model (Mikolov et al., 2013). Secondly, for each modality condition, we modeled the evoked brain activity in each voxel as a weighted linear sum of three categories of features. Regularized linear regression was used to find the weights that best estimate the contribution of each feature space (Nunez-Elizalde et al., 2018). The model prediction accuracies were calculated using held-out test datasets. The two semantic models provided significant predictions for overlapping parts of the cortex including frontal, temporal and parietal areas, showing the validities of the modeling. A direct comparison of the model weights demonstrated that the models acquired similar semantic representations in these areas, although they were estimated completely separately. In addition, the models provided significant inter-modal predictions in the same regions, supporting the notion that the models acquired modality-invariant semantic representation. These results suggest that the brain have a common system for processing semantic information during reading and listening.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-360 自己帰属に関連する脳領域の領域間結合：ラバーハンドイリュージョンとラバーフットイリュージョンを用いて Nanae Matsumoto（松本奈々恵）¹，Ryusuke Nakai（中井隆介）²，Tadashi Ino（猪野正志）³，Akira Mitani（三谷章）¹ ¹京都大院医人間健康 ²京都大学こころの未来研究センター ³音羽病院 The feeling of bodily self-attribution is essential for our interaction with the outside world in our daily life. However, brain areas associated with the bodily self-attribution are not revealed in detail. In experimental studies, subjects have perceived touch sensations as arising from a rubber hand when the rubber hand and our own hand are repeatedly brushed in synchrony with the real hand hidden from view. This rubber hand illusion (RHI) provides a cue for studying the bodily self-attribution. The rubber foot illusion (RFI) is also known to be induced when the paradigm of the RHI is adapted to lower limbs. In the previous study, we hypothesized that brain areas associated with producing the bodily self-attribution would be activated during both RHI and RFI conditions, and investigated the brain areas associated with producing both RHI and RFI to right-handed healthy subjects by using functional magnetic resonance imaging (fMRI). We found that some brain areas were significantly activated when subjects experienced RHI and RFI. In the present study, in order to investigate the relationship between these brain areas, we performed an effective connectivity analysis with Statistical Parametric Mapping 12 (SPM12). It was indicated that the prefrontal cortex had an effective connection with angular gyrus when we set up angular gyrus as volume of interest (VOI). The result suggests that the inferior parietal lobule and prefrontal cortex are associated with producing the bodily self-attribution.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-361 神経回路が人の心のような意識を持った心の状態を持ち得るための条件、およびその探索 Mitsuo Takase（高瀬光雄），Mitsuo Takase（高瀬光雄） LINFOPS有限会社 It is imagined that a neural network with some conditions may create a human-like mind. This research is conducted along with the development of a face robot with emotions. The human-like mind is different from its functions like memorization, remembrance and discovery. It is what we feel its existence of as our minds or ourselves with consciousness. I already showed the case of a neural network example where it chooses the better one of two options with nearly same values and then it may have a human-like mind. Here, another example is shown. The essential conditions there are (1) emotions must be used, and (2) their stimulation is kept for a while to be able to be perceived, and (3) the result of the choice or a solution is obtained. In this situation, a simple mind like that of human beings has a possibility to be created and expected to appear in the neural network. Then, another example is as follows. In the estimation of an important object or a thing through emotions, when the informations used to evaluate are not clear, the estimation result may fluctuate at any time. Then through the repetition, the result will converge by averaging. In the process, the conditions shown above are achieved, the appearance of the mind may be confirmed. There, neural network parts which behave like amygdala can be created by simulations based on natural selection. So these emotion-like signals and the elements expressed by adjectives can be implemented into the total structure of neural networks with the hierarchical structure of memories with episode memories. In the system, the total evaluation of a situation can be conducted from the direct perception of emotion-like signals and by their summation causing seeking or avoidance. Here informations like emotions which came from evolution is combined with memory and intellectual process by PFC. In this setting, it is supposed what we feel as the mind may come out. Neural networks may be able to play the role to give us clues to know the existence of the mind. I have a plan to make the neural network software to simulate these situations.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-362 エーラス･ダンロス症候群原因遺伝子テネイシンX欠損マウスの疼痛解析 Emiko Ashitaka（芦高恵美子）¹，Hiroaki Kakumoyo（角本寛明）¹，Shota Yamanishi（山西翔太）¹，Yuka Kakuchi（角地宥香）¹，Sadahito Uto（宇戸禎仁）¹，Toshiaki Minami（南敏明）²，Kenichi Matsumoto（松本健一）³ ¹大阪工大院工生命工学 ²大阪医大院医麻酔 ³島根大総科支セ生体情報RI Tenascin-X (TNX), a member of the extracellular matrix glycoprotein tenascin family, is implicated in the formation of the extracellular matrix network containing collagen and elastic fibers. The deficiency of TNX leads to Ehlers-Danlos syndrome, a human heritable disorder characterized by skin hyperextensibility and joint hypermobility. Although Ehlers-Danlos syndrome patients present with chronic pain disorders, such as complex regional pain syndrome, it in not clear whether TNX affects pain transmission. Here, we examined the nociceptive behavioral responses of TNX-deficient mice to determine the contribution of TNX to pain transmission. TNX-deficient mice showed increased pain sensitivity to mechanical stimuli in von Frey filament test, and to tactile stimuli in modifying cotton swab assay. In contrast, TNX-deficient mice showed normal pain sensitivity to thermal stimuli in hot plate test. TNX deficiency also induced an increase in pain sensitivity to chemical stimuli and the aggravation of early inflammatory pain elicited by formalin. Nociceptive behavioral responses increased in phase I of the formalin test, especially at 1-5 min, and during the early stage of phase II (16-30 min) in TNX-deficient mice. However, the response during the late stage of phase II (31-50 min) was not significantly different between TNX-deficient and wild-type mice. We further produced a sine-wave electric stimulator and characterized sensory fibers in TNX-deficient mice. TNX-deficient mice were significantly hypersensitized to transcutaneous sine-wave stimuli at frequencies of 250-Hz (Aδ- and Aβ-fiber responses) and 2000-Hz (Aβ-fiber responses), but not to stimuli at frequency of 5-Hz (C- and Aδ-fiber responses). Aβ-fiber responses induced by a 2000-Hz stimulus were inhibited by the intrathecal administration of the AMPA/kainate antagonist CNQX. These results suggest that TNX deficiency contributes the pain responses to mechanical and chemical stimuli, and that these pain responses are mediated by sensory Aβ-fiber.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-363 脊髄後角における疼痛関連タンパクBEGAIN陽性細胞の機能的特徴 Tayo Katano（片野泰代）¹，Kotaro Konno（今野幸太郎）²，Kazuhiko Nishida（西田和彦）¹，Masahiko Watanabe（渡辺雅彦）²，Kenji Sakimura（崎村建司）³，Takuya Kobayashi（小林拓也）¹，Seiji Ito（伊藤誠二）^1,4 ¹関西医大医医化学 ²北海道大院医解剖発生 ³新潟大脳研基礎神経科学細胞神経生物 ⁴大阪医大麻酔 Pathological pain such as allodynia and hyperalgesia is maintained by central sensitization, which is caused by the expression and/or localization change of pain-related molecules and remodeling of the spinal cord circuit. However, the exact pathogenic mechanisms are not fully understood. In our previous study, we identified brain-enriched guanylate kinase associated protein (BEGAIN) as a neuropathic pain-related protein in the PSD fraction of the spinal dorsal horn. BEGAIN protein was localized in the spinal laminae IIi-IIIo and increased after speared nerve injury (SNI). These results indicated that BEGAIN protein or BEGAIN-positive spinal neurons at laminae IIi-IIIo plays an important role in pathological pain transmission in the spinal dorsal horn. Here, to characterize BEGAIN-positive neurons in the spinal dorsal horn, we performed double-labeling RNA in situ hybridization for BEGAIN and VGLUT2 or GAD67. BEGAIN mRNA was expressed in both VGLUT2 and GAD67 positive interneurons. Then we examined the expression of c-fos protein by immunohistochemistry in BEGAIN-positive spinal neurons before and after SNI and found that BEGAIN-positive neurons were activated in the spinal dorsal horn after SNI. Furthermore, pathological pain was attenuated in BEGAIN-knockout mice after SNI. These results suggest that BEGAIN-positive interneurons are related to pathological pain circuits. Further characterization of BEGAIN-positive interneurons before and after SNI may specify a novel spinal cord circuit for pathological pain.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-364 Metabotropic Glutamate Receptor 5 in the Brain: Bridging the Mind and Bodily Pain Geehoon Chung（Chung Geehoon）¹，Chae Young Kim（Kim Chae Young）²，Yeong-Chan Yun（Yun Yeong-Chan）⁴，Sang Ho Yoon（Yoon Sang Ho）²，Soonho Shin（Shin Soonho）²，Myoung-Hwan Kim（Kim Myoung-Hwan）²，Yu Kyeong Kim（Kim Yu Kyeong）³，Sang Jeong Kim（Kim Sang Jeong）²，Sun Kwang Kim（Kim Sun Kwang）¹ ¹Department of Physiology, College of Korean Medicine, Kyung Hee University, Seoul, Korea ²Department of Physiology, Seoul National University College of Medicine, Seoul, Korea ³Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, Korea ⁴College of Korean Medicine, Dongshin University, Naju, Korea Chronic pain often accompanies negative mood symptoms such as depression and anxiety. Conversely, patients with emotional distress are more susceptible to pain symptoms compared with normal individuals. The interaction between abnormally amplified bodily pain and negative emotions implies the existence of common or interacting neural pathways. However, the neural circuits relevant to the interactions and underlying mechanisms are not fully understood. We studied the mechanisms of chronic bodily pain and ensuing negative affections in an animal model of neuropathic pain. The various alterations in neural circuits of pain animals were investigated using brain imaging, electrophysiological recording, pharmacological and genetic manipulation and animal behavior analysis. Metabotropic glutamate receptor 5 was a particular target, as this molecule plays a pivotal role in the plastic changes in neural circuits. We scanned the brain of chronic neuropathic pain model rats using positron emission tomography (PET) technique with an mGluR5-selective radiotracer [11C] ABP688 and found various brain regions with higher or lower level of mGluR5 compared to control rats. We found clusters with such an altered mGluR5 availability levels localized in brain areas involved in sensory, cognitive, and affective aspects of pain processing. A pharmacological blockade of upregulated mGluR5 in the neuropathic pain animal ameliorated the negative symptoms including bodily pain and negative affections, which relieved the subjects from the unpleasant state of chronic neuropathic pain condition. Conversely, lentiviral overexpression of the mGluR5 in the brain structure successfully induced comorbid pain and negative moods in naive rats. Our data provide deeper insight into the shared mechanism of pain perception and negative emotions, identifying a therapeutic target for the treatment of chronic pain and mood disorders.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-365 パクリタキセル誘発末梢神経障害の発症におけるシュワン細胞依存的免疫反応の関与 Madoka Koyanagi（小柳円花），Satoshi Imai（今井哲司），Mayuna Matsumoto（松本真有奈），Yuki Iwamitsu（岩満優輝），Takashi Ogihara（荻原孝史），Ren Hiraiwa（平岩怜），Mpumelelo Ntogwa（Ntogwa Mpumelelo），Yuki Sato（佐藤夕紀），Shunsaku Nakagawa（中川俊作），Tomohiro Omura（大村友博），Atsushi Yonezawa（米澤淳），Takayuki Nakagawa（中川貴之），Kazuo Matsubara（松原和夫）京都大学医学部附属病院薬剤部 Chemotherapy-induced peripheral neuropathy (CIPN) is a dose limiting side effect caused by chemotherapeutic agents such as taxanes. However, the mechanisms underlying CIPN pathogenesis are not fully understood. We previously showed that paclitaxel preferentially impairs Schwann cells, rather than dorsal root ganglia neurons, and induces dedifferentiation of Schwann cells. In present study, we further investigated the functional roles of dedifferentiated Schwann cells in the development of CIPN. We found that mRNA expression of an inflammatory factor, X, was increased in dedifferentiated rat primary cultured Schwann cells after paclitaxel (0.01 μM) treatment. Likewise, repeated i.p. injection of paclitaxel (20 mg/kg) caused the upregulation of the factor specifically within the mouse sciatic nerve compared to other organs at the onset of mechanical allodynia. The upregulation of the factor also coincided with macrophage infiltration into the sciatic nerve in paclitaxel injected mice. Furthermore, murine macrophage cell line (RAW 264.7) showed a chemotaxis response toward the conditioned medium containing an inflammatory factor released from paclitaxel-treated Schwann cells. Consistent with this, we revealed that the perineural application of recombinant protein of this inflammatory factor elicited infiltration of macrophages into the sciatic nerve and mechanical hypersensitivity in mice. Taken together, these findings allow us to conclude that, in response to paclitaxel treatment, an inflammatory factor is released from dedifferentiated Schwann cells to chemoattract macrophages. These Schwann cell-dependent peripheral immune responses in conjunction with direct disturbance of sensory neurons may be associated with paclitaxel-induced CIPN pathogenesis.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-366 一次感覚神経に発現する新規長鎖非コードRNAの神経障害性疼痛における関与 Atsushi Sakai（坂井敦）¹，Motoyo Maruyama（丸山基世）^1,2，Yoshitaka Miyagawa（宮川世志幸）³，Takashi Okada（岡田尚巳）³，Hidenori Suzuki（鈴木秀典）¹ ¹日本医科大医薬理 ²日本医科大実験動物管理室 ³日本医科大医生化・分子生物（分子遺伝） Damage to the somatosensory systems leads to intractable neuropathic pain. MicroRNA, which is small non-coding RNA inhibiting diverse gene expressions, has been shown to contribute to neuropathic pain. In contrast, involvement of long non-coding RNA (lncRNA), which is longer than 200 nucleotides, in neuropathic pain remains poorly understood. Here, we show the involvement of newly identified long non-coding RNAs expressed in the primary sensory neurons in neuropathic pain. A neuropathic pain model was produced by lumbar fifth spinal nerve ligation (SNL) in rats. RNA sequence identified a number of potentially novel transcripts whose expression was changed in the injured dorsal root ganglion 14 days after SNL. Among them, the whole nucleotide sequences of three abundant transcripts were determined by 5' and 3' rapid amplification of cDNA ends and were found to have a low protein-coding potential. Plausible human homologues of these transcripts were also expressed in the primary sensory neurons differentiated from human induced pluripotent stem cells. In the rat, the expression level of lncRNAs was highest in the dorsal root ganglion among key peripheral organs and brain regions examined. In the dorsal root ganglion, expression of the lncRNAs was persistently changed after SNL. In situ hybridization showed that the lncRNAs were mainly present in the cytoplasm of primary sensory neurons. To examine the involvement of the lncRNAs in pain-related behaviors, their expression was modulated specifically in the primary sensory neurons through microinjection of adeno-associated viral vector. Modulation of the lncRNAs expression changed a withdrawal threshold and latency in response to mechanical and thermal stimuli, respectively. Therefore, clarifying a function of the lncRNAs expressed in the primary sensory neurons will provide further insight into the molecular basis of neuropathic pain.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-367 慢性鼻腔炎症と嗅覚遮断が嗅球の層特異的変性に及ぼす異なる影響 Sanae Hasegawa-Ishii（石井さなえ）¹，Fumiaki Imamura（今村文昭）²，Atsuyoshi Shimada（島田厚良）¹ ¹杏林大学保健学部 ²ペンシルベニア州立大学医学校、ハーシー、米国 Chronic nasal inflammation in mice induces gross atrophy of the olfactory bulb (OB) as well as loss of olfactory sensory neurons (OSNs) that convey the odor information to the OB. The OB atrophy may be caused by two mechanisms; inflammatory responses of the OB and odor deprivation due to loss of OSNs. In fact, odor deprivation through naris closure (NC) causes OB atrophy in neonatal and adult mice. Here we investigated whether different mechanisms underlie inflammation-induced and NC-induced OB atrophy. Adult male C57BL/6 mice received repeated administration of LPS into unilateral nostril every other day for 10 weeks, representing a nasal inflammation model (LPS:10w) or saline in the same manner as control (saline:10w). In the other group, unilateral naris was closed for 10 weeks (NC:10w). OB size, area of each OB layer, glial responses and cytokine expression were compared among these 3 groups. OB size was significantly smaller in LPS:10w and NC:10w compared to saline:10w. The outer 3 layers of the OB, including olfactory nerve layer (ONL), glomerular layer (GL) and external plexiform layer (EPL), significantly shrank in LPS:10w, whereas the inner 3 layers, including EPL, mitral cell layer (MCL) and granule cell layer (GCL), shrank in NC:10w. Within EPL, superficial layer was more vulnerable in LPS:10w and NC:10w. In LPS:10w, microglia became activated and the amount of IL-1b mRNA was elevated in the ipsilateral OB. In contrast, these changes did not occur in NC:10w. Next, we examined whether the OB that had atrophied by nasal inflammation returned to normal after inflammation subsided in the presence or absence of odor input. Mice received LPS to unilateral nostril for 10 weeks and housed for another 10 weeks with no additional treatment (LPS:10w+NT:10w) or with NC (LPS:10w+NC:10w). In both groups, nasal inflammation and glial activation subsided and the ONL tended to recover. The OB size, areas of GL and EPL returned to normal in LPS:10w+NT:10w, but not in LPS:10w+NC:10w. Rather, EPL shrinkage was more prominent and MCL and GCL additionally shrank in LPS:10w+NC:10w. These results indicated that different mechanisms underlie nasal inflammation-induced and NC-induced OB atrophy; ONL shrinkage, glial activation and elevated IL-1b; were inflammation-dependent whereas MCL and GCL shrinkage was NC-dependent. GL and EPL shrinkage resulted from nasal inflammation and NC. Differential effects on more caudal parts of the olfactory system awaits further studies.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-368 内包へのリゾレシチン投与による新規脱髄モデルマウスの作製 Reiji Yamazaki（山崎礼二），Jeffrey K Huang（ハングジェフリー）ジョージタウン大学, 生物学部 Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the CNS characterized by progressive remyelination failure and accumulated motor disability. However, whether remyelination promotes motor recovery remains unclear. Here, we compared the effect of experimental demyelination with focal ischemia induction in the internal capsule (IC), a white matter region associated with motor impairment in MS and stroke, on motor behavior in mice. First, to induce the demyelination, we injected lysophosphatidylcholine (LPC) into right side IC using stereotaxic technique. At 7 days after injection, demyelination was observed in the IC by immunofluorescence staining, FluoroMyelin staining and our new method to track CNS demyelinated lesions by neutral red. Next, to examine whether this model affects motor function, we performed adhesive tape removal test, cylinder test and wire hang test. IC-demyelinated mice reduced motor function in mice at 7 days post lesion (dpl). Further, we demonstrated that demyelination of right IC significantly impaired left forepaw motor function. These results suggest that this demyelinated-mice is an asymmetric disease model. Moreover, these mice exhibited motor deficit until 14dpl, but regained motor function by 28dpl, corresponding with reduced inflammation, decreased axonal dystrophy, and increased oligodendrocytes in lesions. By contrast, injection of endothelin-1 (ET1) into the IC, which is known to induce white matter infarct, displayed lasting motor deficit, which is accompanied by persistent inflammation and axonal dystrophy, and reduced oligodendrocytes in lesions. These results demonstrate that IC demyelination induces acute motor deficit and subsequent motor recovery through remyelination, and suggest that inflammation resolution and the restoration of axonal integrity may be required for successful remyelination and motor recovery. Therefore, IC demyelination is a tractable model for assessing the influence of remyelination on motor behavior, and may be used to complement future drug screens for the identification of compounds for promoting remyelination.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-369 マカクザルの脊髄損傷後機能回復における抗RGMa抗体治療と組み合わせた反復経頭蓋磁気刺激の効果 Hajime Yamanaka（山中創）¹，Yu Takata（高田裕生）¹，Hiroshi Nakagawa（中川浩）²，Toshihide Yamashita（山下俊英）²，Masahiko Takada（高田昌彦）¹ ¹京都大霊長研統合脳システム ²大阪大院医分子神経科学 Repetitive transcranial magnetic stimulation (rTMS) has been used not only for basic research, but also for clinical application. On the other hand, it has recently been reported that treatment with antibody against repulsive guidance molecule-a (RGMa) ameliorates impairments in motor functions after spinal cord injury (SCI) in rodents and nonhuman primates. Here we analyzed the possible effect of rTMS combined with anti-RGMa antibody treatment on functional recovery after SCI in macaque monkeys. The rTMS system applicable to a monkey under conscious condition was prepared by using a thermoplastic mask and a laser distance meter. The movement of the conscious monkey head was suppressed, and three-dimensional positioning measurement became possible. Therefore, a better reproducibility of the stimulation site was obtained with this rTMS system. The rTMS trials in the primary motor cortex (especially its forelimb region) started four to five weeks after the SCI surgery and continued for about 10 weeks. To prepare a monkey model of SCI, large hemi-transection lesions were made at the border between the C6 and the C7 segment of the spinal cord. The anti-RGMa antibody was simultaneously delivered around the lesioned site over four weeks through an osmotic infusion pump. The rTMS sessions were performed more than three times a week and, then, behavioral assessments were made on the same day. Forelimb movements were assessed by two behavioral tests consisting of a reaching & grasping task and the conventional Brinkman board test. Especially in the latter case that requires a higher motor skill, a monkey group who underwent combined antibody treatment and rTMS showed greater recovery from motor impairments than a control group with the antibody treatment alone. In the former easier case, however, no distinct effect of rTMS was observed. The present data indicate that rTMS combined with anti-RGMa antibody treatment provides an increased therapeutic effect in a monkey model of SCI.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-370 自動パッチクランプシステムQubeおよびQPatchを用いたヒトiPS細胞由来神経細胞からの電気生理学的測定 Kazuya Tsurudome（鶴留一也）¹，Kadla Roskva Rosholm（Rosholm Røskva Kadla）²，Daniel Sauter（Sauter Daniel）³，Melanie Schupp（Schupp Melanie）²，Rasmus Bjorn Jacobsen（Jacobsen Bjørn Rasmus）² ¹ソフィオンバイオサイエンス株式会社 ²Sophion Bioscience A/S, Ballerup, Denmark ³Sophion Bioscience Inc., Woburn, MA, United States Human induced pluripotent stem cells (hiPSCs) can be differentiated into many cell types, including neurons and cardiomyocytes, and therefore constitute a novel way to model human diseases for drug-testing in vitro. Ion channels represent highly attractive therapeutic targets in the nervous systems, rendering electrophysiological studies of hiPSCs critical for their usage in drug-discovery. However, such studies have traditionally been limited by the labour-intensive and low-throughput nature of patch-clamp electrophysiology. Automated patch clamp (APC) techniques constitute a powerful tool for high-throughput investigation of ion channel properties and excitable cell functions. Here we report our recent study on hiPSC derived neurons from multiple cell vendors using our APC platforms, Qube384 and QPatch. Our results include a measure of channel expression versus time in culture, the pharmacological dissection of endogenous ion channels (e.g. Na_V and K_V), identification of ligand-gated receptors, and recordings of action potentials using current clamp. Na_V channel currents showed high sensitivity to tetrodotoxin. Outward currents were blocked by tetraethylammonium with different sensitivities suggesting the hiPSC derived neurons tested in this study expressed several types of K_V channels. The major challenge when investigating neurons using APC platforms is the requirement to dissociate the cells from their neuronal network while maintaining cell viability and membrane integrity. By optimization of the harvest- and whole-cell protocols we have overcome this obstacle resulting in success rates of up to 50 % or 60% using QPatch or Qube 384, respectively.Our results demonstrate the feasibility of conducting electrophysiological characterization and drug-screening on hiPSC derived neurons on APC platforms like Qube 384 and QPatch. We envision that this combination of iPSCs and APC will pave the way for high-throughput ion channel-targeted screening of drugs for neurological disorders.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-371 Distinct age-dependent subcellular changes in a model mouse of Parkinson's disease as revealed by volumetric FIB/SEM imaging of striatum Laxmi Kumar Parajuli（Parajuli Kumar Laxmi）¹，Laxmi K Parajuli（Parajuli K Laxmi）¹，Ken Wako（Wako Ken）¹，Suiki Maruo（Maruo Suiki）¹，Ryosuke Takahashi（Takahashi Ryosuke）²，Masato Koike（Koike Masato）¹ ¹Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine, Tokyo, Japan ²Department of Neurology, Kyoto University Graduate School of Medicine, Japan Although the behavioral abnormalities in Parkinson's disease is well- characterized, much less is understood about the subcellular changes that accompanies this disease. In particular, pre-clinical morphological features and the characteristics of morphological changes at distinct ages remains of great interest in this field. A longitudinal study with volumetric EM imaging of neuropil at different developmental stages is undoubtedly the most straightforward way to reveal precise ultrastructural changes at distinct ages. Here, using striatum tissues from A53T mutant human synuclein BAC transgenic mice (A53T-BAC- Tg mice), we performed focused ion beam scanning electron microscopy (FIB/SEM) imaging of 101015 μm3 of neuropil volume in striatum and manually segmented dendrites and their spines to obtain measurement of parameters that shape synaptic transmission. We found that at 6 months of age, A53T-BAC-Tg mice display higher spine density. This abnormality in spine density increase is maintained even in 22-months-old mutant mice. However, intriguingly, in addition to the abnormality in spine density, the spine head volume in mutant mice become smaller than the control only in 22 months old mouse. This suggests that initial burst of spinogenesis and higher spine turnover is compensated by concomitant homeostatic reductant in spine head volume and PSD area, thereby maintaining constant synaptic strength between mutant and control mice at 22 months of age. Furthermore, an age dependent increase in spine head volume is observed only in the control mice. Taken together, our data implies that there is an abundance of immature spine morphology in mutant mice, suggesting an abnormality in the molecules that control synapse growth and maturity.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-372 感覚入力を備えたブレインコンピューターインターフェースへの要素技術 Fumiaki Yoshida（吉田史章）^1,2,3 ¹佐賀大医生体構造機能 ²九州大 ³JSTさきがけ Position or touch sense is important for clinical applications of the BMI because ideal prosthetic limbs should be perceived as natural extensions of the users' bodies. We have started to design a cortical modulator using optogenetics-a new method for the manipulation of neurons to dial in potential sensory input in a bi-directional manner. Optogenetics is based on genetically modified ion channels that respond directly to light. These light-gated ion channels, such as Channelrhodopsin-2 (ChR2), allow precise, millisecond control of specific neurons. This technique reduces most of the key problems associated with electrical brain stimulation: there is no associated electrical artifact to interfere with the electrophysiological recordings, nor any tissue damage from the current injection. It also allows for precise control of the spatial pattern of stimulation. A prototype optogenetic implant is presented, that will simultaneously record the activity of cortical neuronal activities and bring complex modulation patterns through optogenetic stimulation of cortical sensory areas. Our newly-invented optogenetic devices consist of both ECoG and LED for optical stimulation. Here we report data from initial bench testing and implantation for the ECoG in both the rat and non-human primate. We have show that the ECoG is effective as a chronic implant in rats, providing high fidelity neural recordings for up to 8 weeks. The initial results suggest that the new ECoG array can be successfully translated from rodents to accommodate the technological challenges associated with successfully interfacing with the non-human primate brain.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-373 プルキンエ細胞特異的Nna1 cKOマウスは、細胞自律的なプルキンエ細胞変性を示した Hirohide Takebayashi（竹林浩秀）¹，Li Zhou（周麗）¹，Hossain M Ibrahim（イブラヒムホサイン）¹，Manabu Abe（阿部学）²，Rie Natsume（夏目里恵）²，Kohtaro Konno（今野幸太郎）³，Masahiko Watanabe（渡辺雅彦）³，Kenji Sakimura（崎村建司）² ¹新潟大学医歯学部神経生物・解剖学 ²新潟大学脳研究所モデル動物開発分野 ³北海道大学医学部解剖発生学 Purkinje cell degeneration (pcd) mutant was first identified as a spontaneous mouse mutant showing ataxic behavior in an autosomal recessive manner. The causative gene is Nna1, also known as Agtpbp1 or CCP1. We generated a conditional allele of Nna1, in which carboxypeptidase domain-encoding exon 21 and 22 are floxed. The floxed Nna1 mice were crossed with beta-actin-iCre mice, Grid2-Cre mice or Grin2-Cre mice to generate Nna1 KO mice, Purkinje cell conditional KO (cKO) or granule cell cKO separately. In this study, we report phenotype analyses of Nna1 KO mice and the conditional KO mice. Nna1 KO mice showed cerebellar ataxia with smaller body size than wild-type. Loss of Nna1 mRNA and Nna1 protein in the Nna1 KO brain was confirmed by in situ hybridization and Western blot analyses. Cerebellar atrophy and degeneration of Purkinje cell (PC) and granule cells were observed in the cerebellum of Nna1 KO mice. Apoptosis-related markers were detected not only in the PC layer, but also in the granule cell layer and the molecular layer. PC-specific Nna1 cKO mice also showed the similar phenotypes (cerebellar ataxia and PC degeneration) with the Nna1 KO mice except for the ones of testis and retina. Our data indicate that loss-of-function of Nna1 in PCs results in the cell-autonomous PC degeneration.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-374 Neuronal avalanches patterns in Parkinson’s Disease relate to clinical disability Pierpaolo Sorrentino（Sorrentino Pierpaolo）¹，Rosaria Rucco（Rucco Rosaria）²，Fabio Baselice（Baselice Fabio）²，Carmine Granata（Granata Carmine）³，Alessandro Tessitore（Tessitore Alessandro）⁴，Rosita Di Micco（Di Micco Rosita）⁴，Leonardo Gollo（Gollo Leonardo）¹，Giuseppe Sorrentino（Sorrentino Giuseppe）² ¹QIMR Berghofer institute for Medical Research ²University of Naples Parthenope, Naples, Italy ³Istitutodi Scienze Applicate e Sistemi Intelligenti , CNR, Pozzuoli, Italy ⁴University of Naples Luigi Vanvitelli, Naples, Italy Background. In many complex systems, such as avalanches, when a unit exceeds a threshold, other units follow it, thus giving rise to a cascade that spreads across the entire system. The avalanche dynamics in critical, and is characterized by the branching ratio. A branching ratio σ=1 indicates that the activity of one unit triggers the activity of another unit. In this critical regime, cascades have sizes and durations varying over orders of magnitude. Neuronal activity can be modeled as avalanches in which the activity of a single brain area propagates in cascade to other brain areas. In this study, we hypothesized that the clinical picture of Parkinsons disease patient might relate to the flexibility of brain activity, and that such feature would relate to the observed clinical disability. Methods. We studied thirty-nine PD patients (18 males and 5 females, age 64.87 ± 9.12 years), matched with thirty-eight controls (14 males and 9 females, age 62.35 ± 8.74 years). UPDRS was used as clinical scale. Both groups underwent five minutes of closed eyes resting-state MEG. The patients were in the off-state. The source level time series of neuronal activity were reconstructed using a beamformer approach based on the native MRI and the AAL atlas, and then filtered in the classical frequency bands. An avalanche was defined as a continuous sequence of time bins in which there was an activation above threshold on any ROI. The pattern of an avalanche was defined as the regions active at in any moment within an avalanche. The number of patterns, in each frequency bands, were compared between groups using FDR corrected permutation testing. The number of times a given region switched between groups was also compared. The correlation between the number of states in each patient and the UPDRS was assessed using linear correlation. Results.Patients show a reduced number of visited states as compared to controls. The number of switching between stated did not differ. The correlations showed an inverse linear relationship between the number of states each patient visited and its UPDRS score. Discussion. Our results show that the capability of the brain to flexibly activate areas in a relevant feature that is related to the clinical state in Parkinsons disease.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-375 脳出血後の早期リハビリテーションは樹状突起の退縮を抑制し，運動機能回復を促進させる． Keigo Tamakoshi（玉越敬悟），Hayao Keishi（早尾啓志），Hideaki Takahashi（髙橋英明），Hiroyuki Tamaki（田巻弘之）新潟医療福祉大学　医療技術学部　理学療法学科 The present study focused on the effect of early exercise on brain damage and the recovery of motor function following intracerebral hemorrhage (ICH) in rats. Under deep anesthesia, the animals were placed inside stereotaxic apparatus, and ICH was induced by injecting a solution of collagenase in 0.9% saline into the left striatum. Subjects were randomly assigned to no training ICH (ICH), no training placebo surgery (SHAM), early treadmill exercise (ICH + ET), and late treadmill exercise (ICH + LT) groups. The SHAM group was injected with 0.9% saline using the same procedure. The ICH + ET, and ICH + LT groups were trained for 7 days from day 2 to day 8, and from day 9 to day 15, respectively, after surgery. At days 16 post surgery, brain was removed, and lesion volume, cortical thickness, and neuronal number were analyzed. Dendritic complexity and length were analyzed using Golgi-cox staining. IL-1b, TGF-b1, and IGF-1 mRNA expressions of the sensorimotor cortex were analyzed using RT-PCR. Motor function was assessed using forelimb placing and horizontal ladder tests. Significant improvement in motor function was noted in the ICH + ET group, but not in the ICH and ICH+LT groups. Cortical thickness and neuronal number of the ICH + ET group were significantly higher than those of the ICH and ICH+LT groups. Dendritic complexity and length of ICH+ET group improved significantly compared with ICH and ICH+LT groups. IL-1b mRNA expression of the ICH + ET group was significantly lower than that of the ICH group. These results suggest that after cerebral hemorrhage, early treadmill exercise may promote the recovery of sensorimotor function by inhibiting cortical atrophy, cell death and dendritic atrophy compared with late treadmill exercise. Early exercise may decrease cell death by inhibiting inflammation following ICH.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-376 Neuroprotective effects of Luteolin-7-O-β-D-glucuronide against cerebral ischemic injury in vivo and in vitro Shengqun Hou（Hou Shengqun）^1,3，Fang Lin（Lin Fang）¹，Ting Li（Li Ting）¹，Yuling Dou（Dou Yuling）¹，Jingtao Zou（Zou Jingtao）²，Dan Ohtan Wang（Wang Dan Ohtan）³，Fang Wang（Wang Fang）¹，Chunfu Wu（Wu Chunfu）¹ ¹Shenyang Pharmaceutical University, Shenyang, China ²Tonghua huaxia Pharmaceutical Co., Ltd.,Tonghua, China ³Institute for Integrated Cell-Material Science, Kyoto University Institute for Advanced Study Cerebral ischemia, caused by a blockade in the artery, reduces the blood flow and oxygen to the brain, leading to damage or death of brain cells, and permanent loss of brain functions in severe cases. Luteolin-7-O-β-D-glucuronide (LGU) is a pharmacologically active flavonoid compound extracted from Ixeris sonchifolia. In the present study, we investigate the neuroprotective effect of LGU against cerebral ischemic injury and potential underlying mechanisms. First, protective effect of LGU against oxygen-glucose deprivation (OGD)-induced death of primary cortical neurons was assessed using MTT metabolic activity assay and LDH cytotoxicity assay. The two tests consistently showed improved survival rate and decreased LDH leakage of primary cortical neurons upon LGU treatment, in a concentration-dependent and time-sensitive manner. To estimate protective effect of LGU in vivo, we generated a cerebral ischemic injury model in rats with permanent middle cerebral artery occlusion (MCAO) and applied LGU. Compared to untreated animals, LGU-treated animals showed improved motor and sensory performances (neurological severity score), less tissue damage (infarct volume ratio), and less edema (brain water content). We estimated that the therapeutic time window of LGU following cerebral ischemia is at least 6 hours for cerebral infarction and brain edema, and 12 hours for neurological dysfunctions. In order to identify the underlying mechanisms, we measured intracellular [ATP], [Ca²⁺], mitochondrial membrane potential (MMP), and expression of necroptosis-related proteins in affected neurons. Parameters associated with cell death, such as decreased ATP, increased Ca²⁺, and decreased MMP were all significantly reverted upon LGU treatment. LGU treatment also reduced the expression level of RIP3 and MLKL and inhibited the translocation of MLKL from cytoplasm to nuclear envelope, indicating that LGU may convey its therapeutic effect by inhibiting one of the necroptosis pathways (caspase8-RIP1-RIP3-MLKL). Collectively, our results show that LGU is a neuroprotective flavonoid therapeutically targeting necroptosis pathways.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-377 行動評価を用いた脳卒中高次脳機能障害患者における因子構造の研究 Yaoko Iwasaki（岩崎也生子）¹，Ken Gyoryo（御領謙）² ¹杏林大 ²千葉大 Introduction: Post-stroke cognitive dysfunction is a common symptom and can have a significant effect on a patient's activities of daily living (ADLs). Neuroimaging studies have identified the cortical association areas related to cognitive function. However, it is difficult to adequately evaluate how post-stroke cognitive dysfunction affects ADLs. The purpose of this study is to more effectively extract clinical characteristics of patients with cognitive dysfunction based on their performance of ADLs and to verify their reliability and validity. Method: Thirty occupational therapists selected 245 evaluation items from the Functional Independence Measure (FIM) relevant to difficulties with ADLs. The items were analyzed using a multidimensional scaling method and cluster analysis, resulting in six groups consisting of 66 items. On the basis of the items, 180 stroke patients with cognitive dysfunction were divided into five grades. Moreover, coma grades (Japan Coma Scale: JCS), FIM, neuropsychological examination (Mini-Mental State Examination; MMSE) and attention scales were checked. Analytical methods were performed as follows: 1) factor structure: factor analysis of the 66 items; 2) reliability: confidence coefficient (Cronbach's α); 3) validity: correlation between the extracted factors and days after onset, FIM, and MMSE; and 4) multiple regression analysis with extracted factor scores, the days after onset, JCS, FIM, MMSE and attention score. Result: Factor analysis revealed five relevant main factors with a total of 30 subsystem items: 1) situation judgment (8 items), 2) memory (7), 3) communication (6), 4) behavioral regulation (5), and 5) perseveration (4). The confidence coefficient ranged 0.840-0.980 (Cronbach's α). Factors 1 to 5 were strongly correlated with consciousness and FIM (r=0.215-0.886, p<0.001). In multiple regression analysis, factors 2 and 4 correlated to ADL independence (p<0.05), and Factor 4 correlated to attention (p<0.01). Conclusion: We identified five main factors in the assessment of cognitive dysfunction. The reliability and validity of these factors were verified. These factors are important in efficaciously evaluating stroke patients with cognitive dysfunction. Observational evaluation could identify the clinical features of patients. We could provide better therapy for cognitive dysfunction by the strengthening relationships between cognitive dysfunction assessment factors and neuroimaging studies.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-378 一夫一婦制を呈するプレーリーハタネズミの脳卒中後うつモデル作製の試み Shinichi Mitsui（三井真一）¹，Yuna Tanaka（田中侑奈）²，Misa Kobayashi（小林未紗）²，Hinano Matsudaira（松平ひなの）² ¹群馬大院保リハビリ ²群馬大医作業療法 Over 30% of stroke survivors are suffered from post-stroke depression that leads patients to cognitive deficits, social withdrawal, anhedonia, and sexual dysfunction. Although social supports have health benefits on humans and other mammals, the underlying neurobiological mechanisms are still obscure. Recently, mouse models of post-stroke depression have been developed and it is shown that pair housing ameliorates post-stroke depressive behaviors in a mouse model. It is necessary to establish a model for clarifying the effects of social bond between spouses on post-stroke depression, since traditional model animals such as rats and mice are promiscuous. Prairie vole (Microtus ochrogaster ) is a monogamous rodent which prefers the partner of the opposite sex after they cohabit each other for over 24 hours. A post-stroke depression model of prairie voles may be a useful tool to analyze the efficacy of a pair bond on this disorder. Male prairie voles were applied by photothrombotic stroke of the left cerebral cortex and followed by chronic mild stresses, such as continuous lighting for 17 h, damp bedding for 5 h, water deprivation for 10 h, restraining for 3 h, tilting cages for 15 h, food deprivation for 21 h, for 14 days. Paralysis and motor deficit were assessed by a corner test and a foot fault test. Anxiety-like and depressive behaviors were assessed by sucrose preference, novelty suppressing feeding, open field, tail suspension, and forced swimming tests. Effects of photothrombosis and chronic mild stresses on partner preference was also analyzed. Immunohistochemical analyses showed cortical ischemic infarct in the left forebrain and microglial activation around the infarct site. Preliminarily, behavioral analyses revealed hemiplegia and increased anxiety-like behaviors. Detailed data will be shown on site.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-379 豊かな環境飼育が新生児低酸素虚血性白質障害モデルラットに与える効果 Atsunori Hattori（服部篤紀），Yoshitomo Ueda（上田佳朋），Naoki Tajiri（田尻直輝），Shino Ogawa（小川紫野），Akimasa Ishida（石田章真），Takeshi Shimizu（清水健史），Hideki Hida（飛田秀樹）名古屋市大医脳神経生理学 Hypoxia-ischemia (H-I) in preterm infants occasionally results in white matter injury (WMI) associated with neurodevelopmental disabilities such as paralysis and cognitive dysfunction. We made a rat model of neonatal WMI with H-I at prenatal day 3 (P3). To investigate the effect of environmental enrichment (EE) on the recovery of disturbed function in the neonatal WMI model, male model rats were divided into two groups and grown in either condition of EE or standard environment (SE) from P25. Behavioral tests (hindlimb retraction, beam walk ability, horizontal ladder) were performed at P70. Although EE has no effects on behavioral tests except for ladder test in sham-operated control, significant effects of EE on behavior were shown in the WMI model at P70. EE effect was shown as early as P35 in the model that exhibited more severe motor deficits in SE condition compared to sham-controls. In the WMI model, less thickness of the ipsilateral sensorimotor cortex was shown at P70. However, the thickness of the cortex in the model of EE group was comparable to the opposite side, and the thickness was also similar level to sham-controls. To know if cortical motor map and electrical responsiveness in the sensory-motor cortex of the WMI model are changed by EE, intracoritical microstimulation was performed at P70. It revealed that the thresholds of hip joint and hindlimb movement were significantly reduced and the cortical area of hip joint and hindlimb was significantly increased by EE. These data suggest that EE during the period of development has effects on motor function in the WMI model, accompanied with significant changes of the cortical motor map and the responsiveness in the sensorimotor cortex.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-380 プロプラノロールは片頭痛モデルマウスの脳血流変化および痛み関連行動を抑制する Yuki Kurauchi（倉内祐樹）¹，Makito Haruta（春田牧人）²，Risako Tanaka（田中理紗子）¹，Kiyotaka Sasagawa（笹川清隆）²，Jun Ohta（太田淳）²，Takahiro Seki（関貴弘）¹，Hiroshi Katsuki（香月博志）¹ ¹熊本大院・生命・薬物活性 ²奈良先端大・物質創成・光機能素子 Propranolol, a β-adrenergic receptor blocker, is one of the most commonly used prophylactic drugs for migraines. Cortical spreading depression (CSD) is the propagation wave of neuronal excitation along with cerebral blood flow (CBF) changes over the cerebral cortex and has been implicated in the pathological process of migraine auras and its pain response. However, the effect of propranolol on CSD-related CBF changes and behavioral responses remains poorly understood. Here, we measured CSD-related CBF responses using a micro-device with a green light emitting diode (LED) and micro-complementary-metal-oxide-semiconductor (CMOS) image sensor and evaluated pain-related reduced locomotor activity in mice. An injection of KCl into the cortex caused CSD-related CBF changes; however, propranolol prevented the increase in CBF as well as delayed the propagation velocity in KCl-induced CSD. Furthermore, KCl injection reduced locomotor activity and induced freezing behavior in awake and freely moving mice, which were prevented by propranolol treatment. These results suggest that the modulation of CSD-related CBF responses by the blockade of β-adrenergic receptor contributes to its prophylactic effects on migraines.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-381 脳虚血環境下での肥満細胞脱顆粒におけるGPR3の役割 Shigeru Tanaka（田中茂）¹，Yuhki Yanase（柳瀬雄輝）²，Yuhki Hamakawa（浜川雄輝）¹，Masahiro Yamamoto（山本真弘）¹，Hiroko Shiraki（白榊紘子）¹，Kana Harada（原田佳奈）¹，Izumi Hide（秀和泉）¹，Norio Sakai（酒井規雄）¹ ¹広島大院医歯薬神経薬理 ²広島大院医歯薬皮膚科学 The G protein-coupled receptor 3 (GPR3) is a member of the class A rhodopsin-type GPCR family that is highly expressed in various neurons. GPR3 is unique in its ability to constitutively activate Gαs protein without the addition of ligands, thereby elevating the basal level of intracellular cAMP. We previously reported that neuronal expression of GPR3 enhances neurite outgrowth and neuronal survival. However, the physiological functions of GPR3 following brain ischemia remain unknown. In the current study, we investigated whether GPR3 expression was modulated following brain ischemia. GPR3 mRNA expression was found to be transiently upregulated in the ischemic hemisphere as early as 4 h after transient middle cerebral artery occlusion in Wistar rats and C57BL/6 mice. However, GPR3 promoter activity was decreased in neurons in the ischemic hemisphere in mice. Analyses of Percoll density gradient fractions from an ischemic brain homogenate indicated that mast cells are the source of GPR3 in the ischemic brain. It has been suggested that ATP from damaged cells can activate brain-resident mast cell degranulation, and released histamine and proteases from mast cell alter the permeability of the blood-brain barrier, thereby modulating vasogenic edema and immune cell infiltration. We then investigated the expression of GPR3 in mast cells and its role in degranulation using mouse bone marrow-derived mast cells (BMMCs). Indeed, degranulation of BMMCs was significantly induced by IgE-mediated DNP-BSA stimulation or ATP and strongly activated by the addition of both ATP and adenosine. In addition, degranulation of BMMCs was significantly inhibited by upregulation of intracellular cAMP. GPR3 mRNA expression in BMMCs was hardly detectable in the unstimulated condition, however it was highly upregulated as early as 1-2 h after BMMC stimulation. Finally, the effect of GPR3 expression on degranulation of BMMCs was investigated. BMMCs from GPR3 knockout mice showed significantly increased degranulation in response to various degranulation stimuli compared with that from the wild-type mice. Taken together, these results indicate that GPR3 has a potential role in inhibiting the degranulation of mast cells, thereby may modulate inflammatory responses triggered by brain ischemia.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-382 脳梗塞領域における胆汁酸合成メカニズムの解析 Souichi Oe（大江総一），Yukie Hirahara（平原幸恵），Saori Wada（和田早織），Hisao Yamada（山田久夫）関西医大医第一解剖 Cerebral infarction is a disease caused by cerebrovascular function failure and subsequent ischemic conditions. We have found that bile acid appears in abundance in the ischemic region in the permanent middle cerebral artery occlusion (pMCAO) model mice by employing imaging mass spectrometry analyses. Furthermore, we revealed the existence of bile acid synthases in neurons by immunohistochemistry. To clarify the molecular basis of bile acid biosynthesis in the ischemic brain, we focused on the transcriptional regulation of Cyp7a1, the rate-limiting enzyme of bile acid synthesis. Bioinformatic analyses using the transcription factor database revealed that Hif1alpha and Stat3 can bind to the Cyp7a1 promoter region. Immunohistological analyses of the distribution of the transcription factors in these ischemic regions showed that the expression levels of Stat 3 and Cyp7a1 increased in neurons but the expression of Hif1alpha was not. Furthermore, we performed a Cyp7a1 promoter assay using neuroblastoma Neuro2a cells. The results showed the little promoter activity under normal condition, but significant increase was detected by ischemic condition. Deletion of GAS element, which is the binding motif of Stat3, in Cyp7a1 promoter reduced the promoter activity under the ischemic condition. Furthermore, this increased activity was suppressed by administration of the selective inhibitor of Stat3. These results suggest that bile acid biosynthesis in the ischemic region is promoted via Stat3-dependent up-regulation of Cyp7a1 expression, and imply that it could function as a response mechanism for environmental changes in the acute phase of cerebral infarction.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-383 リハビリテーション促進薬の開発に向けた機能代償過程の分子イメージング Hiroki Abe（阿部弘基）¹，Susumu Jitsuki（実木亨）¹，Waki Nakajima（中島和希）¹，Yumi Murata（村田弓）²，Aoi Jituski-Takahashi（実木-高橋葵）¹，Tomoyuki Miyazaki（宮崎智之）¹，Hirobumi Tada（多田敬典）¹，Akane Sano（佐野亜加根）¹，Kumiko Suyama（須山紅美子）¹，Nobuyuki Mochizuki（望月修征）^1,3,4，Takashi Komori（小森隆司）^1,3,4，Hitoshi Masuyama（増山仁）^1,3,4，Tomohiro Okuda（奥田智博）^3,4，Yoshio Goshima（五嶋良郎）⁵，Noriyuki Higo（肥後範行）²，Takuya Takahashi（高橋琢哉）¹ ¹横浜市立大学大学院医学研究科生理学 ²産業技術総合研究所人間情報研究部門 ³富山化学工業株式会社 ⁴富士フィルム株式会社 ⁵横浜市立大学大学院医学研究科分子薬理神経生物学 Functional recovery in rehabilitation involves neuroplastic changes in non-injured brain regions, which compensate for the functional loss induced by the brain damage(Nudo, et al., Science, 1996). One basic mechanism underlying neural plasticity is the trafficking of AMPA receptors to synapses(Takahashi et al., Science, 2003). Recently, it is reported by our team that a small compound, edonerpic maleate (T-817MA), which facilitate AMPA receptors trafficking have the potential to accelerate motor function recovery after brain damage in rodent and in non-human primate(Abe et al., Science, 2018). Hence, there arises the needs to reveal the relationship between the AMPA receptors dynamics and functional recovery. Unfortunately, current brain imaging techniques cannot detect the neuroplastic changes occurring during rehabilitation. To overcome this limitation, the AMPA receptor imaging technique has been developed by our team. It has been evaluated that the novel positron emission tomography (PET) tracer [11C] K-2 has a high specificity to AMPA receptors. Using this tracer, which allows for the imaging of AMPA receptors in the living brain, we have found that regions of high AMPA receptor accumulation play an important role in compensating for brain damage-induced functional loss in rodents and humans. In rodent model, AMPA receptors accumulation during the recovery process after brain damage detected by AMPA PET imaging especially in peri-injured region, in which focal injection of AMPA receptors' antagonist could inhibit the recovered-function. In stroke patients in subacute phase, AMPA receptor accumulation increased in motor related cortex, basal ganglia and cerebellar during the rehabilitation process for one month. Thus, the AMPA receptor imaging technique would allow us to reveal the compensative brain regions after brain damage in a novel manner.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-384 アルツハイマー病モデルマウスにペランパネルを急性投与すると間質液中のAβ値が減少する Sakiho Ueda（上田紗希帆）¹，Akira Kuzuya（葛谷聡）¹，Masakazu Miyamoto（宮本将和）¹，Yoshitaka Tashiro（田代善崇）³，Ayae Kinoshita（木下彩栄）²，Ryosuke Takahashi（髙橋良輔）¹ ¹京都大学医学研究科　脳病態生理学講座 ²京都大学医学部人間健康学科 ³国立研究開発法人国立長寿医療研究センター認知症先進医療開発センター Amyloid-β protein (Aβ) is a key initiator in the pathogenesis of Alzheimer's disease (AD). Chronic disruption of Aβ metabolism accompanied by high production and/or low clearance could increase regional Aβ levels in interstitial fluid (ISF), finally leading to local Aβ deposition in AD brains. Recent studies shed light on hippocampal network hyperexcitability as a very early event in AD patients. Since enhanced neuronal/synaptic activity can promote presynaptic Aβ production and release into interstitial space, hippocampal hyperexcitability is considered as one of the promising therapeutic targets to prevent Aβ deposition in AD. A recent study of an AD transgenic mouse model reported aberrant Ca2+-permeable AMPA receptor expression in young mice prior to Aβ deposition. Thus, we asked whether antagonizing AMPA receptors could reduce ISF Aβ levels in an AD mouse model. Using in vivo brain microdialysis that allows for a real-time sampling of ISF, we investigated hippocampal ISF Aβ levels of APP transgenic J20 mice in which microdialysis probes were implanted in the hippocampus. To characterize age-dependent changes in ISF Aβ levels, we measured ISF Aβ₄₀ and Aβ₄₂ levels by ELISA in either young J20 mice without Aβ deposition (2- to 5-month-old) or old J20 ones with Aβ deposition (8- to 11-month-old). Next, young J20 mice were subjected to a single oral administration of perampanel (PER), a selective AMPA receptor antagonist, and evaluated dynamic changes in ISF Aβ levels before and after the treatment. A significant reduction in ISF Aβ levels concomitant with a high ratio of Aβ₄₀/Aβ₄₂ was observed in the old mice, as compared to young ones. In the young mice treated with a single oral administration of PER (2mg/kg), ISF Aβ levels were decreased by 50-60% in approximately 2 hours after the treatment, followed by 4 hours of recovery to baseline levels. Reduction of ISF Aβ levels with a high ratio of Aβ₄₀/Aβ₄₂ in the old J20 mice compared to young ones could represent age-dependent progression in sequestering soluble oligomeric Aβ species leading to Aβ deposition. Acute administration of PER can rapidly reduce ISF Aβ levels in the young J20 mice, possibly in an activity-dependent manner.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-385 アメフラシ中枢へのヒトアミロイドβ投与に伴う摂食障害の発現 Tatsumi Nagahama（長濱辰文）¹，Tomoya Minagawa（皆川友哉）²，Hiroshi Nakajima（中島啓）²，Miki Tokoro（所美樹）²，Mizuki Wakuta（涌田みずき）²，Yuki Enomoto（榎本由貴）²，Setsuko Nagahama（長濱節子）³ ¹帝京平成大学健康メディカル学部 ²東邦大学薬学部 ³帝京平成大学ヒューマンケア学部 Alzheimer's disease (AD) is a neurodegenerative disorder affecting cognitive functions. In addition, eating disorder is often reported in the AD patients. It is widely accepted that the accumulation of amyloid-β peptide (Aβ) is a major cause of the pathogenesis of disease. In our preliminary experiments, Aβ 1-42 monomer (m-Aβ) was injected into the Aplysia body cavity at a final concentration of 300 nM in body fluids and explored the effect of m-Aβ on the food intake. We explored amount of food consumed by measuring the seaweed that remained in the cage at a given time (2 or 4 hrs) after placing the seaweed in an individual animal cage every morning. Food intake was quantified as the percentage of body mass. The beginning of stable seaweed consumption determined the start of experiments. M-Aβ was applied on the fourth day (single application), or the fourth and sixth days (double application) after the start of experiments. In a single application, the amount of food intake decreased only on the next day after application. On the other hand, double application significantly diminished the amount of the food intake for at least five days from the next day after the first application. These results suggest that sufficient amount of m-Aβ may aggregate on the cell surface of Aplysia brain and reduced amount of food intake. In order to ascertain it, we next applied Aβ 1-42 oligomer (o-Aβ) directly on the surface of the buccal ganglia in the surgical treatment on the fourth day after the start of experiments. Then small pieces of filter paper soaked in 1 or 3 μM o-Aβ solution (monomer concentration) were placed on the bilateral buccal ganglia for 1 hour, with exchanging the pieces for the fresh ones every 10 min. Application of 3 μM o-Aβ significantly diminished the amount of food intake for at least five days from the next day after the treatment although application of 1 μM o-Aβ scarcely affected it. These results suggest that application of sufficient amount of o-Aβ to the buccal ganglia caused decline of the food intake by affecting the feeding neural circuit and the synaptic dysfunction induced by o-Aβ may be useful for understanding of the behavioral disorder in AD. Acknowledgment: We wish to thank Misses M. Hayashi and K. Imai for their helpful preliminary experiments for techniques of Aβ application to the buccal ganglia in the surgical treatment.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-386 アクアポリン４欠損に感受性のアミロイド沈着・神経炎症に続いて起こる第二の神経変性過程 Yoichiro Abe（阿部陽一郎）¹，Natsumi Ikegawa（池川夏実）²，Minetaka Murakami（村上峰宇）²，Takumi Tanaka（田中拓海）²，Takako Niikura（新倉貴子）²，Kyosuke Muramatsu（村松恭祐）³，Kaoru Yamada（山田薫）³，Tadafumi Hashimoto（橋本唯史）³，Motohito Goto（後藤元人）⁴，Keitaro Yamamoto（山本大地）⁴，Kenji Kawai（川井健司）⁴，Taichi Goda（合田和香美）¹，Keitaro Yoshida（吉田慶多朗）⁵，Satoko Hattori（服部聡子）⁶，Tsuyoshi Miyakawa（宮川剛）⁶，Kenji Tanaka（田中謙二）⁵，Masaru Mimura（三村將）⁵，Takeshi Iwatsubo（岩坪威）³，Jun-ichi Hata（秦順一）⁴，Masato Yasui（安井正人）¹ ¹慶應大医薬理 ²上智大理工情報理工 ³東京医大院医神経病理 ⁴実中研 ⁵慶應大医精神・神経科学 ⁶藤田医大総医研システム医科学 Aquaporin-4 (AQP4) is the most abundant water channel in the CNS specifically expressed in the astrocytic end-feet around blood vessels and is thought to contribute to water and ion homeostasis in the brain. It has been reported that reactive astrocytosis and microglial activation in acute brain damages are diminished in AQP4 knockout mice. It is well known that reactive astrocytes and activated microglia are accumulated around amyloid plaques, leading to chronic neuroinflammation in brains of Alzheimer's disease (AD) patients as well as AD model mice. Therefore, it is conceivable that AQP4 function is also required for neuroinflammatory responses in AD brains. In addition, it is suggested that brain lymphatic drainage systems play a role in amyloid β (Aβ) clearance, in which AQP4 may be involved. To investigate the implication of AQP4 in Alzheimer's disease (AD) pathology, we crossed an AD model, 5xFAD with AQP4 knockout mice. Age-dependent accumulation of both soluble and insoluble Aβ was observed and consistently, some genes related to neuroinflammation were upregulated in 5xFAD, regardless of AQP4 expression up to 10 months of age. These findings suggest that AQP4 is not involved in Aβ deposition or chronic neuroinflammatory responses in 5xFAD mice. In contrast, nighttime activity of AQP4-deficient 5xFAD was reduced from 5 months of age and was sharply deteriorated after 7 months of age. At this time point, daytime activity was also reduced and sometimes convulsions were observed in these mice. AQP4-deficient 5xFAD also showed loss of grip strength, suggesting that the reduction of activity in AQP4-deficient 5xFAD is accompanied by motor dysfunction. These phenotypes were not observed in 5xFAD expressing intact AQP4 up to 9 months of age. Taken together, these observations suggest that there is another neurodegenerative phase sensitive to AQP4 deficiency other than Aβ deposition or neuroinflammation in 5xFAD mice.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-387 D-β-ヒドロキシ酪酸はアミロイド β オリゴマー誘導性記憶障害を改善する Narumu Oya（大弥成夢），Yu Otagaki（太田垣優），Tsuyoshi Inoue（井上剛）岡山大院医歯薬生体物理化学 Alzheimer's disease (AD) is the most common disease in dementia. It is well known that the memory impairment of AD patients is caused by the aggregation of amyloid β (Aβ) in the brain. Aβ oligomer, a low aggregated form of Aβ, is neurotoxic, and previous studies have shown that the intracerebroventricular (i.c.v.) injection of Aβ oligomer induces the memory impairment in mice. In this study, we examined the effect of D-β-hydroxybutyrate on the Aβ oligomer-induced memory impairment in mice. D-β-hydroxybutyrate is a ketone body, which increases by taking the ketogenic diet and acts as an energy substrate available in the brain. The ketone bodies and ketogenic diet have been well studied in epilepsy, but less studied in AD. We used a novel object recognition test to evaluate long-term memory of mice, and first confirmed that the i.c.v. injection of Aβ oligomer induced the memory impairment in mice. We then found that intraperitoneal (i.p.) injection of D-β-hydroxybutyrate (600 mg/kg) improved the Aβ oligomer-induced memory impairment. In addition, the improvement of the memory impairment was observed by the injection of D-β-hydroxybutyrate 30 min before memory acquisition, but not 120 min before memory acquisition. We then examined the concentration of D-β-hyroxybutyrate in the blood and cerebral spinal fluid (CSF) increased by the i.p. injection of D-β-hydroxybutyrate. The i.p. injection of D-β-hydroxybutyrate (600 mg/kg) increased its blood concentration to about 4 mM and its CSF concentration to more than 1 mM. The CSF concentration peaked at about 20 min after the injection, and was back to the basal level 120 min after the injection. This kinetics of D-β-hydroxybutyrate in CSF is consistent with the time window of the memory improvement by D-β-hydroxybutyrate. These results show that D-β-hydroxybutyrate improves Aβ oligomer-induced memory impairment in mice.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-388 タウオパチーモデルマウス脳における非凝集型リン酸化タウの組織学的検出 Tomohiro Miyasaka（宮坂知宏）^1,2，Yuto Higashi（東優人）¹，Satoko Kakuda（角田聡子）¹，Nobuto Kakuda（角田伸人）² ¹同志社大学生命医科学部 ²同志社大学神経変性疾患研究センター Tauopathies are neurodegenerative diseases characterized by the deposition of abnormal tau protein in the affected neurons. Although, the mechanisms of the pathogenesis of tauopathy is not fully revealed, it is considered that the tau abnormalities, including hyperphosphorylation or other posttranslational modifications and aggregation, is involved. Based on the cumulative analyses, currently, the focus of the studies in toxic species of tau shifts from insoluble tau-inclusions to soluble abnormal species of tau, including tau oligomer or granule that are formed by tau and other molecules. AT8 is an antibody specifically label tau phosphorylation at Ser202 and Thr205. Because its high specificity and selectivity, AT8 is used for the first choice antibody for detection of pathological tau in the brains of AD and model animals in biochemical and histochemical procedures. However, in contrast to the phospho-tau-inclusions, phosphorylated soluble tau, that did not constitute inclusions like neurofibrillary tangles, was hardly labeled by AT8 in immunohistochemistry by unknown reasons. This issue makes it difficult to show the full mapping of the distribution of AT8 positive tau in tissues, and limit to analyze the toxic tau molecules in tauopathy research. In this study, we tried to develop a new method to enable us tissue staining of AT8 positive tau that did not form inclusions. Tau phosphorylation in AT8 site is reversibly enhanced by hypothermia induced by anesthesia in mice. Conventional handling of the tissues using various fixative and antigen retrieval solutions did not achieve tau staining in phosphorylated tau in the brains of hypothermia mice model. However, we found that AT8 staining is possible by a combination of fresh frozen section, formalin vapor fixation, and ionic detergents treatment. This suggests that the low antigenicity of AT8 in soluble tau is caused by its steric hindrance. Using this technique, now we truck the distributions of soluble phosphorylated tau in tau-transgenic mice brains.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-389 ペリサイト、オリゴデンドロサイト前駆細胞、血管内皮細胞間の相互作用に対するアミロイドβの病的意義 Masaru Toyokawa（豊川勝）¹，Takakuni Maki（眞木崇州）²，Seiji Kaji（梶誠兒）²，Yoshitaka Tashiro（田代善崇）³，Megumi Asada（浅田めぐみ）¹，Ryosuke Takahashi（高橋良輔）²，Ayae Kinoshita（木下彩栄）¹ ¹京都大院医　人間健康科学 ²京都大院医臨床神経 ³国立長寿医療セ研究所アルツハイマー [Background] Recent research has been revealing that neuron-centric view cannot explain all aspects of Alzheimer's disease (AD) pathology. Disruption of blood brain barrier (BBB) has been shown to play crucial roles on the onset and progression of AD. Although recent studies show that the dynamic interactions among pericytes (PCs), oligodendrocyte precursor cells (OPCs) and endothelial cells (ECs) may contribute to the regulation of BBB integrity and brain homeostasis, the pathogenic roles of these cell types and their interactions in AD remain to be elucidated. [Purpose] We investigated the behaviors of PCs, OPCs, and ECs in AD pathology and the influence of Aβ oligomers, one of pathogenic protein of AD, on these cell types and their interactions in vitro. [Material and Methods] Aβ42 oligomers were administered to each cell, and incubated for 48 hours, subsequently washed twice with fresh media and incubated for 24 hours. The conditioned media (CM) were then collected. WST-8 assay was performed for the evaluation of cell viability. BBB integrity was evaluated by measuring the trans endothelial electrical resistance score and the endothelial permeability of FITC-dextran plated on transwells. For the media transfer experiments, the CM from one cell type were diluted with the basal media for another cell type at a ratio of 50:50. Control media were prepared from empty wells and diluted in the same way as the CM collection. [Results] The viability of PCs treated with Aβ42 oligomers was significantly decreased in a concentration-dependent manner. The viability of OPCs also showed a trend to decrease. The viability of ECs was not decreased when treated with 5 and 10µM Aβ42 oligomers, while the BBB integrity was decreased. These results suggest that Aβ42 oligomer reduces the viability of BBB-composing PCs and OPCs, and that Aβ42 oligomer dampens BBB integrity with increased EC permeability. As for CM transfer experiments, we found that normal OPCs- and PCs- derived CM reinforce BBB integrity. However, the CM derived from Aβ-treated OPCs and PCs aggravate BBB functions. [Discussion and future plan] As one of underlying mechanisms for the above findings, MMP-9 may mediate the Aβ-induced disruption of BBB integrity in a non-cell autonomous manner via OPCs and PCs, which needs further studies for confirmation.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-390 場所および時間をコードする海馬の細胞はアルツハイマー病モデルマウスにおいて異なる脆弱性を有する Risa Takamura（高村理沙）^1,2,3,4，Kotaro Mizuta（水田恒太郎）^2,4，Yukiko Sekine（関根友紀子）^3,4，Tanvir Islam（イスラムタンビル）^3,4，Takashi Saito（齊藤貴志）^3,4，Takashi Takekawa（竹川高志）⁵，Masamichi Ohkura（大倉正道）⁶，Tomoki Fukai（深井朋樹）^3,4，Junichi Nakai（中井淳一）⁶，Toshio Ohshima（大島登志男）¹，Takaomi C Saido（西道隆臣）^3,4，Yasunori Hayashi（林康紀）^2,4,6 ¹早稲田大院先進理工生医, 東京, 日本 ²京都大院医, 京都, 日本 ³理研CBS, 埼玉, 日本 ⁴理研BSI, 埼玉, 日本 ⁵工学院大情報, 東京, 日本 ⁶埼玉大BBSSI, 埼玉, 日本 Functional breakdown of neuronal microcircuit due to the deposition of pathogenic proteins leads to the cognitive dysfunctions in Alzheimer's disease (AD). AD patients exhibit both spatial and temporal cognitive dysfunctions as early symptoms, in which the hippocampus plays an essential role. However, it is still unclear how the breakdown of these functions progresses under the AD pathology. Most studies of AD used transgenic mice highly overexpressing amyloid precursor protein (APP), which might cause artificial phenotypes. To address these issues, chronic two-photon calcium imaging was performed under the virtual spatial-temporal recognition task by using a new AD model mouse line (App^NL-G-F (Saito et al., 2014)) with a fluorescent calcium protein G-CaMP7 with calcium-insensitive DsRed2. We aim to elucidate the entire process of functional breakdown in hippocampal microcircuit of AD model mice. In AD mice, G-CaMP7- and DsRed2-positive aggregates were observed near toxic Aβ, which allowed us to detect location and growth of toxic Aβ in vivo. We detected the activity of 400-700 hippocampal CA1 pyramidal neurons over months with two-photon microscopy from head-fixed mice behaving under a virtual reality system, where running laps on a track (120 cm) are interleaved by 4-second wait period. In AD mice, the rate of stable place cells started declining at 4 months before the percentage of place cells declined and hyperactive cells (activity rate: ≥20%) increased at 7 months. These abnormalities occurred significantly at the vicinity of the aggregates. On the other hand, the number and the stability of time-encoding cells observed during the 4-second waiting time between laps, did not show any detectable impairment until 10-month. These results suggest that the place cells are more vulnerable to the pathological changes of AD, while time-encoding cells are more robust, likely due to the difference in circuitry mechanisms forming these cells. The establishment of simultaneous visualizing AD pathology and cell activity over months will also lead to effective drug evaluations.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-391 アルツハイマー病モデルとしての脳の糖尿病ラットにおける入力障害について Akiko S. Shingo（新郷明子）¹，Shozo Kito（鬼頭昭三）² ¹冲中記念成人病研究所 ²医療法人長谷川会湘南ホスピタル To confirm the intraventricularly streptozotocin-injected ""brain diabetes"" rat is a definite model of Alzheimer's disease, morphological studies of the dentate gyrus granular cell dendrites of the hippocampus were done. In our previous studies, it has been known that this ""brain diabetes"" rat shows spatial cognition disorders, increase of hippocampal amyloid beta protein with decreased immunoreactivities of IDE, somatostatin, Akt in the hippocampus. To more confirm that this rat is a definite model of Alzheimer's disease, dendrites of hippocampal granular cells were analysed by Sholl's method. As the results, in the ""brain diabetes"" rat, branching of the dendrites was increased with decreased spine densities. An intraventricular injection of an insulin analogue did not cause of total spine number. This has been considered due to overall null changes since new spine formation due to insulin stimulation has been compensated by the loss of old spines. It is concluded that cognitive decline in the ""brain diabetes"" rat is primarily due to impaired intracerebral insulin signalling and the ultimate results were injured excitatory inputs through the perforant pathway.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-392 Mnイオンの神経細胞内取り込みを利用したアルツハイマー病モデルウスのMn-MRI法による機能解析 Yuriko Inoue（井上由理子）¹，Hiromitsu Ezure（江連博光）¹，Hiroshi Moriyama（森山浩志）¹，Jyunji Ito（伊藤純冶）²，Chika Sawa（澤智華）³，Koichi Shiraishi（白石貢一）⁴，Yoshinobu Manome（馬目佳信）⁵，Akio Inoue（井上明男）⁶，Naruhito Othuka（大塚成人）¹ ¹昭和大学　医学部　解剖学講座　肉眼解剖学部門 ²昭和大学　保健医療学部　理学療法学科 ³昭和大学医学部　解剖学講座　顕微解剖学部門 ⁴慈恵会医科大学　医用エンジニアリング研究室 ⁵慈恵会医科大学　基盤研究施設　分子細胞生物学 ⁶京都大学医学部　附属脳機能総合研究センター We studied the cause of Alzheimer's disease using transgenic mouse expressing a large amount of amyloid. The transgenic mouse shows the decrease of memory by the Y-maze test. However, no clear anatomical difference was observed between transgenic and control mice. As nerve cells uptake Mn ions depending on nerve activity, We injected Mn ions into the abdominal cavity of mouse and examined Mn in the brain using Bruker 9.4T MRI machine with cryoprobe. Magnetic resonance imaging (MRI), uses a strong magnetic field and radio waves to generate images of parts of the body. The contrast of image is produced by the difference of stability of activated state produced by radio wave. Nerve cells uptake Ca ions during neuronal excitement. Ca ions were monitored using Ca indicator, Fluo4 which binds with Ca ions and the complex is fluorescent. Fluo4 is soluble, and does not enter into cells. Then, Fluo4-AM which is lipid soluble and pass through the cell membrane was used. Fluo4-AM is hydrolyzed into Fluo4 in the cytoplasm and stay in the cell. Mn ions also pass through the Ca channel and enter into nerve cells. Mn ions bind more strongly with Fluo4, but the Mn-Fluo4 complex is not fluorescent. Hippocampal neurons were isolated from Wistar rats at 18 days of gestation (E18). The cells after DIV(Days in Vitro)21 are used for the experiments. Activated state of proton in free water is rather stable compared to that in lipid and proteins. As Mn ions destabilize the activated state, T1 intensity is activated by Mn ions.The transgenic mouse shows the decrease in Mn incorporation to the dentate gyrus of hippocampus and also the cerebrum including cingulate cortex, motor and sensory area.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-393 アストロサイトにおける新規アミロイド前駆体タンパク質発現シグナル Ayaka Komatsu（小松彩夏），Sari Kishikawa（岸川咲吏），Miho Watanabe Iida（飯田渡辺和泉），Fumiko Harada（原田史子），Miho Terunuma（照沼美穂）新潟大学大学院　医歯学総合研究科　口腔生化学分野 Ammonia, which is a byproduct of cellular metabolism, has been reported as a potent neurotoxin that causes several negative effects in the central nervous system (CNS). Excessive ammonia levels have been found in the brain of Alzheimer's disease (AD) patients, therefore, ammonia could be a factor which contributes to the progression of AD. Astrocytes, the most abundant cells in the CNS, takes up excess ammonia and glutamate and convert it into glutamine. Previous studies have shown that hyperammonemia, which can result in hepatic encephalopathy, causes astrocyte swelling whereby a disturbance in neurotransmission. To examine the molecular mechanisms by which ammonia alters astrocyte function and neurotransmission, we used rat cultured astrocytes and treated with ammonium chloride (NH₄Cl). A 72-hour treatment with NH₄Cl significantly increased the expression of amyloid precursor protein (APP), which is a source of amyloid beta (Aβ) peptides. Interestingly, the amount of APP mRNA remained unchanged, suggesting that the constitutive degradation of APP maybe stopped by NH₄Cl. The expression of two enzymes, β- and γ- secretase, which cleave APP to produce Aβ were also unchanged, suggested that NH₄Cl treated astrocytes have an ability to produce Aβ. Finally, we used a mouse model of hyperammonemia to examine the expression of APP and Aβ in vivo. A thioacetamide treatment which causes severe liver failure in mice significantly increased blood ammonia levels. The expression of APP and Aβ were also increased in the hippocampus of these mice suggesting that ammonia induces the expression of Aβ. Our results demonstrated for the first time that ammonia contributes to the progression of AD by enhancing the production of APP in astrocytes.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-394 アルツハイマー病モデルマウスにおけるP2Y1受容体欠損の影響 Ami Tamada（玉田あみ），Tatsuhiro Hisatsune（久恒辰博）東京大院新領域創成科学先端生命科学細胞応答化学 Alzheimer's disease (AD) is one of the neurodegenerative diseases, and genes involved in neuroinflammation may affect symptoms in cognitive function in AD. We used a transgenic mouse (Tg) as an AD model mouse (APPswe / PS 1 dE 9) and investigated the role of P2Y1 receptor (P2Y1R) that associated with inflammatory response. To evaluate the influence on memory function under P2Y1R deficiency (KO), tests were conducted on Morris water maze (MWM) in mice at 12 months old with 2 groups of mice (Tg and Tg/KO). In MWM, mice swam in a circular pool for 60 seconds and then searched the underwater platform for 5 days (4 trials / day) (Tg: n = 4, Tg/KO: n = 6). From the 7th day, we evaluated the reverse learning for 3 days. There was a significant improvement tendency with Tg/KO over Tg (p = 0.0024). We also evaluated the amount of Aβ (42) contained in serum of Tg and Tg/KO mice (n = 3 each). By ELISA, we found significantly larger amount in Tg/KO (p = 0.0101). This may be due to the possibility that the metabolism of amyloid plaques, which is a characteristic in AD, was promoted when P2Y1R was deficient. In summary, both the behavioral experimental data showing signs of improvement and the difference in the amount of amyloid plaques in blood suggest that P2Y1 receptor is involved in pathophysiological processes in AD. We are in progress to increase the number of mice to verify these findings and to confirm the expression of P2Y1R and the activity of glial cells to deepen our understanding of the role of P2Y1R in AD pathogenesis.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-395 アルツハイマー病関連遺伝子、リン酸化タウ変異およびアミロイド前駆体タンパク質変異による認知機能低下 Ayaka Higuchi（樋口文香），Tatsuhiro Hisatsune（久恒辰博）東京大院新領域創成科学先端生命科学細胞応答化学 Alzheimer's disease (AD) is one of the major causes of dementia. In the brain of the AD patients, the neuropathological characteristics, such as the nerve fibril change due to the phosphorylation of tau protein and senile plaques formed by accumulation of amyloid β (Aβ), are seen, but it is not known the causal correlation between Aβ and tau. In this study, we tested cognitive function of 4 groups of the six months old mice, APP/Tau, APP, Tau, wild type (WT), in order to understand the synergistic effects between tau mutation and Aβ accumulation in cognitive decline. To evaluate their abilities of spatial memory function, we performed Morris water maze (MWM) (APP/Tau; n=10, APP; =10, Tau; n=10, WT; n=10) In MWM, we performed 4 trials per day and days of acquisition were divided into the initial 5 days and then the reversal session (the position of platform was changed) for 3days. As a result, the difference was not seen in initial learning between the four groups of mice (F= 1.4166, p = 0.2587: Multi-way repeated measures ANOVA), but in reversal learning task we detected significant difference between four groups. (F= 5.1012, p = 0.0061: Multi-way repeated measures ANOVA). Their abilities of spatial memory function of APP/Tau mouse were lower significantly than of WT mouse. We could detect a sign of the cognitive functional decline in the APP/Tau mouse, but could not specify the effect of Aβ or tau which influences cognitive decline. Apparent difference may be seen by evaluating it with the older mouse. We performed the same memory test in the eight months old mice. As a result we detected significant difference between WT and APP/Tau in reversal learning test (APP/Tau; n=4, WT; n=4) (F= 9. 8462 p = 0.0061: Multi-way repeated measures ANOVA). In addition, we performed the experiment with the contextual fear conditioning test. We also processed brain slices of theses mice for immunostaining in order to evaluate neuro- inflammation by comparing the quantity of phosphorylation tau protein and the amount of Aβ accumulation.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-396 若齢アルツハイマー病モデルマウス(3xTg AD)における自発走行障害 Koji Usui（臼井弘児）¹，Munenori Ono（小野宗範）²，Nobuo Kato（加藤伸郎）²，Shigenori Kawahara（川原茂敬）¹ ¹富山大学大学院理工学研究部 ²金沢医大医生理 Alzheimer's disease (AD) is a progressive cognitive disease and the prominent deficits usually appear on the later stage in life. Early detection and treatment of the deficits would prevent them from becoming worse. Therefore, it is important to find out a sensitive behavioral assay to the early abnormalities. For this purpose, we examined the usefulness of a modified version of wheel running task, in which animals learned to voluntarily run in a wheel. We used 2- and 4-month old triple-transgenic (3xTg) AD mice and their background 129/Sv mice as controls, n=8 each. Mice were enclosed in a plastic running wheel of 10 cm diameter and allowed to freely run for five minutes every day for five days. Running data were recorded by a rotary encoder connected to the axis of wheel and evaluated by two parameters: average and maximum speed of running. The average and maximum running speed of 4-month old control mice increased from 6.6 to 20 cm/s and from 24 to 41 cm/s, respectively, during five days of training. By contrast, the running parameters of 4-month old 3xTg mice did not change so much: the average and maximum speed increased from 2.0 to 6.5 cm/s and from 11 to 18 cm/s, respectively. There were significant differences between 3xTg mice and control mice (average speed: p < 0.01, max speed: p < 0.005, two way repeated measures ANOVA). There were similar impairments in 2-month old 3xTg AD mice, compared with the control mice. In this work, we found that 3xTg-AD mice have an impairment in voluntary wheel running training, compared with the control mice, already at 2 months old. Although the responsible sites or mechanisms for this impairment are not clear yet, the results suggested the usefulness of this test, which is easy to perform and might be sensitive to some deficits associated with AD.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-397 プレセニリン１/２キメラ体を用いたガンマセクレターゼ活性調節機構に対する検討 Shinsuke Matsuzaki（松﨑伸介）^1,4，Melissa Eccles（Eccles Melissa）²，Hironori Takamura（高村明孝）^3,4，Miheer Sabale（Sabale Miheer）²，Taiichi Katayama（片山泰一）⁴，David Groth（Groth David）²，Imran Khan（Khan Imran）²，Mark Agostino（Agostino Mark）^2,5，Giuseppe Verdile（Verdile Giuseppe）^2,6,7，Paul Fraser（Fraser Paul）^3,8 ¹和歌山県立医大医薬理 ²Sch. of Pharmacy and Biomedical Sci., Curtin Health Innovation Research Institute, Curtin University, Bentley, Australia ³Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, Toronto, Canada ⁴大阪大院連合小児発達分子生物遺伝 ⁵Curtin Institute for Computation, Curtin University, Bentley, Australia ⁶Sch. of Medical Sci., Edith Cowan University, Joondalup, Australia ⁷Sch. of Psychiatry and Clinical Neurosciences, University of Western Australia, Crawley, Australia ⁸Department of Medical Biophysics, University of Toronto Presenilin-1 (PS1) is essential for em-bryonic development and missense mutations are a primary cause in familial Alzheimer's disease. In contrast, loss of presenilin-2 (PS2) has only limited consequences and has been shown to have reduced proteolytic activity as compared to PS1. However, it is unclear if this is due to the inherent properties of PS2 or the particular γ-secretase complex into which it is assembled. We investigated chimeric proteins where PS1 domains, with demonstrated sequence divergence compared to PS2, were swapped into a PS2 backbone. Catalytic activities were assessed using cell-free processing assays for the amyloid precursor protein (APP) proteolytic fragments that constitute the amyloid β (A β) peptide, amyloid intracellular domain (AICD), and the Notch receptor fragment, Notch intracellular domain (NICD). The results indicate the PS1 transmembrane helices 3-4 (TM3-4) combined with PS1 C-terminal region (Ct) are the minimum requirement to increase the ability of PS2 to process APP. However, any chimera containing the PS1 TM3-4 increased Notch processing. Molecular dynamic simulations of models created based on cryoEM structure of γ-secretase, revealed changes in substrate binding energies associated with PS1 TM3-4 and Ct. These findings suggest that certain combinations of PS1 domains are functionally important and that different combinations of these are key to its activity on different substrates.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-398 SV2BはSV2AよりもBACE1を抑制的に制御する Masakazu Miyamoto（宮本将和）^1,2，Akira Kuzuya（葛谷聡）²，Yasuha Noda（野田泰葉）¹，Kengo Uemura（植村健吾）²，Megumi Asada-Utsugi（浅田ー宇津木めぐみ）^1,2，Shinji Ito（伊藤慎二）³，Yoshiyasu Fukusumi（福住好恭）⁴，Hiroshi Kawachi（河内裕）⁴，Ryosuke Takahashi（髙橋良輔）²，Ayae Kinoshita（木下彩栄）¹ ¹京都大院医人間健康 ²京都大院医臨床神経 ³京都大院医学研究支援セ ⁴新潟大学腎研究センター腎分子病態学分野 Recent studies shed light on hippocampal network hyperactivity as a very early event in AD patients. Since amyloidβ protein (Aβ) is produced and released locally at presynapses in response to synaptic activity, synaptic hyperactivity is considered as one of the promising therapeutic targets to prevent Aβ deposition in AD. We previously identified synaptic vesicle protein 2B (SV2B) as a novel binding partner of BACE1 to inhibit amyloidogenic APP processing. Interestingly, the anti-epileptic drug levetiracetam targeting SV2A, another homologous isoform of SV can restore impaired synaptic transmission and cognitive function in AD model mice and patients. In the present study, thus, we aimed to compare the binding affinity of BACE1 and effects on BACE1-mediated APP processing between SV2A and SV2B. Co-transfection of wt BACE1 with either SV2A, SV2B or empty vector was performed in HEK293 cells. The conditioned media were subjected to ELISA assay to detect secreted levels of Aβ40, Aβ42 and sAPPβ, respectively. The lysates were used for co-immunoprecipitation assay followed by Western blot. SV2B overexpression led to significant reduction in the levels of Aβ40 and Aβ42 as well as sAPPβ in the conditioned media, compared with SV2A or control overexpression. Further, BACE1 was preferably co-immunoprecipitated with SV2B as compared to SV2A. SV2B rather than SV2A preferably interacts with BACE1 and exerts significant inhibitory effect on BACE1-mediated APP processing.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-399 SCA36モデルショウジョウバエの樹立 Tomoya Taminato（田港朝也）¹，Morio Ueyama（上山盛夫）¹，Toru Yamashita（山下徹）³，Yoshio Ikeda（池田佳生）²，Koji Abe（阿部康二）³，Yoshitaka Nagai（永井義隆）¹ ¹大阪大院医神経難病治療学 ²群馬大院医脳神経内科学 ³岡山大院医歯薬学総合研脳神経内科学 Non-coding repeat expansion disorder (NRD) is inherited group of diseases such as C9orf72-linked amyotrophic lateral sclerosis (ALS)/frontotemporal dementia (FTD), myotonic dystrophy and several types of spinocerebellar ataxia (SCA), which are caused by abnormal expansions of repeat sequences located in the non-coding region of causative genes. A pathological hallmark of NRDs is the accumulation of expanded repeat RNA as RNA foci, which sequestrates RNA-binding proteins, resulting in their disfunction. Furthermore, it is known that the expanded repeat RNA is translated into proteins by unconventional repeat associated non-ATG (RAN) translation. Although these RNA foci and RAN proteins are suggested to be involved in the neurodegeneration, the precise molecular mechanisms are still unknown. SCA type 36 (SCA36) is an NRD which is caused by an GGCCTG repeat expansion (>25) in the first intron of the NOP56 gene. Interestingly, SCA36 patients develop not only progressive cerebellar ataxia but also motor neuron symptoms similar to ALS. To elucidate the molecular mechanisms of SCA36, we established novel Drosophila models expressing GGCCTG repeat RNA, because Drosophila is a powerful genetic model system for the identification of genetic modifier or drug candidates. Firstly, (GGCCTG)₄₃ repeat sequence with flanking region of NOP56 gene from SCA36 patient was successfully subcloned into plasmid vectors. To obtain a longer GGCCTG repeat sequence, we performed the repeat expansion technique. Briefly, we carried out two PCR reactions to amplify the repeat sequence using usual primers and either 5' or 3' special primers introduced with BsgI restriction sites, which cleaves just before or after the GGCCTG repeat sequence. Then amplicons digested with BsgI were ligated to double the repeat length, and transgenic flies were made using these constructs. When we expressed GGCCTG repeats in the compound eyes, we observed obvious eye degeneration in flies expressing (GGCCTG)₁₁₈, while (GGCCTG)₇ expressing flies showed almost normal eye phenotype. Moreover, we detected intranuclear RNA foci in salivary glands of flies expressing (GGCCTG)₁₁₈ by fluorescent in situ hybridization. The expression of RAN proteins in our model flies is under investigation. Elucidating molecular mechanisms using our model files will promote understanding on the pathogenesis and developing potential therapy of SCA36 and other NRDs.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-400 Very-low-density lipoprotein from metabolic syndrome patient-facilitated heart dysfunction, brain microglia expression and cognitive dysfunction in mice- from heart to brain Shiou Lan Chen（Chen Shiou Lan）¹，Ying Shao Lin（Lin Ying Shao）¹，Hsiang-Chun Lee（Lee Hsiang-Chun）^2,3,5，Mei-Chuan Chou（Chou Mei-Chuan）⁴，Ching-Kuan Liu（Liu Ching-Kuan）⁴，Chu-Huang Chen（Chen Chu-Huang）^3,5，Liang-Yin Ke（Ke Liang-Yin）³，Yun Fan（Fan Yun）¹ ¹College of Medicine, Kaohsiung Medical University ²Division of Cardiology, Department of Internal Medicine, KMU Hospital, Kaohsiung, Taiwan. ³Lipid Science and Aging Research Center, College of Medicine, KMU, Kaohsiung, Taiwan. ⁴Department of Neurology, KMU Hospital, Kaohsiung, Taiwan. ⁵Vascular and Medicinal Research, Texas Heart Institute, Houston, Texas, USA. Introduction: Epidemiological studies indicate that dyslipidemia might facilitate the progression of cognitive dysfunction in the elderly. Many studies claim that very-low-density lipoprotein (VLDL), one major lipoprotein in the blood, is important for developing of dyslipidemia. In addition, using anion exchange chromatography, we previously classified VLDL into different negatively charged grades based on its degree of surface anodization. We found that the highest electro-negative VLDL was the most cytotoxic and that it upregulated endothelial cell apoptosis. Patients with metabolic syndrome (MetS) had significantly higher concentration of plasma electro-negative VLDL than did healthy controls (15.2 vs 5.5 mg/dL, respectively). However, the effects and mechanisms of long-term exposure to electro-negatively charged VLDL in neuronal function needs further study. Methods: B6 mice were injected with VLDL extracted from humans with normal VLDL (nVLDL) or MetS (metVLDL). The Y Maze test was used to assess the cognitive function of mice whose brains were then dissected and analyzed. And the cardiac function were measured by echocardiography. Results: After three weeks of exposure to metVLDL, the mice showed early cognitive dysfunction: they did not recognize novel arm in Y Maze, which became even more evident after six weeks of exposure. Mice injected with nVLDL were also significantly affected after six, but not three, weeks of exposure to nVLDL. Microglia were activated in the medial prefrontal cortex (mPFC) and hippocampus both in nVLDL and in metVLDL groups after 6 weeks of exposure. The expression of microglia and TNF-α in mPFC were significantly correlated with serum cholesterol levels and with the cardiac chamber size and function. Moreover, the time spent in novel arm was significantly negatively correlated to the levels of microglia in mPFC, serum cholesterol levels and cardiac chamber size. Conclusion: Chronic exposure to VLDL, especially to metVLDL, induced cardiac dysfunction, elevated microglia and TNF-αexpression in the brain, and contributed to the early cognitive dysfunction.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-401 Optineurinの欠損はTDP-43陽性封入体形成とGolgiの断片化を生じる． Takashi Kurashige（倉重毅志）¹，Ryosuke Ohsawa（大澤亮介）²，Yui Yamashita（山下結衣）^3,4，Go Shioi（塩井剛）⁴，Hiroyuki Morino（森野豊之）²，Hidefumi Ito（伊東秀文）⁵，Yusuke Sotomaru（外丸祐介）⁶，Hirofumi Maruyama（丸山博文）⁷，Hideshi Kawakami（川上秀史）² ¹国立病院機構呉医療セ・中国がんセ脳神経内 ²広島大原医研分子疫学 ³理研CLST生体モデル開発ユニット ⁴理研CLST生体ゲノム工学研究チーム ⁵和歌山県立医大脳神経内 ⁶広島大自然科学研究支援開発セ ⁷広島大院医歯薬脳神経内 [Introduction]Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease caused by the loss of both upper and lower motor neurons. Approximately 10% of ALS patients have a family history of the disease, and 22 genes were already reported to be implicated in ALS. In addition, some of these genes were associated with various dysfunctions of multiple organs. However, the pathogenesis of ALS still remains unclear. We previously showed that optineurin (OPTN) mutations lead to the development of ALS. In this study, we investigate the mechanism underlying its pathogenesis by using Optn knockout (KO) mice. [Materials and methods] We generated Optn knockout (KO) mice and evaluated clinicopathologically at the age of 12, 18, and 24 months. [Results]The Optn (-/-) mice were normal in survival time, body weight, and motor functions. However, we found dot-like TAR-DNA binding protein 43 (TDP-43) -positive inclusions and fragmentation of Golgi apparatus, hallmarks of sporadic and familial ALS, in cell bodies of spinal cord motor neurons of the Optn (-/-) mice at the age of 24 months. Autophagic adaptor protein sequestosome1/p62 (p62) -positive inclusions were also detected in spinal cord motor neurons, and TDP-43 and p62 inclusions were partly detected in the same cells. These protein inclusions were also observed at earlier stages in sciatic nerve fibers. Epon-embedded sections revealed that large myelinated nerve fibers decreased at the age of 24 months. [Discussion]Our findings suggest that OPTN deletion results in dysfunction of protein homeostasis, thereby leading to neurodegeneration of motor neurons from distal axons. The Optn (-/-) mice recapitulate aspects of sporadic and familial ALS cases and may serve as a mouse model for invention of therapeutic strategies.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-402 Neurodegeneration and neuroinflammation study of viral encephalitis Chun-Jung Chen（Chen Chun-Jung），Su-Lan Liao（Liao Su-Lan），Cheng-Yi Chang（Chang Cheng-Yi），Yu-Ting Hung（Hung Yu-Ting） Taichung Veterans General Hospital Background: Neuroinflammation has a central role in common pathologies of brain diseases. Japanese encephalitis, which is caused by infection with mosquito-transmitted Japanese encephalitis virus (JEV), is an example of an immunopathological disease closely associated with pathological disruption of the BBB and severe neuroinflammation. Those immunopathological changes correlate well with the onset and disease progression of Japanese encephalitis and represent promising targets for therapeutic control. Methods: Cultured neuron, astrocytes, and microglia were prepared from cerebral cortices of Sprague-Dawley rats. JEV NT113, a JEV strain isolated from mosquito, was propagated in C6/36 cells and used to infect cultured cells. Results: Through the application of primary culture of neurons, astrocytes, and microglia establishment from neonatal rat brain, we found that JEV infection selectively caused cytotoxicity towards neurons and had negligible effect on astrocytes and microglia. In addition to direct neurotoxicity, alternative indirect neurotoxicity caused by soluble factors released by JEV-infected neurons and microglia also played critical roles. In response to JEV infection, neurons, astrocytes, and microglia activated the expression of several pattern recognition receptors, including TLR2, PKR, MAD5, RIG-I, IRF1, IRF3, IRF7, and NLRP3 leading to production of diverse bioactive molecules. Those molecules contributed to differential infectivity, cytotoxicity, direct/indirect neurotoxicity, and microglial activation. Microglia and neurons, particularly the former, expressed and released neurotoxic molecules. Together with relevant studies, released NO, TNF-alpha, IL-1 beta, glutamate, and HMGB1 showed crucial roles in JE-associated neurotoxicity during the course of JEV infection. Conclusion: The data advance our understanding about the neuroinflammation and damaging mechanisms of JEV regarding the involvement of selective pattern recognition receptors and indicate that pattern recognition receptors might represent alterative targets for the alleviation of JEV infection-induced neurodegeneration and neuroinflammation.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-403 Effect of H. erinaceus mycelium (HEM) against MPTP induced neurotoxicity and protective neuronal cell in vitro and in vivo via ROS-dependent and PAK1/cdc42/PI3K/AKT/LIMK2/MEK pathway Hsing Chun Kuo（Kuo Hsing Chun）¹，Kam‑Fai Lee（Lee Kam‑Fai）²，Shui-Yi Tung（Tung Shui-Yi）³，Li-Ya Lee（Lee Li-Ya）⁴，Wan-Ping Chen（Chen Wan-Ping）⁴，Chin-Chu Chen（Chen Chin-Chu）⁴ ¹Chang Gung University of Science and Technology ²Department of Pathology, Chang Gung Memorial Hospital at Chiayi ³Department of Hepato-Gastroenterology, Chang Gung Memorial Hospital, Chiayi ⁴GRAPE KING BIO Ltd, Taoyuan Background: Hericium erinaceus is well known as a valuable pharmaceutical and edible mushroom, that can produce mycelia and biological metabolites. Our previous study showed that physiological and brain histological changes provide H. erinaceus mycelium neuron-protective insights into the progression of Parkinson's disease. However, the mechanism by which its isolated diterpenoid derivative, erinacine A, therapeutic efficacy from MPTP-induced neurotoxicity remains poorly understood. Methods: Cell viability was determined by Annexin V FITC, propidium iodide staining and generation of ROS using a DCFDA staining assay to determine the effects of erinacine A treatment on the neuronal cell fate and plasticity of N2a cells. The efficacy of neurological deficits and pathogenesis are measured with H. erinaceus administration by histological examination and proteomic approaches. Results: Our results demonstrate that erinacine A treatment of N2a and neurons from C57BL6 mouse substantia nigra decreased cell cytotoxicity and reactive oxygen species production as well as increased cell proliferation, can act as an anti-neuroinflammatory agent to bring about therapy using an MPTP in animal models of oxidative stress. Furthermore, these modulation of MPP+ injury factors in the neurons treated with erinacine A seems to result in the mechanisms of ER stress-sustained inactivation of the IRE1, TRAF2, ASK1, p21, GADD45 and neuronal survival or plasticity, associated with sustained phosphorylation of the PAK1, AKT, LIMK2, ERK, Cofilin pathway as well as efficacy in vitro and in vivo. Taken together, our proposal indicated that HEM provide a potential agents and improvement neuronal differentiation. The findings provide further insight into the molecular targets through which erinacine A exerts its beneficial effect on neurodegenerative insults and suggests erinacine A may be a promising agent for aging disease in complementary and alternative medicine.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-404 多系統萎縮症モデルマウスにおけるオリゴデンドロサイト前駆細胞のα-synuclein蓄積制御 Asuka Sasaki（佐々木飛翔）¹，Chenghua Jin（金成花）¹，Tamaki Iwase（岩瀬環）^1,2，Ikuru Yazawa（矢澤生）¹ ¹国立長寿医療セ研究所バイオリソース ²名古屋市厚生院 Multiple system atrophy (MSA) is a neurodegenerative disease pathologically characterized by glial cytoplasmic inclusions (GCIs) and neuronal α-synuclein (α-syn) accumulation in the central nervous system. GCIs are confined to oligodendrocytes and are composed α-syn in the lesions. We previously generated a transgenic (Tg) mice model in which human wild-type α-syn was overexpressed in oligodendrocytes under the control of murine CNPase promoter as a model of MSA. Our studies revealed that oligodendrocytic insoluble α-syn inclusions induced neuronal α-syn accumulation, leading to neuronal degeneration. We then established primary culture cells derived from the brain of Tg mice, and showed that cystatin C (Cys C) was released by the oligodendrocytes expressing human α-syn. Cys C causes endogenous mouse α-syn accumulation in neurons. We demonstrated that Neuro2a cells treating with purified recombinant Cys C uptook the Cys C into the cells and that recombinant Cys C enhanced α-syn accumulation in the cells. In contrast, Neuro2a cells expressing endogenous Cys C did not exhibit an increase in the α-syn accumulation. Thus, we concluded that it was the exogenous Cys C released by the oligodendrocytes that induced neuronal α-syn accumulation. In the present study, we show that Tg mouse primary cultured cells contain less number of oligodendrocyte precursor cells (OPCs) than those in non-Tg mouse primary cultured cells. To assess the influence of OPCs on neuronal α-syn accumulation, we treated Tg mouse primary cultured cells with OPC transplantation or growth factors to increase OPCs. Our data suggest that OPCs play an important role in the neuronal α-syn accumulation in the Tg mouse CNS, which could be a novel therapeutic target for MSA.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-405 Interleukin-13 improved poststroke hyperglycemia and inflammation in permanent cerebral ischemia rat model Wen-Ying Chen（Chen Wen-Ying），Keng-Ying Liao（Liao Keng-Ying） National Chung Hsing University Background: Cerebral ischemia (stoke) is one of the leading causes of death and disability worldwide, whereas hyperglycemia is known as a risk factor for stroke prognosis. However, with the unknown mechanism, there has not been a protocol for treating post-ischemic hyperglycemia. On the other hand, research found that acute stroke secretes counter-regulatory hormones, pro-inflammatory cytokines, and other regulatory factors, which enhances insulin resistance. Interleukin-13 (IL-13), as a member of the Th2 anti-inflammatory cytokine family, was reported to be involved in glucose homeostasis. Studies showed that IL-13 is capable of enhancing insulin sensitivity and suppressing hepatic gluconeogenesis, and that overexpressing IL-13 reverses abnormal glucose metabolism and insulin resistance in obese mice. Methods: Stroke animal model was induced by permanent middle artery occlusion in male Sprague-Dawley rats and explored the mechanism of post-ischemic hyperglycemia and the role IL-13 may play. Results: Our results showed that post-ischemic hyperglycemia was accompanied by elevated insulin resistance index, elevated serum insulin concentration, elevated hepatic gluconeogenesis, and decreased insulin signaling. Meanwhile, systemic pro-inflammatory cytokine was also found significantly elevated and anti-inflammatory cytokine decreased. With IL-13 treatment, the above pathological state was reversed due to the JAK1/STAT6 signaling activation, which shifted the inflammatory state towards Th2 immunity. Conclusion: Our findings suggest that IL-13 treatment may be a new perspective for the prevention and control of post-ischemic hyperglycemia.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-406 球脊髄性筋萎縮症におけるSrcシグナル異常に対する治療法開発 Madoka Iida（飯田円）^1,2，Kentaro Sahashi（佐橋健太郎）¹，Naohide Kondo（近藤直英）¹，Hideaki Nakatsuji（中辻秀朗）¹，Genki Tohnai（藤内玄規）¹，Masahisa Katsuno（勝野雅央）¹ ¹名古屋大学大学院医学系研究科神経内科学 ²日本学術振興会特別研究員RPD [Objective] Spinal and bulbar muscular atrophy (SBMA) is a slowly progressive neuromuscular disease caused by the expansion of the polyglutamine tract in the androgen receptor (AR) gene. We aimed to clarify the upstream molecular changes involved in the pathological process of SBMA in neuronal and muscular tissues. [Methods] We measured the expression levels of 17 phosphorylated proteins using Bio-rad Bio-plex assays, in the spinal cord and skeletal muscle specimen of the mouse model of SBMA (AR-97Q mice) at three stages: pre-onset, early symptomatic, and advanced. Based on the findings in the assays, we analyzed the activation levels of Src pathway in cellular and mouse models of SBMA, and assessed the efficacy of Src kinase inhibitor (SKI) on cellular and mouse models of SBMA. We also investigated the alteration of phosphorylation levels of Src effector molecules using SKI-treated cellular and mouse models of SBMA. [Results] In Bio-plex assays, the expression level of phosphorylated Src (p-Src) was markedly up-regulated both in the spinal cords and skeletal muscles of AR-97Q mice prior to the onset of neurological symptoms. In spinal cord, the activation of Src sustained until the advanced stage of the disease. Src pathway was also up-regulated in neuronal and muscular cellular models of SBMA, induced pluripotent stem cell-derived motor neurons from patients with SBMA and human myogenic cells overexpressing AR-97Q. The intraperitoneal administration of A419259 trihydrochloride, an SKI, improved the behavioral and histopathological phenotypes of AR-97Q mice. We identified p130Cas as an effector molecule of Src; the suppression of the phosphorylation level of p130Cas attenuated the toxicity of polyglutamine-expanded AR. [Conclusions] Src pathway plays a causative role in the pathophysiology of SBMA. SKI has the potential to exert neuroprotective effects on patients with SBMA.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-407 SYNE1のC末端部変異とSCAR8患者における運動ニューロン病の関連 Kodai Kume（久米広大）¹，Hiroyuki Morino（森野豊之）¹，Osamu Komure（小牟禮修）²，Yukiko Matsuda（松田由喜子）¹，Ryosuke Ohsawa（大澤亮介）¹，Takashi Kurashige（倉重毅志）³，Yuhei Kanaya（金谷雄平）¹，Yui Tada（多田有似）¹，Hideshi Kawakami（川上秀史）¹ ¹広島大学原爆放射線医科学研究所分子疫学研究分野 ²尼崎だいもつ病院神経内科 ³独立行政法人国立病院機構呉医療センター神経内科 Background: Spinocerebellar ataxias (SCAs) are clinically and genetically heterogeneous disorders, classified as autosomal dominant, autosomal recessive, X-linked and mitochondrial. Spinocerebellar ataxia, autosomal recessive 8 (SCAR8) is caused by a mutation in SYNE1 and originally reported as late-onset cerebellar ataxia with few extracerebellar symptoms in French-Canadian families. Subsequent reports have expanded the clinical spectrum including cases associated with extracerebellar symptoms such as motor neuron disease (MND). Method: We evaluated two siblings from a consanguineous Japanese family. To identify the causative gene, we performed exome sequencing on the proband and validated the mutation in the proband and the proband's little sister using Sanger sequencing. We review the previously reported mutations and phenotype in patients with SCAR8. Results: We identified a novel homozygous nonsense mutation in the SYNE1 gene (NM_033071: c.19138C>T, p.R6380). In our cases and previously reported SYNE1* cases, significantly more mutations causing MND were located in C-terminal regions lower than the 5406th methionine at which a short isoform of nesprin-1, transcripts of SYNE1 starts (MND and cerebellar ataxia group were respectively 14/20 [70%] and 10/27 [37%]; chi-square test p = 0.025). Conclusions: We reported a novel nonsense mutation of SYNE1, p.R6380, causing SCAR8 associated with MND. Mutations in the C-terminal region of SYNE1* could be associated with lower motor neuron signs in patients with SCAR8, affecting the expression of the short isoforms of nesprin-1.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-408 遺伝的リスク因子を導入したパーキンソン病モデルマウスの作成 Tomoyuki Taguchi（田口智之）¹，Masashi Ikuno（生野真嗣）¹，Maiko Uemura（上村麻衣子）¹，Mari Hondo（本堂茉莉）²，Yusuke Hatanaka（畑中悠佑）¹，Norihito Uemura（上村紀仁）¹，Hodaka Yamakado（山門穂高）¹，Masashi Yanagisawa（柳沢正史）²，Ryosuke Takahashi（高橋良輔）¹ ¹京都大学大学院医学研究科　臨床神経学 ²筑波大学国際統合睡眠医科学研究機構 Objective Parkinson's disease (PD) is the most common movement disorder and is characterized by dopaminergic (DA) cell loss and the accumulation of pathological α-synuclein (α-syn), but its precise pathogenetic mechanisms remain elusive. To develop disease-modifying therapies for PD, an animal model that recapitulates the pathology and symptoms of the disease, especially in the prodromal stage, is indispensable. The aim of this study is to create mouse models reproducing the course of PD pathology and symptoms. Methods We generated bacterial artificial chromosome (BAC) transgenic mice harboring the entire human α-syn gene (SNCA) and its gene expression regulatory regions. In order to enhance the pathological properties of α-syn, we inserted into SNCA an A53T mutation, two single-nucleotide polymorphisms and a Rep1 polymorphism, all of which are causal of familial PD or increase the risk of sporadic PD. The mice were analyzed behaviorally, histologically and biochemically. Results A53T SNCA BAC transgenic mice expressed truncated, oligomeric and proteinase K-resistant phosphorylated forms of α-syn in the regions that are specifically affected in PD and/or dementia with Lewy bodies, including the olfactory bulb, cerebral cortex, striatum, substantia nigra and dorsal motor nucleus of vagus nerve. These mice exhibited rapid eye movement (REM) sleep without atonia, which is a key feature of REM sleep behavior disorder (RBD), at as early as five months of age. Consistent with this observation, the REM sleep-regulating neuronal populations in the lower brainstem, including the sublaterodorsal tegmental nucleus, nuclei in the ventromedial medullary reticular formation and the pedunculopontine nuclei, expressed phosphorylated α-syn. In addition, they also showed hyposmia at nine months of age, which is consistent with the significant accumulation of phosphorylated α-syn in the olfactory bulb. The DA neurons in the substantia nigra pars compacta degenerated, and their number was decreased in an age-dependent manner by up to 17.1% at 18 months of age compared to wild-type, although the mice did not show any related locomotor dysfunction. Interpretation A53T SNCA BAC transgenic mice recapitulated the pathological changes and prodromal symptoms of PD. This novel transgenic mouse model can provide an opportunity to understand the very early changes associated with the disease and to start disease-modifying therapies for PD.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-409 ラット歯周組織から青斑核への三叉神経系内侵入経路(超高磁場核磁気共鳴画像法) Yasuhiro Ooi（大井康浩）¹，Chizuko Inui-Yamamoto（乾-山本千珠子）²，Tadashi Sasai（笹井正思）³，Yoshitaka Nagase（永瀬佳孝）⁴，Yoshichika Yoshioka（吉岡芳親）⁵，Akitoshi Seiyama（精山明敏）⁶，Hiroyuki Ooi（大井博之）⁷，Junji Seki（関淳二）⁸ ¹大阪大学歯学研究科歯科薬物学講座 ²大阪大学歯学研究科第一口腔解剖学講座 ³大阪大学歯学研究科歯科放射線学講座 ⁴大阪大学歯学研究科第二口腔解剖学講座 ⁵大阪大学先導的学際研究機構 ⁶京都大学医学研究科人間健康科学系専攻情報理工医療学講座 ⁷八津統計相談室 ⁸関西大学システム理工学部物理・応用物理学科 Abstract Purpose: The locus coeruleus (LC) is degenerated by neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and multiple sclerosis. Since LC is adjacent to the mesencephalic trigeminal nucleus, it reminds us a possibility that the damage of LC caused by periodontitis triggers the neurodegenerative diseases. In order to verify this possibility, the continuity of cerebrospinal fluid (CSF) channel within the trigeminal nerve was investigated from periodontal tissue to the brainstem including LC by 11.7 T ultra-high field MRI. Methods: T2-weighted MRI with a spatial resolution up to 78 μm (repetition time=5000 ms; echo time=40 ms; slice thickness=250 μm, imaging plane=coronal) was applied to follow the trigeminal nerve both outside and inside the brainstem of normal rats at 10 weeks of age. Mean signal intensity and variance were obtained for twelve regions of interest including cranial nerves (branches of trigeminal nerve, optic nerve, facial nerve), white matter and cerebral cortex, and they were statistically compared with one another. Results: Twelve regions were divided by their mean signal intensity into the low signal intensity group [white matter, optic tract, optic chiasm, optic nerve outside the central nervous system] and the high signal intensity group [cerebral cortex (CC), spinal trigeminal tract (Vst), ophthalmic and maxillary nerve (V1/2), mandibular nerve, inferior alveolar nerve, trigeminal motor nerve outside and inside the brainstem, facial nerve]. Further, the variances of intensity distribution were significantly larger in V1/2 (outside the brainstem) and Vst (inside the brainstem) than in CC, which reflects that the state of water in the V1/2 is similar as in Vst but it is different from the CC. Conclusions: The present experimental results suggest the continuation of water, probably CSF, in the trigeminal nerve from outside to inside the brainstem. The water channel in the trigeminal nerve may offer a path to the pathogens in the patients with periodontitis to contribute the onset of neurodegenerative diseases.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-410 Manganese nitrate induced cytotoxicity and genotoxicity in microglia Yu-Hsiang Kuan（Kuan Yu-Hsiang）¹，Chun-Jung Chen（Chen Chun-Jung）²，Wen-Ying Chen（Chen Wen-Ying）³，Su-Lan Liao（Liao Su-Lan）²，Hui-Wen Lin（Lin Hui-Wen）⁴ ¹Chung Shan Medical University ²Department of Education and Research, Taichung Veterans General Hospital ³Department of Veterinary Medicine, National Chung Hsing University ⁴Department of Optometry, Asia University Manganese (Mn) is an essential trace element maintained at an optimal level in human body. Mn plays the important activator of enzymes in metabolism for protein digestion and utilization. The major exposure source of Mn is from the occupational environment, including welders and miners are the occupational exposure population. Manganese nitrate (Mn(NO3)2), a Mn compound, is the major material in alloy and battery. There are evidence have demonstrated that Mn is a potential risk factor in neurological disorders. Up to now, the evidence for Mn(NO3)2-induced cytotoxicity and genotoxicity and its related mechanism in microglia cells is not supportive. At the present study, we first found that Mn(NO3)2 induced cytotoxicity in BV2 microglia cells in a concentration and time-dependent manners. Genotoxicty, including microcucleus (MN) formation and DNA damage, was induced by Mn(NO3)2 in a concentration-dependent manner via alkaline single cell gel electrophoresis (COMET) and cytokinesis-block MN assays. In addition, apoptosis and related signal molecular mechanism, including caspases (-3, -8, and -9) activation and mitochondria disruption, were induced by Mn(NO3)2. ROS generation play an important role in apoptosis. Here, we also found intracellular ROS generation induced by Mn(NO3)2. More, N-acetyl-cysteine (NAC), a ROS scavenger, decreased the effects of cytotoxicity and genotoxicity induced by Mn(NO3)2.In conclusion, we had demonstrated that Mn(NO3)2-induced cytotoxicity and genotoxicity on BV2 cells were mediated by caspase-3,-8,and -9 activation. This study was supported by research grants from Ministry of Science and Technology of Taiwan (MOST 106-2320-B-040 -022 -MY3).		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-411 ALSにおいて発現が低下するADAR2遺伝子の発現をヒト脊髄運動神経にて制御しうる転写因子の探索 Naoki Hirose（廣瀬直毅）¹，Sayaka Teramoto（寺本さやか）^1,2，Takenari Yamashita（山下雄也）^1,2，Shin Kwak（郭伸）^1,2 ¹東京大院医 ²東京医大神経学分野 Pathogenesis of sporadic amyotrophic lateral sclerosis (ALS), a fatal neurological disease, is closely related to a decrease in the expression of adenosine deaminase acting on RNA 2 (ADAR2), an RNA editing enzyme, in the human spinal motor neurons (MNs). However, mechanisms underlying ADAR2 down-regulation in the ALS-MNs have been yet to be determined. Previously, we determined active promoter regions of the ADAR2 gene (ADARB1) in the MNs by conducting cap analysis of gene expression (CAGE) on laser-captured MNs of two non-ALS humans. Here, we performed in vitro luciferase assay on the promoters of ADARB1 by overexpressing transcription factors (TFs) that are expressed in the human MNs and previously shown to be down-regulated in sporadic ALS-MNs. We hypothesized that a decrease of TFs acting on ADARB1 promoters in the human MNs may lead to ADAR2 down-regulation in the ALS-MNs. First, based on our CAGE data and publicly available RNA-seq data on the laser-captured MNs, we determined TFs that are abundantly expressed in the human MNs. Then, from the published high throughput data of gene expression in laser-captured MNs, whole spinal cords and motor cortices of sporadic ALS patients, we selected TFs that were decreased in the tissues of ALS patients compared to those of non-ALS subjects. Third, in vitro luciferase assay on the ADARB1 promoters was performed by overexpressing each of the selected TFs in HeLa cells, showing that every TF had different activating effects on transcription at the ADAR2 promoters, which were strengthened by concomitant overexpression of a subset of the selected TFs. The present results suggest that different sets of multiple TFs expressed in the human MNs may regulate transcription at each of the ADAR2 promoters in the MNs additively or synergistically. Because these TFs have been shown to be decreased in tissues of ALS patients, altered expression of these TFs may play a role in ADAR2 down-regulation in MNs of sporadic ALS patients.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-412 ザンドホッフ病モデルマウスでのアストロサイトに発現するA_2A受容体の抑制はミクログリアの活性化を抑制する Yasuhiro Ogawa（小川泰弘）¹，Eiri Furusawa（古澤栄梨）¹，Takahiro Saitoh（齋藤貴洋）¹，Hiroki Sugimoto（杉本雄幹）¹，Takumi Omori（大森拓実）¹，Shinya Shimizu（清水伸弥）¹，Mika Yamazaki（山崎美香）¹，Hisatsugu Kondo（近藤久嗣）¹，Hitoshi Sakuraba（櫻庭均）²，Kazuhiko Oishi（大石一彦）¹ ¹明治薬大薬生命創薬・薬理学 ²明治薬大薬臨床遺伝学 Astrocyte-microglia communication influences the onset and progression of central nervous system (CNS) disorders. In this study, we determined how chronic inflammation by activated astrocytes affected and regulated CNS functions in Sandhoff disease (SD), a CNS lysosomal storage disorder. SD triggers intense CNS inflammation such as microglial activation and astrogliosis. It is caused by mutation of the HEXB gene, which reduces β-hexosaminidase (Hex) enzymatic activity in lysosomes, leading to accumulation of the substrate GM2 ganglioside in neuronal cells. Hexb^-/- mice display a phenotype similar to human patients that suffer from chronic inflammation characterized by activation of astrocytes and microglia. In Hexb^-/- mice, tremors and loss of muscle coordination begins at ~12 weeks. Interestingly, we found that reactive astrocytes expressed adenosine A_2A receptor in the cerebral cortices of Hexb^-/- mice at the later inflammatory phase. In cultured astrocytes, expression of A_2A receptor could be induced by astrocyte defined medium, and then the activation of the A_2A receptor induced ccl2 expression. In Hexb^-/- mice, inhibition of the A_2A receptor antagonized by istradefylline decreased the number of activated microglial cells and inflammatory cytokines/chemokines at 13 weeks. Thus, the astrocytic A_2A receptor is an important sensor that regulates microglial activation in the late phase of inflammation.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-413 低酸素による細胞死を抑制する新規低分子化合物はALSの進行を抑制する Tomonori Hoshino（星野友則）^1,2，Shu-ichi Matsuzawa（松澤秀一）^1,2，Ryosuke Takahashi（高橋良輔）² ¹京都大学医学部附属病院創薬インキュベーションラボ ²京都大学大学院医学研究科臨床神経学 Central nervous system (CNS) suffers from oxidative stress during ATP production. Decreased oxygen supply by impairment of blood supply results in critical damage of CNS. Since chronic hypoxia condition causes diverse effects such as cell death and inflammation by inducing excessive production of reactive oxygen species (ROS), protection from hypoxia damage is important for cell survival. Recent studies revealed that various markers of hypoxia have been changed in neurodegenerative diseases, such as amyotrophic lateral sclerosis (ALS), indicating that hypoxia is implicated in ALS. However, any therapeutic methods targeted hypoxia-induced pathway in ALS has not been developed yet. Previously, we screened small-molecule compounds library and identified a novel neuroprotective agent KUDN-1 which inhibit hypoxia-induced cell death. We hypothesized that the modulation of hypoxia signaling by KUDN-1 might protect motor neurons in rodent models of ALS in vivo. In this study, we show that the KUDN-1 prevents neurodegeneration in familial ALS rodent model. Administration of KUDN-1 into familial ALS rodent model (SOD1 G93A Transgenic mice) significantly attenuated disease progression, and improved locomotor dysfunction. We also found that KUDN-1 prevented motor neuron loss and gliosis. Our results indicate that KUDN-1 might be a potential therapeutic tool for cure of ALS. Moreover, these results suggest that modulation of hypoxia signaling pathway provides a good hint for developing therapy for other type of neurodegenerative diseases caused by excessive ROS.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-414 Modification of motor cortex forelimb lateralization preference in a hemiparkinsonian rat model Alain A Rios（Rios Alain A），Satoshi Nonomura（Nonomura Satoshi），Tomohiko Yoshizawa（Yoshizawa Tomohiko），Masanori, Kawabata（Kawabata Masanori,），Tomomi Sakairi（Sakairi Tomomi），Yutaka Sakai（Sakai Yutaka），Yoshikazu Isomura（Isomura Yoshikazu） Tamagawa University Brain Science Institute Motor cortex and basal ganglia undergo a dynamic remodeling of movement representation under parkinsonian state. One of these changes is the loss of the normal contralateral lateralized activity pattern, being evident as a decrease in the forelimb selectivity. The increase of the movement-related neurons responding to ipsilateral or bilateral limbs movements may cause some motor problems, such as impaired balance, decreased bimanual coordination and mirror movements. In order to explore the effect of dopamine depletion on the forelimb lateralization under parkinsonian state, we used a partial hemiparkinsonian model (6-hydroxydopamine lesion) in rats performing unilateral movements in a two pedal task, while recording from primary and secondary motor cortices. We summarize the finding as follow: 1) The unilateral 6-OHDA produce a movement impairment in the contralateral forelimb in the early stage of the task learning. However, the performance was similar in both forelimbs after two weeks of training. 2) The 6-OHDA lesion produced a decrease in the preferred activation during contralateral movements in both M1 and M2. 3) The decrease in the contralateral forelimb preference was different in the identified IT and PT cortical projection neurons depending of the cortical region. The loss of the contralateral biased activity was present in the PT neurons of M1, while in M2 the PT neurons showed no change in forelimb preference. 4) The FS neurons in the non-lesioned hemisphere exhibited an increased activity during contralateral movements than normal, suggesting either a pathological or compensatory change due to the dopamine loss of the contralateral hemisphere.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-415 Pelizaeus-Merzbacher病におけるPLP1遺伝子重複を標的としたAAVによるartificial miRNA遺伝子治療 Heng Li（李コウ）¹，Hironori Okada（岡田浩典）²，Sadafumi Suzuki（鈴木禎史）¹，Kazuhisa Sakai（境和久）³，Hitomi Izumi（泉仁美）¹，Yukiko Matsushima（松島由紀子）¹，Noritaka Ichinohe（一戸紀孝）³，Takashi Okada（岡田尚巳）²，Yu-ichi Goto（後藤雄一）¹，Ken Inoue（井上健）¹ ¹国立精神・神経セ神経研疾病２ ²日本医科大医生化・分子生物（分子遺伝） ³国立精神・神経セ神経研微細構造 Copy number increase or decrease of certain dosage-sensitive genes may cause genetic diseases with distinct phenotypes, conceptually termed genomic disorders. The most common cause of Pelizaeus-Merzbacher disease (PMD), an X-linked hypomyelinating leukodystrophy, is genomic duplication encompassing the entire proteolipid protein 1 (PLP1) gene. Although the exact molecular and cellular mechanisms underlying PLP1 duplication, which causes severe hypomyelination in the central nervous system, remain largely elusive, PLP1 overexpression is likely the fundamental cause of this devastating disease. Here, we investigated if adeno-associated virus (AAV)-mediated gene-specific suppression may serve as a potential cure for PMD by correcting quantitative aberrations in gene products. We developed an oligodendrocyte-specific Plp1 gene suppression therapy using artificial miRNA under the control of human CNP promoter in a self-complementary AAV (scAAV) platform. A single direct brain injection achieved widespread oligodendrocyte-specific Plp1 suppression in the white matter of WT mice. AAV treatment in Plp1 transgenic mice, a PLP1 duplication model, ameliorated cytoplasmic accumulation of Plp1, preserved mature oligodendrocytes from degradation, restored myelin structure and gene expression, and improved survival and neurological phenotypes. Together, we provide evidence that AAV-mediated gene suppression therapy can serve as a potential cure for PMD resulting from PLP1 duplication and possibly for other genomic disorders.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-416 A型およびB型モノアミン酸化酵素による神経保護作用を有する遺伝子発現への関与 Masayo Shamoto-Nagai（永井雅代），Keiko Inaba-Hasegawa（稲葉桂子），Wakako Maruyama（丸山和佳子），Makoto Naoi（直井信）愛知学院心身科学健康栄養 The role of type A and B monoamine oxidase (MAO-A, -B) in the expression of pro-survival genes was investigated in neuroblastoma SH-SY5Y (SH) and glioblastoma U118MG (U) cells. The results were discussed in relation to the neuroprotective property of rasagiline and selegiline, MAO-B inhibitors, Materials and Methods. MAO expression was knockdown by use of siRNA targeting Mao-A and Mao-B mRNA (sc-35847, sc-35849, Santa Cruz biotechnology, siMao-A and siMao-B) in SH cells and U cells expressing MAO-A and MAO-B, respectively. Induced genes were quantitatively determined by real-time RT-PCR method in samples prepared from cells treated with the MAO inhibitor for 24 hours. Bcl-2 protein was quantified by Western blot analysis. Results Mao-A and -B knockdown (KD) decreased the level of Mao-A mRNA in SH cells and Mao-B mRNA in U118MG cells, respectively. In SH cells, rasagiline increased the mRNA and protein levels of Bcl-2 and brain-derived and glial cell line-derived neurotrophic factor (BDNF, GDNF), which Mao-A KD suppressed. In U cells Mao-B KD significantly increased the basal and rasagiline-induced Bcl-2 and mRNA levels of neurotrophic factors, Bdnf, Gdnf, nerve growth factor, and neurotrophin-3. Selegiline synergistically enhanced the expression of Bdnf more markedly than rasagiline. Discussion MAO-A and -B were shown to function either as a mediator or repressor of pro-survival gene expression in SH and U cells, respectively. Further study on cellular mechanism underlying regulation of signal pathways by MAO-A and -B may bring us new insight on the role of MAOs in decision of neuronal fate in neuropsychiatric disorders.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-417 シヌクレイン抑制 AmNA 核酸によるパーキンソン病の治療 Chi Jing Choong（鐘其静）¹，Takuya Uehara（Uehara Takuya）¹，Masayuki Nakamori（Nakamori Masayuki）¹，Hideki Hayakawa（Hayakawa Hideki）¹，Kousuke Baba（Baba Kousuke）¹，Hiroshi Tsuda（Tsuda Hiroshi）¹，Satoshi Obika（Obika Satoshi）^2,3，Hideki Mochizuki（Mochizuki Hideki）¹ ¹Graduate School of Medicine, Osaka University ²National Institutes of Biomedical Innovation, Health and Nutrition ³Graduate School of Pharmaceutical Sciences, Osaka University Purpose Parkinson’s disease (PD) is a neurodegenerative disease characterized by the loss of dopaminergic neurons in the substantia nigra and the presence of Lewy bodies and neurites predominantly composed by aggregated α-synuclein (SNCA). Multiplication of the SNCA gene in familial PD and pathological accumulation of SNCA protein during progression of sporadic PD suggest that elevated levels of SNCA protein increase the risk of PD. Thus, reducing SNCA expression levels could delay PD onset or modify the disease course. Methods We designed and synthesized amido-bridged nucleic acids (AmNA)-modified antisense oligonucleotides (ASOs) that targeted SNCA with improved stability and cellular uptake and screened them for SNCA downregulation in HEK293 cells which express SNCA endogenously. The most potent AmNA-ASO was selected to probe in vivo relevance in SNCA transgenic mice. Results AmNA-ASO with the highest potency emerged from the initial screening efficiently downregulated SNCA at both mRNA and protein level in vitro and in vivo. Notably, AmNA-ASO could be widely distributed throughout the mouse brain by a single intracerebroventricular injection in the absence of any carrier or conjugation and efficiently taken up by neuronal and, to a lesser extent, non-neuronal cells. Furthermore, administration of the chosen AmNA-ASO ameliorated motor deficits in transgenic PD mouse model expressing human wild type SNCA. Taken together, these findings suggest that AmNA-ASO is a promising therapeutic strategy for SNCA-associated pathology in PD.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-418 ゴルジストレスは神経細胞においてβAPPプロセッシングに影響しCaspase3依存性アポトーシスを誘導する Kei Suga（須賀圭）^1,2，Sachiko Yamamoto（山本幸子）¹，Masafumi Nishino（西野将史）^1,2，Yasuo Terao（寺尾安生）²，Kimio Akagawa（赤川公朗）²，Makoto Ushimaru（丑丸真）¹ ¹杏林大医化学 ²杏林大医病態生理 Endoplasmic reticulum (ER) stress and the activation of caspase3 have been implicated in neurodegenerative diseases such as Alzheimer's disease (AD). However, involvement of the dysfunction of neuronal cells caused by Golgi stress during the progression of AD is poorly understood. We have been focusing on the function of ER-Golgi soluble N-ethylmaleimide-sensitive factor-attachment protein receptors (ER-Golgi SNAREs) in βAPP processing under cellular stress. We previously showed that ER stress upregulates de novo synthesis of ER-Golgi SNAREs Syntaxin5 (Syx5), and the reduction of β-amyloid peptide (Aβ peptide) in Neuroblastoma-Glioma hybrid cell line NG108-15 (Exp. Cell Res., 2015) and in hippocampal neurons (Neurosci. Lett., 2015). In addition, the reduction of Aβ secretion by ER stress was significantly suppressed by Syx5 knock down. Conversely, down regulation of Syx5 protein was observed upon apoptosis which was due to the degradation by activated caspase3 (Data in Brief, 2015, 2016). Furthermore, we reported that a chemical chaperone 4-phenylbutyrate (PBA) showed alleviation of caspase3-dependent apoptosis induced by ER stress. In order to know the consequences of Golgi dysfunction in neuronal cells, we examined the effect of multiple stress inducers on the expression of ER-Golgi SNAREs and the processing of βAPP. We used ionophores that disrupt the pH gradient between the cytosol and the acidic compartments, and Golgi disturbing compounds that interfere with secretory pathways. By using time lapse imaging, western blotting, and sandwich ELISA analyses, we show that Golgi stress also promotes upregulation of ER-Golgi SNAREs Syx5, reduction of Aβ peptide secretion, and the caspase3-dependent apoptosis. Furthermore, we will report some of the data showing the effect of chemical chaperone (PBA) and would like to discuss these results.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-419 EAEにおけるmPGES-1誘導の調節について Marumi Kawakami（川上万留実）¹，Chisen Takeuchi（竹内千仙）²，Takako Takemiya（竹宮孝子）¹ ¹東京女子医大総研研究 ²東京都立北療育センター神経内科 Microsomal prostaglandin E synthetase-1 (mPGES-1) is an inducible terminal enzyme forthe production of prostaglandin E2 (PGE2). In experimental autoimmune encephalomyelitis (EAE),an animal model of multiple sclerosis, mPGES-1 is induced in vascular endothelial cells (VECs)around inflammatory foci and facilitates inflammation, demyelination, and paralysis. Therefore,we investigated the role of CD31-positive VECs in mPGES-1-mediated EAE aggravation usingimmunohistochemical analysis and imaging of wild-type (wt) and mPGES-1-deficient (mPGES-1-/-)mice. We demonstrated that EAE induction facilitated vascularity in inflammatory lesionsin the spinal cord, and this was significantly higher in wt mice than in mPGES-1-/- mice.In addition, endothelial interleukin-1β(IL-1β) production was significantly higher in wt mice than in mPGES-1-/- mice. Moreover, endothelial PGE2 receptors (E-prostanoid (EP) receptorsEP1-4) were expressed after EAE induction, and IL-β induced in EP receptor-positive VECs.Furthermore, IL-1 receptor 1 expression on VECs was increased upon EAE induction. Thus, increasedvascularity is one mechanism involved in EAE aggravation induced by mPGES-1. Furthermore,mPGES-1 facilitated the autocrine function of VECs upon EP receptor induction and IL-1β production,modulating mPGES-1 induction in EAE.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-420 ニーマンピック病C型における獲得免疫系による神経変性制御 Toru Yasuda（安田徹）¹，Nobuyuki Watanabe（渡辺信之）¹，Toru Uchiyama（内山徹）¹，Hideki Mochizuki（望月秀樹）²，Masafumi Onodera（小野寺雅史）¹ ¹国立成育医療セ研究所成育遺伝 ²大阪大医神経内科 Niemann-Pick disease type C (NPC) is an autosomal-recessively inherited lysosomal storage disorder affecting an estimated 1 in 120,000 live births worldwide. Mutations in NPC1 or NPC2 gene represent approximately 95% or 5% of total patients with NPC, respectively. Their gene products, NPC1 and NPC2 proteins, function cooperatively in late endosomes and lysosomes to transport unesterifed cholesterol to the plasma membranes and the other organelles in the cell. Typical clinical feature of the disease is neurovisceral accumulation of unesterified cholesterol and several forms of glycosphingolipids. NPC can present with a broad range of clinical manifestation from a neonatal acute fatality to an adult-onset chronic disease associated with neurodegeneration. The development of neurological symptoms, including cerebellar ataxia, laughter-induced cataplexy, dystonia, and progressive dementia, affects quality of life of the patients drastically. Hence, it is essential to explore the pathogenic events that trigger and/or promote the neurodegenerative process for future clinical interventions. In this study, we addressed an involvement of immune system in neuropathogenic process using a murine model of NPC. Breaching of blood-brain barrier and infiltration of monocyte-derived macrophages correlated spatially and temporally with the loss of cerebellar Purkinje cells, which is a hallmark of neurodegeneration in NPC. Reduction of microglial cells and circulating monocytes did not affect and ameliorated Purkinje cell degeneration, respectively. On the other hand, lack of acquired immune system enhanced cerebellar ataxic phenotype and Purkinje cell loss, suggesting that lymphoid cells may have therapeutic effect in NPC. We found that peripheral injection of CD4-positive T cells ameliorated cerebellar ataxia and Purkinje cell loss. Our results disclose a previously unrecognized neuropathogenicity of immune system in NPC and would benefit future remedies for devastating neurological diseases.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-421 シュワン細胞のジストニン異常は末梢神経障害を引き起こす Masao Horie（堀江正男）¹，Nozumu Yoshioka（吉岡望）²，Satoshi Kusumi（久住聡）¹，Hiromi Sano（佐野裕美）³，Hossain Ibrahim（Ibrahim Hossain）²，Izumi Iida-Watanabe（飯田-渡辺和泉）⁴，Satomi Chiken（知見聡美）³，Manabu Abe（阿部学）^4,5，Kenji Sakimura（崎村建司）^4,5，Atsushi Nambu（南部篤）³，Masahiro Shibata（柴田昌宏）¹，Hirohide Takebayashi（竹林浩秀）² ¹鹿児島大院医歯形態科学 ²新潟大院医歯学神経生物解剖学 ³生理研生体システム;総研大生理科学 ⁴新潟大脳研基礎神経科学細胞神経生物 ⁵新潟大脳研モデル動物 Dystonin (Dst) is a causative gene for Dystonia musculorum (dt) in mice, which is an inherited disorder showing dystonia and ataxia with sensory degeneration. Dst is expressed in the variety of tissues including the central (CNS) and peripheral (PNS) nervous system, muscles, and skin, whereas the causative Dst-expressing cells for motor abnormalities have not well characterized. To address the question whether the deletion of Dst in the CNS or PNS induces movement disorders, Dst gene trap (DstGt) mice were mated with P0 (myelin protein zero)-Cre transgenic mice to generate Dst conditional knockout mice in the PNS. First, we checked the P0-Cre dependent reporter gene expression in P0-Cre; Rosa-tdTomato mice and then confirmed the preferential expression of tdTomato protein in the Schwann cells. Next, we analyzed the phenotype of P0-Cre; Dst cKO mice (Dst cKO mice). In the Dst cKO mice, the expression of normal Dst mRNA was significantly decreased in the Schwann cells, but not decreased in the dorsal root ganglion neurons. Dst cKO mice seemed to be normal in the motor phenotype during juvenile periods, thereafter they started to show an ataxic phenotype, but not dystonic movements, around 3 months old. A series of behavior tests showed impaired motor abilities of the Dst cKO mice. Electron microscopic observation on the PNS of the Dst cKO mice showed the significant numbers of hypo-myelinated peripheral nerves and numerous macrophage-like cells surrounding myelin-like structure. These data suggest that disruption of Dst in Schwann cells destructs the myelin structure in the PNS and induces ataxia, but not dystonia and that disruption of Dst in the CNS probably induces dystonia.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-422 遺伝性感覚性自律神経性ニューロパチーVI型モデルマウスにおける末梢神経系選択的なdystonin発現回復の治療効果 Nozomu Yoshioka（吉岡望）^1,2，Hirohide Takebayashi（竹林浩秀）¹ ¹新潟大学・医・神経生物・解剖 ²新潟大学・研究推進機構・超域学術院 The dystonin (DST), also known as bullous pemphigoid antigen1 (BPAG1) encodes the cytoskeletal linker protein. DST is the causative gene for the hereditary sensory autonomic neuropathy VI (HSAN6), which is an inherited human disorder of the nervous system. Original dystonia musculorum (dt) mice carrying mutations in the Dst gene have been characterized as a naturally occurred mutant showing neuronal degeneration in the sensory neurons in the dorsal root ganglia (DRG) and progressive ataxia. We had generated conditional/invertible Dst gene trap mice in which the Dst gene trap allele inverts from the mutant (DstGt) to functional (DstGt-inv) allele and from functional (DstGt-inv) to mutant (DstGt-DO) allele by Cre-mediated recombination. In the Dst gene trap allele, the conditional gene trap construct was inserted in actin-binding domain at the N-terminus of neuronal Dst-a and muscular Dst-b isoforms. Homozygous mice carrying DstGt or DstGt-DO allele showed a deletion of Dst-a mRNA in the neuronal tissue and progressive ataxia, while mice carrying DstGt-inv allele was phenotypically normal. To generate Dst conditional rescue (Res) mice in the peripheral nervous system (PNS) and hindbrain, Dst gene trap mice were crossed with Wnt1-Cre transgenic mice, in which Cre recombinase is expressed in neural crest cells and the hindbrain of the embryos. As expected, Cre-mediated inversion of the Dst gene trap allele occurred in DRG neurons. In homozygous DstGt mice, the stress-induced transcription factor ATF3 was up-regulated in DRG neurons and neurofilament was accumulated. In Wnt1-Cre; Dst cRes mice, Dst mRNA was recovered and pathological remarks were ameliorated in DRG neurons. Wnt1-Cre; Dst cRes mice showed increased life span and recovered from ataxia, compared to DstGt homozygous. These results suggest that complementation with DST in the Wnt1-lineage cells including PNS and hindbrain will be candidate strategy for the HSAN6 treatment.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-423 Krabbe病モデルマウスでのオリゴデンドロサイトの分化・成熟異常を改善する試み Naoko Inamura（稲村直子）¹，Shinji Go（郷慎司）²，Hiroshi Takase（高瀬弘嗣）³，Kiyofumi Asai（浅井清文）³，Hirohide Takebayashi（竹林浩秀）⁴，Junko Matsuda（松田純子）²，Yasushi Enokido（榎戸靖）¹ ¹愛知県コロニー発達障害研究所　病理学部 ²川崎医大・病態代謝学 ³名市大・医学研究科 ⁴新潟大・医歯学総合研究科 Krabbe disease (KD; Globoid cell leukodystrophy) is an autosomal recessive leukodystrophy caused by a defect of lysosomal galacrocerebrosidase (GALC), a catalytic enzyme of galactosylceramide. GALC deficiency results in accumulation of cytotoxic sphingolipid psychosine leading to demyelination. Psychosine has been shown to the cause of neurotoxicity in KD, however, the pathogenic mechanisms of KD remain to be elucidated. We have studied the mechanisms underlying demyelination in KD during development in vivo and in vitro using an authentic mouse model of KD, twitcher mouse and demonstrated that myelination was suppressed in postnatal three weeks of age when the period of active myelination prior to progressive demyelination in KD model mouse (twitcher mouse) brains. In addition, abnormal differentiation and maturation and increase of apoptosis was observed in cultured twitcher mouse oligodendrocytes (OLs). Here we examined therapeutic efficacy of reagents in abnormal differentiation and maturation of twitcher OLs. We examined the effect of L-cycloserine in twitcher OLs development. L-cycloserine, which broadly decreases the synthesis of sphingolipids including psychosine and has been reported to improve lifespan in the twitcher mice. L-cycloserine increased MBP-positive OLs and reduced the level of endogenous psychosine compared with untreated twitcher OLs. However, the number of cleaved caspase-3-positive apoptotic cells was partially reduced, which suggest that the effect of L-cycloserine is limited. We also examined other molecules which may be more effective than L-cycloserine. The distinct effects of these molecules will be discussed.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-424 in vitroおよび細胞内における蛍光性アプタマーを用いたリピートRNAの検出 Yoshihiro Kino（紀嘉浩），Yasuyuki Masaki（正木泰行），Minami Kawai（河合美南），Jun-ichi Satoh（佐藤準一）明治薬大 Expansions of repetitive DNA tracts cause a number of neurological diseases including myotonic dystrophy and amyotrophic lateral sclerosis/frontotemporal dementia. Repeat-containing transcripts produced from the expanded alleles form RNA foci, or ribonuclear inclusions, that are pathological hallmarks of a subset of repeat expansion diseases. RNA foci are thought to sequester RNA-binding proteins that are essential for cellular functions and may mediate disease pathogenesis. In addition, repeat-containing RNAs are subjected to non-canonical translation known as repeat-associated non-ATG translation, in which repetitive, aggregation-prone polypeptides are produced from the expanded repeat RNAs. Although repeat RNAs are important therapeutic targets in these diseases, their intracellular dynamics and regulatory factors have been elusive. Here, we tried to visualize the intracellular localization of repeat RNAs fused with fluorescent aptamers. Spinach2 and dimerized Broccoli (dBroccoli) are RNA aptamers that mimic GFP and emit green fluorescence in the presence of DFHBI-1T. While monomers of both Spinach2 and dBroccoli showed fluorescence in vitro, only dBroccoli showed an enhancement of fluorescence when the number of tandemly arranged aptamers increased. We could detect intracellular fluorescence of dBroccoli and dBroccoli-GGGGCC, but not Spinach2, in COS-7 cells. In addition, dBroccoli fluorescence showed cell-type specificity and was susceptible to photobleaching. Fluorescence in situ hybridization revealed that a fraction of dBroccoli is non-fluorescent and some RNA foci formed by repeat RNAs are not detected by the dBroccoli fluorescence. Unexpectedly, we noticed that a tandemly fused dBroccoli forms intracellular aggregates resembling RNA foci even without disease-associated RNA repeat. Moreover, fusion of repeat RNA with tandem dBroccoli diminished both foci formation and aptamer fluorescence, suggesting certain interactions between the aptamer and the repeat tracts. In conclusion, though tandemly-fused dBroccoli has an enhanced fluorescence, care should be taken for its use in intracellular localization studies of repeat RNAs.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-425 Resident-Intruder系を用いた心理社会的ストレスの１０回連続負荷は背側海馬特異的なBMPシグナルの活性化を引き起こす Masayoshi Mori（森征慶），Naoko Hanakita（花北奈央子），Haruka Yamashiro（山城遥香），Mayu Kikuchi（菊池真由），Hiroyoshi Harada（原田洸秀），Yusuke Murata（村田雄介），Kenji Ohe（大江賢治），Munechika Enjoji（遠城寺宗近）福岡大学薬学部臨床薬物治療学教室 Introduction Our previous study demonstrated that psychosocial stress-induced the decrease in dorsal hippocampal neurogenesis is associated with the pathology of anxiety disorders. However, the mechanisms underlying the stress-mediated changes in dorsal hippocampal neurogenesis are incompletely understood. Here, we examined whether psychosocial stress-induced changes in hippocampal bone morphogenetic protein (BMP)-4 signaling contribute to the decrease in levels of hippocampal neurogenesis using Resident-Intruder paradigm. Methods For the psychosocial stress exposure, Long-Evans rats were used as Resident, and male Sprague-Dawley rats were used as Intruder. Male SD rats were exposed to psychosocial stress for a consecutive ten days. One day after the last stress session, the intruder rats were sacrificed under anesthesia. Then, hippocampal formation was collected, and BMP4 and phosphorylated Smad1/5/9 protein levels in the dorsal and ventral hippocampus were evaluated by western blotting analysis. Results In dorsal hippocampus, BMP4 and phosphorylated Smad1/5/9 protein levels were significantly increased in stressed rats compared to control rats. In contrast, there was no significant difference in BMP4 and phosphorylated Smad1/5/9 expression levels in the ventral hippocampus between control and stressed rats. Discussion Psychosocial stress preferentially activated in the dorsal hippocampal BMP4 signaling. These results suggest that the BMP signaling pathway in the dorsal hippocampus may be a potential target for new therapeutic strategy for anxiety disorders.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-426 ジフェニルアルシン酸によるラット小脳由来アストロサイトの異常活性化とチオール含有重金属キレート剤の保護効果 Takayuki Negishi（根岸隆之），Shoto Sasaki（佐々木翔斗），Tomoka Shibata（柴田朋香），Ayumi Takagi（髙木梓弓），Yuki Ohishi（大石悠稀），Shuya Wakasugi（若杉周弥），Maho Takano（髙野真帆），Yuho Kondo（近藤優帆），Mai Nakajima（中嶋真唯），Ai Kitabeppu（北別府愛），Takamasa Tsuzuki（都築孝允），Kazunori Yukawa（湯川和典）名城大学薬学部生理学 Diphenylarsinic acid (DPAA) is the pentavalent arsenic chemical found in the arsenic poisoning in Kamisu, Ibaraki, Japan 2003, where people using contaminated water suffered from neurological symptoms including cerebellar symptoms. We previously reported that, in cultured rat cerebellar astrocytes, DPAA could induce cell-proliferation and following cell-death, upregulation of anti-oxidative stress proteins including heme oxygenase-1 (HO-1), and heat-shock protein 70 (Hsp70), activation (phosphorylation) of major protein kinases including extracellular signal-related kinases 1/2 (ERK1/2), p38 mitogen-activated protein kinase (p38MAPK), stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), and other kinases such as PKA, PKC, and PKD, and induction/activation of transcription factors including CREB, c-Jun, and c-Fos. Here, in order to propose promising therapeutic drugs against DPAA-induced neurological symptoms, we evaluated and compare the protective effect of four thiol-containing chelators against heavy metals; dimercaprol (BAL), D-penicillamine (DPEN), dimercaptosuccinic acid (DMSA), 2,3-dimercapto-1-propanesulfonic acid (DMPS), and α-lipoic acid (LIP) in astrocytes exposed to DPAA (0, 10, and 50 μM) for 96 h. BAL, the most major antidote against inorganic arsenic poisoning, and DMPS showed no preventive effect against DPAA-induced activation of astrocytes. However, DMSA could completely inhibit high-dose (50 μM) DPAA-induced cell-death and low-dose (10 μM) DPAA-induced biological activations such as MAP kinases activation in DPAA-exposed astrocytes, while DPEN and LIP showed limited preventive effects. These results suggested that DMSA among thiol-containing heavy metal chelators would be a promising drug preventing onset of DPAA-induced astrocyte activation and subsequent neurological symptoms.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-427 SH-SY5Y細胞における低リン酸に対するPDGF-BBの神経保護効果 Naoko Takase（高瀬奈央子），Masatoshi Inden（位田雅俊），Yuki Kaneko（金子由希），Yumeka Yamada（山田夢華），Hisaka Kurita（栗田尚佳），Isao Hozumi（保住功）岐阜薬大・薬・薬物治療 [Purpose] Idiopathic basal ganglia calcification (IBGC) is a rare genetic condition characterized by symmetric calcification in the basal ganglia and other brain regions, a wide spectrum of neuropsychiatric symptoms. In 2012, the first causative gene SLC20A2 has been found which encode a sodium-dependent phosphate cotransporters, PiT2. The type-III sodium-dependent phosphate cotransporter (NaPiTs) contains two subtype, PiT1 and PiT2. PiT2 mutations associated with IBGC were predicted to be unable to transport Pi from the extracellular environment. However, the roles of type-III NaPiTs under low Pi loading is unknown. Platelet-derived growth factor (PDGF) is one of the main growth factors of mesenchymal cells and forms four heterodimers and one heterodimer from four PDGF genes. PDGF-B encoded by PDGFB has the highest affinity for PDGF receptor. PDGF-BB is involved in neuroprotection, cerebral angiogenesis and function regulation of the blood-brain barrier in the cerebral nervous system. In this study, we investigated the effect of low Pi loading on the human neuroblastoma cell line SH-SY5Y. We also investigated whether PDGF-BB protects neurotoxicity by low phosphate loading. [Method] SH-SY5Y cells were incubated overnight in FBS-free / Pi-free DMEM. An appropriated amounts of PDGF-BB in sodium phosphate buffer (0.1 M Na₂HPO₄/NaH₂PO₄, pH 7.4) was administered in the cell culture. Cell Count Reagent SF was used for cytotoxicity assays. The amount of phosphate uptake into cells was measured using radioactive isotope ³²P. Cell Rox regent were used to carry out the evaluation of oxidative stress. To elucidate the signaling pathway of PDGF-BB, ERK proteins and Akt proteins were measured by Western blot. The phosphate transporter which exprssed on the cell membrane surface were evaluated by META screening assay Kit. [Result/Discussion] Long term treatment with low phosphate (for 24, 48, and 72h) caused cell death in SH-SY5Y cells, which increased with treatment time. PDGF-BB inhibited the low Pi loading-induced neurotoxicity, but the expression level of type-III NaPiTs by PDGF-BB treatment did not change. PDGF-BB prevented ROS production caused by low Pi loading. Moreover, our study demonstrated that PDGF-BB inhibited the low Pi loading-induced neurotoxicity via increased PiT1 membrane migration by Akt signaling activation. The findings suggest that PDGF-BB has potential for the therapy in IBGC.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-428 パーキンソン病前駆期モデルマウス運動野におけるシナプス不安定性 Yusuke Hatanaka（畑中悠佑），Ryosuke Takahashi（高橋良輔）京都大院医臨床神経 Recent evidence implicates that the impairment of synapses and neuronal circuitry rewiring as important factors of pathogenesis in neuropsychiatric disorders. The synaptic mechanisms in PD pathogenesis remain to be elucidated, because of the absence of an animal model which replicates neuronal circuitry pathology. We have generated BAC transgenic mice harboring human A53T α-synuclein gene SNCA with PD risk SNPs and its expression regulatory regions. A53T SNCA-BAC mice show progressive neuronal loss of dopaminergic neurons in the substantia nigra and accumulation of phosphorylated α-synuclein in the cerebral cortex. A53T SNCA-BAC mice do not show obvious PD-like motor dysfunctions, however, they exhibit RBD-like behavior at the age of 10 weeks and smell disturbance at 9-month-old. In this study, we focused on presymptomatic synapse pathologies in the primary motor cortex by using A53T SNCA-BAC mice as a model of prodromal PD. By crossing A53T SNCA-BAC mice with Thy1-EGFP mice, layer 5 pyramidal neurons in the cortex of the offspring were labeled. A cranial window was implanted 4 weeks prior to imaging. In vivo two-photon imaging was performed weekly for 4 weeks. A53T SNCA-BAC mice showed a decreasing density of dendritic spines at the age of 12 weeks, and it reached the plateau at 24-week-old. This was due to the imbalance of enhanced spine formation and elimination at the age of 12 weeks, and the imbalance was equalized in 24-week-old. Although spine formation and elimination rate were balanced when they grow up, both formation and elimination rate of the spine remained to be higher than control mice. A53T SNCA-BAC mice also exhibited lower pre-existed spine stability and newly-formed spine survivability at the age of 24 and 48 weeks. Furthermore, colocalization of presynaptic protein marker and dendritic spine was lower in A53T SNCA-BAC mice. These results suggest that a synaptic loss in the primary motor cortex is occurred even in a prodromal PD model, and this synaptic loss is caused by an excess elimination of dendritic spines. After the abnormal synaptic pruning is finished, the instability of dendritic spines still remains as a neuronal circuitry pathology.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-429 大脳基底核および小脳ネットワークでの安静時機能結合の変容はパーキンソン病のすくみ足を反映する Hiroki Togo（東口大樹）¹，Tatsuhiro Nakamura（中村達弘）¹，Noritaka Wakasugi（若杉憲孝）¹，Noriko Nishikawa（西川典子）²，Yuji Takahashi（高橋祐二）²，Takashi Hanakawa（花川隆）¹ ¹国立精神・神経セ脳病態統合イメージングセ ²国立精神・神経セ病院脳神経内科 Freezing of gait (FOG) is a gait disorder that typically affects advanced Parkinson's disease (PD) and related disorders. FOG may result in a fall and reduces quality of life in patients with PD, but it is difficult to develop effective therapy since pathophysiology of FOG is not yet clear. A number of studies have begun to show that a resting-state functional magnetic resonance imaging (rsfMRI) may provide a useful biomarker to help diagnose and/or evaluate patients with PD. First, rsfMRI is suitable for observing functional connectivity (FC), which likely reflects pathophysiology in PD. Second, rsfMRI has advantage over other functional imaging methods since it is task-free and thus is not confounded by differences in task performance across groups or individuals. Therefore, we examined the relationship between FOG severity and FCs in PD patients to explore FCs as a marker of FOG. We examined 60 patients with PD on medication. The study was approved by the Ethics Committee. Severity of FOG was evaluated by using new FOG questionnaire (NFOGQ) score, which is currently the gold standard for assessment of FOG. RsfMRI (TR=2.5 s) were acquired for 10 min with eyes open. Preprocessing included distortion correction, motion correction, slice timing correction and spatial smoothing. Next, we performed individual-level independent component analysis (ICA) and auto-removal of noise components. We performed two types of FC analysis. One is a dual regression analysis with permutation testing for positive or negative correlation between NFOGQ scores and FC. We used ICA template extracted from PD and healthy control subjects in our previous study. The other is to construct a model that can predict FOG from FC, using least absolute shrinkage and selection operator (LASSO). We calculated the correlation among extracted time-courses from rsfMRI data in an atlas-based volumes-of-interest after noise removal. In the dual regression of basal ganglia resting-state network, we found positive correlation between NFOGQ and FC in the putamen and amygdala. Also, we found positive correlation between NFOGQ and FC in vermis in the dual regression of cerebellar resting-state network. In the LASSO analysis, moreover, we found the model which constitute of 36 connectivities including basal ganglia, cerebellum and motor-related FCs. In conclusion, basal ganglia and cerebellum-related FC carries information reflecting severity of FOG in PD.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-430 前頭側頭葉変性症神経細胞を用いたタウ伝播モデル Keiko Imamura（今村恵子）^1,2,3，Haruhisa Inoue（井上治久）^1,2,3 ¹京都大学iPS細胞研究所 ²理化学研究所バイオリサーチ研究センター ³理化学研究所革新知能統合研究センタ― Accumulating evidence strongly implicates cell-to-cell propagation of misfolded proteins as a common mechanism for the pathophysiology of various neurodegenerative diseases including neurodegenerative tauopathies, which are a proteinopathy associated with the pathological aggregation of microtubule-associated protein tau (MAPT) in the brain. Mutations in MAPT gene cause a familial tauopathy, frontotemporal lobar degeneration, which presents dementia and is characterized by atrophy in the frontal and temporal lobes of the brain. Neuron-to-neuron propagation of misfolded proteins is implicated as a central process of neurodegeneration, including hereditary frontotemporal lobar degeneration due to tau pathology (FTLD-tau). However, it is not clear how these proteins propagate their neurotoxicity. We established an FTLD-tau propagation model with induced pluripotent stem cells. The FTLD-tau neurons exhibited accumulation of intracellular and increase in extracellular oligomeric tau as well as hyperexcitability. Furthermore, oligomeric tau released from FTLD-tau neurons caused neuronal hyperexcitability. Regulating neuronal activity with designer receptors exclusively activated by designer drugs (DREADDs) showed that neuronal excitation promoted oligomeric tau propagation and its insults (Imamura et al., 2016). This FTLD-tau propagation model would provide mechanistic insights into tauopathy pathogenesis and potential treatments.		7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-431 Dynactin1とTRAPPC9によるオートファゴソームとリソソームの融合 Hiroaki Adachi（足立弘明），Zhe Huang（黃哲），Tomoko Toyota（豊田知子），Kazumasa Okada（岡田和将），Keiko Ohnari（大成圭子），Tomoyo Hashimoto（橋本智代），Yukio Iwanaka（岩中行己男）産業医科大学 Background and Purposes: Dynactin is a dynein-activator complex required for most of the cellular functions of cytoplasmic dynein. Dynactin is composed of seven to nine polypeptides, including the dynactin-1 (p150Glued) that forms the sidearm of the complex and binds both to microtubules and to dynein; the Arp1 polypeptide that forms an actin-like filament at the base of the complex; and the dynamitin, or p50, the subunit that localizes to the shoulder between the sidearm and the base. Autophagy is essential for neuronal homeostasis, and its dysfunction has been linked to many neurodegenerative disorders. In this report, we show that regulatory relationship between dynactin-1 and fusion of autophagosomes and lysosomes. Methods: In the present study, we used a combination of molecular biological techniques and morphological methods such as western blot, immunofluorescence, RFP-AcGFP-LC3 reporter assay, and immunoelectron microscopy on lentivirus-mediated dynactin-1 knockdown NSC 34 motor neuron cell line, and we determined the autophagosome-lysosomes fusion efficiency. Results: The levels of dynactin-1 protein expression were decreased in the dynactin-1 knockdown cells. The cell viability was decreased, and the expression levels of the mutant SOD1 proteins were increased in dynactin-1 knockdown motor neuron cells. The level of the autophagosome marker LC3-II in cell culture was increased and the autophagosome-lysosome fusion was inhibited in dynactin-1 knockdown NSC 34 motor neuron cells. Conclusions: Our study identifies the dynactin-1 as a regulator that controls the fusion of autophagosomes and lysosomes. These findings suggest that dynactin-1 play important roles in the autophagy and implicate autophagosome-lysosome fusion defects in the pathogenesis of neurodegenerative diseases.

7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-432 Emerging Roles of RNF216/TRIAD3 in the hypothalamic-pituitary-gonadal axis Arlene Joann George（George Arlene Joann），Angela M Mabb（Mabb Angela） Georgia State University Ubiquitin E3 ligases are substrate-specific enzymes in the ubiquitin conjugation pathway that participate in the turnover of proteins and control receptor trafficking. Recessive mutations in the E3 ligase RNF216/TRIAD3 cause Gordon Holmes syndrome (GHS), with symptomologies of hypogonadotropic hypogonadism, cognitive impairment, dysarthria, cerebellar ataxia, and dementia. Individuals diagnosed with GHS have dysfunction at multiple levels of the reproductive endocrine axis also known as the hypothalamic-pituitary-gonadal (HPG) axis. Previous literature demonstrates that RNF216 ubiquitinates activity-regulated cytoskeleton-associated (Arc) protein which is actively involved α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor trafficking. Moreover, removal of RNF216/TRIAD3 was found to increase Arc expression and decrease synaptic expression of AMPA receptors. In support of this research, we found that RNF216/TRIAD localizes to the endosome. Functionally, both knockdown and overexpression of RNF216/TRIAD3 disrupts trafficking of the ubiquitous transferrin receptor in neurons suggesting RNF216/TRIAD3 broadly controls receptor trafficking. HPG axis receptors, particularly kisspeptin G-Protein Coupled Receptor 54 (GPR54) and gonadotropin-releasing hormone receptor (GnRHR), are found in diverse brain regions including the hypothalamus, thalamus, hippocampus, cerebellum, amygdala, cortex, brainstem, and pituitary. To test the role of RNF216/TRIAD3 function in regulating receptor trafficking within the HPG axis, we knocked out RNF216/TRIAD3 using the CRISPR-Cas9 system in hypothalamic (GT1-7) cells and found no significant changes in total expression of GPR54 or GnRHR. Using a high-content live cell antibody feeding assay, we will test if deletion of RNF216/TRIAD3 alters surface expression of GPR54 and GnRHR. Additionally, we will measure the gene expression of GPR54 and GnRHR using real-time quantitative polymerase chain reaction to determine if deletion of RNF216/TRIAD3 changes HPG receptor expression transcriptionally. Moreover, we will determine if RNF216/TRIAD3 deletion alters the production of HPG axis hormones, such as GnRH. Our findings will link ubiquitin E3 ligases to HPG axis function and will provide therapeutic insights into neurological disorders involving HPG axis disruption.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-434 記憶痕跡細胞の樹立不足が結節性硬化症モデルマウスの記憶障害を引き起こす Keiko Moriya-Ito（守屋敬子），Tadayuki Shimada（島田忠之），Hiroko Sugiura（杉浦弘子），Kanato Yamagata（山形要人）東京都医学総合研 Tuberous sclerosis complex (TSC) is one of developmental disorders caused by loss-of-functional mutations in Tsc1 or Tsc2 gene, and the main pathogenesis of TSC is growth of hamartomas in the brain and other organs. The TSC1 and TSC2 protein complex inhibits the activation of a Ras-like small G-protein Rheb. TSC1 or TSC2 protein dysfunction causes the hyperactivation of Rheb that regulates protein synthesis, cell growth and differentiation. TSC patients are accompanied with neuropsychiatic disorder, such as epilepsy, autism and intellectual disability. We have found that Tsc2^+/- mice exhibit the impairment of fear conditioning memory. However, its mechanism still remains unknown. To clarify the mechanisms of memory impairment in Tsc2^+/- mice, we focused on the memory engram cell theory, that is, the memory is encoded by some specific neurons. We labeled the activated neurons in the dentate gyrus during memory encoding, with AAV9-RAM-tTA-TRE-GFP using doxycycline (Dox) on-off system. The RAM promoter, an artificial immediately-early gene promoter, can strongly label activated neurons with GFP under the Dox-off condition. On the next day, the activated neurons during memory recall were labeled by c-Fos immunohistochemistry. In the WT mice, the engram cells, expressing both GFP and c-Fos, were clearly observed. By contrast, many c-Fos positive cells had no or weak GFP expression in Tsc2^+/- mice, indicating that Tsc2^+/- mice failed to establish memory engram cells. Next, we compared the dendritic spine morphology of engram cells and non-engram cells between WT and Tsc2^+/- mice. In the WT engram cells, large dendritic spines were frequently observed, whereas they rarely appeared in the Tsc2^+/- engram cells. Furthermore, to confirm the Rheb activation is critical for these abnormalities, we administrated a Rheb inhibitor to Tsc2^+/- mice before contextual fear conditioning. In these mice, the engram cells significantly increased, and the dendritic spines were moderately enlarged. These results indicate that the memory impairment of Tsc2^+/- mice may be partly caused by insufficient establishment of engram cells through abnormal Rheb activation.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-435 SynGap1ヘテロノックアウトマウスの網羅的行動解析 Ryuichi Nakajima（中島龍一）¹，Satoko Hattori（服部聡子）^1,2，Hirotaka Shoji（昌子浩孝）^1,2，Keizo Takao（高雄啓三）^3,4，Noboru H. Komiyama（Komiyama Noboru）⁵，Seth G.N. Grant（Grant Seth）⁵，Tsuyoshi Miyakawa（宮川剛）^1,2,4 ¹藤田医科大総医研システム医科学 ²科学技術振興機構CREST ³富山大学研究推進機構研究推進総合支援センター生命科学先端研究支援ユニット ⁴生理研行動様式解析室 ⁵Genes to Cognition pgrm, Cent for Clinical Brain Sci, Univ of Edinburgh, Edinburgh, UK Synaptic Ras GTPase-activating protein 1 (SynGap1) regulates synaptic plasticity through AMPA receptor trafficking. SynGap1 mutations have been found in human patients with intellectual disability (ID); approximately half of the SynGap1-related ID patients have autism spectrum disorder (ASD). SynGap1 mutations are estimated to account for 0.7 to 1 percent of all ID cases. In mouse models with SynGap1 mutation, strong cognitive and emotional dysfunctions have been reported, but some behavioral phenotypes have not been consistently observed in all studies. Therefore, we systematically evaluated behavioral alterations in a mouse model with a heterozygous knockout of the SynGAP gene. We subjected male mice heterozygous for a null mutation of the SynGAP gene to a comprehensive behavioral test battery. This battery consists of a general health and neurological screen, and open field, elevated plus maze, light/dark transition, social interaction, prepulse inhibition, Porsolt forced swim, tail suspension, T-maze, Y-maze, Barnes maze, contextual and cued fear conditioning, and home cage locomotor activity tests. Heterozygous SynGap1 knockout mice showed increased locomotor activity, decreased prepulse inhibition, and impaired working and reference spatial memory, consistent with previous studies, with study-wide statistical significance. Preceding studies have reported impaired contextual fear memory and increased startle reflex in SynGap1 mutant mice. In this study, we failed to reproduce these phenotypes. We show, for the first time, with study-wide significance, that these mice have decreased sensitivity to painful stimuli and that the mice show impaired motor function. We also detected seemingly-decreased anxiety- and depression-like behavior, although increased locomotor activity could potentially be a confounding factor of these phenotypes. Significantly increased home cage locomotor activity indicated that these mice show hyperlocomotor activity not only in novel, but also in familiar environments. In these heterozygous SynGap1 knockout mice, we reproduced most of the cognitive and emotional dysfunctions that have been previously reported. The decreased sensitivity to painful stimuli, and impaired motor function, which we have found in this study, are consistent with common characteristics of patients with SynGap1-related ID. We further confirmed that heterozygous SynGap1 knockout mice recapitulate symptoms of ID and ASD patients with SynGap1 mutations.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-436 One carbon metabolism 関連遺伝子変異体を用いたDOHaDモデルの開発 Tamio Furuse（古瀬民生）¹，Ikuko Yamada（山田郁子）¹，Tomoko Kushida（串田知子）¹，Ikuo Miura（三浦郁生）¹，Ai Ozaki（尾崎藍）¹，Akane Nagase（永瀬茜）¹，Shigeharu Wakana（若菜茂晴）²，Masaru Tamura（田村勝）¹ ¹理研BRCマウス表現型解析開発チーム（日本マウスクリニック） ²神戸医療産業都市推進機構先端医療研究センター老化機構研究部 The developmental origins of health and disease paradigm (DOHaD) is a concept that fetal environmental factors affect the adult phenotypes. The purpose of the present study is to develop a new mouse model of the DOHaD. Methionine and folate are metabolized in one carbon metabolism (OCM) and the OCM supports multiple physiological processes including biosynthesis (purines and thymidine), amino acid homeostasis (glycine and except for methionine), epigenetic maintenance, and redox defense. The OCM is consisted of methionine cycle and folate cycle. The folate cycle is involved in DNA synthesis and methionine cycle is involved in DNA methylation by donating methyl group. It is speculated that during development, maternal environmental and nutrient influences by their effects on one carbon transfer can impact the health of the mother, impair growth and reprogram metabolism of the fetus, and cause long term morbidity in the offspring. In the present study, we aim to evaluate effects of maternal OCM deficit on neurobehavioral phenotypes of progeny. We transferred C57BL/6N embryo to uteruses of wild type and heterozygous Mat2a knockout (Mat2a mutant) females. The offspring from the Mat2a mutants exhibited increased locomotor activity and deficits of fear conditioned learning in the comprehensive behavioral phenotyping system in the Japan Mouse Clinic (http://ja.brc.riken.jp/lab/jmc/mouse_clinic/en/pipeline_en/ pipeline_02.htm#p2-00). In order to evaluate the effects of Mat2a mutation on nutritional environment of the fetuses, we performed LC-MS measurements of OCM metabolites in plasma of heterozygote of the Mat2a mutants and wild type mice. The pre-pregnancy females of the Mat2a mutants exhibited decreased concentration of S-adenosyl methionine (SAM). In addition, Mat2a mutants exhibited decreased concentration of the SAM on 14th day of pregnancy whereas not on 19th day of pregnancy. These results suggest that the exposure to decreased SAM level during early to middle pregnancy affects brain function of progenies. We are now investigating histological and functional change of brain of the progenies that exposed to maternal decrement of the SAM. Acknowledgements This research is supported by KAKENHI (Grant Number, 17K07144) and partially supported by the Strategic Research Program for Brain Sciences from Japan Agency for Medical Research and development, AMED (Grant Number, JP19dm0107080).

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-437 Gad1ヘテロ接合体への胎生期ストレスは神経発達及び行動関連遺伝子にエピジェネティックな変化をもたらし表現型にも影響する Tianying Wang（王天英）¹，Adya Saran Sinha（シンハアジャ）¹，Hiroki Mutoh（武藤弘樹）¹，Tenpei Akita（秋田天平）¹，Yuchio Yanagawa（柳川右千夫）²，Tomoko Kawai（河合智子）³，Kenichiro Hata（秦健一郎）³，Atsuo Fukuda（福田敦夫）¹ ¹浜松医大・医・神経生理 ²群馬大院・医・脳神経発達統御・遺伝発達行動 ³国立成育医療研究センター研究所・周産期病態研究部 Exposure to prenatal stress (PS) and mutations in Gad1, which encodes the GABA synthesizing enzyme glutamate decarboxylase (GAD) 67, are both risk factors for psychiatric disorders. Using GAD67-GFP knock-in heterozygous (HT) mice subjected to PS from embryonic day 15.0 to 17.5, we previously reported disruption of GABAergic neurogenesis in the MGE. Postnatally, the density of parvalbumin (PV)-positive GABAergic interneurons was significantly decreased in the mPFC of HT-PS mice, which is a common trait shared by different psychiatric diseases. These result suggested that these two key genetic and environmental susceptibility factors could specifically disturb the proliferation of PV-positive neurons. By contrast, these findings were not observed in wild type offspring. Although our evidence indicate that morphological changes in the brain of offspring, the specific molecular targets and the pathogenic pathways involved in brain alterations induced by a combination of genetic and environmental risk factors are still far from being elucidated. In this study, we found multiple genes functionally associated with neurogenesis and behavior were hyper- or hypo-methylated in HT-PS mice. Some differentially expressed genes detected by high-throughput microarray expression were identical with epigenetic regulation induced by DNA methylation and further validated by real-time PCR. In addition, we examined the behavioral phenotypes and found that HT-PS mice demonstrated significant deficits in sensorimotor gating, cognitive processes, social engagement, without change in anxiety and exploratory behaviors. When assessing the function of interneurons in the mPFC of HT-PS mice, we found that frequency of miniature inhibitory postsynaptic currents was decreased and tonic inhibition was significantly enhanced. Finally, electrocorticogram recording showed reduction in power spectrum density at gamma-frequency range in mPFC of HT-PS mice, indicating GABAergic dysfunction could underlie the behavioral deficit. These findings may provide new insights into mechanisms of the pathogenesis of psychiatric disorders.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-438 Alteration of social dominance behavior in rats after developmental exposure to secondary organic aerosol Win-Shwe Tin-Tin（Tin-Tin Win-Shwe），Yuji Fujitani（Fujitani Yuji），Seishiro Hirano（Hirano Seishiro） National Institute for Environmental Studies, Tsukuba, Japan Background and Aim: Secondary organic aerosol (SOA) is a major component of PM 2.5 and formed in the atmosphere by oxidation of products from anthropogenic and biogenic volatile organic compounds. The importance of SOA formation in urban areas is well-recognized, not only in the atmosphere but also in indoor environments. In this study, we aimed to examine the effects of gestational and lactational exposure to SOA on social dominance behavior and gene expression of neuroimmune biomarkers in male rats. Methods: Sprague-Dawley pregnant rats were purchased from Charles River Laboratories Japan, Inc., Japan and were exposed to clean air, DE and SOA in the whole-body exposure chamber (Sibata) for 5 h per day (from 22:00 h to 03:00 h) on 5 days of the week from gestational day 14 to postnatal day 21. The male offspring rats at PND 21 were allocated into three different groups (n = 6 per group) as follows: 1) rats exposed to clean filtered air; 2) rats exposed to DE; 3) rats exposed to SOA. Social dominance behavior was investigated at 10-week-old rats using a social dominance tube test (Muromachi Kikai Co. Ltd., Japan). Prefrontal cortex was removed under deep anesthesia to examine neuroimmune biomarkers using real-time RT-PCR. Results: Social behavior was assessed using a social dominance tube test which measures social dominance without injuring each other. The socially dominant rats push the counterpart rats to the end of the tube. We found that SOA exposed rats were more socially dominant compared with the control and DE-exposed rats. The mRNA expression levels of serotonin receptor, brain derived neurotrophic factor (BDNF) were downregulated and interleukin (IL)-1 β and heme oxygenase (HO)-1 were upregulated in the prefrontal cortex compared to the control rats. We suggest that, although the potential toxic substances contained in SOA have not yet been identified, they may reach the brain via placenta of pregnant mice during fetal period and via the olfactory nerve route or systemic circulation during neonatal period and induce neurotoxicity. Discussion and Conclusion: Our results indicate that developmental exposure of SOA may affect social dominance of male rats by modulating social behavioral-related genes and proinflammatory markers in the prefrontal cortex. Among the constituents of SOA, organic carbon is the potential candidate to induce neurotoxicity.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-439 Investigation into the role of Glra4, a functionally unknown subunit of glycine receptors, in brain function and neurological disorders Mohamed Ibrahem Darwish（Darwish Mohamed Ibrahem）^1,2，Takaho A Endo（Endo Takaho）³，Kyosuke Uno（Uno Kyosuke）⁴，Keizo Takao（Takao Keizo）^1,5，Hirofumi Nishizono（Nishizono Hirofumi）⁵ ¹Department of Behavioral Physiology, Graduate School of Innovative Life Science, University of Toyama, Japan. ²Department of Biochemistry, Faculty of Pharmacy, Cairo University, Egypt. ³Laboratory for Integrative Genomics, RIKEN Center for Integrative Medical Sciences, Japan. ⁴Graduate school of Medicine and Pharmaceutical Sciences, University of Toyama, Japan. ⁵Life Science Research Center, University of Toyama, Japan. Human glycine receptor α4 (GLRA4) was thought to be a pseudogene due to lack of transmembrane domain, but surprisingly it was recently reported that microdeletion at Xq22.2 including GLRA4 shows symptoms similar to Pelizaeus-Merzbacher disease (PMD). PMD is a hereditary disease that develops due to abnormalities in myelination in the central nervous system and the patients die in infancy in most cases. Mouse orthologous gene Glra4 is also located on chromosome X, however, its function is not fully understood. To understand the biological roles of GLRA4 in cognitive brain functions and whether it is involved in X-linked syndromic intellectual disability, we developed a modified CRISPR/Cas9 genome editing protocol to generate Glra4-KO mice. Our protocol combines a modified method for cryopreserving 1-cell C57BL/6J embryos with optimized electroporation conditions. Using our protocol, we generated several lines of mutant mice: knockout mice via non-homologous end joining (NHEJ) and knock-in mice via homology-directed repair (HDR) with high-efficient mutation rates (100%, 75%, respectively) and a low mosaic rate within 4 weeks. Transcriptomic analysis using public database and immunohistochemical experiments demonstrated that Glra4 was expressed in the midbrain; 5N (motor trigeminal nucleus), 7N and 7n (facial nerve and facial nucleus). To elucidate the function of Glra4 in vivo, we subjected Glra4-KO mice to behavioral analysis. The mutant mice exhibited anxiety-like behavior as demonstrated by the decrease in the number of entries and time spent on the open arm. Taken together, Glra4 is not supposed to be a pseudogene but a protein-coding gene expressed at midbrain and other nervous systems, which affects phenotypes in mouse behavior. Further molecular and behavioral analyses on the mouse should be conducted to examine the function of GLRA4 and its consequences on diseases.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-440 ヒト脳スライス標本を用いたてんかん病態の可視化 Hiroki Kitaura（北浦弘樹）¹，Masafumi Fukuda（福多真史）²，Yukihiko Fujii（藤井幸彦）³，Akiyoshi Kakita（柿田明美）¹ ¹新潟大学脳研究所　病理学 ²西新潟中央病院　脳神経外科 ³新潟大学脳研究所　脳神経外科 [Introduction] Mesial temporal lobe epilepsy (MTLE) is the most frequent focal epileptic syndrome in adults, and the majority of seizures originate primarily from the hippocampus. The resected hippocampal tissue often shows severe neuronal loss as that referred to hippocampal sclerosis (HS). Accordingly, there is a paradox between the clinical and pathological features: why should epilepsy be derived from such degenerated tissue? Here we investigated epileptiform activities ex vivo using living hippocampal tissue taken from patients with MTLE. [Methods] We prepared acute brain slices from patients with MTLE within 45 min after resection, and optical imaging or local field potential recordings (LFP) was performed ex vivo. We also used a brain block corresponding to the mirror surface of each slice and performed histopathological examination. [Results] We revealed that epileptiform activities developed from the subiculum, regardless of the existence of HS. We found spontaneous rhythmic activities in the subiculum and detected discrete component of high frequency oscillations (HFO), a clinical biomarker of the ECoG suggesting the epileptogenic regions. Immunohistochemistry of the HS tissue revealed loss of inwardly rectifying K+ channel 4.1 (Kir 4.1) in astrocytes in the subiculum, indicating failure of the extracellular K+ buffering and possible association with neuronal hyperexcitability. [Conclusion] These results indicate that pathophysiological alterations involving the subiculum could be responsible for epileptogenesis in patients with MTLE.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-441 hTERT RPE-1細胞においてTULP3は繊毛膜タンパクの局在に必要である Sarina Han（韓薩日娜）^1,2，Ko Miyoshi（三好耕）¹，Sho Shikada（鹿田星）¹，Genki Amano（天野元揮）¹，Yinshengzhuoma Wang（王銀生卓瑪）¹，Takeshi Yoshimura（吉村武）¹，Yasuomi Ouchi（尾内康臣）²，Taiichi Katayama（片山泰一）¹ ¹大阪大院・連合小児発達・分子生物遺伝学 ²浜松医大・光先端医学教育研究・生体機能イメージング The primary cilia are known as biosensors that receive extracellular signals and transduce them to the cell body in vertebrate cells. In humans, impairment of cilia-mediated signaling causes ciliopathies characterized by cognitive deficits, obesity, renal abnormalities, retinal degeneration, and so on. Tubby-like protein 3 (TULP3), a member of the tubby family, has been shown to interact with the intraflagellar transport-A complex (IFT-A) that is required for the assembly and function of cilia. A previous study has revealed that mice with disrupted Tulp3 are embryonically lethal and display neural tube closure defect, while the biological role of TULP3 in development is poorly understood. In order to better understand TULP3 function, we generated and analyzed TULP3-knockout (KO) hTERT RPE-1 (RPE1) cells. Formation of cilia and localization of ciliary membrane proteins in the TULP3-KO cell lines and parental RPE1 cells were observed by an immunofluorescent study. TULP3 ablation affected the number and length of the cilium. Moreover, the ciliary membrane proteins, ADP-ribosylation factor-like 13B, inositol polyphosphate-5-phosphatase E and GPR161, an orphan G-protein-coupled receptor, failed to localize to the primary cilia in TULP3-KO cells. These defects in the localization of membrane proteins in TULP3-KO cells were rescued by the exogenous expression of wild-type TULP3, but not that of mutant TULP3 lacking the ability to bind IFT-A. Our findings indicate that TULP3 engages in ciliogenesis, and acts on the membrane proteins to the primary cilia by binding to IFT-A.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-442 バルプロ酸自閉症モデルマウス胎児の超音波発声異常の検出 Takahiro Tsuji（辻隆宏）^1,2，Tomoaki Fujisaku（藤咲友朗）²，Chiharu Tsuji（辻知陽）² ¹福井大医眼科学 ²金沢大子どものこころ発達研究センター Autism spectrum disorder (ASD) is a lifelong condition which is diagnosed by the age of 8. The characteristic symptoms of ASD are deficits in the social skills, impairment of speech and nonverbal communication, and the repetitive behaviors. However, ASD symptoms are shown in the wide variety of range with combination of other symptoms which makes the ASD not a uniform disorder. The accumulating evidences have shown that the cause of ASD is due to the different combinations of genetic and environmental influences. Further studies are necessary to understand the ASD, because there is almost no pharmacological treatment to cure the ASD symptoms. Valproic acid (VPA) is an antiepileptic drug widely used to treat epilepsy and migraine, but shown to increase the risk of malformation, ASD, and mental retardation in the children by the use of VPA during pregnancy. According to these reports, researchers use VPA to create ASD model rodents. The siblings (VPA pups) born from VPA-exposed-dams show anxiety-like behavior, social behavior deficit, and cognitive dysfunction which are common symptoms of the ASD. However, all of the behavioral studies have been conducted during adulthood and not during childhood. Here, we examined the ultrasonic vocalization of the pups induced by separation from dam. VPA pups did not show any difference in the frequency and the duration of USV calls, however the number of USV calls decreased in VPA-pups compare to the wild type after postnatal day 11. Since the difference was observed in the later stage of the lactation period, our data suggested that VPA pups may have altered emotional stage or less sensitivity to the social condition compare to the wild type pups.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-443 FoxG1因子量による自閉症病態の発症機構 Goichi Miyoshi（三好悟一）¹，Yoshifumi Ueta（植田禎史）¹，Yuki Yagasaki（矢ケ崎有希）¹，Hironobu Osaki（尾崎弘展）¹，Rob Machold（Machold Rob）²，Gord Fishell（Fishell Gord）²，Mariko Miyata（宮田麻理子）¹ ¹東京女子医科大学 ²ニューヨーク大学　医学部 The mammalian cerebral cortex is composed of a sophisticated neuronal network that processes higher order information such as perception, consciousness and memory. Thus, mutations in genes involved in the specification and migration of neurons as well as the formation of the correct synapses within the six-layered neocortex often lead to neurological diseases. Both up-regulation (duplication) and down-regulation (haploinsufficiency) of the transcription factor FoxG1 have been identified in syndromic forms of autism spectrum disorders (ASD). Furthermore, studies on idiopathic ASD patients have implicated FoxG1 dysregulation more broadly in ASD etiology. Here, we elucidate the mechanisms underlying these disease outcomes by demonstrating that dynamic regulation of FoxG1 expression is essential during cortical development. At the cellular level, dynamic changes in FoxG1 levels at key transition points during differentiation assure proper circuit formation. In maturing circuits, either increased or decreased FoxG1 levels result in animals developing ASD-like social behavior impairments but only when changes are matched between excitatory/inhibitory populations. Unexpectedly, at the circuit level, we find that the excitation/inhibition balance within the juvenile prefrontal cortex is particularly sensitive to perturbations in FoxG1 dosage. Our findings illuminate the critical dosage and timing dependent requirements for FoxG1 in brain development and social behavior.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-444 母体免疫活性化により産まれたマウスの脳神経イメージング Hiroshi Mizuma（水間広）¹，Keigo Hikishima（疋島啓吾）²，Nami Hosaka（保坂奈美）¹，Yoshino Matsumoto（松本佳乃）¹，Norio Takata（高田則雄）³，Hirotaka Onoe（尾上浩隆）⁴ ¹理研生命機能科学研究セ ²沖縄科学技術大学院大 ³慶應大医精神・神経科学 ⁴京都大院医脳機能総合研究セ Pregnant bacterial or viral infection has been considered as one of the risk factors of neurodevelopmental disorder in offspring, such as autistic spectrum disorder (ASD) and schizophrenia. Animal studies indicated that maternal immune activation (MIA) by pseudoviral infection in pregnant mice using polyinosinic:polycytidylic acid (poly(I:C)) elevated pro-inflammatory cytokines in maternal blood, displaying structural abnormalities in fetal brain and ASD-like behavior in postnatal offspring. However, the brain function in MIA model underlying cause of the behavioral phenotype in MIA offspring remained undetermined. To investigate the functional connectivity and activity in brain of MIA-affected offspring, we performed resting state functional magnetic resonance imaging (rs-fMRI) using ultra-high tesla (11.7T) and regional cerebral glucose metabolism using positron emission tomography (PET) with [¹⁸F]fluorodeoxyglucose (FDG) under the awake condition which we have established previously. MIA model was generated by intraperitoneal injection of Poly(I:C) (20 mg/kg) in pregnant mice on gestational day 12.5. The MIA-affected offspring showed a significant social deficit in the three-chamber approach test. Rs-fMRI study revealed that MIA-affected offspring with social deficit had hyper connectivity of functional networks with over and local preferred connectivity, which were similar to those of human studies in ASD patients reported previously. In addition, [¹⁸F]FDG-PET imaging showed low glucometabolism in the thalamus, midbrain, and central gray matter of the MIA-affected offspring. These results suggest that abnormal functional network connectivities associated with glucometaboic activity may be crucial in the appearance of behavioral phenotype in the MIA offspring.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-445 自閉スペクトラム症に関係するCaspr3のマウス神経系におけるオートファジーへの関与 Reika Takeda（武田嶺花）¹，Tsuyoshi Koide（小出剛）²，Yasushi Shimoda（霜田靖）¹ ¹長岡技科大工生物 ²国立遺伝研マウス開発研究室 Autophagy is a process of disassembling and reusing cellular components such as damaged cell organelles or unused proteins. Abnormalities of the autophagy in the nervous system were implicated in autism spectrum disorder (ASD). However, ASD is one of the neurodevelopmental disorders whose pathogenic mechanisms have not been elucidated. Recently, genetic variation of contactin associated protein like 3 (Caspr3, also known as Cntnap3) was found in ASD patients. Caspr3 encoded by CNTNAP3 gene is a transmembrane protein belonging to neurexin superfamily. Caspr3 deficient mice showed ASD like behavioral phenotypes such as abnormalities in social interactions and increase of repetitive behaviors. These facts suggested that Caspr3 is associated with ASD. In addition, Caspr3 expression was upregulated in the intestinal tissue of the patients with Crohn's disease and may be involved in regulating autophagy in cultured cells. We previously showed that Caspr3 was localized inside primary cultured neurons, though it remains unclear why and how Caspr3 resides inside the neurons. Taken together, we hypothesized that Caspr3 may play a role in autophagy in the nervous system, and Caspr3 mutation might be involved in the onset of the ASD by causing autophagic abnormalities. In this study, we investigated whether Caspr3 is associated with autophagy in the nervous system. First, we examined the localization of Caspr3 and LC3B, a marker for autophagy, using mouse brain sections. We found that expression of Caspr3 was partially overlapped with that of LC3B. Next, we assessed LC3B expression level in Caspr3 deficient mouse brain, indicating abnormal expression of LC3B expression in the mutant mouse. These results suggested that Caspr3 is involved in autophagy in the mouse brain.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-446 自閉スペクトラム症で同定されたコピー数多型と相同なCntn5変異マウスの行動解析 Yu Ishida（石田悠）¹，Ayako Tanaka（田中絢子）¹，Yuji Imai（今井悠二）²，Kazuto Yoshimi（吉見一人）^2,3，Tsuyoshi Koide（小出剛）²，Yasushi Shimoda（霜田靖）¹ ¹長岡技科大工生物 ²国立遺伝研マウス開発研究室 ³大阪大院医 Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder characterized by impaired social communication and restricted, repetitive behavior. Besides these behavioral phenotypes, patients with ASD often suffer from hypersensitivity to sensory stimuli, especially auditory hypersensitivity. However, the pathological mechanism of ASD symptoms including auditory hypersensitivity remains to be clarified. Among hundreds of genes associated with the onset of ASD, CNTN5 gene encodes the cell adhesion molecule that has been implicated in the development of the auditory nervous system. Very recently, copy number variation of CNTN5 gene lacking the flanking region of exon 4 was found in an ASD patient. It was shown that there are two major splicing isoforms of CNTN5, long and short isoforms, in the presence and absence of amino acid sequence encoded by exon 4, respectively. These led us to hypothesize that imbalance in long/short isoforms of CNTN5 could possibly cause the onset of ASD. In this study, we generated knockout mice in which the flanking region of exon 4 of Cntn5 gene (Cntn5-ex4 KO mice) using CRISPR/Cas9 system and performed behavioral analyses on Cntn5-ex4 mice. First, Western blot analysis of Cntn5-ex4-deficient brain revealed that the short isoform of Cntn5 was expressed, while the long isoform was not. Then, we conducted some behavioral tests to evaluate ASD-related behaviors of Cntn5-ex4 mice. While no significant difference was observed between Cntn5-ex4 KO and WT mice in the open field test, the social exploration test indicated that Cntn5-ex4 KO males, not females, may exhibit a deficit in social interaction. There is a possibility that defect of the long isoform of Cntn5 protein might affect social behavior.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-447 自閉症モデルマウスBTBRと高社会性マウスC57BL/6Jの遺伝子発現の包括的解析 Shota Mizuno（水野翔太）¹，Junna Hirota（弘田淳奈）¹，Hirohide Iwasaki（岩崎広英）²，Shigeo Okabe（岡部繁男）²，Yoshitake Sano（佐野良威）¹，Teiichi Furuichi（古市貞一）¹ ¹東京理科大院理工応用生物科学 ²東京大院医細胞生物・解剖 Autism spectrum disorder (ASD) is a neurodevelopmental disorder whose prevalence rate is approximately 1%, and that is characterized by two core symptoms: 1) impaired social interaction and communication, and 2) restricted and repetitive behavior. Genetic factors strongly contribute to the risk for ASD. Recent studies reported ASD have high degree of genetic heterogeneity and are probably caused by complex interaction between multiple genes. Over a thousand genes have been reported as ASD-associated genes and candidate genes, so far. However, it is unclear how a combination of different gene variation causes a common mind symptom. Here, we compared brain gene expression between BTBR T+tf/j mouse model for ASD and C57BL/6J (B6) mice, which show high levels of social behavior (n=4 for each strain). For this purpose, we applied DNA microarray (Agilent, 27,122 genes, 4,578 lncRNAs) analysis for genome-wide expression profiling in the cerebral cortex and striatum because of the possible involvement of the corticostriatal pathway in ASD core symptoms. As a result, we could identified 1,081 downregulated genes and 359 upregulated genes. Among these genes, 45 genes (34 downregulated and 11 upregulated) have already been reported as ASD gene candidates, and 268 genes (189 downregulated and 79 upregulated) are non-coding RNA(ncRNA). First, we compared different expressed ASD candidate genes to previous reported gene lists targeted by Fmr1 and Cpeb4, ASD candidate RNA binding protein which interact with several ASD gene candidates. we identified 4 and 15 targeted genes, suggested important genes. Second, to investigate different expressed ncRNA functional interaction, we collected 15 gens RNA interaction data from database and described interaction network. Therefore, network was composed of 15 ncRNA and 135 target genes including 6 ASD candidate genes. Lastly, Gene ontology (GO) enrichment analysis using DAVID (https://david.ncifcrf.gov/) highlighted 96 annotation including synapse and immune related that previous human study reported. Interestingly, two novel GO terms were extracted: "extracellular exosome" and "metal ion binding", suggesting that co-expression pathways related to exosome and/or metal ion binding genes may be affected in the corticostriatal pathway of BTBR mice. These comprehensive study shows transcriptional feature between BTBR and B6, may contribute to identify novel molecular targets for pharmacological studies.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-448 ラパマイシン投与によるバルプロ酸曝露マウスの社会性行動障害と遺伝子発現への影響 Hiroko Kotajima（古田島浩子）^1,2，Toshiyuki Kobayashi（小林敏之）³，Hirofumi Kashii（柏井洋文）¹，Atsushi Sato（佐藤敦志）⁴，Yoko Hagino（萩野洋子）¹，Miho Tanaka（田中美歩）⁵，Yasumasa Nishito（西藤泰昌）⁶，Yukio Takamatsu（高松幸雄）⁶，Shigeo Uchino（内野茂夫）^1,2，Kazutaka Ikeda（池田和隆）¹ ¹東京都医学総合研究所 ²帝京大学理工学部バイオサイエンス学科 ³順天堂大学大学院医学研究科分子病理病態学 ⁴東京大学医学部附属病院 ⁵国立精神神経医療研究センター精神保健研究所知的・発達障害研究部 ⁶東京都医学総合研究所基盤技術研究センター Introduction Autism spectrum disorder (ASD) is a neurodevelopmental disorder that is characterized by deficits in social interaction. Various genetic and environmental factors have been investigated in studies for ASD. However, the medical treatment for impairment of social interaction in ASD has not been established. Valproic acid (VPA) is used as an anti-epileptic drug, a mood stabilizer and a migraine. Pregnant women treated with VPA deliver their children with ASD. Exposure to VPA prenatally in animals has been used to ASD model. Recent studies showed that aberrant mTOR signaling pathway causes ASD-like behaviors in VPA-exposed animals. The mTOR signaling pathway plays a crucial role in neuronal cell proliferation and synaptogenesis. The overactivation of mTOR has been implicated in the pathogenesis of syndromic ASDs, such as tuberous sclerosis complex (TSC). Treatment with rapamycin, mTOR complex inhibitors, improves social deficits in Tsc heterozygous mice. A recent study reported that an aberrant mTOR signaling pathway causes ASD-like behaviors in VPA-exposed animals. These studies suggest that overactivation of mTOR signaling pathway is associated with ASD and mTOR inhibitor is a potential therapeutic drug for ASD. Therefore, our aim is to clarify the effect of rapamycin treatment in VPA-exposed animals. Methods Pregnant female mice received a subcutaneous injection of 600 mg/kg VPA on day 12.5 after conception. The pups were injected with rapamycin or vehicle once daily for 2 consecutive days and social interaction test was conducted after administration of rapamycin in both ages 5-6 weeks (adolescence) and 10-11 weeks (adult) mice. The mouse whole brains were extracted after the social interaction test on adult, and microarray and western blots analysis were performed. Results and Conclusion We found that rapamycin treatment in VPA-exposed mice improved social deficits in both adolescence and adult. Mice that were prenatally exposed to VPA and treated with vehicle exhibited the aberrant expression of genes in the mTOR signaling pathway, and rapamycin treatment recovered changes in the expression of some genes, including Fyb and A330094K24Rik. Moreover, rapamycin treatment suppressed S6 phosphorylation in VPA-exposed mice. These results suggest that rapamycin treatment is effective for treatment of non-syndromic ASD that is caused by aberrant mTOR signaling pathway.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-449 妊娠中に食事制限したラットの仔における小脳の構造と代謝状態の変化 Kana Miyamoto（宮本佳菜）¹，Kwong Soon Thomas Tiong（Thomas Tiong Kwong Soon）¹，Seta Sato（佐藤誠太）¹，Yoko Nomura（Nomura Yoko）²，Yasunari Kanda（諫田泰成）³，Sachiko Yoshida（吉田祥子）¹ ¹豊橋技科大院工　環境・生命工学 ²ニューヨーク市立大学 ³国立医薬品食品衛生研究所 Embryonic environment plays some important roles in healthy condition after birth. The Hunger Winter (Dutch Famine from 1944 to 1945) has been studied much and it was reported that offspring under malnutrition condition suffered from depression, schizophrenia and metabolic diseases. It would be a result of epigenetic gene regulation. In modern developed countries including Japan, there is a tendency for low birth weight children to increase due to self-decided food restriction (FR). Stresses such as FR and exposure of chemical substances during embryonic period could affect offspring to lifelong effects. Recently, we have reported that valproate (VPA)-administrated rat offspring show the developmental alteration of Purkinje cells and excess folds in some cerebellar lobules. Developing cerebellum could be very sensitive to embryonic chemical stress. In this study, we report the alteration in cerebellar structure in rat offspring under FR. From the embryonic day 3 (E3) to E17, pregnant mother rats in a FR group were restricted meals to 75% of the control group. Pregnancy period of FR mothers were elongated about 1 day. Harder food restriction, 50 % or 60 % restriction, resulted failure of pregnancy continuation. In the cerebellum of the FR rat pups at postnatal day 14, excess folding between the lobule V and VI was observed as same as VPA-administrated pups. Moreover, some FR rat offspring at over 12 weeks after birth showed glucose tolerance, over 200 mg / dL blood glucose level even at 2 hours after glucose intake. This is a numerical value of the onset of diabetes, suggesting the development of pediatric diabetes due to food restriction during pregnancy. We suggest that one of the prenatal stresses of food restriction would affect the cerebellar structures of offspring.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-450 自閉スペクトラム症と統合失調症における眼球運動の比較 Tomoko Shiino（椎野智子）^1,2，Kenichiro Miura（三浦健一郎）^3,4，Michiko Fujimoto（藤本美智子）^1,4，Noriko Kudo（工藤紀子）¹，Hidenaga Yamamori（山森英長）^1,5，Yuka Yasuda（安田由華）^1,6，Manabu Ikeda（池田学）⁴，Ryota Hashimoto（橋本亮太）^1,2,4 ¹国立精神・神経セ精神保健研精神病態 ²大阪大院連合小児発達 ³京都大院医認知行動脳科学 ⁴大阪大院医精神 ⁵地域医療機能推進機構大阪病院 ⁶生きる・育む・輝くメンタルクリニック Eye movement abnormalities are often associated with psychiatric illness. Subjects with schizophrenia has been reported to have different eye movement characteristics compared to healthy controls. It has been reported that subjects with schizophrenia can be distinguished from healthy individuals by using eye movements during standard eye movement tasks. On the other hand, subjects with Autism Spectrum Disorder (ASD) has also been reported to show eye movement abnormalities. However, it is still unclear whether the eye movement abnormalities in ASD involve common features with those in schizophrenia or not. In this study, we aimed to understand similarity/difference of eye movement abnormalities of subjects with ASD and those with schizophrenia. We studied eye movement characteristics of 17 patients with ASD, 84 patients with schizophrenia and 256 healthy controls with no history of psychiatric illness. Eye movements of the subjects were gathered during fixation, smooth pursuit and free viewing tasks, and 35 eye movement characteristics representing ocular motility or performance of task completions were extracted from the data obtained using these three eye movement tasks. This study was performed in accordance with the World Medical Association's Declaration of Helsinki and was approved by the Research Ethics Committee of Osaka University. In comparisons between eye movement characteristics of subjects with schizophrenia and healthy controls, we found significant differences in 19 eye movement characteristics during fixation, smooth pursuit and free viewing tasks (Bonferroni corrections, p<0.05/35), of which 6 eye movement characteristics had large effect-size (Cohen's d>0.8). However, after Bonferroni corrections, eye movement characteristics of the subjects with ASD were not significantly different from healthy controls. All the eye movement characteristics showed intermediate effect-size or less. Furthermore, compared with schizophrenia, the subjects with ASD showed a significant difference in the number of saccades during the free viewing task (p=6.1*10-4, Cohen's d=0.90). The present results suggest that the subjects with ASD did not show eye movement abnormalities similar to patientsh schizophrenia during free viewing tasks. Furthermore, the three eye movement tasks used here may be usable to distinguish subjects with schizophrenia from those with ASD.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-451 統合失調症における環境要因のエピゲノム解析と分子病態の解明 Zhiqian Yu（兪志前），Mai Sakai（坂井舞），Chiaki Ono（小野千晶），Yuta Takahashi（高橋雄太），Hiroaki Tomita（富田博秋）東北大災害研災害精神医学 Background: Epidemiological evidence indicates that female offspring exposed to maternal infection during pregnancy are significantly more susceptible to schizophrenia. We examined sex differences in the effect of polyriboinosinic-polyribocytidylic acid [Poly(I:C)] administration, which mimics maternal viral infections, to pregnant mice on offspring behavior. Transcriptional profiles in the brain were investigated as candidate mechanisms underlying the phenomena. Methods: Pregnant mice were treated with Poly(I:C) in late gestation. Following behavioral characterization of the adult male and female offspring (n=10/group), gene expression and DNA methylation in the prefrontal cortices (PFC)s were characterized using microarray and methylated-DNA binding domain-sequencing (MBD-seq). The findings were validated with replicated mouse experiments (n=10/group) using real time-PCR and pyrosequencing. Postmortem samples from schizophrenic or healthy subjects (n=9/group) were assessed to validate the present findings in the human homologous locus. Results: Female offspring of Poly(I:C)-treated dams predominantly express schizophrenia-linked behavioral changes, profound alterations in gene expression profiles, hypermethylation of genomic DNA, and increased Dnmt1 transcription in the PFCs. Gene Ontology term enrichment analysis indicated that schizophrenia-relevant genes were significantly overrepresented among Poly(I:C)-affected genes in the female offspring. Of these genes, the hypermethylation of the Acsbg1/ACSBG1 locus and decreased gene transcription were validated in the PFCs of the female offspring of Poly(I:C)-treated dams and postmortem brains of female patients with schizophrenia.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-452 神経発達障害モデルマウスの行動異常に対する海馬内リーリン補充の影響 Daisuke Ibi（衣斐大祐）¹，Nayu Koide（小出菜優）¹，Genki Nakasai（中齋玄紀）¹，Masahito Sawahata（澤幡雅仁）²，Takayoshi Mamiya（間宮隆吉）¹，Kiyofumi Yamada（山田清文）²，Masayuki Hiramatsu（平松正行）¹ ¹名城大薬薬品作用 ²名古屋大院医医療薬学 Although chronic atypical antipsychotic administration produces significant reduction or even complete remission of psychosis such as hallucinations and delusions, the majority of schizophrenia patients have cognitive and emotional deficits, which often do not improve upon antipsychotic drug treatment. Given that atypical antipsychotic drugs all have in common a high affinity for the dopamine D2 receptor, as well as a modest affinity for the serotonin 5-HT2A receptor, cognitive and emotional deficits is hypothesized to involve neural networks beyond the classical dopaminergic mesolimbic pathway, including serotonin systems as well. It is reported that mutation of RELN gene encoding Reelin, an extracellular matrix protein involved in the neural development and synaptic plasticity, is associated with neurodevelopmental disorders such as schizophrenia and autism spectrum syndrome. Further, hippocampal Reelin level was reportedly down-regulated in the brain of both schizophrenics and preclinical rodent models. In this study, we investigated the effect of Reelin supplementation into the hippocampus on behavioral phenotypes in a preclinical mouse model to evaluate the validity of Reelin signaling activation as a new treatment strategy to mental disorders beyond the classical monoamine target. We employed offspring of pregnant mice exposed to immune activation as a preclinical model exhibiting cognitive and emotional deficits as well as histological decrease in Reelin-positive cell number and synaptoporin levels, a synaptic vesicle protein, in the hippocampus. Microinjections of Reelin protein into the hippocampus improved the object cognitive memory impairments, anxiety-like behavior as well as tended to recruit synaptoporin in the hippocampus. These results suggest that Reelin supplementation therapy has a potential to cure cognitive and emotional impairments and synaptic disturbances in patients with neurodevelopmental disorders including schizophrenia.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-453 Orleans reelerマウスのリン酸化プロテオミクスにより同定された新規reelin関連リン酸化シグナルTWF1 Taku Nagai（永井拓），Takaaki Matsui（松井崇晃），Ayato Sato（佐藤礼都），Norimichi Itoh（伊藤教道），Kiyofumi Yamada（山田清文）名古屋大院医医療薬学 Reelin plays pivotal roles in CNS such as radial migration of cortical neuron in developmental stage and synaptic plasticity in adulthood. We previously found behavioral alterations in Orleans reeler (Reln^rl-Orl) mice. However, it remains largely unknown what molecules in downstream signaling of Reelin contribute to its phenotypes. To clarify the molecular mechanism, we performed comprehensive phosphoproteomic analysis using Reln^rl-Orl mice. Brain lysate from WT, hetero (Reln^rl-Orl/+) or homo (Reln^{rl-Orl/rl-Orl}) mice was immunoprecipitated with anti-phosphotyrosin antibody followed by LC-MS/MS analysis. We identified twinfilin-1 (TWF1) as one of the decreased phosphoproteins in Reln^rl-Orl mice. To examine whether Src and Fyn phosphorylate TWF1, Myc-TWF1 was cotransfected with either control GFP, GFP-Src or GFP-Fyn in HEK cells, and measured the level of tyrosinphosphorylated TWF1. Cotransfection of TWF1 with GFP-Src or GFP-Fyn increased tyrosine phosphorylation of TWF1, suggesting that TWF1 is substrate for both Src and Fyn kinase. To investigate the phosphorylation site of TWF1 by Src and Fyn, we performed phosphorylation assay with phosphorylation-defective mutants of TWF1 (TWF1 Y137F and Y309F). Src-mediated phosphorylation of TWF1 was significantly decreased in TWF1 Y309F mutant, but not TWF1 Y137F. These results indicate that Src phosphorylates TWF1 at tyrosine 309. The role of TWF1 in spine morphology was examined with primary cultured neurons. The spine head size tended to decrease in TWF1 knockdown neuron. These results suggest that TWF1 may be associated with spine morphology. The tyrosine phosphorylation of TWF1 by Src family kinase might be one of the crucial downstream signaling of Reelin.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-454 新生仔期VPAC2受容体作動薬投与マウスの認知機能障害はクロザピン特異的に改善される Shuto Takeuchi（竹内修斗）¹，Ryosuke Yamauchi（山内良介）¹，Takuya Kawanai（河内琢也）¹，Tatsunori Miyaoka（宮岡辰典）²，Lu Chen（Chen Lu）²，Atsuko Hayata（早田敦子）^1,3，James A. Waschek（Waschek A. James）⁴，Takanobu Nakazawa（中澤敬信）^1,5，Kazuhiro Takuma（田熊一敞）^1,3,5，Shinsaku Nakagawa（中川晋作）²，Hitoshi Hashimoto（橋本均）^1,3,6,7，Yukio Ago（吾郷由希夫）² ¹大阪大・院薬・神経薬理 ²大阪大・院薬・薬剤 ³大阪大・院連合小児発達・子どものこころセンター ⁴カリフォルニア大ロサンゼルス校・精神医学 ⁵大阪大・院歯・薬理 ⁶大阪大・データビリティフロンティア機構 ⁷大阪大・先導的学際研究機構 Rare microduplications at 7q36.3, containing VIPR2, have been known to confer a risk for schizophrenia and autism spectrum disorder. VIPR2 encodes the VPAC2 receptor that binds two homologous neuropeptides, VIP and PACAP. Lymphocytes from patients with these microduplications exhibited higher VIPR2 gene expression and VIP responsiveness, indicating the functional significance of the microduplications and VPAC2 receptor overactivation. We previously found that treatment of neonatal mice with Ro25-1553, a selective VPAC2 receptor agonist, caused anhedonia, sensorimotor gating deficits, cognitive impairments, and alterations in dendritic morphology of prefrontal and perirhinal cortical neurons at adulthood. Here we aimed to clarify the effects of several antipsychotic drugs on behavioral and dendritic abnormalities in Ro25-1553-treated mice. Chronic treatment with clozapine, but not haloperidol, risperidone, or aripiprazole, during adulthood improved cognitive impairments in the novel object recognition test. Clozapine, but not risperidone, also ameliorated dendritic structural abnormalities in prefrontal and perirhinal cortical neurons of Ro25-1553-treated mice. Chronic clozapine treatment upregulated several immediate early response genes in the prefrontal cortex of Ro25-1553-treated mice, implying the increased baseline neuronal activity. Clozapine is the only antipsychotic agent licensed for treatment-resistant schizophrenia (TRS); thus the findings from this study suggest that mice neonatally treated with Ro25-1553 might provide some aspects of neural and behavioral endophenotypes relevant to TRS. Further study using this mouse model would offer new insights into the mechanism underlying the effects of clozapine and an alternative therapeutic strategy for TRS.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-455 健常者と統合失調症者の重り負荷課題における前腕の動きと筋電図の特性 Yui Kikuchi（菊池ゆひ），Sumika Matsushima（松島純香），Tetsuo Ota（太田哲生），Mitsugu Yoneda（米田貢），Takako Ohno-Shosaku（少作隆子）金沢大・医薬保健・リハビリテーション科学 Prediction-based, feed-forward motor control, which involves the cerebellum, is important for performing movements smoothly and skillfully. Patients with schizophrenia have been reported to show motor symptoms and morphological abnormalities of the cerebellum. We previously demonstrated that prediction-based motor control is impaired in patients with schizophrenia by measuring anticipatory response in a simple loading task with a haptic device named the Space Interface Device for Artificial Reality (SPIDAR). In that study, anticipatory response, an upward movement just before the onset of loading in the predictable condition, was shown to be smaller in most patients than in normal subjects, but comparable in some patients, suggesting the heterogeneity in motor control of patients. In the present study, we analyzed hand movement and electromyogram during a loading task in the real world, without using the device SPIDAR, for future clinical use. Loading was started by putting a weight (500 g) on the right palm of a subject by himself/herself ("predictable condition") or by someone else ("unpredictable condition"). Loading was repeated more than five times under each condition. Movements of the weight and the right hand were monitored by accelerometer (AC). Surface electromyograms (EMG) were recorded from two flexor muscles and one extensor muscle of forearm. The timing of onset of loading was detected from the AC signal of the weight. Under the unpredictable condition in normal subjects, the EMG activity of flexor muscles increased immediately after onset of loading, followed by the extensor muscle activity. Under the predictable condition in the normal subjects, the EMG activity of flexor muscles increased before onset of loading, reflecting anticipatory motor response. Some of the patients with schizophrenia showed no differences in the timing of the EMG activity increase of flexor muscles between the predictable and unpredictable conditions. We also found that some patients showed tonic activity of both flexor and extensor muscles. These patterns of EMG activity during a loading task could not be predicted simply from the amplitude of anticipatory response measured by SPIDAR. These data, taken together with previous results, suggest that the impairment of prediction-based motor control in the patients with schizophrenia is complex and heterogeneous. Whether these properties can be predicted from AC signal of hand will be discussed.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-456 ケタミン投与統合失調症モデルマウスの海馬におけるパルブアルブミン陽性ニューロンとペリニューロナルネットの変化 Risako Fujikawa（藤川理沙子），Jun Yamada（山田純），Shozo Jinno（神野尚三）九州大院医神経解剖学 Schizophrenia is a severe mental disorder that is characterized by delusions, hallucinations, and other cognitive difficulties. To date, dopamine antagonists are widely used for treatment of schizophrenia based on the dopamine hypothesis. However, recent post-mortem studies have indicated a loss of parvalbumin-positive (PV⁺) GABAergic neurons and perineuronal nets (PNNs) in the hippocampus and cortex of the patients of this disease. In the present study, we aimed to elucidate the potential involvement of hippocampal PV⁺ neurons and PNNs recognized by monoclonal antibody Cat-315 in the pathophysiology of a ketamine model of schizophrenia. Mice received one-week ketamine administration exhibited the hyperactivity in the open-field test and the reduced prepulse inhibition of startle response. The densities of total PV⁺ neurons and Cat-315⁺ PNNs in the hippocampus were generally decreased in ketamine-treated mice. The excitatory/inhibitory ratios of synaptic boutons covering PV⁺ neurons were elevated after ketamine administration. Unexpectedly, we found that the densities of Cat-315^-/PV⁺ neurons remained unchanged in ketamine-treated mice. In addition, Cat-315⁺/PV^- GABAergic neurons appeared after ketamine administration, while they were never observed in vehicle-treated controls. Because Cat-315⁺ PNNs are formed around specific subclasses of PV⁺ neurons, our findings suggest that subclass-dependent changes in PV⁺ neurons may be essential for understanding the pathophysiology of schizophrenia.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-457 Reelinの前頭前皮質内投与がMK801誘発性行動障害およびc-Fos発現に与える影響 Masahito Sawahata（澤幡雅仁）¹，Hiroki Asano（浅野裕樹）¹，Yumi Tsuneura（常浦祐未）¹，Taku Nagai（永井拓）¹，Takao Kohno（河野孝夫）²，Toshitaka Nabeshima（鍋島俊隆）^3,4，Mitsuharu Hattori（服部光治）²，Kiyofumi Yamada（山田清文）¹ ¹名古屋大院医医療薬学 ²名古屋市大薬病態生化学 ³藤田保衛大総医研システム医科学 ⁴藍野大 Reelin is a huge secretory glycoprotein and plays an essential role in brain development. Recently, Reelin has attracted research attention in schizophrenia (SCZ). It has been reported that SCZ patients show reduced levels of reelin mRNA and protein in medial prefrontal cortex (mPFC) and other brain areas. Previous our study indicated that intra-cerebroventriclular reelin has preventive effects on phencyclidine-induced cognitive and sensory-motor gating deficits. However, it remains unclear how Reelin prevents cognitive dysfunction. In this study, we examine the effect of reelin in the mPFC on memory impairment induced by NMDA receptor antagonist MK801. We used Expi293FTM expression system to prepare the recombinant Reelin. Expi293FTM cells were transfected with a full-length mouse WT Reelin or PDDK mutant which is degradation-resistant form of Reelin. For purified Reelin, cultured medium was incubated with anti-PA tag antibody coated-beads and concentrated by spin column. Recombinant Reelin was bilaterally injected into mPFC 5 days before the beginning of behavioral tests. MK801 (0.15 mg/kg) was administered in a single intraperitoneal injection 30 min before behavioral test or training. To examine the effect of reelin on SCZ like behavior, we performed pre-pulse inhibition (PPI) test, novel object test (NORT) and Y-maze test. As a result, microinjection of Reelin into the mPFC had no effect on MK801-induced impairment of sensorimotor gating and short-term memory. However, microinjection of Reelin significantly attenuated the long-term memory dysfunction in NORT. Next, we performed c-Fos staining to examine the mechanism of this reelin effect. Microinjection of Reelin improves the MK801-induced c-Fos expression. These results suggest that Reelin prevent MK801-induced long-term memory impairment and c-Fos expression in the mPFC.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-458 Environment enrichment ameliorates synaptic and behavioral impairments in a NMDA receptor hypofunction model Cora Sau Wan Lai（Lai Cora Sau Wan）^1,2，Yuhua Huang（Huang Yuhua）¹，Albert Hiu Ka Fok（Fok Albert Hiu Ka）¹，Xiaoyang Li（Li Xiaoyang）¹ ¹LKS Faculty of Medicine, The University of Hong Kong, HKSAR, China ²State Key Laboratory of Cognitive and Brain Research, The University of Hong Kong, HKSAR, China Schizophrenia is a neurodevelopmental psychiatric disorder with an age of onset in late adolescence and early adulthood. Although the precise pathogenesis and etiologies of the disease are remaining elusive, hypofunction of N-methyl-D-aspartate (NMDA) receptor has been proposed to underlie the emergence of symptoms. NMDA receptor antagonists such as MK-801, phencyclidine (PCP), and ketamine have been shown not only to induce schizophrenia-like symptoms in healthy subjects but also exacerbated existing psychosis in schizophrenia patients, suggesting they can be used as appropriate agents to study schizophrenic symptoms. Structural imaging of postmortem studies shows a decrease of dendritic spine density on pyramidal neurons (PNs), a structure for receiving the majority of excitatory synaptic input, in the prefrontal cortex (PFC) of schizophrenia patients. Dendritic spines undergo activity-dependent morphological changes over the lifetime and subtle changes in dendritic spines may have marked effects on synaptic function and connectivity in neuronal circuits and even cognition and behavior. The exposure to enriched environments (EE) during early life influences brain development and leads to altered behavior. In animal studies, EE has been demonstrated to reverse key schizophrenia-like behaviors such as hyperactivity and sensorimotor gating deficits. However, the effect of EE on dendritic spine plasticity in animal schizophrenic models is still unclear. In this study, we aim to use in vivo two-photon imaging of adolescence fluorescent mice to investigate the structural plasticity of PNs' dendritic spine in a chronic MK801-treated schizophrenic mouse model. We found that repeated exposure to the NMDA receptor antagonist MK801 induced schizophrenia-like behaviors and deficits of dendritic spine plasticity; while chronic treatment of EE ameliorated the synaptic and behavioral deficits. Further studies will focus on better understanding environmental modulation and NMDAR-mediated glutamatergic system interactions on synaptic and behavioral levels through manipulation of neuronal activity.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-459 早期精神症における白質radial diffusivityと幻覚の関係：拡散テンソル画像研究 Yutaro Sato（佐藤祐太朗）¹，Atsushi Sakuma（佐久間篤）²，Noriyuki Ohmuro（大室則幸）²，Masahiro Katsura（桂雅宏）²，Koichi Abe（阿部光一）¹，Kazuho Tomimoto（冨本和歩）¹，Kunio Iizuka（飯塚邦夫）²，Hiroo Matsuoka（松岡洋夫）^1,2，Hiroaki Tomita（富田博秋）^1,2,3，Kazunori Matsumoto（松本和紀）^1,2 ¹東北大学大学院医学系研究科精神神経学分野 ²東北大学病院精神科 ³東北大災害研災害精神 Recent studies using diffusion tensor imaging (DTI) have revealed reduced white matter integrity in individuals with hallucinations. However, these studies have focused mainly on fractional anisotropy in individuals with chronic schizophrenia. It remains unclear whether hallucination development is associated with alterations in radial diffusivity (RD). We aimed to investigate the relationship between RD and hallucination severity in individuals with at-risk mental state (ARMS) and first-episode psychosis (FEP). In total, 109 participants comprising 59 individuals with ARMS and 50 with FEP underwent magnetic resonance imaging. Hallucination severity was assessed using the Positive and Negative Syndrome Scale (PANSS) item P3. Correlation analysis of hallucination severity and white matter RD was performed using tract-based spatial statistics. Increased RD in the bilateral superior longitudinal fasciculus, left inferior longitudinal fasciculus, left inferior fronto-occipital fasciculus, corpus callosum, right cortical spinal tract, and bilateral optic radiation was associated with more severe hallucinations. Our results suggest that the development of hallucinations is accompanied by altered structural connectivity within brain networks associated with language processing, interhemispheric connection, motor control, and visual perception. These findings are consistent with those of previous DTI studies implying that hallucinations are associated with white matter abnormalities in multiple brain networks. Further neuroimaging studies are required to examine the role of these networks in the development of hallucinations.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-460 統合失調症における脳内意味ネットワーク異常：エンコーディングモデルとグラフ理論解析を用いて Yukiko Matsumoto（松本有紀子）¹，Satoshi Nishida（西田知史）²，Shuraku Son（孫樹洛）¹，Akio Murakami（村上晶郎）¹，Naganobu Yoshikawa（吉川長伸）³，Ryusuke Hayashi（林隆介）⁴，Shinji Nishimoto（西本伸志）²，Hidehiko Takahashi（高橋英彦）⁵ ¹京都大学大学院医学研究科脳病態生理学講座（精神医学） ²情報通信研究機構脳情報通信融合研究センター ³大阪大学医学部 ⁴産業技術総合研究所　人間情報研究部門 ⁵東京医科歯科大学大学院　医歯学総合研究科　精神行動医科学 Introduction Thought disorder, clinically characterized by loose association, is considered as a hallmark of schizophrenia. Several lines of researches have reported that disorders of concept formation and categorization ability are implicated in this symptom. In treatment for schizophrenia, it is important to know how concepts are organized in the individual patients' mind and assess the level of disorder based on some sort of objective metrics. In the previous study, our group have developed a new analytical method for fMRI using encoding modeling approach to evaluate semantic representation in the human brain. Here, we measured the fMRI activity from schizophrenia patients and predicted the semantic network using the encoding modeling approach. We then applied graph theoretic analyses to the predicted semantic network to characterize the disfunction of concept formation in the patients. Method fMRI signals were measured during presentation of natural movies from schizophrenia patients as well as healthy controls. Movies are annotated by Japanese captions every 1 second and we calculated the time-series scene vector of movies as the sequence of ""bag of words"" or the word vector space (word2vec) extracted from captions. We then estimated the model weights of regularized linear regression that best predicted fMRI signals from scene vector, and obtained semantic vector representations of the words for each subject. To quantify the structure of concept formation, we calculated pair-wise distance between word vector representations as a measure of connectivity in a semantic network for each subject. Results Several network metrics in the schizophrenia brain showed significant difference from those of the control group. The computational approach demonstrated in the present study offers novel perspectives on clinical assessment for psychiatric diseases.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-461 間欠・連続的な社会的敗北ストレスはうつ、不安/恐怖関連行動に異なる影響を示す Hiroyoshi Harada（原田洸秀），Masayoshi Mori（森征慶），Yuri Kouno（河野友里），Shunsuke Kawanabe（川邊隼輔），Yuna Hachou（八丁優奈），Yusuke Murata（村田雄介），Kenji Ohe（大江賢治），Munechika Enjoji（遠城寺宗近）福岡大学　薬学部　臨床薬物治療学 Introduction) Psychosocial stress is a crucial factor for developing psychiatric disorders including depression and anxiety disorders. Previous studies indicated that social defeat has opposite effects on the reward behaviors in rodents depending on the pattern of stress paradigm. Intermittent social defeat paradigm induced escalation of drug abuse behaviors and neuroadaptation, whereas continuous exposure induced blunted responses to sweet rewards and cocaine. However, it is poorly understood whether the pattern of stress paradigm affects differentially the fear response, anxiety- and depression-like behaviors. Thus, the present study was conducted in order to elucidate the effect of intermittent and continuous social defeat stress on behavioral and neurobiological changes. Methods) For the psychosocial stress exposure using Resident-Intruder paradigm, Long-Evans rats were used as ""Resident"", and male Sprague-Dawley rats were used as ""Intruder"". Intruder rats were divided into two groups depending on the pattern of stress paradigm. One group was intermittently exposed to 10 times social defeat stress over three weeks. The other group underwent continuous 10 times social defeat stress for ten consecutive days. Social defeat paradigm was carried out one time per day. After the termination of social defeat stress sessions, intruder rats in both groups were sequentially submitted to a battery of behavioral tests for assessing fear response, anxiety- and depression-like behaviors. Then, the brain of intruder rats was collected and subjected to the neurobiological assessment. Results and Discussion) Intermittent defeat group exhibited the significant increases in anxiety- and depression-like behavior assessed by the novelty suppressed feeding test and the forced swim test, respectively. On the other hand, continuous defeat group showed a trend to increase in the fear response in contextual conditioned fear test. These results suggest that psychosocial stress induces divergent psychiatric symptoms depending on the pattern of stress exposure. We are currently analyzing the neurobiological changes induced by intermittent and continuous defeat paradigm.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-462 Effect of dopamine reuptake inhibitor on depressive-like behaviors in mice Meina Zhu（Zhu Meina），Kanako Nozaki（Nozaki Kanako），Hidenori Aizawa（Aizawa Hidenori） Dept Neurobiol, Sch Biomed Health Sci, Hiroshima Univ, Hiroshima, Japan Depression is a prevalent disorder and one of the leading causes of disability. Despite the first line drug targeting to the serotoninergic transmission, some of the core symptoms such as anhedonia could be resistant to the treatment. Previous studies implicated that dopamine plays critical roles in motivation and rewarding behaviors. To address the effect of dopamine metabolism on hedonic behavior in mice, we developed a cost-effective method using intracranial self-stimulation (ICSS) paradigm. We applied the open-source operant training system Bpod (Sanworks) to the ICSS in which the mice were trained to poke the port to obtain the stimulation of the medial forebrain bundle (MFB) including the dopaminergic fibers. Results showed that the mice quickly learned nose poking behavior within several days, allowing us to analyze quantitatively the threshold frequency of the self-stimulation as a measure of the hedonic behavior. Descending orders of stimulation frequency were consecutively applied to the session to find the minimal threshold frequency for the hedonic behavior. Since the depressive-like phenotypes were associated with increased threshold in ICSS, we focused on nomifensine (NOM), dopamine reuptake inhibitor, on the hedonic behavior in ICSS. Our voltammetric recording revealed that systemic injection of NOM resulted in significant increase of released striatal dopamine induced by MFB stimulation, corroborating its effect on upregulation of dopamine metabolism. To investigate a role of NOM in hedonic behavior, we administered 3 and 7 mg/kg NOM intraperitoneally twice, in ascending and then descending order. Vehicle doses were given between each dose test as well as on the first and last test days. The pretreatment with NOM consistently enhanced locomotor activity with increased velocity in the open field (OFT), elevated plus maze (EPM) and novelty suppressed feeding (NSF) tests, and longer total distance in OFT and EPM. It also modified the anxiety-like behavior with less time spent in the open arm and longer latencies to consume the pellet in the arena in EPM and NSF, respectively. In tail suspension test (TST), NOM facilitated the struggling behavior with longer time spent in freezing and greater body movement. Taken together, our results demonstrated that NOM improved motivation with increased locomotor activity, which resulted in the amelioration of despair behavior under acute stress.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-463 PCR-free method reveals a highly brain region-specific mtDNA mutation spectrum in CaMKIIα-Polg mice Emilie Kristine Bagge（Bagge Emilie Kristine），Mie Kubota-Sakashita（Kubota-Sakashita Mie），Noriko Fujimori-Tonou（Fujimori-Tonou Noriko），Takaoki Kasahara（Kasahara Takaoki），Tadafumi Kato（Kato Tadafumi） Lab. for Molecular Dynamics of Mental Disorders, RIKEN CBS The involvement of mitochondria in neuropsychiatric diseases has recently become an area of increased focus. Interestingly, patients with bipolar disorder display an increased prevalence of mtDNA deletions compared to non-symptomatic controls and have a higher rate of disease-associated mutations of the POLG gene, the mitochondrial DNA polymerase. However, the link between accumulation of mitochondrial DNA mutations and changes in neuronal function is not clearly understood especially on a brain region-specific level. The CaMKIIα-Polg mouse model express proof-reading deficient Polg in forebrain neurons thereby accelerating the accumulation of mtDNA mutations. Behavioural analysis has shown these mice to experience spontaneous recurring episodes of depressive behaviour. In this study, we implemented a PCR-free method to enrich mtDNA from very limited tissue pieces from mouse brain. Using next-generation sequencing, we sequenced mtDNA from six different brain regions from aged Polg-mice and littermate controls (WT) to map the mutation spectrum. We find that the paraventricular thalamic nucleus (PVT) and nucleus accumbens (NAC) are hotspots of both deletions and SNVs whereas substantia nigra show no or limited changes. Interestingly, breakpoint sites are reused excessively in Polg-mice, though we find no specific motif pinpointing such breakpoints and no specific usage of direct-repeats. On the other hand, SNVs in Polg-mice lose their sequence specificity compared to WT, assuming a much more random motif. We find that SNV profiles for cortex, NAC and PVT are highly overlapping in an intra individual-independent manner. Surprisingly, we identify a brain region-specific accumulation of control region multimers, tandem repeats of D-loop region, adding further complexity to the Polg phenotype. These data indicate that there is a heterogeneous response of mitochondria to transgene expression across the mouse brain that may underlie differences in mitochondrial dynamics and maintenance of mtDNA integrity, highlighting the importance of brain-region specific mechanisms in mitochondria-based neuropsychiatric disorders and their research.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-464 MAGE-D1遺伝子欠損マウスにおける行動変化におけるノルアドレナリン作動性神経系の関与 Akihiro Mouri（毛利彰宏）¹，Mami Hirakawa（平川茉実）¹，Ken Watanabe（渡辺研）³，Kazuo Kunisawa（國澤和生）^1,4，Yuko Mori（森優子）⁵，Hisayoshi Kubota（窪田悠力）¹，Moe Niijima（新島萌）¹，Yasuko Yamamoto（山本康子）⁵，Yukihiro Noda（野田幸裕）²，Kuniaki Saito（齋藤邦明）⁴，Toshitaka Nabeshima（鍋島俊隆）⁴ ¹藤田医大院医療科学レギュラトリーサイエンス ²名城大院薬病態解析 ³長寿研運動器疾患 ⁴藤田医大院医療科学先進診断システム ⁵藤田医大院医療科学病態制御解析 Major depressive disorder (MDD) is a common mental disorder characterized by reduced motivation, diminished interest and pleasure, and anhedonia. We have proposed melanoma-associated antigen D1 (MAGE-D1) KO mice as depression model mice, and which is involved in the serotonergic hypofunction. Not only serotonin but also noradrenaline malfunctions are deeply involved in depressive behaviors. In this study, we investigate the involvement of noradrenergic nervous system in depression-like behaviors of MAGE-D1 KO mice. MAGE-D1 KO mice showed a decrease in locomotor activity, social interaction time and sucrose preference, increase in immobility time in the forced swimming test (FST), and extension in feeding latency in the novelty suppression feeding test. Noradrenaline (NA) tissue content in the prefrontal cortex, hippocampus, and amygdala, and potassium-evoked noradrenaline release in the prefrontal cortex and hippocampus were decrease in MAGE-D1 KO mice. The protein expression of noradrenaline transporter (NAT) was increased in the prefrontal cortex of the MAGE-D1 KO mice. Phosphorylation of NAT at threonine and protein expression of its kinase protein kinase C (PKC) were decreased, but ubiquitination or mRNA expression of NAT was not changed. Acute administration of NA reuptake inhibitors (desipramine and atomoxetine) attenuated increase in immobility time in the FST and decrease in sucrose preference, but not the other behavior changes in MAGE-D1 KO mice. These results suggested that depressive behavior in MAGE-D1 KO mice might be associated with noradrenergic hypofunction due to NAT overexpression through decrease in PKC-dependent phosphorylation.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-465 反復拘束ストレス負荷によるうつ様行動と生理学的変化に対するマウス系統の違い Shoko Tsuchimine（土嶺章子）¹，Hitomi Matsuno（松野仁美）¹，Kazunori Ohashi（大橋和徳）¹，Shuichi Chiba（千葉秀一）^1,2，Aya Yoshimura（吉村文）^1,3，Shingo Nakajima（中島進吾）¹，Sayuri Ishiwata（石渡小百合）¹，Shintaro Ogawa（小川慎太郎）¹，Kazuhiro Sohya（惣谷和広）¹，Hiroshi Kunugi（㓛刀浩）¹ ¹国立精神・神経セ神経研疾病３ ²岡山理科大学獣医学部獣医生理学講座 ³藤田医科大学研究支援推進センター疾患モデル教育研究施設 The chronic restraint stress (CRS) model in rodent is often used as a simple model of mood disorder. C57BL6/J (B6) and BALB/c mice are the most frequently used strains in genetics and behavioral studies. Since depression model studies have been developed mainly in rats, there are only a few studies on stress susceptibility in mice. In this study, we evaluated the differences between B6 and BALB/c mice in behavioral tests and physiological changes after chronic restraint stress in our experimental condition. B6 and BALB/c mice were exposed to CRS for 6 hours a day for 21 days, respectively. In both B6 and BALB/c mice, CRS induced several physiological changes, such as decreased body weight gain, thymic involution, increased adrenals weight and hair corticosterone levels, and fur state. In the sucrose preference test (SPT), stressed BALB/c mice significantly reduced the sucrose preference compared to control mice. However, such anhedonia-like behavior was not observed in stressed B6 mice. No significant difference was observed in both strains between stressed and control mice in the forced swim test (FST) and tail suspension test (TST). In conclusion, we revealed differences in sensitivity to CRS between mouse strains. Our data suggested that CRS model using BALB/c mice would be more appropriate to study the pathophysiology of depression.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-466 神経障害性疼痛病態形成における時計遺伝子Period2の役割 Naoki Kusunose （楠瀬直喜）¹，Yuta Yamaguchi（山口悠太）¹，Naoya Matsunaga（松永直哉）^1,2，Satoru Koyanagi（小柳悟）^1,2，Shigehiro Ohdo（大戸茂弘）¹ ¹九州大学薬学研究院薬剤学分野 ²九州大学薬学研究院グローカルヘルスケア分野 Neuropathic pain hypersensitivity is often caused by peripheral nerve damage accompanied by diabetes, viral infection, or cancer invasion, but the threshold of pain hypersensitivity changes in a circadian (approximately 24-h) time-dependent manner. Circadian rhythms in physiological functions are regulated by clock gene which are components of an internal self-sustained molecular clock. Recent studies have revealed that clock gene participate in pathogenesis of diseases such as sleep disorder, obesity, and cancer. In this study, we used clock gene Period2 mutant (Per2^{m/m^{) mice to clarify the role of clock gene in the development of neuropathic pain hypersensitivity. The sciatic nerve-ligated mice were prepared as neuropathic pain hypersensitivity model. Paw withdrawal threshold was evaluated by von Frey filament and up-down method. The paw withdrawal threshold of nerve injured side of wild-type mice was significantly decreased after sciatic nerve ligation, indicating the development of neuropathic pain hypersensitivity. Although behavioral responses of Per2^{m/m^{mice against acute and inflammatory pain were comparable to those observed in wild-type mice, but neuropathic pain hypersensitivity was not developed in Per2^{m/m^{mice. Spinal expression of P2ry12 gene, which encodes P2Y12 receptor, has been indicated to be associated with neuropathic pain hypersensitivity in sciatic nerve-ligated mice. The levels of P2ry12 mRNA in spinal cord of wild type mice significantly elevated after sciatic nerve ligation, but the elevation was not detected in Per2^{m/m^{mice. Furthermore, Down-regulation of Period2 expression by miRNA resulted in the reduction of P2ry12 mRNA levels in MG6 cells, a mouse cell line of microglia. These data suggested that Period2 is involved in P2ry12-dependent pathogenic mechanisms of neuropathic pain hypersensitivity.}}}}}}}}

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-467 EGFシグナルによるモルヒネ鎮痛効果の増強 Mitsushi J Ikemoto（池本光志）^1,2 ¹産総研バイオメディカル ²東邦大院理生物分子科学 Morphine activates epidermal growth factor (EGF) signaling by transactivating epidermal growth factor receptor (EGFR) via μ opioid receptor (MOR) in astrocytes and human lung cancer cells. Furthermore, both extracellular and intracellular epidermal growth factor receptor (EGFR) inhibitors dramatically improve neuropathic pain in human patients. These reports raise a possibility that EGF signaling may lead to develop MOR-dependent analgesic effect. Here, to verify this hypothesis, we evaluated morphine-induced analgesic effect of ddY (6-week old) mice by hot plate test. First, we examined effect of AG1478, a specific EGFR tyrosine kinase inhibitor, on morphine-induced analgesic effect. Pretreatment of AG1478 (5 mg/kg, i.p.) drastically attenuates acute anti-nociceptive effect when a single morphine (10 mg/kg, s.c.) was injected to mice (p < 0.01). Furthermore, AG1478 (1 or 5 mg/kg twice a day, i.p.) significantly suppressed the development of tolerance to the anti-nociceptive effect in a dose dependent manner when mice were given in repeated morphine (10 mg/kg, s.c. twice a day) for 4 days. Next, we investigated whether activation of EGF signaling in the periaqueductal gray (PAG) in midbrain modulate the acute morphine-induced analgesic effect because PAG highly expresses MOR and plays a main control center for descending pain modulation. Interestingly, mice infused recombinant EGF (8 pmoles) into PAG showed significant enhanced analgesic effect compared than that of vehicle when a single morphine (10 mg/kg, s.c.) was given (p < 0.01). Meanwhile, mice received AG1478 (50 pmoles) into PAG had a drastic reduction of acute morphine analgesic effect compared than control mice (p < 0.01). Furthermore, intra-PAG infusion of GM6001 (5 nmoles), a strong wide range MMP inhibitor, had no effect on acute morphine-induced anti-nociceptive effect, suggesting that MMP-induced EGFR transactivation system does not participate in this phenomenon. These evidences reveal that EGF signaling may modulate morphine-induced analgesic effect.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-468 側頭葉てんかん新規バイオマーカーの提案 Keiko Kato（加藤啓子），Manami Ota（太田真菜美），Akiko Fujita（藤田明子）京都産大生命科学,　京都 Epilepsy is a chronic noncommunicable disease of the brain that affects people of all ages and is characterized as the seizure, a transient behavior caused by disordered, synchronized, and rhythmic firings of neuronal groups in the brain. The epilepsy is uncontrolled pharmacologically in up to 30% of patients and the uncontrolled epilepsy is associated with poorer quality of life, increased accidents, and sudden death. Sometimes, patients of drug-resistant epilepsy receive alternative treatment through dietary manipulation, such as a ketogenic diet, leading to improvement of seizures. Conversely, it is known that epileptic seizures themselves and antiepileptic drugs lead to metabolic changes in body weight and hormonal alterations. Hence, as epilepsy and metabolism seem to be correlated, metabolites could be used as biomarkers of epilepsy. In the present study, we used amygdala-kindled mouse model, in which conscious unrestrained mice received a biphasic square wave pulse into the basolateral amygdala once a day (almost for 3 weeks). This model was established using rat first in 1969, then using canine, feline, ape, and mouse. Then, symptoms of these animals that received amygdala-kindling stimulations were as like as symptoms of human temporal lobe epilepsy (TLE). Hence, for finding biomarkers for TLE in human and companion animals, we focused on profiling urinary volatile organic compounds (VOCs) in the mouse model of TLE using gas chromatography mass spectrometry (GC-MS) and detected one hundred thirty-five metabolites including a various of chemical structures, such as aldehyde, ketones, nitrogen compounds, terpenes, acids, alcohols, benzene derivatives, furan and sulphur, and so on. Thirteen urinary VOCs exhibited differential abundance between epileptic and control mice, and the corresponding areas under the receiver operating characteristic (ROC) curve were greater than 0.8. Principal component analysis (PCA) based on these 13 VOCs separated epileptic from sham operated-mice, suggesting that all these 13 VOCs are epilepsy biomarkers. Promax rotation and dendrogram analysis concordantly separated the 13 VOCs into three groups. Stepwise linear discriminant analysis extracted methanethiol; disulfide, dimethyl; and 2-butanone as predictors. Based on known metabolic systems, the results suggest that TLE induced by amygdala stimulation could affect both endogenous metabolites and the gut flora. Finally, we provide candidates of biomarkers in TLE.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-469 Effects of the anti-migraine drug Sumatriptan on larva zebrafish seizure-like behaviour Mayeso Naomi Victoria Gwedela（Gwedela Mayeso Naomi Victoria），Hidenori Aizawa（Aizawa Hidenori） Hiroshima University, School of Biomedical and Health Sciences, Department of Neurobiology Epilepsy is one of the most common disorders of the brain and affects at least 70 million people worldwide. Comorbidity and the prophylactic effect of anti-epileptic drugs (AEDs) on migraine led to the hypothesis that a sub-population of those diseases might share pathophysiology. We therefore aimed at investigating the effects of sumatriptan (SUM), a 5-HT_1B/1D agonist used most frequently in the treatment of migraine, on seizure-like behaviour induced by pentylenetetrazole (PTZ) as a chemoconvulsant in zebrafish. Larvae were incubated in diazepam (DZP), SUM, or embryo medium before adding PTZ and video recording. PTZ-induced seizure-like behaviour ranged from thigmotaxis and rapid whirlpool-like movement to stiffening and loss of posture. Temporal analyses showed typical PTZ-induced behaviour patterns of cyclic high and low velocity episodes evident during ictal-like and post-ictal-like periods, respectively. Mean velocity, defined as the total larval displacement over the recording time, was significantly increased by PTZ. This effect of PTZ was suppressed by SUM and DZP pre-treatment, suggesting anti-seizure activity. The percentage number of frames showing larval displacement in the recording, termed % time moving, significantly increased PTZ-treated larvae relative to the control baseline levels. The PTZ group predictably spent a higher percentage of time in motion, which was suppressed by pre-treatment with SUM and DZP. Prolonged exposure to PTZ led larvae to spend longer time in the most severe seizure-like stage with loss of posture and impaired locomotor activity,as represented by the temporal loss of statistical contrast between the control and PTZ groups. Collectively, using zebrafish as a model, we found that seizure-suppressive effects of SUM were comparable to DZP, making it a candidate for a novel AED.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-470 うつ・不安症を示すST3Gal IV欠損マウスにおける尿中揮発性有機化合物の解析 Akiko Fujita（藤田明子），Takaya Okuno（奥野貴也），Mika Oda（織田美伽），Keiko Kato（加藤啓子）京都産業大生命科学 Psychiatric disorders including depression and anxiety comprise a broad range of conditions with different symptoms. We have developed a mouse model of depression/anxiety that knockout of the ST3 beta-galactoside alpha-2,3-sialyltransferase gene, St3galIV. ST3GalIV KO mice showed downregulation of growth hormone (GH) and insulin-like growth factor 1 (Igf1) mRNA in cerebral cortex (J. Neurochem. 2014, 131:675-87). Furthermore, we found changes in emotional behaviors generated by consumption of a diet containing oil differed between ST3GalIV-KO mice and WT mice (PLoS One, 2015, 10: e0120753). These data suggested that ST3GalIV was involved in lipid metabolism and neuropsychiatric disorders. We used a gas chromatography-mass spectrometry (GC-MS) to study the metabolic profiles of urinary volatile organic compounds (VOCs) from male, wild-type mice and ST3GalIV KO mice. Eighteen putative VOCs were identified as significantly changed in middle-aged mice (20-35 weeks of age). Comparative analysis between middle-aged mice and young mice (10-16 weeks of age) revealed six VOCs were increased in St3GalIV-KO mice, regardless of aging. The levels of these six VOCs (pentanoic acid, 4-methyl, ethyl ester, 3-heptanone, 6-methyl, 3-heptanone, 5-methylene/5-hepten 3-one, 5-methyl, benzaldehyde, 5,9-undecadien-2-ol, 6,10-dimethyl and unknown compound RI1237) were correlated with the startle response (r = 0.620) which was related to an auditory emotional response. Beta-farnesene and alpha-farnesene were known as pheromone and 6-methyl 3-heptanone and 5,9-undecadien-2-ol, 6,10-dimetyl were pheromone related compounds. We investigated the correlation between VOC production and social behavior (naïve male mice encountering female mice). We found that the six VOCs including 3-heptanone, 6-methyl and 5,9-undecadien-2-ol, 6,10-dimethyl were not changed, whereas levels of the two farnesenes were increased following the encounter. Additionally, the levels of nonpheromonal VOCs, including texanol, texanol isomer, and styrene, were altered regardless of St3galIV deficiency. These results indicated that VOC production was linked to the emotional reaction via different pathways. Taken together, our findings revealed that urinary VOCs were associated with emotional response and that emotional behavior could lead to changes in urinary VOCs.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-471 Distributions of epilepsy/ anxiety responsive, alpha2,3-sialyltransferase ST3Gal IV mRNA and visualization of mCherry in CRISPER/CAS mediated mCherry knock-in at St3gal4 locus of mouse Siriporn Tangsudjai（Tangsudjai Siriporn）¹，Akiko Fujita（Fujita Akiko）¹，Masahito Ikawa（Ikawa Masahito）²，Keiko Kato（Kato Keiko）¹ ¹Division of Life Sciences, Kyoto Sangyo University, Kyoto, Japan ²Research Institute for Microbial Diseases, Osaka University, Osaka, Japan Alpha 2,3 - sialyltransferase (ST3Gal4) is a sialyltransferase which generates Siaα2,3Gal linkages in the sensory relay neurons of the thalamus. In mouse, while increase of ST3beta-galactoside alpha-2,3-sialyltransferase 4 (St3gal4) is involved in development of epileptic seizures, the St3gal4 deficiency causes anxiety and depression. While the St3gal4-KO mice also decrease plasma triglyceride level, when mice fed pellets made from AIN93G powder containing 18% of a mixture of 1-palmitoyl-2-oleoyl-3-palmitoyl glycerol (POP) and 1-stearoyl-2-oleoyl-3-stearoyl glycerol (SOS), plus 2% soybean oil, St3gal4-deficient mice decrease tone fear and the littermate wild-type mice decrease context fear in delay fear conditioning test. It shows that extracranial metabolism affects emotional memory differentially between St3gal4 deficiency or not. In human, genome-wide association study has displayed the associations of the ST3GAL4 gene variant and of the single nucleotide polymorphism (SNP) with lipid trait including high ApoB level, total cholesterol, and triglyceride levels, and coronary artery disease. Hence, the involvement of ST3GAL4 with lipid metabolism might affect development of psychiatric disorders in both mice and human. To know interactions between intra- and exrtra-cranial metabolism, we investigated the localization of mRNAs encoding St3gal4 using in situ hybridization. It is known that the expression of St3gal4 is region specific and stage specific in brain. Among tissues of peripheral organs, we presently found that basal cells of stratified squamous epithelium in the esophagus, vagina, and cervix, and epithelial cells of crypts surround the basal regions of the colon and rectal, and gland cells in the salivary and preputial glands expressed St3gal4 mRNA strongly. It provides that St3gal4 expressing in the alimentary system might be involved in interactions between intra- and exrtra-cranial metabolism. Recently, we established CRISPR/Cas9 mediated knock-in of mCherry-poly A fragment at ST3Gal4 exon 3 locus in mouse. Forty-four F0 mice were born and 21 mice had mCherry-poly A on target site. Finally, we got 53 F1 knock-in mice and proceeds to generate the next generation by backcross with C57BL/6. Now, we visualize the expression of St3gal4 to find where and when St3gal4 acts on glycoprotein(s).		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-472 先天性高ビリルビン血症モデルラットの前頭葉におけるセロトニン伝達障害 Shouko Miura（三浦章子）¹，Keiko Tsuchie（土江景子）¹，Ryosuke Arauchi（荒内亮輔）¹，Tsuyoshi Miyaoka（宮岡剛）¹，Masatoshi Inagaki（稲垣正俊）¹，Arata Oh-Nishi（大西新）^2,3 ¹島根大医精神医学 ²島根大医免疫精神神経学 ³株式会社　RESVO Recent studies have shown that congenital hyeprbilirubinaemia, including neonatal Jaundice, is associated with an increased risk of developing psychiatric disorders or any psychotic symptoms in adulthood. Our previous studies have reported that the Gunn rat, which is known as an animal model of Crigler Najjar syndrome type1 , its show congenital hyeprbilirubinaemia, has abnormal behavior similar to schizophrenia and neurodevelopmental disorder. In addition, we shown that the behavior abnormalities in Gunn rats were improved by treatment of antipsychotics such as serotonin-dopamine antagonist (Tsuchie et al. 2013). It may explain the association between the abnormality of the serotonergic and/or dopaminergic functions and its abnormal behavior. Here we report the results of serotonergic dysregulation in the frontal cortex of rat model of Gunn rats. The amounts of serotonin and its metabolites in 5-hydroxyindoleacetic acid(5-HIAA) in the 4 brain regions (Frontal cortex, ACC, Striatum, hippocampus) of mature Gunn rats were measured by High performance liquid chromatography. Moreover, serotonin related behaviors were investigated. We found that there were significantly higher serotonin and 5-HIAA at the Frontal cortex and abnormal serotonin related behaviors in Gunn rats. These findings indicate that the congenital hyeprbilirubinaemia produces serotonergic dysregulation in the frontal cortex. Serotonergic function in the frontal cortex seems to play an important role in regulating psychotic state and associated behaviors. Our study suggests that congenital hyeprbilirubinaemia is one of the psychosis risk factors in the adulthood.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-473 半側空間無視患者における主観的視覚性垂直の特徴 Kimihiko Mori（森公彦）^1,2，Kae Nakamura（中村加枝）²，Shingo Hashimoto（橋本晋吾）¹，Masanori Wakida（脇田正徳）³，Kimitaka Hase（長谷公隆）⁴ ¹関西医科大学附属病院リハビリテーション科 ²関西医科大学生理学講座 ³関西医科大学香里病院リハビリテーション科 ⁴関西医科大学リハビリテーション医学講座 Background: A distorted perception of verticality affects upright posture. Such errors are represented by tilt of subjective visual perception of verticality - visual vertical (VV) and its variability. Although patients with unilateral spatial neglect (USN) with deficits of cognitive processes or attention to the contra-lesional space displayed deviation and variability of the VV, details have not been reported. Specifically, we investigated how patients with USN judge the VV subjectively. Methods: Forty-three acute stroke patients and 33 age-matched healthy subjects (Healthy) were examined. Seventeen and 26 patients were judged as patients with (USN+) and without (USN-) neglect by three psychological tests. To measure VV, the visual line, tilted to the left or right in 30° from vertical (0°), was presented on the screen in front of the subject. The line then rotated toward the vertical at the speed of 1°/ 1.2 sec. Subjects judged the line as vertical verbally. VV was the discrepancy between judged tilt and real vertical. Five trials in each direction of tilt was measured. Mean (VVm) and standard deviation as variability (VVv) of VV from 10 trials were then computed, and analyzed by a 1-way ANOVA with group (USN+, USN-, Healthy). The effect of group and the initial tilt (left or right) for 5 each trial was judged by a 2-way ANOVA. Results: VVm in USN+ often deviated to the contra-lesional direction compared with USN- (P=0.075) and Healthy (P=0.055). VVv in USN+ was significantly larger than those in USN- and Healthy (P<0.001, respectively), indicating unstable VV. Considering initial tilt, the mean of VV for 5 each trial in USN+ (contra-lesional tilt -8.9 ± 5.9°, ipsi-lesional tilt -4.7 ± 6.1°) deviated from the vertical to the direction of initial tilt than USN- (contra-lesional tilt -1.6 ± 3.0°, ipsi-lesional tilt -0.8 ± 3.7°, P<0.001) and Healthy (contra-lesional tilt -1.5 ± 1.9°, ipsi-lesional tilt -1.2 ± 2.4°, P<0.001). The standard deviation of VV for 5 trials in USN+ varied more within trials from each side than USN- and Healthy (P<0.001, respectively). Conclusions: Majority of subjects, particularly with hastiness in USN+, showed vertical estimation biased to the initial tilt. In the space without visual information, the VV in USN+ would deviate to neglected side due to spatial attention deficits, whereas unstable due to non-spatial attention deficits.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-474 ドパミンD₂受容体反復刺激による強迫性障害様症状の発現とアデノシンA_2A受容体遮断による症状改善機構 Nozomi Asaoka（浅岡希美）^1,2，Naoya Nishitani（西谷直也）¹，Haruko Kinoshita（木ノ下晴子）¹，Yuma Nagai（永井佑茉）¹，Hikari Hatakama（幡鎌輝）¹，Kazuki Nagayasu（永安一樹）¹，Hisashi Shirakawa（白川久志）¹，Takayuki Nakagawa（中川貴之）³，Chihiro Yabe-Nishimura（矢部千尋）²，Shuji Kaneko（金子周司）¹ ¹京大院薬・生体機能解析 ²京都府立医大・薬理 ³京大病院・薬剤部 Obsessive-compulsive disorder (OCD) is a neuropsychiatric disorder characterized by repeatedly rising concern (obsessions) and repetitive behaviors to get rid of obsessions (compulsions). Previously, OCD was classified as an anxiety disorder caused by serotonergic dysregulation, whereas recent clinical evidences suggest that cortico-striatal dysfunction-induced abnormal decision-making and behavioral choices are involved in the OCD symptoms. However, precise neurological mechanisms of OCD symptoms remain to be elucidated. In this study, we showed that repeated injection of a dopamine D₂ receptor agonist, quinpirole elicited two distinct OCD-related behaviors; cognitive inflexibility and repetitive behavior. Consistent with the distinct therapeutic efficacies for selective serotonin reuptake inhibitors (SSRIs), the only approved anti-OCD drugs, between subtypes of OCD symptoms, chronic SSRI administration rescued the cognitive inflexibility, while no improvement was observed for the repetitive behavior. Chronic SSRI administration also ameliorated the hyperactivity in the lateral orbitofrontal cortex (lOFC) in quinpirole-treated mice. Short-term administration of a 5-HT_2C antagonist exhibited similar effects to chronic SSRI administration. Additionally, microinjection study showed that D₂receptor and A_2A receptor signaling in the central striatum (CS) was involved in SSRI-resistant repetitive behavior. An adenosine A_2A antagonist, istradefylline rescued abnormal excitatory synaptic function in the CS indirect pathway medium spiny neurons of quinpirole-treated mice and also alleviated both of the OCD-related behaviors with only short-term administration. These results provide a new insight into neurological and therapeutic mechanisms of OCD symptoms and indicate the potential of A_2A antagonists as a rapid-acting anti-OCD drug.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-475 側頭葉てんかんモデルマウスにおける代謝変化 Yasuhiro Togawa（利川泰博），Manami Ota（太田真菜美），Akiko Fujita（藤田明子），Keiko Kato（加藤啓子）京都産業大学 Epilepsy is a chronic disorder of the brain that affects approximately 50 million people of all ages in the worldwide. Approximately 30% of people with epilepsy is under inadequate control of their seizures and refractory to treatment with drugs. One-half of patients with refractory epilepsy are characterized as having mesial temporal lobe epilepsy (TLE), and showing comorbidity such as depression, anxiety and high risk of obesity. It is known that antiepileptic drugs are associated with weight gain. Furthermore, high blood glucose concentration including diabetes decrease seizure threshold. Ketogenic diet has been used for treatment of drug-resistant epilepsy. It is suggested that sugar / lipid metabolic pathway is related with severity or improvement of epilepsy. We have used an amygdala-kindled model, in which conscious unrestrained mice received a biphasic square wave pulse once a day. The amygdala-kindled mice showed up-regulation of growth hormone (GH) along the neural circuit. It is known that GH involves the stimulation of lipolysis in the adipose tissue and has antagonism of insulin action. Therefore, we investigated body weight and tissue weights of kindling model mice and measured total triglycerides, total cholesterol, lipoproteins, and total protein, in plasma on more than 40 days after development of epilepsy, to study the relationship of epilepsy with peripheral metabolisms. Body weight, white adipose tissue and brown adipose tissue were increased in kindling mice (x1.1, p = 0.02, x3.4, p < 0.01, x3.4, p < 0.01, Mann-Whitney U-test), whereas food intake was not changed. Five species of fatty acids (myristic acid, palmitoleic acid, stearic acid, oleic acid and eicosapentaenoic acid (EPA)) from glycerides were increased in plasma following kindled-seizures in mice and showed strong correlation with body weight (Pearson's correlation coefficient: 0.7 < r < 0.9). Plasma cholesterol, total protein and lipoprotein had no difference between kindling mice and sham-operated mice. Kindling mice indicated glucose tolerance after fasting for 16 hours and increased plasma insulin levels. Taken together, kindling model mice is a useful model to elucidate the mechanism of obesity risk as a comorbidity of temporal lobe epilepsy.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-476 母体免疫活性化(MIA)のセロトニン神経形成に対する影響 Jinmin Li（李金旻）¹，Fumihiro Shutoh（首藤文洋）^1,2，Tetsuya Sasaki（佐々木哲也）^1,2，Yosuke Takei（武井陽介）^1,2 ¹筑波大院　人間総合科学感性認知脳科学 ²筑波大　医・解剖・神経科学 Serotonergic system widely project in the central nervous system. Most of their anterior fibers are from the dorsal and median raphe nuclei that pass through diencenphalon and arrive at basal nuclei and major cerebral areas. On the way, they make critical projections with nuclei that concerned to mental health, like ventral tegmental area, globus pallidus, hippocampus, olfactory nucleus and septal nuclei. Then, abnormal development of serotonergic system is considered as major reasons of autism (ASD) and other psychiatric disorders. Recently, maternal immune activation (MIA), which induced by viral infection in duration of pregnancy, can be a high-risk factor of psychiatric diseases in offspring such as ASD and schizophrenia. Serotonergic system has many important functions. A study reports that the mouse embryos which exposed immune activation on E13 showed abnormal development in serotonergic systems. In this study, we tried to examine the influences of MIA to serotonergic system in different developmental stages using MIA model mice that made with Poly(I:C) injection at different pregnant stages (E10.5, E12.5 and E16.5). The raphe nuclei development starts on E10, and serotonergic fibers enter diencephalon and frontal areas and cortices on E12-17. The offspring were sacrificed at 9 and 12 weeks old, the extracted brain samples were analyzed by immunohistochemical method. The results of observation analyses of quantity and shaping of cell and fibers reveal some changes in serotonergic neurons in MIA offspring.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-477 MONETシミュレータにおける神経回路モデル構築の並列化 Jun Igarashi（五十嵐潤）¹，Hiroshi Yamaura（山浦洋）²，Tadashi Yamazaki（山﨑匡）^1,2 ¹理化学研究所情報システム本部 ²電気通信大学大学院情報理工学研究科 Human-scale whole-brain simulations are expected to realize using supercomputers in the 2020s'. In such large-scale simulations, in addition to numerical calculations of neural network models, building the neural networks before simulation tend to take a lot of time. In the current study, we investigated an efficient way to parallelize the processing of building spiking neural networks using our in-house simulator, MONET (Millefeuille-like Organization NEural neTwork) which has been being developed for large-scale simulation of spiking neural networks with layered sheet structure, such as cortex and cerebellum. To test computational performance, we used layered sheet type of cortical and cerebellar models developed based on anatomical and electrophysiological data including information of neural density and spatial extent of connections. We used a leaky integrate-and-fire neuron model for all neuron types and conductance-based synaptic models for all synapses. We used K computer for all numerical calculations. In programming of parallel computing, we used C language, Message Passing Interface (MPI) and, Open Multi-Processing (OpenMP) libraries. We parallelized the processing of building neural networks by hybrid parallelization using a combination of MPI process-parallel and OpenMP thread parallel. In MONET simulator, each MPI process takes charges of one partitioned tile of layered sheet model and information of connections are stored on a process with a postsynaptic neuron. Most of the processing time in building networks are spent for wiring of connections between neurons. We assigned the processing for all in-degree connections to the postsynaptic neurons within one tile to MPI process responsible for the one tile. The wiring processing in one MPI process was further divided to be assigned to OpenMP threads in the one MPI process. We assigned processing of connections from one presynaptic neuron type to all postsynaptic neuron types to one OpenMP thread. The parallelization realized speeding up the processing of building networks inc cortical and cerebellar models. When we increased the total size of the network and the numbers of compute nodes together with keeping the assigned tile size per compute node, the processing time almost kept constant because of load balancing in tile partitioning. This result suggests the possibility of further scaling-up of layered sheet type of neural networks by the next-generation supercomputer.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-478 MONETシミュレータを用いたスーパーコンピュータ「京」上での大規模小脳神経回路モデルのシミュレーション Hiroshi Yamaura（山浦洋）¹，Jun Igarashi（五十嵐潤）²，Tadashi Yamazaki（山崎匡）¹ ¹電気通信大学 ²理化学研究所 Owing to the Moore's law on the performence of supercomputers, it is becoming possible to simulate a spiking neural network of the human brain on a supercomputer. However, it is difficult to harness the performance, since users need to optimize source codes to a target supercomputer. In this study, we evaluated the performance of developing neural network simulator called MONET (Mille-feuille like Organization NEural neTwork). We built a spiking neural network model of the cerebellum based on electrophysiological and anatomical data using the MONET simulator. We investigated the network dynamics of the cerebellar network model. We could reproduce the activity pattern similar to the previous simulation study. Next, we analyzed weak scaling property of the cerebellar network model. In the weak scaling, number of neurons per compute node is fixed while the number of compute nodes is varied, which lead to increasing the size of the cerebellar network model. We carried out the simulation while changing the number of compute nodes on K computer from 1024 to 82944 nodes, and measured the computational time. We obtained a good scaling property. Further, we investigated breakdown of the computational time. The computational time consists of (1) calculation of membrane potentials, (2) calculation of synaptic inputs, and (3) communication via network to exchange spike information among different computational nodes. We carried out two simulations. In one simulation, mossy fibers issue spikes in a high frequency, as if external stimulus is fed. In the other simulation, mossy fibers emit spikes in a low frequency that simulates spontaneous activity. We found that in the low frequency condition, (3) communication time was negligible. This is because the amount of spikes and so the communication among nodes was reduced substantially. These results suggest that the MONET simulator implements efficient communication mechanisms that scales well depending on the number of emitted spikes.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-479 神経回路の脱同期のための深部脳刺激の電流波形最適化 Shigeru Kubota（久保田繁）¹，Rubin E Jonathan（Jonathan E Rubin）² ¹山形大学 ²ピッツバーグ大 Deep brain stimulation (DBS) of localized brain regions with repetitive current pulses is widely used for the treatment of Parkinson's disease (PD). To prolong the battery life for the implanted stimulators, it is important to reduce the electrical energy required for DBS. A promising technique for improving the efficiency of stimulation and decrease the energy consumption is the coordinated reset (CR) neuromodulation, which acts to desynchronize network by the delivery of spatially distributed currents using multiple electrodes. In this study, we propose and apply a new optimization method for improving the energy efficiency of CR stimulation to achieve desynchronized state. We construct a network model composed of leaky integrate-and-fire neurons with multiple electrodes, to simulate the network dynamics in the presence of CR stimulation. We describe the waveform of electrode currents using Fourier series and conduct the optimization of the Fourier coefficients as well as the period, to minimize the energy necessary for decreasing the synchronization level. In our method, the optimized stimulation pattern is directly searched for by modulating the current waveform and evaluating the change in the order parameter, which quantifies the degree of synchrony. This direct optimization framework has a remarkable advantage that basically arbitrary dynamical changes in the network, which could affect the effectiveness of DBS, can be taken into account in the optimization search. The simulation results show that the energy consumed by the proposed method is considerably lower than that required for the conventional method. In addition, we examine the effects of externally applied inputs and non-random network topology (small-world topology) on the energy efficiency of the proposed stimulation technique.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-480 脳波による手の把握動作の識別 Koki Okada（岡田幸樹）¹，Natsue Yoshimura（吉村奈津江）^1,2，Yasuharu Koike（小池康晴）^1,2 ¹東工大院情報通信 ²東工大科学技術創成研究院 Brain Machine Interface (BMI) is the system which controls a robot or a computer by human brain activity. BMI using electroencephalogram (EEG) has been researched for many years, because the electroencephalograph is portable and non-invasive. Current EEG-based BMI controls computer by distinguishing the brain site which occurs Event Related Desynchronization (ERD) during motor imagery. One of these researches controls a target in the display. The user can moves the target up and down when he/she imagines right and left hand movement relatively. This method can decode the imagined movements with high accuracy, if the sites are far away each other like right and left hand motor imagery. However, since imagined movements don't map to the command to perceive, the system is not intuitive. Additionally, it is difficult to decode if the active sites are near. Our research purpose is to make a decoder which distinguishes hand grasping and opening. During hand grasping and opening, brain active sites are so near that it is difficult to decode using ERD. To solve this problem, we use Variational Bayesian Multimodal EncephaloGraphy (VBMEG). VBMEG is one of the current source estimation methods. This method estimates high spatial resolution current source signals from EEG using pre-defined brain activity measured by fMRI experiment. Estimated current source signals are used to make a decoder. Sparse Logistic Regression are used as a decoder and leave-one-out cross validation are used to calculate its accuracy. We expect this research is applicable to artificial arm which moves intuitively.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-481 捕食者被食者系における攻撃／防御行動形質の共進化：分断選択と表現型可塑性の個体ベースモデル Hiroyuki Ichijo（一條裕之）¹，Tomoya Nakamura（中村友也）¹，Masato Yamamichi（山道真人）² ¹富山大医解剖 ²東京大学大学院総合文化研究科広域システム科学系 Innate behaviors are genetically instructed, and they are differential phenotypes in the same individual that respond to environmental changes without changing genotype; thus, innate behaviors are phenotypically plastic traits and their neuronal circuits are regarded as genetic machineries for phenotypic plasticity. Defenses: flight and freeze, are well-known innate behaviors. It is hypothesized that selection pressure arising from predation promotes evolution of neuronal circuits for defense behaviors of preys against predators. Its evolution modifies the predator-prey interaction, which further changes the selection pressure for evolution of behaviors not only in preys but also in predators. Here we investigated eco-evolutionary feedbacks to understand coevolution of behaviors between predator and prey species in an individual-based model (IBM). In the IBM, it is assumed that agents randomly move in a constant speed (v) in the two-dimensional space, and that a predator eats a prey when they are in the same place. Each genotype consists of plasticity-sensitivity (ps) and plasticity-potential (pp) to represent phenotypic plasticity; ps affects sensitivity to environment, and pp determines the velocity of agents after the encounter. Agents detect interacting agents in an area with radius ps and change their velocity from v to v + pp although ps and pp are initially set at zero. They reproduce asexually with random mutations, and there is a cost of phenotypic plasticity. The IBM was implemented in NetLogo (version 6.02) and its results were analyzed by R (version 3.5.1). With the predator-prey interaction, ps and pp coevolved in both predator and prey species. Moreover, disruptive selection promoted bifurcation of pp into two discrete phenotypes in both species: increasing and decreasing velocity after the encounter. They corresponded to behaviors of attacks in the predator (chase/waylay) and defenses in the prey (flight/freeze). The results indicate that behaviors of attacks and defenses can emerge with simple predator-prey interactions. Pairs of behaviors in attacks (chase and waylay) or defenses (flight and freeze) stem from the same origin. The disruptive selection is likely because the minorities in both species have more chances to survive and reproduce with higher degree of fitness.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-482 Bayesian regularization of undersampled cortical dynamics during natural behaviour Dmitry Lyamzin（Lyamzin Dmitry），Mohammad Abdolrahmani（Abdolrahmani Mohammad），Ryo Aoki（Aoki Ryo），Andrea Benucci（Benucci Andrea） RIKEN During behaviour, the brain receives neural signals that convey information about body movements, which allows animals to distinguish between self-generated and environment-generated sensory stimuli. The effects of these neural signals on sensory processing have been studied extensively, but largely involved individual limb movements in isolation - a condition that is behaviourally unnatural and gives an overly simplistic understanding. One of the reasons for this reductionist approach is the high-dimensionality of body movement data, which prohibits an extensive sampling of neural activity for the full parameter space. Here, we propose a generalized linear model with Bayesian regularization to overcome these limitations and analyse neural activity in the occipital cortex of the mouse brain originating from diverse near simultaneous movements while an animal is engaged in a visual discrimination task. Using the model we increase the interpretability of found response characteristics, specifically in conditions when several movements occur near simultaneously, and where the parameter space is comparatively undersampled. We found nonlinear interaction properties that depend on the sequence of the occurring movements. Our work shows the utility of Bayesian regularization methods for the research of movement-related neural signals during natural behaviour, and demonstrates their complex nonlinear interactions that have the potential to affect visual processing and perceptual decision-making.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-483 膜電位時系列データに基づくHodgkin-Huxley型Regular-Spikingニューロンモデルのパラメータ最適化 Daiki Futagi（二木大樹）¹，Ryota Kobayashi（小林亮太）²，Katsunori Kitano（北野勝則）³ ¹立命館大立命館グローバル・イノベーション研究機構 ²国立情報学研究所情報学プリンシプル研究系 ³立命館大情報理工 In order to reveal nervous systems, it is effective to analyze mathematical models of neurons and understand neuronal properties quantitatively. Hodgkin-Huxley (HH) equation is often used as physiologically reasonable model of neuron. This is because HH-type model could describe various firing properties based on ion channel gating kinetics. However, HH-type model often includes many parameters that are not independent of each other and determine the model behavior. Therefore, it is necessary to optimize parameter values for fitting the model behavior to experimental data and knowledge. A simple method for model parameter optimization is to evaluate all possible combinations of values for all parameters. However, the exhaustive method does not often work out in practice. In this study, we propose a hybrid method for model parameter optimization that is designed to complement population-based and gradient-brad algorithms. In order to assess efficiency of the proposed method, we conducted numerical experiments that fit HH-based model of regular spiking neuron to multiple sets of artificial membrane potential data. The neuron model consists 5 types of ion channels: voltage-dependent sodium, potassium and calcium channels, calcium-dependent potassium channel and leakage channel. The optimized parameters (39 in total) are the follows: maximum conductance and kinetic parameters of the ion channels, decay time constant of calcium transient, etc. By using the neuron model, we simulated current clamp and generated time-series data of membrane potentials. In the optimization task, the artificial data is used to resemble actual experiment data for parameter tuning, and we confirmed how the proposed method could search for the parameter values used for the artificial data generation. As a result, the proposed method could search for optimal values of the model parameters that are not described in differential equation for membrane potential. In addition, the proposed method shows higher performance than the non-hybrid methods that use the population-based and gradient-based algorithms separately. Although the proposed method is based on the simple algorithms, it would be useful in terms of efficiency and accuracy.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-484 NPY-Creマウスを利用した側坐核NPYニューロンの形態学的解析 Shunji Yamada（山田俊児）¹，Nienke Van Kooten（Van Kooten Nienke）^1,2，Katsutoshi Taguchi（田口勝敏）¹，Takuma Mori（森琢磨）³，Atsushi Tsujimura（辻村敦）⁴，Masaki Tanaka（田中雅樹）¹ ¹京都府立医科大学　大学院医学研究科　生体構造科学 ²School for Mental Health and Neuroscience (MHeNS), Maastricht Univ, Maastricht, Netherlands ³信州大学　医学部　分子細胞生理学 ⁴京都府立医科大学　大学院医学研究科　基礎老化学 Neuropeptide Y (NPY) is a neural peptide distributed widely in the brain and has various functions in each region. We previously analyzed mice expressing an unedited RNA isoform of 5-HT_2C receptor. The mice showed behavioral despair and decreased NPY gene expression in the nucleus accumbens (NAc). This result implies that NAc-NPY neurons are involved in emotional behavior. Moreover, overexpression of NPY, but not injection of NPY or NPY receptor agonist, in the NAc ameliorated behavioral despair, suggesting that the NPY neurons are projecting neurons. Obtaining more knowledge about NAc-NPY neurons, in the present study, we investigated neural output and input of the NAc-NPY neurons using NPY-Cre mice. To investigate projection of NAc-NPY neurons, we injected AAV-FLEX-mCherry into the NAc in NPY-Cre mice. We found many mCherry-positive fibers in the lateral hypothalamus (LH), and paraventricular nucleus of the thalamus (PVT). Next, we investigated neural input to the NAc-NPY neurons using a first-infected cell specific monosynaptic retrograde tracing. Two cre-dependent AAVs expressing TVA-mCherry and glycoprotein (G) were injected into the NAc in NPY-Cre mice. TVA and G are required for infection of genetically-modified rabies virus and subsequent transsynaptic spread, respectively. Two weeks after AAVs injection, we injected EnvA-enveloped HEPdG-GFP (High egg passage-fluey (HEP) strain rabies virus possessing GFP gene instead of gene for G) into the same region and examined the brain one week later. We detected GFP and mCherry double positive cells only in the NAc, probably showing first-infected NPY neurons. We also found that GFP- but not mCherry-positive cells in the PVT, LH, the bed nucleus of the stria terminalis, and midbrain of lateral dorsal tegmental nucleus. These results indicate a possibility that a part of NAc-NPY neurons project to the LH and PVT and the neurons are reciprocally regulated by the neurons in these nuclei.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-485 光遺伝学によるコモン・マーモセット前肢運動の誘発 Teppei Ebina（蝦名鉄平）¹，Keitaro Obara（小原慶太朗）¹，Yoshito Masamizu（正水芳人）^1,2，Shin-Ichiro Terada（寺田晋一郎）¹，Akiya Watakabe（渡我部昭哉）³，Ryota Matoba（的場涼太）¹，Nobuhiko Hatanaka（畑中伸彦）⁴，Atsushi Nambu（南部篤）⁴，Hiroaki Mizukami（水上浩明）⁵，Tetsuo Yamamori（山森哲雄）³，Masanori Matsuzaki（松崎政紀）^1,6,7 ¹東京大学大学院医学系研究科　機能生物学専攻　細胞分子生理学 ²JSTさきがけ ³理化学研究所　脳神経科学研究センター　高次機能分子解析チーム ⁴自然科学研究機構　生理学研究所生体システム研究部門 ⁵自治医科大学　分子病態治療研究センター　遺伝子治療研究部 ⁶東京大学　国際高等研究所　ニューロインテリジェンス国際研究機構 ⁷理化学研究所　脳神経科学研究センター　脳機能動態学連携研究チーム Optogenetics has been a fundamental tool to reveal the relationship between neuronal activity and behavior. However, unlike other small model animals, optogenetic stimulation of cortical neurons in non-human primates failed to induce or modulate any upper-limb movements. This may be because the number of photoactivated neurons in the non-human primate might not reach the sufficient level to elicit hand/arm movements. In this study, to solve this problem, we used an adeno associated virus with a tetracycline-inducible gene expression system, which amplifies neuronal expression of induced genes. Using this technique, we enhanced channelrhodopsin-2 expression in excitatory neurons in the motor cortex of common marmosets. Marmoset has the smaller motor cortex than macaque and squirrel monkeys, so it would not be difficult to photostimulate a relatively large proportion of motor cortical neurons dominating the hand/arm movements. In an awake state, blue laser photostimulation through the cranial glass window induced stable upper-limb movements with earlier responses of upper-limb muscles. Long-duration photostimulation induced discrete upper-limb movements, which could be markerlessly tracked with a deep-learning algorithm. During performance of a upper-limb movement task, weak photostimulation around the onset of task-relevant movement modified movement trajectories. Thus, in the marmoset, optogenetic stimulation of cortical neurons successfully induces and modulates upper-limb movements.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-486 単発経頭蓋磁気刺激が一次運動野局所神経活動に与える影響と運動誘発電位との関係 - サルにおける硬膜下皮質表面電位と運動誘発電位の同時計測による検討 Yasutaka Honda（本田保貴）¹，Shinya Nakamura（中村晋也）¹，Kentaro Ogawa（小川顕太郎）¹，Toshio Iijima（飯島敏夫）¹，Yukio Nishimura（西村幸夫）²，Ken-Ichiro Tsutsui（筒井健一郎）¹ ¹東北大院生命科学脳神経システム ²東京都医学総合研脳機能再建プロジェクト Transcranial magnetic stimulation (TMS) is a widely used, non-invasive method for stimulating the nervous system. It has been reported that TMS is effective treatment for various types of psychiatric and neurological disorders, such as major depression, obsessive compulsive disorder, Tourette syndrome, and schizophrenia. However, its mechanisms of effect are still not well understood. In order to investigate how TMS affects the local neural activity, we recorded electrocorticogram (ECoG) in monkeys while single-pulse TMS was applied to the primary motor cortex (MI). We implanted an ECoG electrode array sab-durally over the MI cortex of a Japanese monkey. We then simultaneously recorded ECoG and motor evoked potential (MEP) induced in abductor pollicis brevis muscle contralateral to the stimulated hemisphere. TMS was given with a figure-of-eight coil with single biphasic pulses every minute and an intensity of 100% of the resting motor threshold. (The motor threshold was defined as the minimal stimulation intensity which elicit MEPs in at least 50% of trials.) We observed a positive wave with a peak around 5~7 ms after a TMS pulse. The occurrence/absence of MEP coincided completely with the occurrence/absence of this positive wave. Furthermore, the amplitude of this positive wave was significantly correlated with that of the MEP evoked. Previous human studies report that the single-pulse TMS elicits a corticospinal positive wave, recorded by an epidural electrode placed over the high cervical cord, around 5 ms after TMS. Thus, we infer that the positive wave with a peak around 5~7 ms after a TMS pulse observed in this study is likely to be the population firing of the pyramidal neurons of layer V.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-487 電気的外部ノイズ下における培養神経回路の活動パターン Kei Wakimura（脇村桂）¹，Hideaki Yamamoto（山本英明）²，Ayumi Hirano-Iwata（平野愛弓）^1,2 ¹東北大学　電気通信研究所 ²東北大学　材料科学高等研究所 At network-level neuroscience, a large gap separates the in vivo and in vitro systems, which is, e.g., apparent in spontaneous activity patterns. Previously, we focused on the difference in cellular connectivity between in vivo and in vitro cortical networks and showed that the globally synchronized bursting activity that defines the spontaneous activity of in vitro networks can be suppressed by controlling network structure by micropatterning. This was effectivity achieved by inducing modular organization, that characterizes cortical connectivity to in vitro networks [Yamamoto et al., Sci. Adv. 4, eaau4914 (2018)]. However, neural correlation was still excessive in the in vitro cortical networks. Recently, an in vivo study showed that deep anesthesia, that reduce thalamocortical input, increases synchrony in the spontaneous activity in mouse cortex. As in vitro cortical cultures are devoid of external noise sources, this led us to hypothesize that the excessive synchrony should be further reduced by applying external noise. Here, we investigated how external noise applied from two platinum wires positioned in the culture chamber modulates the degree of synchrony in the spontaneous activity of cultured cortical networks. Primary neurons were obtained from embryonic rat cortices at day 18 and cultured on a poly-D-lysine coated coverslip. Neuronal activity was measured by fluorescence calcium imaging. A gaussian white noise was generated using a waveform generator. We found that application of external gaussian noise to cultured neuronal networks reduced the occurrence of epileptic network bursts and the fraction of neurons involved in the bursting events. The response could be confirmed within 5 min after the noise application. Analyses of spontaneous activity under noise application in modular neuronal networks will also be reported and compared with random networks to address the synergetic effect of connectivity and noise in regulating the network activity.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-488 ドーパミン検出用マイクロダイアリシスと組み合わせたLED光刺激デバイス Yasumi Ohta（太田安美），Mamiko Kawahara（河原麻実子），Mark Christian Guinto（Guinto Christian Mark），Tasuku Kawasaki（川崎祐久），Makito Haruta（春田牧人），Kiyotaka Sasagawa（笹川清隆），Takashi Tokuda（徳田崇），Jun Ohta（太田淳）奈良先端科学技術大学院 Optogenetics has been remarkably developed and contributes to the variety fields of neuroscience. We have developed a new device that can simultaneously perform optical stimulation by micro LEDs and microdialysis. It is important to actually detect the release of neurotransmitters, especially when neural pathways and networks are unknown. For the following two reasons we use LEDs for optical stimulation instead of an optical fiber, which is conventionally used in optogenetics. The first reason is that a micro LED has a wider light output distribution than that of an optical fiber and thus can stimulate a wider range of cells. The second reason is that a micro LED is compact and lightweight so that arbitrary light stimulation pattern can be formed by arranging them. We have developed four types of devices utilizing this advantage of a micro LED. The first device comprises a ring-shaped optical pattern where four micro LEDs are arranged so as to produce a uniform light output and a hole for microdialysis probe insertion placed in the center of the ring. This type was used with placing on the rodent's brain surface. The second device arranged four micro LEDs linearly along the dialysis probe and was used to implant into the rodent's brain. The third device was developed to be placed on the brain surface of a monkey so as to stimulate a part of the cerebral cortex. It was equipped with 48 micro LEDs in the range of 6.5 mm × 9 mm and made nine holes for inserting the dialysis probe between the LEDs so as to be able to measure the detailed part of neurotransmitter release. The last type was developed to photo-stimulate the deep areas of the brain such as VTA of monkeys. Actually, using the animals expressing ChR2 or ChrimsonR in nerves, dopamine concentration changes and distribution of electroencephalograms depending on the positions in the prefrontal cortex of a monkey was confirmed in response to optical stimulation by the micro LED-based devices. We also evaluated the device by means of temperature measurement inside brain during device working, power measurement and light distribution simulation. Currently we are developing a system that can study the neural activity in more detail by using it together with our developed brain-implanted imaging device.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-489 精密CLEM~エポン樹脂包埋薄切を用いて Isei Tanida（谷田以誠）¹，Souichiro Kakuta（角田宗一郎）^2,3，Yasuo Uchiyama（内山安男）² ¹順天堂大医神経生物 ²順天堂大院医神経疾患 ³順天堂大院医形態イメージ Correlative light-electron microscopy (CLEM) is a combination of fluorescent microscopy and electron microscopy. In classic CLEM, fluorescent images of cells are obtained after chemical fixation using either paraformaldehyde, glutaraldehyde, or a mixture of the two. Thereafter, the cells are post-fixed with osmium tetroxide, dehydrated, and embedded in epoxy resins. Thin sections of the samples are prepared and observed using transmission electron microscopy. The fluorescent image is then computationally correlated with an electron microscopic image, because its morphology is chemically and mechanically distorted by the post-fixation, dehydration, and sectioning of the samples. This indicates the presence of an unavoidable limitation of the correlation in classic CLEM. One of the best ways to overcome this limitation is to obtain both fluorescent and electron microscopic images from the same section following the embedding in Epon resins. However, the fluorescent intensity of most fluorescent proteins is significantly weakened during post-fixation with osmium tetroxide, but this post-fixation is essential in order to preserve the membranous structures. We found that the fluorescence of CLEM-Red, a variant of Katushka, had improved with only a one-step treatment from a TUK solution after the treatment with osmium tetroxide. To investigate whether this recovery is applied to a CLEM on the same Epon-embedded sections, we expressed a mitochondria-targeting CLEM-Red protein in COS1 cells. The fluorescent image of mitochondria-targeting CLEM-Red protein in the Epon-embedded sections showed mitochondrial localization, and an electron microscopic image of the same sections showed representative mitochondrial structures. These results indicated that this method led to a real correlation of light-electron microscopy with only a minimal modification of the protocol for preparation of the biological sample of TEM.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-490 マーモセット前頭前皮質の結合マッピング Akiya Watakabe（渡我部昭哉）¹，Henrik Skibbe（Skibbe Henrik）²，Ken Nakae（中江健）²，Jian Wang（Wang Jian）¹，Masafumi Takaji（高司雅史）¹，Hiroaki Mizukami（水上浩明）³，Alexander Woodward（Woodward Alexander）⁴，Rui Gong（Gong Rui）⁴，Yoko Yamaguchi（山口陽子）⁴，Junichi Hata（畑純一）⁵，Hideyuki Okano（岡野栄之）⁵，Shin Ishii（石井信）²，Tetsuo Yamamori（山森哲雄）¹ ¹理研CBS 高次脳機能分子解析 ²京都大学論理生命学 ³自治医科大学 ⁴理研CBS神経情報基盤センター ⁵理研CBS マーモセット神経構造 The prefrontal cortex (PFC) of primates consists of orbitomedial areas that has homology with rodents as well as of dorsolateral areas that lack apparent homology. Whereas the former has close ties with ""limbic"" system that is associated with emotional control, the latter has extensive cortical connectivity. It is anticipated that the network of such conserved and specialized brain areas forms the basis of high cognitive functions of primate PFC. The aim of the current study is to elucidate such a network by ""mesoscale connectomic"" mapping. For this purpose, we implemented the Serial Two photon tomography technique (Ragan 2012) for marmoset brains, which allows detection of axonal signals in an accurate 3D coordinate. We are now finalizing our informatics pipeline, which automatically stitches image data to section series, registrates each sample brain to the standard coordinates, and segments the tracer signals out of the tissue background for quantitative analyses. We have so far finished injections into the dorsolateral PFC (A9, A8aV, A8aD, A46) and continuing injections into the orbitomedial PFC (A32, A14, A25, A11, A13 etc). These PFC areas have tight connections with the parietal, temporal and cingular areas, while bypassing the motor and auditory areas. The topological and modular nature of connectivity is being analyzed. At a finer level of analysis, PFC projections often converged in columns of approximately 300 μm. Interestingly, when we co-injected retrograde and anterograde tracers, we generally observed the convergence of axonal and cell signals at the same columns, suggesting reciprocal connection at the columnar level. In some notable exceptions, however, the retrograde cells spread wider, suggesting directional connectivity. We are also analyzing the topological connections of PFC areas to the subcortical regions, such as thalamus, striatum, superior colliculus and amygdala. We are hoping that such detailed mapping will provide insights to the elucidation of functional architecture of the PFC network.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-491 成獣ラット脳におけるBcas1遺伝子発現細胞の細胞内DiI染色による形態的研究 Masahiko Kase（加瀬政彦），Tetsuo Sugimoto（杉本哲夫）関西医科大学　解剖学 Bcas1 is a tumor-associated gene expressed at high levels in human breast cancers but, also expressed in the normal brain. The morphology of Bcas1 gene-expressing cells were investigated through in situ hybridization (ISH) in the adult normal rat brain. ISH revealed that these cells were scattered throughout the brain and that the cell bodies were similarly round or oval in shape. Co-existence with several neuronal or glial cell marker proteins in these Bcas1 ISH-labeled cells were searched but could not be found through the double staining of ISH for Bcas1 mRNA and immunohistochemistry (IHC) for those marker proteins. After ISH, additional intracellular DiI staining for Bcas1 ISH-labeled cells was performed using lightly fixed (1.5% paraformaldehyde in 0.1 M phosphate buffer by transcardial perfusion) and thick sections. Fluorescent dye DiI was injected into Bcas1 ISH-labeled cells through glass micro pipette by natural diffusion. Intracellular DiI staining for Bcas1 ISH-labeled cells enabled us to see their processes, which exhibited some characteristic morphological forms. One of these was a thick process extending tangentially away from the cell body surface. Another was a bundle of short linear fibers extending conically from one site of the cell body. In addition to it, some other findings were observed. On the basis of ISH and DiI staining following ISH, we presumed that Bcas1 ISH-labeled cells might represent a population of morphologically immature cells in the adult rat brain.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-492 線虫の全頭部神経の活動イメージングにより明らかにする神経ネットワークの動態 Yuichi Iino（飯野雄一）¹，Yu Toyoshima（豊島有）¹，Hirofumi Sato（佐藤博文）¹，Manami Kanamori（金森真奈美）¹，Stephen Wu（Wu Stephen）²，Moon-Sun Jang（Jang Moon-Sun）¹，Yuko Murakami（村上悠子）³，Suzu Oe（大江紗）³，Terumasa Tokunaga（徳永旭将）⁴，Osamu Hirose（広瀬修）⁵，Sayuri Kuge（久下小百合）³，Takayuki Teramoto（寺本孝行）³，Yuishi Iwasaki（岩崎唯史）⁶，Ryo Yoshida（吉田亮）²，Takeshi Ishihara（石原健）³ ¹東京大院理生物科学 ²情報・システム研究機構統計数理研究所 ³九州大理生物 ⁴九工大情報工学研究院システム創成情報工学研究系 ⁵金沢大理工研究域生命理工学系 ⁶茨城大工学部機械システム工学科 The neural network is the basis of information processing and generation of behaviors. How information flows through the network in a stereotyped, plastic or probabilistic manner is a central question in neuroscience. To answer these questions by observing the real nervous system, we chose the model organism Caenorhabditis elegans, whose nervous system consists of exactly 302 individually named neurons. We developed a 4D imaging system to observe the activity of all head neurons at the same time. Fluorescent calcium indicators were expressed in the nuclei of all neurons, along with three other fluorescent proteins to facilitate annotation of neurons (naming each neuron). We developed algorithms to precisely detect each neuron in the tightly packed head ganglia and track them through time series 4D images. Annotation of neurons was performed by experienced annotators with a help of automatic annotation algorithm that we have developed. Animals were trapped in a small channel in a microfluidic device and observed by 4D imaging system while being stimulated by concentration changes of NaCl, which in the natural environment causes behavioral responses of C. elegans that causes chemotaxis to either higher or lower concentration of NaCl. Obtained time-series data of neuronal activities show various activity patterns, most of which are not directly correlated with the sensory input. Two groups of neurons were regularly found whose activities were highly correlated within each group, while anti-correlated between groups. These groups included forward- and backward-locomotion related neurons and therefore likely correspond to this type of behavioral switch. However, member neurons included in each group seemed to differ between individuals. Independent component analysis of all neurons revealed component signals, some of which corresponded to the putative forward-backward behavior and others corresponded to the NaCl sensory stimuli, with other activities whose identities are unknown. Coss-embedding analysis also revealed that some of the neurons are related to the sensory input and others to motor output. These analyses, along with Granger causality and partial least square regression, revealed putative information flow, and revealed some neurons that seemed to combine sensory and motor activities. Through these techniques we hope to understand how behavior is generated by an interaction of sensory inputs and spontaneous neuronal activities.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-493 ドレブリンナノボディの開発 Hiroyuki Yamazaki（山崎博幸）¹，Kenji Hanamura（花村健次）¹，Narutoshi Tsukahara（塚原成俊）²，Banri Segawa（瀬川万里）¹，Akikazu Murakami（村上明一）²，Tomoaki Shirao（白尾智明）¹ ¹群馬大学大学院医学系研究科　神経薬理学 ²琉球大学大学院　医学研究科　寄生虫・免疫病因病態学 Drebrin functions as an F-actin stabilizing factor localized at the cell-cell communication subcelluar regions of several cell types. In matured neurons, drebrin is concentrated in dendritic spines, where it regulates spine morphogenesis and is involved in synaptic plasticity. Drebrin decreases not only in the brain of Alzheimer's disease but also mild cognitive impairment. Moreover, drebrin is involved in cancer metastasis, immune response and stress-resistance to oxidation. In the course of drebrin research, a variety of monoclonal and polyclonal antibodies have been developed from over 30 years ago, including M2F6 clone, and they can be used for various assays such as immunostaining, enzyme-linked immunosorbent assay, immunoblotting and immunoprecipitation. On the other hand, GFP-fused drebrin (GFP-drebrin) has been used for several studies, which is very useful for real-time imaging such as fluorescent recovery after photobleaching. However, there are some problems in use of GFP-drebrin. For example, it is difficult to control the expression level in the transfection assay. When GFP-drebrin is expressed at higher level in neurons, the spines and filopodia abnormally change their shapes by F-actin-remodeling activity of drebrin. Furthermore, the observation relating to GFP-drebrin, which is not identical with native drebrin, may be at risk of experimental artifacts. To address these issues, we tried to make a novel tool for drebrin-imaging from camelidae antibody that consists of a single monomeric variable antibody domain. In this study, we isolated some "variable domains of heavy chains of camel heavy-chain antibody " (VHH) with reactivity to drebrin by affinity selection from VHH domain-phage library. All VHH clones recognized exogenous and endogenous drebrin in western blotting. One of drebrin-VHH clone (3E3) showed a similar immunoreactivity to a typical drebrin monoclonal-antibody (clone M2F6) in the immunocytochemistry using cultured neurons. Next, we fused the VHH to "IgG-Fc with GFP " in order to observe the endogenous drebrin in living cells. When GFP-3E3 expression vector was transfected to cultured neurons, the GFP signals were visualized mainly in dendritic spines, indicating that the GFP-3E3 nanobody binds to endogenous drebrin in the neurons. We are going to test if the nanobody has some toxicity to cells during long expression period in vitro and in vivo.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-494 電解銅箔テープを使ったATUM-SEM大規模高解像度脳組織電子顕微鏡画像 Yoshiyuki Kubota（窪田芳之）^1,2，Yasuo Kawaguchi（川口泰雄）^1,2 ¹生理研大脳皮質機能大脳神経回路論 ²総研大院生命科学生理 The electron microscopy (EM)-based reconstruction of neuronal circuits from serial ultrathin sections has attracted considerable recent attention, because EM is a reliable method to identify synaptic contacts in brain microcircuits. Until recently, the electron microscopic analysis of a neural circuit was conducted manually, requiring a huge amount of labour. However, in the past decade, new automated/semi-automated systems for the acquisition of serial electron micrographs have been developed and adopted for neuroscience research, in parallel with rapid technological innovations. In particular, the automated tape-collecting ultramicrotome (ATUM) - scanning electron microscope (SEM) system has several valuable features; for example, the same serial ultrathin sections can be observed multiple times and the imaging area can be very large, including entire cortical layers. The ATUM-SEM method improves current research approaches using the electron microscope to analyse the functional architecture of brain microcircuits. However, the serious problem of finding a suitable section-collecting tape for this approach remained. We recently introduced carbon nanotube (CNT) polyethylene terephthalate (PET) tape for the ATUM-SEM (Nature Communications 2018, 9: 437). It is optimal due to its extremely high surface conductivity and low endogenous signal, and it can provide high quality images of tissue sections with SEM. However, it is not commercially available yet and is expensive. We therefore now propose an alternative optimal tape, an electrodeposited copper foil (EDCF) tape which has an extremely high surface conductivity and a very smooth surface, and most importantly the price is about 5-10 times cheaper than the CNT-PET tape. Electron micrographs of brain tissue on the EDCF tapes captured with a SEM are comparable with those on the CNT-PET tape with low acceleration voltage < 2.5 keV. At higher acceleration voltage endogenous signal from the EDCF interferes with the tissue image and consequently the signal-to-noise ratio gets worse. Recent FE-SEM models can overcome this. We confirmed that the new machines could capture excellent electron micrographs of brain tissue on EDCF tape. Thus EDCF tape can be used as an alternative optimal tape for the ATUM-SEM.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-495 生体埋植蛍光イメージングデバイスの高性能化 Kiyotaka Sasagawa（笹川清隆），Yasumi Ohta（太田安美），Mamiko Kawahara（河原麻実子），Erus Rasutami（Rustami Erus），Makito Haruta（春田牧人），Takashi Tokuda（徳田崇），Jun Ohta（太田淳）奈良先端大物質創成 Implantable image sensor devices are very small imager by using CMOS image sensor technology. With its small dimensions and low weight, it can be inserted into a living tissue with very low invasiveness. Thus, this technique is suitable for observation of deep area and long-term in-vivo experiments. The implantable imaging device has lensless setup in order to reduce invasiveness as long as possible. However, it was difficult to achieve high excitation reduction. In this study, we developed a high-performance emission filter that can be applied to an implantable image sensor. The filter is composed of interference and absorption filters. This composite filter shows excitation transmission of approximately 10^-8 in a lensless imaging device (Sasagawa, K. et al., Biomed. Opt. Express 9, 4329, 2018). However, in our previous work, the filter was fabricated on a thick fiber optic plate, which is a plate of optical fiber bundle. Thus, it was not able to use in implantable sensors. To solve this problem, we adopt a filter transfer technique using laser removing process. In this technique, an interference filter deposited on a fused silica substrate is removed by irradiating UV laser pulses (266 nm) from the backside. Thus, the composite filter of bandpass interference and absorption filter was successfully fabricated on a small image sensor chip. The proposed technique improves fluorescence imaging performance of implantable image sensor. By the combination of thin excitation light source, in-vivo fluorescent imaging of weak fluorescent signal would be realized.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-496 淡蒼球内節・外節を分ける内側随板の自動検出に向けた拡散MRI指標の検討 Shiho Okuhata（奥畑志帆）¹，Ryusuke Nakai（中井隆介）²，Tetsuo Kobayashi（小林哲生）¹ ¹京都大院工 ²京都大こころの未来研究セ Introduction: The significance of advanced analysis of basal ganglia based on MRI has been pointed out in recent clinical neuroimaging research. Automatic segmentation algorithms such as FSL-FIRST enable us to obtain desired subcortical ROIs based on structural MRIs (sMRI). Major challenge of the existing methods is to separate complex subdivisions such as internal and external globus pallidus (GPi and GPe). Previous study reported the parameter-optimized sMRIs gave a sufficient contrast to detect the medial medullary lamina (MML) of globus pallidus, a white matter structure that divides GPi and GPe. However, it is preferable to use MRI data acquired within the conventional clinical protocol, considering participants' physical and psychological loads in clinical settings. Present study aims at investigating the feasibility of fiber orientation distribution (FOD) derived from diffusion MRI (dMRI) as an effective index for detecting MML. Methods: 3T-MR scanner (SIEMENS, Verio) was used to acquire sMRIs and dMRIs from 8 healthy adults (mean age 25.37 ± 5.65, 6 male). sMRIs (T1 MPRAGE and FGATIR) were acquired with voxel size of 0.5 × 0.5 × 1.0 mm. The dMRIs were acquired with the voxel size of 2.0 mm^3, b value of 1000/ 2000 s/mm², and MPG of 30/ 64. Firstly, automatic subcortical segmentation was done by FSL-FIRST. Secondly, GP and MML ROIs were manually determined by superimposing FIRST result onto FGATIR image. Thirdly, after preprocessing, dMRIs were resliced to match the voxel size of sMRIs. The FOD was obtained from multi-shell dMRIs. The 1st peak vector and its amplitude were obtained from FOD of each voxel. The distribution of the angle of the 1st peak vector and the amplitude sampled from GP and MML were analyzed with the Kolmogorov-Smirnov test. Results and discussion: Distribution of FOD amplitudes of x and y components of the 1st FOD peak were significantly different between GP and MML ROIs (p<0.01), indicating the applicability of the FOD amplitudes for segmentation of GPi and GPe by detecting MML. Based on the result, we aim at development of the automatic segmentation method based on the indices derived from dMRIs acquired with the conventional clinical protocol. Acknowledgement: This work was supported by AMED (JPdm0207009) and by a Grant-in-Aid for Research (15H01813, MEXT), Japan. This study was conducted using the MRI scanner and related facilities of Kokoro Research Center, Kyoto University.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-497 イオン性高浸透圧造影剤を用いた造影CTによるラット・マウス脳のex vivo短時間撮像 Jun Ogura-Aoki（小倉-青木淳）¹，Miho Ota（太田深秀）¹，Susumu Jitsuki（實木亨）²，Aoi Jitsuki（實木葵）²，Takuya Takahashi（高橋琢哉）²，Hiroshi Kunugi（功刀浩）¹ ¹国立精神・神経セ神経研疾病３ ²横浜市立大医生理 Background: Computed tomography (CT) has been considered to be difficult to obtain high-contrast image of brain structures covered with the skull. However, previous studies have suggested that ex vivo imaging with iodinated contrast agent could enable us to do so. A drawback in the method of previous studies is that CT instrument with a lower rated output requires long time for acquisition. Here we report a novel method which can obtain high contrast image in a short time. Methods: After two rats or two mice brains perfused fixed with 4.0% paraformaldehyde were soaked in ionic iodinated contrast agent diluted with phosphate buffered saline (PBS) for a period of 7 days, images were obtained by Clairvivo-CT (Shimadzu Co., Tokyo Japan). Scan parameters of CT were examined two or more conditions based on previous studies. After CT scanning, frozen sections were produced using the brain and were processed for Kluver-Barrera (KB) staining (double staining for myelin and Nissl body). We compared obtained ex vivo images to brain atlas and stained slides. Results: Good contrast images were obtained in tube voltage peak of 50 kVp and tube current of 200 μA. In resolution, images generated by the acquisition of 1000 projections to previous studies had many noises, therefore we changed this parameter to 3600 projections. Using this condition, we were able to obtain high contrast brain images in approximately 30 minutes. There seemed to be no problem in staining pattern for KB staining of the brain soaked in contrast agent. When the ex vivo images were compared with brain atlas, we confirmed that the morphology of the brain structures were closely matched each other. Conclusions: We could obtain high contrast brain image in brief CT scanning by increasing the number of projections. Further studies will enable us to obtain more detailed brain images of the brain, which will be useful in the research of neuropsychiatric diseases.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-498 Investigation of neural activity related to feeding behavior in GCaMP6 transgenic mice using an implantable imaging device Mark Christian San Gabriel Guinto（Guinto Mark Christian San Gabriel），Yasumi Ohta（Ohta Yasumi），Mamiko Kawahara（Kawahara Mamiko），Makito Haruta（Haruta Makito），Kiyotaka Sasagawa（Sasagawa Kiyotaka），Takashi Tokuda（Tokuda Takashi），Jun Ohta（Ohta Jun） Nara Institute of Science and Technology Developing new tools for visualizing neural activity within deep brain regions at a cellular scale is critical for advancing our knowledge on how the brain processes information and controls decision-making that is eventually manifested by animal behavior. As a visceral behavior displayed by all organisms, the mechanism of feeding and its aspects relating to motivational salience are of prime interest to neuroscience and remain an active area of research. Feeding behavior is regulated by neural circuits that are known to be involved in the processing of satiety, reward and pleasure. At present, available methods for calcium imaging in deep brain areas are still a challenge to execute without interfering with the normal behavior of an awake mouse. Our current approach centers on an implantable micro-imaging device that can visualize neural activity from calcium dynamics in the deep brain developed in our laboratory [1]. Here we report a monitoring scheme for the feeding behavior using our device and discuss noteworthy results from the experiments. The micro-imaging device consists of a CMOS-based image sensor chip embedded on a flexible substrate and a μ-LED that serves as an excitation light source [1]. Deep brain implantation of the device in GCaMP6 transgenic mice demonstrated successful detection of neuronal activity in regions related to feeding behavior, including the lateral and the arcuate nucleus of the hypothalamus, sections of the striatum, and the amygdala [2]. The increase in fluorescence intensity observed in putative somas expressing GCaMP6, a genetically-engineered calcium-binding protein whose fluorescence is modulated by changes in intracellular [Ca²⁺], is used as an indicator for neuronal activity. Distinct patterns of somatic fluorescence were recorded from the sampled regions over a range of behaviors (e.g., grooming, eating, resting) exhibited by the animal. Moreover, in contrast to presently available imaging systems, our design offers several advantages: (1) streamlined design results in reduced invasiveness, (2) light weight (~0.02 g), and (3) the miniature size (0.5-mm device width). Our ultra-light, compact device presents a useful paradigm for exploring neural dynamics in freely moving setups, cueing potential for simultaneous imaging of multiple sites across neural circuits. References [1] J. Ohta et al., Proc. IEEE, 105(1), 158, 2017. [2] M.C. Guinto et al., Annual Meeting of Society for Neuroscience 2018

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-499 Implantable needle-type imaging sensor device for simultaneous detection of neuronal activity in nuclei involved in nociception Romeo B Rebusi（Rebusi Romeo B），Mark Guinto（Guinto Mark），Yasumi Ohta（Ohta Yasumi），Mamiko Kawahara（Kawahara Mamiko），Makito Haruta（Haruta Makito），Kiyotaka Sasagawa（Sasagawa Kiyotaka），Takashi Tokuda（Tokuda Takashi），Jun Ohta（Ohta Jun） Nara Institute of Science and Technology The lateral capsular subdivision of the central nucleus of the amygdala (CeLC) is known as the nociceptive center of the amygdala. It receives and relays nociceptive signals and is responsible for the emotional sensation and response to pain. This amygdalar region receives serotonergic input from the dorsal raphe nucleus of the midbrain (DRN) which have Substance P-secreting neurons. This substance is an important element for pain perception. It is not fully understood what role the DRN has in nociception and its processing in relation to its downstream amygdalar counterparts. In this study, we aim to elucidate the direct nociception neural pathway bridging the DRN to the CeLC by the use of an implantable needle-type device developed by our laboratory. This device is composed of a CMOS-based image sensor chip embedded on a flexible substrate with a μ-LED light source. Its small size allows for a minimally intrusive implantation that only affects brain regions immediate to the path and target. Also because of its size, successful simultaneous implantation are most possible. Its lightweight also allows the mouse to move freely without stress. The combination of these features permits for high output in vivo studies unrestricted by bulky sensors and extensive injury to the animal subject. For the methodology, two devices are simultaneously implanted next to the CeLC and the DRN of GCaMP6 mice. As of now, the implantation method for these two nuclei are being refined. The DRN is an especially problematic site for access because the region immediately above it is the location of large blood vessels, complicating straightforward implantation. Angular implantation trajectories have demonstrated good accuracy with relatively less injuries. It is currently planned to proceed with in vivo pain-induction experiments with the devices implanted. Pain induction will be via the formaldehyde test wherein 5% formaldehyde will be injected subcutaneously to induce a bi-phasic pain response. Nociception will be detected by the devices recording neuronal activities in the CeLC and the DRN. The results will help further understand the neural network underlying pain perception and pinpoint the exact participating subsections of the nuclei.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-500 Direct measurement of cerebral capillary blood flow across the sleep/wake cycle in mouse Chia-Jung Tsai（Tsai Chia-Jung）^1,2，Takeshi Nagata（Nagata Takeshi）²，Chih-Yao Liu（Liu Chih-Yao）^1,2，Takehiro Miyazaki（Miyazaki Takehiro）¹，Takeshi Kanda（Kanda Takeshi）¹，Kaspar Vogt（Vogt Kaspar）¹，Masashi Yanagisawa（Yanagisawa Masashi）¹，Yu Hayashi（Hayashi Yu）¹ ¹WPI-IIIS, Univ of Tsukuba, Tsukuba, Japan ²Ph.D Program in Human Biology, SIGMA, Univ of Tsukuba, Tsukuba, Japan Cerebral blood flow (CBF) is critical in maintaining energy-dependent processes, clearing metabolic byproducts generated by neuronal activity, and predicting neurotransmitter activities (Dukart et al., Sci Rep, 2018). Impaired CBF regulation can affect protein and ATP synthesis, disrupt action potential generation, or cause ischemic neuronal death (Hossmann et al., Ann Neurol, 1994). CBF dysregulation is associated with neurodegenerative disorders such as Alzheimer's Disease (AD) (Kassandra et al., Nature Rev Neurosci, 2017). CBF is regulated strictly and independently from the peripheral circulation. Here, I aimed to investigate the role of sleep in the regulation of CBF. Till now, various approaches have been taken to investigate how CBF changes during wake, NREM sleep and REM sleep. However, conflicting conclusions have been drawn from multiple approaches. According to studies using positron emission tomography, CBF is decreased during NREM sleep and comparable to wake during REM sleep (Madsen et al., Cerebrovasc Brain Metab Rev, 1991), whereas according to studies using Doppler flowmetry, CBF is high during REM sleep and comparable to wake during NREM sleep (Grant et al., J Physiol, 2005). These conflicts likely stem from differences in data processing and normalization procedures as well as from differences in the type of blood vessels that were analyzed. We hypothesized that the problem with the above approaches is that they cannot directly observe blood flow in individual capillaries, where the actual substances exchange between blood and glias/neurons occurs. Therefore, we developed an alternative approach using two-photon microscopy imaging to directly measure the velocity of red blood cells in capillaries while monitoring sleep/wake stages. This technique would help us draw a final conclusion on how CBF changes in sleep/wake. Analyzing CBF via two-photon microscopy is not only able to achieve high spatio-temporal resolution, but also provides a novel method for studying other physiological activities.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-501 大気圧プラズマ照射後ラット大脳皮質の広域電顕撮像による微細構造解析 Masanori Yamato（大和正典）^1,2，Yasuhisa Tamura（田村泰久）^1,2，Mitsuyo Maeda（前田光代）^1,2，Asami Eguchi（江口麻美）^1,2，Satoshi Kume（久米慧嗣）^1,2，Yosky Kataoka（片岡洋祐）^1,2 ¹理化学研究所BDR 細胞機能 ²理研-JEOL連携センター　マルチモダル微細構造解析 Electron microscopy is a powerful tool for observing tissue and cellular microstructure, however imaging area on the tissue is very limited due to super-high spatial resolution. Recently, we have developed a scanning electron microscopy (SEM) system realizing serial and automatic capturing more than thousands of ultra-fine morphological images in a wide range of biological tissue. The stitching procedure of all the images into a single tiling image allowed us to obtain large-scale electron microscopic images covering several square millimeters even at high resolutions. In order to advance the efficiency of imaging data acquisition, functions for automatic focusing and automatic astigmatism adjustment were added to the SEM system. In this study, we analyzed tissue microstructure in rat cerebral cortex after irradiation of atmospheric pressure plasma, using the large-scale SEM system. Plasma is an ionized gas composed of electrons, photons, charged particles and reactive oxygen species. Atmospheric pressure plasma is being developed for a wide range of medical applications, including wound healing, blood coagulation, malignant cell apoptosis, cell proliferation etc. However, effect of the atmospheric-pressure plasma on the central nervous system of mammals remains unclear. To date, we have observed characteristic histological changes in the cerebral cortex after irradiation of atmospheric-pressure plasma to the parietal cortex of adult rats. At 3-7 days after the irradiation, immunohistochemical observation with a light microscopy revealed that many cells were aligned to encircle the irradiated region, and that a large number of those cells were proliferating. Around such a tissue structure, rod-like cells expressing immature cell markers also appeared. Using our newly-developed SEM technique, we could acquire large-scale electron microscopic images covering entire cortical regions affected by plasma irradiation. In this meeting, we will demonstrate the time series variation of microstructure (cell nuclei and cytoplasmic organelles, such as mitochondria) in all the cells including neurons and glial cells after plasma irradiation. Further, we will demonstrate the appearance of plasma irradiation-derived immature cells even in the cerebral cortex of adult rats.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-502 特定の神経回路を光顕と電顕で広域イメージングする方法の開発 Shinsuke Shibata（芝田晋介）¹，Tomoko Shindo（信藤知子）^1,2，Ryo Ihara（井原諒）^1,2，Taro Iseda（伊勢田太郎）^1,2，Nobuko Moritoki（盛一伸子）¹，Toshihiro Nagai（永井俊弘）¹，Hideyuki Okano（岡野栄之）² ¹慶應大医電顕室 ²慶應大医生理 Recently the correlative light and electron microscopy (CLEM) and scanning electron microscopy technologies were drastically improved for observing biological specimens. In order to identify the localization of the specific molecules in the specific neural circuitry, fluorescence labeling with the antibody or with the genetic modification was usually used for visualization. In this study, we developed a novel CLEM method using multi-beam scanning electron microscopy, which enabled us to carry out the large area observation at the resolution of electron microscopy level with extraordinary high speed. Here, we described the detailed procedure how to carry out this novel large-area CLEM (named as LA-CLEM) with the central nervous system from primates and rodents. The marmoset and mouse neural tissue were labeled with several kinds of antibodies, and a large-area images were obtained both from fluorescence microscopy and from multi-beam scanning electron microscopy with the same brain. The LA-CLEM approach allows us to understand the detail of neural circuitry through correlative observation of both specific molecules' localization and synapse-level neural connection in the millimeter order large-area. It is known that visualizing the detailed neural circuitry is quite important to understand the precise connection of the neural circuit. We hope this novel procedure would help to detect the abnormality of the brain suffering from the various neuropsychiatric diseases in near future.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-503 分子インプリント高分子をプローブとした神経伝達物質分泌のリアルタイムイメージング Yasuo Yoshimi（吉見靖男）¹，Yuto Katsumata（勝俣湧斗）¹，Narumi Teshima（手島成洋）¹，Kenta Endo（遠藤健太）¹，Kenji Umeta（梅田健司）¹，Risako Mori（森莉紗子）¹，Tatsumi Nagahama（長濱辰文）²，Takashi Inutsuka（犬塚隆志）³，Maya Okamoto（岡本摩耶）³ ¹芝浦工業大学工学部応用化学科 ²帝京平成大学　健康メディカル学部 ³日本薬理評価機構 Analysis of the secretion of neurotransmitters in nervous system is important for elucidating the mechanism of neural network in nervous system. Thus, development of a probe which can track a specified neurotransmitter in real time with high selectivity has been required. Molecularly imprinted polymer (MIP), which is a molecular recognition material obtained by polymerization with a template-effect of the target molecule may be applicable for the probe. In this study, nanoparticle of MIP including fluorescent group (fMIP-NP) were developed as the probe of serotonin. The serotonin, as the template, was immobilized on glass beads by using mixed anchor of 3-aminopropyltrimethoxysilane (shorter anchor) and 3- (2-aminoethylamino) propyltrimethoxysilane (longer anchor) (1:1 in weight) with glutaraldehyde. The template-immobilized beads were fluidized in a mixed solution of a fluorescent monomer, a template-affinity monomer, and a crosslinking monomer under UV irradiation. The colloidal fMIP-NP was collected from the surface of the beads by washing with dimethylformamide. And the dispersion medium of the colloidal fMIP-NP was replaced with 0.15 M phosphate buffer. The fluorescent intensity and the radius of the fMIP-NP was increased by addition of serotonin but was insensitive to L-tryptophan which is the analogue of serotonin. The buccal ganglion of Aplysia sea snail was stained by the fMIP-NP by soaking in the dispersion. Spike oscillation in the fluorescent intensity were observed at the sensory neurons in the ganglion immediately after the taste of Nori seaweed was given to the radula connected with the ganglion. The oscillation was enhanced by addition of an inhibitor of monoamine oxidase which collects serotonins from the synapse. The results indicate that the serotonin secreted from neurons can be detected by imaging technology using the fMIP-NP templated by serotonin as a probe. Most of transmitters can be targets of MIPs, thus imaging with probes of fMIP-NPs would enable analysis of dynamism of neurotransmitters in central nerve system.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-504 全脳レベルから超微細構造レベルまでをつなぐ神経回路形態マルチスケール解析を目指すScaleS法の活用 Takahiro Furuta（古田貴寛）¹，Kenta Yamauchi（山内健太）²，Shinichiro Okamoto（岡本慎一郎）²，Yoko Ishida（石田葉子）²，Aya Takenaka（竹中綾）¹，Jungwon Hwang（黄晶媛）²，Kaoru Isa（伊佐かおる）³，Tadashi Isa（伊佐正）^3,4，Hiroyuki Hioki（日置寛之）² ¹大阪大院歯高次脳口腔機能学講座 ²順天大院　医神経生物・形態 ³京都大院医神経生物 ⁴京都大構成的ヒト生物学研究拠点 An optical clearing method, "ScaleS", provides stable tissue preservation and thus can be combined with electron microscopy (EM). However, there is still room for improvement on membrane integrity in the cleared samples. In this project, we aim to set up a pipeline optimized for multi-scale imaging from macro- to nano-levels based on ScaleS technology. We established a rapid tissue clearing procedure, "ScaleSF", by modifying the ScaleS method. It was confirmed that brain tissue treated by ScaleSF showed comparable transmittance with that of tissues treated by original ScaleS method. Even after fixation with glutaraldehyde (GA) of a high concentration (2%), the tissue transmittance was retained. Furthermore, by using EM, we observed that addition of GA contributed to preservation of ultrastructures in cleared brain tissue. The effect of fixation with GA on ultrastructures was investigated not only in mice but also in marmosets. Then, aiming at realizing successive LM/EM imaging, we developed an AAV vector expressing both EGFP and APEX2. It was noted that APEX2 peroxidase activity was declined with time after fixation. To circumvent this problem, we deposited biotin molecules in APEX2 expressing cells by using tyramine-signal-amplification (TSA) method before optical clearing. The dual expression of GFP and APEX2 is expected to contribute to identifying locations of highly-magnified ultrastructures (labeled by APEX2) in large-scale light microscopic data (by GFP). Now, we are trying successive LM/EM imaging by combining ScaleSF technique with the dual expressing AAV to achieve multi-scale imaging in large scale 3D structure.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-505 N6-メチルアデノシン脱メチル化酵素ALKBH5に特異性を示す阻害剤の探索 Yoshie Fujiwara（藤原芳江）¹，Yume Kato（加藤結女）²，Kanako Hori（堀加奈子）³，Yohei Katoh（加藤洋平）⁴，Kazuhisa Nakayama（中山和久）⁴，Tatsuya Takagi（高木達也）²，Yu-Shi Tian（田雨時）²，Dan Ohtan Wang（王丹）¹ ¹京都大iCeMS ²大阪大院薬情報計量薬学 ³大阪大薬 ⁴京都大院薬生体情報制御学 Gene expression is regulated both at translation and at transcription level in response to external signals/stimulations. Neurons establish complicated neuronal circuit and use local proteomes at synapse to regulate synaptic activities that can occur independently from nuclear regulation. We have previously shown that in this context, N6-methyladenosine (m⁶A), a reversible and prevalent modification in mRNA, is deposited on synaptic transcripts that encode regulatory proteins of synaptic activities. We have shown that the protein expression level of APC, a highly modified transcript, was regulated through m⁶A recognition and m⁶A contributes to the development and function of synapses. In contrast to APC, whose translation requires m⁶A activity, demethylation has been shown to activate translation in developing axons. ALKBH5 (AlkB homolog 5) and FTO (fat mass and obesity-associated) are the m⁶A demethylases that belong to Fe²⁺/2-oxoglutarate-dependent dioxygenase family. FTO deletion impairs neurogenesis, and learning and memory in adult mice. ALKBH5 deficiency disturbs cerebellar development in addition to spermatogenesis. Thus demethylases are required for brain development and function. To further understand the function of these proteins and for potential therapeutic applications, we have searched for FTO or ALKBH5-specific inhibitors, especially ALKBH5-specific inhibitor that has not been found. Since FTO and ALKBH5 have similar structures of the catalytic sites, inhibitor compounds for ALKBH5 with target specificity is hard to obtain. We have performed the biochemical assay using pharmacologically/physiologically active substance libraries. Several compounds are found to preferentially inhibit ALKBH5 versus FTO. Now we are validating their cytotoxicity and in vivo activities, using ALKBH5 or FTO knocked-out cell line.		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-506 掌サイズの生体アンプを使用した携帯可能な脳波利用型意思伝達支援装置 Nobuyuki Yamawaki（山脇伸行）近畿大生物理工医用工学 Patients who have difficulty moving their muscles freely due to amyotrophic lateral sclerosis (ALS) require support for various situations that occur in everyday life. Brain-computer interface (BCI) systems using motor imagery or event-related potential provide a means of communication for the patients. Since the auditory BCI is suitable for the patients in later stages of ALS who lose all voluntary muscular control, including voluntary control of gaze, the application of P300 event-related potential for auditory stimulation to the communication has been studied. However, the auditory BCI using the P300 requires long time to communicate with high accuracy. Therefore, in order to shorten the required time, the motor imagery is used in the proposed system. The EEG signals changes by the motor imagery that does not need visual stimuli or instructions. The system uses the temporal and spatial change of the EEG signals by the motor imagery. The system is controlled by only EEG depend on two kinds of brain activities with closed eyes and allows of expression of ""yes"" or ""no"". The system was evaluated by three healthy subjects in each a hundred tasks and 88% accuracy was achieved. In the present study, the system was made portable by using a palm-size biological amplifier (Hanwa Electronic Ind. Co., Ltd.). The biological amplifier is possible to measure 4 -channel electroencephalogram signals and there are analog signals and Bluetooth signals as its output format. Therefore, in the system using the analog output of the conventional medical electroencephalograph as in our EEG-based communication support system, it is easy to replace the electroencephalograph with the biological amplifier. Furthermore, by using the Bluetooth, wireless EEG signals measurement becomes possible. However, since the performance of a medical electroencephalograph and the biological amplifier are slightly different, the influence is reported.

7月26日（金）17:15～18:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-507 Large-Scale Automatic Tracer Segmentation in Brain Section Fluorescence Images Using Artificial Intelligence Woodward Mark Alexander（Alexander Woodward Mark）¹，Rui Gong（Gong Rui）¹，Hiroshi Abe（Abe Hiroshi）²，Masahide Maeda（Maeda Masahide）¹，Noritaka Ichinohe（Ichinohe Noritaka）^2,3，Yoko Yamaguchi（Yamaguchi Yoko）¹ ¹Neuroinformatics Unit, RIKEN Center for Brain Science ²Ichinohe Neural System Group, Laboratory for Molecular Analysis of Higher Brain Function, RIKEN Center for Brain Science ³Department of Ultrastructural Research, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Tokyo, Japan We have developed an artificial intelligence solution for fast and accurate anterograde tracer signal segmentation for common marmoset brain connectomics. Brains injected with a viral tracer were sectioned and imaged with a batch slide scanner (Nanozoomer 2.0 H-T). Images are 16-bit, with an in-plane resolution of 0.46 μm/pixel and can reach sizes of up to 50,000×50,000 pixels and 3 GB in file size. The tracer signal is a challenge to segment due to images having varying brightness and contrast offsets and the fine-scale tracer signal having similar pixel intensities to the background. Conventional approaches such intensity thresholding, independent component analysis (ICA) [1], and Gaussian mixture unsupervised learning [2] struggle to accurately and consistently segment such signals. Along with the huge spatial size, these images pose a challenge for fast and accurate tracer segmentation. The U-Net [3] neural network architecture, a popular approach for biomedical image segmentation, was used for this work. We selected representative 1000×1000 pixel subimages and manually annotated the tracer signal to create binary masks for training. All masks were validated by a neuroanatomist and intensity normalization and data augmentation was applied. Our neural-network approach is parameterless, handles brightness/contrast offsets, and can distinguish between tracer signals and background features. We found that the network generalizes well to unseen images and this approach provides consistent segmentation of tracer signals that can then be used for quantitative analysis of axonal projection patterns. 1. Hiroshi Abe, Toshiki Tani, Hiromi Mashiko, Naohito Kitamura, Naohisa Miyakawa, Koki Mimura, Kazuhisa Sakai, Wataru Suzuki, Tohru Kurotani, Hiroaki Mizukami, Akiya Watakabe, Tetsuo Yamamori, Noritaka Ichinohe, 3D reconstruction of brain section images for creating axonal projection maps in marmosets, Journal of Neuroscience Methods, Vol.286, 102-113, 2017. 2. Alexander Woodward, Carlos Enrique Gutierrez, Hiromichi Tsukada, Hiroshi Abe, Noritaka Ichinohe, Kenji Doya, Yoko Yamaguchi (2017) A Connectomics Pipeline for Tracer Injection Studies of the Marmoset Monkey Brain. Neuroinformatics 2017. 3. Olaf Ronneberger, Philipp Fischer and Thomas Brox. U-Net: Convolutional Networks for Biomedical Image Segmentation. Medical Image Computing and Computer-Assisted Intervention (MICCAI), Springer, LNCS, Vol.9351: 234--241, 2015		7月27日（土）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-508 NMDA受容体の活性を阻害する精神作用活性物質を効率的に検出するハイスループットシステムの確立 Toshinari Mitsuoka（光岡俊成）¹，Kenji Hanamura（花村健次）¹，Noriko Koganezawa（小金澤紀子）¹，Ruri-Kikura Hnajiri（花尻（木倉）瑠璃）²，Tomoaki Shirao（白尾智明）¹，Yuko Sekino（關野祐子）³ ¹群馬大院医神経薬理 ²国立医薬品食品衛生研生薬 ³東京大院薬ヒト細胞創薬 In recent years, New Psychoactive Substances (NPS) have been widely distributed for abuse purpose in many nations. NPS have raised the public health threat. The effective measures to counter the spread of these non-controlled substances is to grasp the distribution route or the cross-border transaction route, and to legislate promptly through the risk assessment process by the early warning system. Moreover, the generic scheduling on NPS, which is able to regulate exhaustively a group of substances having similar structure by accumulating concerning data in structure and activity correlation, was founded to have a great effect on the control of NPS. In few years, instead of decreasing cannabinoid derivatives and synthetic cathinone, phencyclidine and ketamine analogues having the N-methyl-D-aspartate type glutamate receptor (NMDAR) inhibitory effects have emerged in Japan. Therefore, it will be important to establish high-throughput system for efficiently detecting NPS that can inhibit the activity of NMDAR. We have shown that NMDAR activation reduces the amount of an actin-binding protein drebrin in postsynaptic sites. We utilized the characteristics of drebrin to detect the inhibitory effects of phencyclidine-like substances on NMDAR activity. After 3 weeks in vitro, cultured neurons were preincubated with phencyclidine (PCP) or PCP-like substances, such as 3-Methoxyphencyclidine (3-MeO-PCP) and 4-[1-(3-methoxyphenyl)cyclohexyl]morpholine (3-MeO-PCMo), and then treated with 100 μM glutamate for 10 min. After fixation, cultured neurons were immunostained with anti-drebrin and anti-MAP2 antibodies. Drebrin cluster density along dendrites were automatically quantified by originally-developed protocol. Preincubation with PCP or PCP-like substances significantly reduced the glutamate effect. Maximal inhibitory rate of 3-MeO-PCMo was lower than that of PCP or 3-MeO-PCP. Thus, it is indicated that our high-throughput imaging system using cultured neurons is useful for detecting the effects of PCP-like substances on NMDA receptor activity.

7月26日（金）12:55～14:15　ポスター会場（朱鷺メッセ　1F　展示ホール） PB-510 Ａ11神経細胞と入力線維の神経化学的特徴付け Tsuyoshi WA Yamaguchi（山口わ剛）¹，Hidechika Ozawa（小澤継史）²，Shuichi Ueda（上田秀一）¹ ¹獨協医大解剖（組織） ²獨協医大麻酔 The A11 dopaminergic cell group is the only group that includes neurons innervating the spinal cord among the A8-A16 dopaminergic cell groups and a decrease of A11 dopaminergic transmission at the spinal cord is thought to contribute to the pathogenesis of restless legs syndrome. However, the mechanisms regulating the neuronal activity of A11 dopaminergic neurons remain to be elucidated. Unraveling the neuronal composition, distribution and connectivity of A11 neurons would provide insights into the mechanisms regulating the spinal dopaminergic system. To address this, we performed immunohistochemistry for calcium-binding proteins such as calbindin (Calb) and parvalbumin (PV), in combination with the retrograde tracer Fluorogold (FG) injected into the spinal cord. Immunohistochemistry for Calb, PV, or tyrosine hydroxylase (TH), a marker for dopaminergic neurons, revealed that there were at least three types of neurons in the A11 region: neurons expressing Calb, TH, or both TH and Calb, whereas there were no PV-immunoreactive (IR) cell bodies. Both Calb- and PV-IR processes were found throughout the entire A11 region, extending in varied directions depending on the level relative to bregma. We found retrogradely labeled FG-positive neurons expressing TH, Calb, or both TH and Calb, as well as FG-positive neurons lacking both TH and Calb. In addition, we performed double immunohistochemistory for calcitonin gene-related peptide, androgen receptor, or estrogen receptors in combination with anti-Calb antibody for neurochemical characterization of A11 Calb positive cells, and performed double immunohistochemistry for dopamine β-hydroxylase or corticotoropin-releasing factor with TH to characterize neurochemical innervation of A11 neurons. These findings indicate that the A11 region is composed of a variety of neurons that are distinct in their neurochemical properties, and suggest that the diencephalospinal dopamine system may be regulated at the A11 region by both Calb-IR and PV-IR processes, and at the terminal region of the spinal cord by Calb-IR processes derived from the A11 region.