ポスター（Poster）

Poster ポスター

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-001 大脳皮質神経幹細胞におけるグリコーゲン代謝の機能と調節機構 Hitoshi Gotoh（後藤仁志），Tadashi Nomura（野村真），Katsuhiko Ono（小野勝彦）京都府立医大院医　神経発生生物学 Glycogen is a polysaccharide formed by glucose and serves as an energy store for energy deprivation. Glycogen metabolism is regulated by several enzymes, such as glycogenin (Gyg), glycogen synthase (Gys), or glycogen phosphorylase (GP). Gyg and Gys synthesize glycogen whereas glycogen phosphorylase (GP) degrates glycogen for energy supply. Glycogen metabolism is involved in the synaptic transmission through astrocytic release of lactate into synaptic area or clearance of synaptic neurotransmitter. Although glycogen appears in early embryonic brain, less is known about its role in development as compared with that the role in adult. Especially, physiological significance of glycogen metabolism in the radial glial cells remains unknown, in spite of the fact that the glycogen is rich in this cell type. We examined the distribution of glycogen during cortical development. Glycogen appears around E14.5 and increase until E18.5. Glycogen particles were present specifically in Pax6+ radial glial cells but not in Tbr2+ intermediate progenitor cells or differentiated neurons. We examined the expression of genes involved in glycogen metabolism, such as Gys, GP, or Gyg, and found that these genes were expressed in the ventricular zone radial glial cells. We introduced shRNA vectors against Gyg and found that knockdown of Gyg induced premature differentiation of radial glial cells, suggesting that the glycogen metabolism is important for maintaining radial glial cells. In addition, we found that mRNA of ppp1r3c, another essential regulator of glycogen synthase/phosphorylase activity, is accumulated in radial glial endfeets. Recently, it was reported that transcripts of several radial glia-specific genes were transported to the radial glial endfeet and translated locally dependending on 3'-UTR of mRNA. Therefore, we tagged nuclear localizing Venus (nls-Venus) with 3'-UTR of ppp1r3c, and electroporated the construction into embryonic cortex. The Venus fluorescent is observed in samples electroporated with nls-Venus without 3'-UTR. However, nuclear GFP with 3'-UTR transported to the basal endfeet of radial glial cells and locally translated in 3'-UTR dependent manner, suggesting that 3'-UTR of ppp1r3c might be fundamental mechanism of radial glial cell-specific glycogen metabolism. These data suggest that glycogen metabolism is important for maintaining neural stem cells, and the localization of ppp1r3c regulates radial glial cell-specific glycogen metabolism.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-002 The compensatory redundancy between Prdm16 and Mecom regulate stem cell maintenance during cortical development Fatma Rabia Urun（Urun Fatma Rabia）^1,2，Adrian W Moore（Moore Adrian）¹ ¹Laboratory for Neurodiversity, RIKEN Center for Brain Science, Saitama, Japan ²Graduate School of Science and Engineering, Saitama University, Saitama, Japan The transcriptional repressor Hes1 is an important regulator of neuronal development as it promotes Neuronal Stem Cell (NSC) self-renewal. Our lab has shown that Mecom, a Prdm family transcription factor, amplifies the Notch-mediated Hes1 transcriptional output for self-renewal, and thus maintains the stem cell identity in embryonic olfactory NSCs. We previously showed that Prdm family members are present in NSC domains in different parts of developing nervous system. Recently Prdm16, the closest homolog of Mecom, has been shown to be required for NSC maintenance in developing cortex, as well as the production of intermediate precursors. Given these roles of Prdm16, it is surprising that knockout of Prdm16 in stem cell compartment in cortex does not lead to serious developmental defects or lethality. Considering the role of Mecom in NSC self-renewal and close relationship between Mecom and Prdm16, we asked if there may be a compensatory functional redundancy occurring between Prdm16 and Mecom that rescues the cortical development defects by supporting Hes1 transcriptional output in cortical NSCs when Prdm16 is lost. We found that loss of Prdm16 leads to Mecom upregulation in cortical NSCs, where Mecom expression is normally found to be in very low levels after the onset of neurogenesis. For further investigation of compensatory redundancy between Prdm16 and Mecom, WT and Prdm16/Mecom double knockout cortices are now being compared in terms of patterning and expression levels of cells expressing the marker genes associated with cortical neurogenesis and NSC self-renewal. Fine regulation and maintenance of the NSC self-renewal and neuronal differentiation is important for the proper development of the nervous system. For this reason, identification of the compensatory functional redundancy going between transcription factors that are required for proper occurrence of neural development may be crucial for identifying the causes of the subtle changes in neurogenesis that are causative of neurodevelopmental disorders such as ASD or ADHD.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-003 幹細胞性を司る遺伝子の転写活性によるヒト脳巨大化への役割 Hidenori Tabata（田畑秀典）^1,5，Tsuyoshi Hachiya（八谷剛史）^2,3，Kouji Shimoda（下田耕治）⁴，Kanehiro Hayashi（林周宏）⁵，Koh-ichi Nagata（永田浩一）¹，Yasubumi Sakakibara（榊原康文）³，Kazunori Nakajima（仲嶋一範）⁵ ¹愛知県心身障害者コロニー発達障害研 ²岩手医大メガバンク ³慶應大理工生命情報 ⁴慶應大医動物実験センター ⁵慶應大医解剖 During evolution, humans have acquired a huge brain. One of the reasons for this is thought to be higher production of neurons. The cortical neurons of mammals are produced from apical progenitors in the ventricular zone (VZ) or from progenitors in the subventricular zone (SVZ). Recent studies revealed the existence of self-renewing progenitors in the primate SVZ called "outer subventricular zone radial glia like cells" (oRG cells). Repeated cell divisions of oRG cells increase the size of SVZ and form the histologically defined outer SVZ (OSVZ). The expansion of oRG cells enables enormous production of neurons during the cortical development of humans. On the other hand, we have previously reported two distinct populations migrating out of the VZ in mice. One population exits the VZ using the somal translocation mode and divides in the SVZ. The other population becomes postmitotic in the VZ, stays there for about 10 hours and then transforms into the multipolar cells to accumulate just above the VZ or lower SVZ, which we named the multipolar cell accumulation zone (MAZ). Because the former exits the VZ earlier than the latter, we named it "rapidly exiting population" (REP). REP cells have several common features with oRG cells, namely they have a long ascending process, proliferate, and distribute broadly in the SVZ, suggesting that REP contains the oRG-equivalent cells in mice. We have also reported that the production rate of REP cells is higher in the lateral cortical VZ than in dorsomedial. We, therefore, supposed that the molecular mechanisms for the regulation of the REP production rate in mice might also be involved in the oRG production in humans. To identify such molecular mechanisms, we conducted microarray analyses and identified genes that were differentially expressed between the lateral and medial VZs. We then introduced these candidate genes into the VZ by in utero electroporation, and found one gene that strongly enhanced production of the REP cells. Interestingly, the transcriptional activity of human regulatory region of this gene is higher than that of mouse, suggesting that this difference is the key to determine the size of brains in these species. The molecular mechanisms for this will be discussed.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-004 Cohen症候群の原因遺伝子であるVps13bは大脳皮質神経細胞の移動に重要な役割を果たしている Kei-ichi Katayama（片山圭一），Norimitsu Inoue（井上徳光）和歌山県立医科大学医学部分子遺伝学講座 Cohen syndrome is an autosomal recessive disorder characterized by microcephaly, intellectual disability, typical facial features, progressive retinal dystrophy and intermittent congenital neutropenia, which is caused by loss-of-function mutations in the VPS13B (also known as COH1) gene. Mutations in the VPS13B gene were also found in patients suffering from autism spectrum disorders. All these reports suggest that VPS13B is indispensable for the neurodevelopmental processes. Previous studies showed that Vps13b is a scaffold protein of the Golgi complex and has important roles in vesicle trafficking through the Golgi apparatus, and depletion of Vps13b in cultured neurons resulted in impaired neurite outgrowth. However, Vps13b is expressed in the developmental central nervous system from early embryonic period, indicating its role in much earlier neurodevelopmental events, such as neurogenesis, neuronal migration and differentiation. In the present study, we knocked down Vps13b in cortical neurons by utilizing in utero electroporation to investigate its role in migration of cortical neurons. RNAi-mediated silencing of Vps13b in cortical neurons significantly impaired their migration. Morphologically, Vps13b-depleted neurons have longer leading processes compared to control neurons, suggesting its role in somal/nuclear movement. This result suggests that defects in neuronal migration caused by the mutations in the VPS13B gene may be involved in the pathogenesis of Cohen syndrome and autism spectrum disorders.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-005 ECMを再編成するADAMTS2は大脳新皮質の放射状神経細胞移動を制御する Noe Kaneko（金子乃愛）^1,2，Kei Yura（由良敬）³，Nobuaki Maeda（前田信明）^2,3，Chiaki Maruyama（丸山千秋）¹ ¹都医学研神経回路形成 ²お茶大院ライフサイエンス ³早稲田大先進理工 In the developing neocortex, newborn neurons initially show multipolar (MP) morphology, moving slowly with no particular direction (MP migration). When MP neurons reach the subplate (SP) layer, they change into bipolar (BP) shape (MP-to-BP transition), and initiate fast radial glial fiber-dependent locomotion (locomotion) toward pial surface. This radial neuronal migration process occurs in a birth- date-dependent inside-out manner, leading to the generation of the six layered structure of neocortex. Previously, we performed gene expression profiling of migrating excitatory neurons, and found that the expressions of genes related to extracellular matrix (ECM) are significantly changed during migration. Among these genes, we focused on ADAMTS2 (a disintegrin and metalloproteinase with thrombospondin motifs 2), the expression of which remarkably increased during migration. Both siRNA-mediated knockdown and overexpression of ADAMTS2 in migrating neurons disturbed MP-to-BP transition, suggesting that the proper levels of ADAMTS2 are required or the initiation of locomotion. Recently, it has been reported that TGF-β signaling-related proteins are potential proteolytic substrates of ADAMTS2. It has also been reported that TGF-β initiates signaling pathways to fate naive neurites into axons in the early neocortical neurons. However, the involvement of TGF-β signaling in radial neuronal migration remains to be clarified. Immunohistochemistry of pSmad3, which is one of the activated downstream protein of TGF-β signaling, revealed that pSmad3 is expressed just under the subplate, Thus, we hypothesized that ADAMTS2 regulates neuronal migration through transient activation of TGF-β signaling at the SP. In order to investigate the involvement of TGF-β signaling in neuronal migration, we monitored TGF-β/ pSmad signaling during neuronal migration using cultured cortical slices by luminescent imaging. The results suggested that TGF-β signaling is up-regulated in the migrating MP neurons just before switching to the locomotion mode. We are now analyzing the downstream pathways of TGF-β signaling in radial migration process including the regulation of F-actin dynamics.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-006 大脳皮質発生におけるニューロン移動停止の制御機構 Yuki Hirota（廣田ゆき），Kazunori Nakajima（仲嶋一範）慶應大医解剖 During neocortical development, excitatory neurons generated in the ventricular zone or subventricular zone migrate towards the pial surface. Neurons stop migrating when they reach just beneath the marginal zone and form a cortical plate in a so-called "inside-out" pattern, that is, the newly arriving neurons pass through their predecessor neurons to settle in the most superficial position in the cortical plate. This birthdate-dependent positioning (layer formation) is regulated by several signaling cascades, including the Reelin signaling. Reelin is a glycoprotein mainly secreted by Cajal-Retzius neurons in the marginal zone, and known to function via its receptors, apolipoprotein E receptor 2 (ApoER2) and very low density lipoprotein receptor (Vldlr), and the cytoplasmic adaptor protein disabled 1 (Dab1). The Vldlr single mutant shows invasion of migrating neurons into the MZ without defect in the radial migration, whereas reeler mice lacking the Reelin protein, Apoer2/Vldlr double knockout mice and Dab1 mutants show severe migration defects with roughly inverted formation of the cortical layers, suggesting that Vldlr has specific roles for termination of neuronal migration beneath the MZ. However, precise mechanisms by which Reelin signaling controls the termination of neuronal migration remains unclear. Here, to gain insight into the role of Vldlr-mediated Reelin signaling during cortical development, we examined the migratory behavior of Vldlr-deficient neurons in the developing brain. Stage-specific labeling of newborn neurons revealed that both the earlier-born and later-born neurons ectopically invaded the MZ in the Vldlr mutants and that Vldlr-deficiency significantly impaired dendrite outgrowth in the MZ. Rescue experiments showed that Vldlr has a cell-autonomous function and that adhesion molecules function downstream of Vldlr. These results suggest that Vldlr controls the proper termination of radial migration during cortical development.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-007 成体脳内を移動する新生ニューロンにおける一次繊毛の時空間的な動態 Mami Matsumoto（松本真実）¹，Masato Sawada（澤田雅人）¹，Huy Bang Nguyen（Nguyen Bang Huy）^2,3，Vicente Herranz-Perez（Herranz-Perez Vicente）^4,5，Truc Quynh Thai（Thai Quynh Truc）²，Keishi Narita（成田啓之）⁶，Natsuko Kumamoto（熊本奈都子）⁷，Shinya Ugawa（鵜川眞也）⁷，Yumiko Saito（斎藤祐見子）⁸，Sen Takeda（竹田扇）⁶，Naoko Kaneko（金子奈穂子）¹，Jose Manue Garcia-Verdugo（Garcia-Verdugo Jose Manue）⁴，Nobuhiko Ohno（大野伸彦）^2,9，Kazunobu Sawamoto（澤本和延）^1,10 ¹名古屋市大院医再生医学 ²自然科学研究生理学研究所分子神経生理研究部門 ³Dept. Anatomy, Faculty of Medicine, Univ of Medicine and Pharmacy, Ho Chi Minh, Vietnam ⁴Laboratorio de Neurobiologia Comparada, Instituto Cavanilles, Univ de Valencia, CIBERNED, Valencia, Spain ⁵Predepartamental Unit of Medicine, Faculty of Health Sciences, Universitat Jaume I, Castello de la Plana, Spain ⁶山梨大院医工総合研解剖学生物学 ⁷名古屋市大医院機能組織学 ⁸広島大院総合科学行動科学 ⁹自治医大医解剖学組織学 ¹⁰自然科学研究生理学研究所神経発達・再生機構研究部門 In the adult rodent brain, neural stem cells still reside in the ventricular-subventricular zone (V-SVZ) and continuously produce new neurons. These new neurons form chain-like cell aggregates and migrate through the rostral migratory stream toward the olfactory bulb, where they differentiate into olfactory interneurons and contribute to various olfactory functions. Primary cilium is a microtubule-based tiny protrusion, and involved in the processing of extracellular signals for the regulations of various facets of adult neurogenesis. However, it remains unknown whether the adult V-SVZ-derived migrating new neurons have primary cilium. Here we show the morphology and dynamics of primary cilium in the V-SVZ-derived migrating new neurons. We found that the V-SVZ-derived migrating new neurons express Arl13b, a ciliary small GTPase, in the close vicinity of centrioles. To investigate detailed ultrastructural morphology of primary cilium in migrating new neurons in vivo, we used transmission electron microscopy. Our observations suggest that primary cilium submerges by deep membrane invagination, but still has access to the extracellular environment. Furthermore, by using serial-block face scanning electron microscopy in vivo and time-lapse imaging of primary cilium in cultured new neurons, we found that submerged state of the primary cilium in new neurons is dependent on the migration phase of saltatory movement. Our results provide the basis to understand the regulation of primary cilium dynamics in neuronal migration in the adult brain.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-008 神経系におけるHITS-CLIP解析に基づいた新しいRNA制御機構の解明と創薬への応用の可能性 Yoshika Yano（矢野佳芳）¹，Akihide Koyama（小山哲秀）²，Hirohide Takebayashi（竹林浩秀）¹，Kenji Sakimura（崎村建司）³，Hideyuki Okano（岡野栄之）⁴，Masato Yano（矢野真人）¹ ¹新潟大院医歯神経生物解剖 ²新潟大院医歯法医 ³新潟大脳研モデル動物開発 ⁴慶應大医生理 In general, RNA binding proteins (RNABPs) participate in multiple forms of post-transcriptional gene regulation, such as maturation, transport, stability and translation of coding and non-coding RNAs (ncRNAs), generating a complicated transcriptomic network. Only 2% of RNABPs are tissue-specific and thought to play a role in tissue-specific gene regulatory networks. We have been focusing on roles of tissue- and cell- specific expressing RNABPs to understand how they generate cell-specific transcriptomics and function in brain. Here we show the molecular mechanism and cellular functions of one such a RNABP, Qki5, a member of STAR family proteins, specifically expressed in a particular type of cell during the brain development. By using several comprehensive approaches, such as whole RNAseq and HITS-CLIP, and mouse genetics, we discovered a mechanistic rule of Qki5-dependent alternative splicing events and a crucial biologic role for the maintenance of the early embryonic neural stem cells and proper neurogenesis through beta-catenin signaling pathway. Further to investigate new roles of Qki5 in RNA regulation, including an enigmatic class of ncRNAs, we used uncharacterized Qki5 HITS-CLIP sites, named orphan CLIPed clusters, which are not related to alternative splicing events. An Analysis of orphan CLIPed clusters containing Qki5 binding code (ACUAAy) combined with whole transcriptome data sets revealed direct functions of Qki5 on biogenesis of two enigmatic RNA classes of circular RNAs and DoGs-like transcripts. We will discuss a novel RNA regulatory loop in gene regulation and a possibility of application to design RNA-switching oligonucleotides for a therapeutic approach in neurological diseases. Hayakawa-Yano et al, Gene Dev 2017. Hayakawa-Yano et al, in revison 2019.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-009 RORγt 過剰発現マウスにおける脳内・行動表現型への影響 -新規精神疾患モデルの可能性- Saki Tome（當銘幸貴）¹，Rei Nagata（永田玲）³，Tetsuya Sasaki（佐々木哲也）²，Yosuke Takei（武井陽介）² ¹筑波大院　人間総合科学感性認知脳科学 ²筑波大　医　生命医科学・解剖学・神経科学 ³国際基督教大 Th17 cells, which produce the proinflammatory cytokine IL-17, play important roles in the induction of inflammation. A previous study has reported that increase in IL-17A contribute to disruption of blood brain barrier integrity, which leads to memory disturbance in mice. Furthermore, elevated levels of IL-17A have been detected in the serum of a subset of ASD patients. However, to date, the role of IL-17A in pathogenesis of neuropsychiatric disorders remains to be determined. To evaluate the effect of long-term upregulation of IL-17A to function and development of CNS in vivo, and elucidate the significance of IL-17A in pathogenesis of neuropsychiatric diseases, we used transgenic mice with C57BL/6 background in which retinoic acid related orphan receptor gamma-t (RORγt) is overexpressed under the control of human CD2 gene (RORγt Tg mice). RORγt is known to be a transcription factor required for differentiation of Th17 cells. In this study, we confirmed that serum IL-17A level is markedly upregulated in the RORγt Tg mice compared to WT mice. To obtain a first clue of the effect of IL-17A upregulation in CNS, we performed immunohistochemical analysis of RORγt Tg mice using various neuronal and glial markers. We also carried out analysis of behaviors that could relate with higher brain functions. In this presentation, we will show the results of histological, behavioral, and other data of the RORγt Tg mice, and discuss about their significance in pathogenesis of neuropsychiatric disorders such as Autism Spectrum Disorder and schizophrenia. Since it has been suggested that IL-17 play important role in dysregulation of fetal brain development caused by maternal immune activation, we examined pregnant female RORγt Tg mice, and found a significant increase in abortion rate after maternal immune activation induced by poly(I:C) administration. This suggests an alteration of intrauterine environments in RORγt Tg mice, which could alter fetal brain development.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-010 自閉症感受性遺伝子Auts2のマウス大脳皮質構築における役割 Kazumi Shimaoka（嶋岡可純）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，Mikio Hoshino（星野幹雄）¹ 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 Autism susceptibility candidate 2 (AUTS2) has been implicated as the gene associated with numerous psychiatric disorders such as autism spectrum disorders (ASDs), intellectual disabilities (ID) and schizophrenia. In mouse developing CNS, AUTS2 is highly expressed at several brain regions responsible for the higher brain functions such as cerebral cortex and hippocampus. We have previously demonstrated that cytoplasmic AUTS2 regulates the neuronal migration and neurite formation in the developing cerebral cortex. Furthermore, nuclear AUTS2 has been reported to be involved in the transcriptional regulation of multiple genes for neural development by interacting with the Polycomb group protein complex 1 (PRC1). However, there remain many questions about the physiological roles for AUTS2 in the brain development. In this study, we analyzed the function of AUTS2 in the cerebral corticogenesis, especially focusing on the neurogenesis. In the developing forebrain, AUTS2 is broadly expressed at the cerebral cortex. Moreover, we found that loss of Auts2 in mice leads to reduction in the thickness of cortical plate. Pulse-labeling assay exhibited that the number of EdU-positive cortical neurons was significantly decreased in the Auts2 mutant mice, and most notably in the Cux1-positive upper layer neurons. On the other hand, the number of neural progenitor cells at the ventricular zone was not different between wild-type (WT) and Auts2 mutant mice whereas the number of cells with immunopositive for pH3, a specific marker for the mitotic phase of the cell cycle as well as the S-phase neural progenitor cells pulse-labeled with EdU were dramatically decreased in the Auts2 mutants compared to WT mice, implying that defects of AUTS2 induces the cell cycle delay of neural progenitor cells and results in a decrease in the production of cortical neurons in Auts2 mutant mice. Furthermore, quantitative real-time PCR analysis revealed that cyclin D1 mRNA level was decreased whereas p57kip2 was increased in Auts2 mutant cerebral cortex. Taken together, these results suggest that AUTS2 plays a key role for the control of transition from neural progenitor to postmitotic cells via regulation of the expression of cell cycle-related molecules during later cerebral corticogenesis.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-011 小脳顆粒細胞の発生におけるNhlh1/2の機能解析 Mana Shimoda（霜田真奈）¹，Satoshi Miyashita（宮下聡）¹，Toma Adachi（足立透真）^1,2，Mikio Hoshino（星野幹雄）¹ ¹国立精神神経医療研究センター ²早稲田大学 Neuronal precursor cells first proliferate continuously to expand the precursor pools, then exit the cell cycle and initiate the differentiation programs during the development of the mammalian central nervous system (CNS). However, it is still elusive what molecules regulate the timing of cell cycle exit and the initiation of differentiation programs. Cerebellar granule cells (GCs), which occupy the majority of cerebellar neurons, are one of the useful models to study the molecular mechanisms regulating the proliferation and differentiation of neurons. From embryonic day 13 (E13), granule cell precursor (GCPs), which are born in rhombic lip (RL), form the external granule layer (EGL). From the later embryonic stage to the first two postnatal weeks, GCPs actively proliferate in the outer layer of EGL (oEGL), exit the cell cycle, and then move to the inner layer of EGL (iEGL), where they initiate differentiation. After that, GCs move through the molecular layer (ML) to the internal granule cell layer (IGL) to become mature GCs. These developmental processes from GCPs to GCs are well known, but the molecular mechanisms to stop proliferation and initiate differentiation of GCPs are still unknown. When cells differentiate from progenitor cells, it is generally considered that the cell fates are determined by switching of transcription factors. Especially in CNS development, it has been indicated that the proper order switching of the basic helix-loop-helix (bHLH) family, which is the family of the transcription factors, controls the cell proliferation, determination, and differentiation. Nescient helix-loop-helix 1 (Nhlh1) and Nhlh2 are the bHLH transcription factors. As Nhlh1 and 2 are known to be expressed in differentiated neurons in several regions of the CNS, it is suggested that both genes are involved in differentiation of neurons. In this study, we investigated the functional role of the Nhlh1/2 during the development of GCs. We first performed the immunohistochemistry to estimate the protein expression of NHLH1/2 and revealed that NHLH1/2 showed complementally expressions in EGL. We next carried out the gain-of-function and loss-of-function experiments to assess the function of Nhlh1/2 in EGL. We also identified several down stream targets of NHLH1. These results shed light on the novel molecular mechanisms involved in the cell cycle exit and the initiation of differentiation mediated by the proper order switching of Nhlh1 and Nhlh2.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-012 Early-Generated Interneurons Regulate Neuronal Circuit Formation during Early Postnatal Development Yongchun Yu（Yu Yongchun）¹，Chang-Zheng Wang（Wang Chang-Zheng）¹，Jian Ma（Ma Jian）^1,2，Shao-Na Jiang（Jiang Shao-Na）¹，Tian-Qi Chen（Chen Tian-Qi）¹，Ye-Qian Xu（Xu Ye-Qian）¹，Zu-Liang Yuan（Yuan Zu-Liang）¹，Xiao-Yi Mao（Mao Xiao-Yi）¹，Ying-Hui Fu（Fu Ying-Hui）¹，Shu-Qing Zhang（Zhang Shu-Qing）¹，Lin-Yun Liu（Liu Lin-Yun）¹，Yong-Chun Yu（Yu Yong-Chun）¹ ¹Institutes of Brain Science, State Key Laboratory of Medical Neurobiology and Collaborative Innovation Center for Brain Science, Fudan University ²School of Life Sciences, Tsinghua-Peking Joint Center for Life Sciences, IDG/McGovern Institute for Brain Research, Tsinghua University, Beijing The pioneer interneurons that are generated earliest are a unique subpopulation of interneurons in the neocortex. It has long been postulated that early-generated interneurons are essential for the proper development of neural circuits. However, whether they can help to precisely shape the emergence of synaptic transmitting neuronal connections, as well as spontaneous network oscillations in the developing neocortex remains largely unclear. To address these issues, we used an inducible genetic fate-mapping approach to selectively label early-generated interneurons (EGIns) and pseudo-random interneurons (pRIns) in the developing neocortex. We found that sparse EGIns are mainly comprised of SST-positive interneurons and predominantly located in deep neocortical layers. These EGIns exhibited more mature electrophysiological and morphological properties and higher synaptic connectivity as compared to pRIns at early postnatal stages. Moreover, we found that about 1/6 of EGIns in neocortical layer 5 can single-handedly influence network dynamics, whereas very few pRIns displayed a similar capability. Notably, ablation of EGIns significantly altered network synchronization and reduced inhibitory synaptic transmission of layer 5 pyramidal neurons at early postnatal stages. Our findings thereby demonstrate that pioneer interneurons in the neocortex, by contributing to neuronal synchrony at early postnatal stages, could play an important role in the wiring of immature cortical circuits. In summary, our study complements and expands on previous works by providing new insights into EGIns that regulate network oscillations and are critical for shaping the development of precise synaptic circuits in the neocortex during early postnatal stages. Given that disruptions of GABAergic circuitry at several points can contribute to neurodevelopmental disorders, results from this study may be particularly important for our understanding of cell-type-specific network dysfunctions in these disorders.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-013 STAR RNA 結合タンパクQuakingのミューラーグリアにおける機能解析 Kako Takahashi（高橋果子）¹，Ayane Matsubara（松原綺音）¹，Takahiro Sogo（十河孝浩）^3,4，Teruhisa Kawamura（川村晃久）^2,4，Satoru Moritoh（森藤暁）^1,4，Chieko Koike（小池千恵子）^1,4 ¹立命館大学薬学部 ²立命館大学生命科学部 ³立命館グローバル・イノベーション研究機構 ⁴システム視覚科学研究センター Quaking (QKI) is RNA-binding proteins which belongs to the STAR family of KH domain-containing. QKI functions in pre-mRNA splicing, microRNA regulation, and formation of circular RNA. QKI has three isoforms, QKI-5, QKI-6 and QKI-7. The amount of expression of QKI-5 in mouse retina is the most of the three isoforms. QKI-5 is expressed in nuclear and QKI-6, QKI-7 are expressed in cytoplasmic. QKI plays critical roles in myelinogenesis in the central and peripheral nervous systems and has been implicated neuron-glia fate decision in the brain. QKI is also involved in cancer, liver disease, and intellectual disability. However, the function of QKI in retina remains unclear. Previously we reported that QKI, especially Qki-5, is strongly expressed in Muller glia cells (MG) in the adult retina¹. QKI is also expressed in progenitors and differentiating neurons during retinal development, and their expressions were eventually weakened or diminished during maturation¹. Our previous data raise the possibility that QKI may function in retinal cell fate determination and maturation on both glia and neurons. In this study, we examined the effects of reducing QKI expression in mouse MG. We introduced lentivirus QKI shRNA vector into primary cultured mouse retinal MG², and confirmed the reduction of QKI mRNA and protein levels. We extract RNA and are analyzing changes of marker gene expressions of retinal cells by real time PCR. We will present the result of real time PCR and immunocytochemistry when QKI is knocked down in mouse retinal MG. ¹Suiko et al. (2016) Expression of Quaking RNA-Binding Protein in the Adult and Developing Mouse Retina. PLOS ONE 11 (5): e0156033. ²Ueki et al. (2012) p53 is required for the developmental restriction in Muller glial proliferation in mouse retina. Glia 60 (11): 1579-1589.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-014 転写因子Meis1による小脳顆粒細胞とバーグマングリアの発生制御 Tomoo Owa（大輪智雄）¹，Ryo Shiraishi（白石椋）¹，Satoshi Miyashita（宮下聡）¹，Toma Adachi（足立透真）¹，Takuro Nakamura（中村卓郎）²，Ryo Goitsuka（五飯塚僚）³，Shinichiro Taya（田谷真一郎）¹，Mikio Hoshino（星野幹雄）¹ ¹国立精神・神経セ神経研病態生化学 ²がん研究所　発がん研究部 ³東京理科大学　生命医科学研究所　発生及び老化研究部門 In the nervous system, there are a wide variety of neuronal cell types that have morphologically, physiologically, and histochemically different characteristics and this diversity may enable us to elicit higher brain function. However, the molecular mechanisms underlying the developmental brain are largely unknown. The cerebellum is a good model system to study this issue because a variety of types of neurons are produced. In this study, we focused on Myeloid ecotropic viral integration site 1 (Meis1) that was expressed in the cerebellum during development. We found that Meis1 was expressed in granule cells, Bergmann glia and neuroepithelial cells in the cerebellar primordium. To understand the function of Meis1, we generated two types of Meis1 conditional knockout (cKO) mice by crossing Meis1-flox mice with Atoh1-Cre line and with En1-Cre line. In the former cKO mice, Meis1 gene was specifically deleted in the granule cell lineage and in the latter cKO, Meis1 gene was deleted in the whole cerebellum. In Meis1fl/fl; Atoh1-Cre-Tg, the size of the cerebellum was smaller than that of wild type littermates, yet the number of cerebellar lobules was increased. Interestingly, many Atoh1-positive cells were ectopically observed in deeper regions (parenchyma) of the postnatal cerebellum. These cells showed mitotic features and therefore we suspected they contributed to generating extra lobules. In Meis1fl/fl; En1-Cre mice, the size of the cerebellum was much smaller than that of wild type or Meis1fl/fl; Atoh1-Cre-Tg mice. Moreover, vermis and cerebellar lobules disappeared. In contrast to wild type or Meis1fl/fl; Atoh1-Cre-Tg mice, granule cells could not migrate tangentially into the inner cerebellum. Interestingly, position and morphology of Burgmann glia was disorganized, which was never observed in Meis1fl/fl; Atoh1-Cre-Tg. As Bergmann glia is known to regulate migration of granule cells, dysfunction of Brugmann glia may account for migration disorder of granule cells in Meis1fl/fl; Atoh1-Cre-Tg. These results suggest that Meis1 plays important roles in differentiation of both granule cells and Bergmann glia, thereby controlling coordinated development of the cerebellum.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-015 神経幹細胞増殖能のハイスループットスクリーニング方法の確立 Yoshihiko Morisue（森末良彦），Hitoshi Inada（稲田仁），Noriko Osumi（大隅典子）東北大学大学院医学系研究科附属創生応用医学研究センター発生発達神経科学分野 Brain is an organ enriched with fatty acids including polyunsaturated fatty acids such as docosahexaenoic acid (DHA) and arachidonic acid (ARA). Extensive studies have reported essential roles of the fatty acids in brain development and adult neurogenesis. The effect of fatty acids on the neurogenesis, however, has yet to be fully understood. To quantitatively analyze roles of the fatty acids and identify novel functional fatty acids affecting neurogenesis, we established a high-throughput screening method using mouse neural stem cell (NSC) culture (i.e., neurosphere culture). Briefly, the subventricular zone, a niche of neurogenesis, was dissected from the brain of 10-week-old male mice, and subsequently incubated with papain (20 U/mL) and DNaseI (100 U/mL) at 37ºC for 60 min. Enzymatic activity was inactivated by adding ovomucoid solution, and the tissue was finally dissociated into single cells by passing a needle. The cells were seeded in culture medium containing B27/N2 supplements, penicillin/streptomycin, epidermal growth factor (20 ng/mL), and fibroblast growth factor (20 ng/mL). After 5 days in culture, primary spheres were dissociated with 0.025% Trypsin-EDTA, seeded at 4 × 10⁴ cells/mL, and cultured for 5 days to form secondary spheres. The secondary spheres were dissociated at 37ºC for 5 min, seeded at 4 × 10⁴ cells/mL, and cultured for 5 days to form tertiary spheres. The tertiary spheres were similarly dissociated and seeded in 96-well plate at 5 × 10⁴ cells/mL in the presence of different fatty acids for 3 days. After 3 days culture, the cells were fixed with 4% PFA-PBS and subsequently stained with Hoechst33342, and, the number and size of neurospheres were measured with a cell image analyzer (Cellomics, Thermo Scientific). In our pilot study, 10^-10 M to 10^-4 M of DHA or ARA was applied to the cultured neurospheres. The results were similar to our previous study (Sakayori et al., 2011) showing that both DHA and ARA affect proliferation of NSCs in a concentration dependent manner, although we need further investigation to draw a conclusion. We are planning to screen functional fatty acids by using a commercial fatty acid library.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-016 ゲノム編集を用いた効率的エピトープタグ挿入による低抗原性遺伝子産物のマウス生体内可視化 Takayoshi Inoue（井上高良），Junko Asami（浅見淳子），Takahiro Morita（森田卓宏），Yuki Morimoto（森本由起），Yukiko U Inoue（井上上野由紀子）国立精神・神経セ神経研 It would be a mandatory task in neuroscience studies to generate high-affinity antibodies against given gene products since such antibodies allow rigorous evaluation of molecular localization, interacting partners and functions for those products in vivo. Here we aim to establish an efficient and less time/money-consuming knock-in protocol for short yet hyper-immunoreactive epitope tag based on CRISPR/Cas9 system. As the model, we select type II classic cadherin cell-cell adhesion molecules, whose antigenicity is extremely low due to their critical roles in cell-cell recognition and elaborating multicellular organization. We knock-in two tandem copies of HA-tag (YPYDVPDYA) for Cdh6 and two tandem copies of PA-tag (GVAMPGAEDDVV) for Cdh8 in frame at the C-terminus by means of simple electroporation mediated delivery of commercially available Cas9 protein and custom-made cr/tracrRNA as well as single strand oligonucleotides carrying the homology arms and coding sequences for tags into mouse fertilized eggs. Consequently, we quickly obtain epitope tag knock-in founder mice as designed and the tag staining for stable knock-in homo mouse lines clearly reveal precise protein localization patterns of Cdh6 and Cdh8 in the developing brain, which could have never been noticed from the results of these mRNA in situ hybridization. Our results accordingly demonstrate the value of systematic epitope tag knock-in strategy in promoting molecular mechanism-oriented neuroscience studies in vivo.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-017 汎用除草剤グリホサートの発達期神経毒性 Ken Futagami（二神健）¹，Lin Yong Huang（Huang Yong Lin）²，Yoko Nomura（Nomura Yoko）²，Yasunari Kanda（諫田泰成）³，Naohiro Hozumi（穂積直裕）⁴，Sachiko Yoshida（吉田祥子）¹ ¹豊橋技科大工 ²Queens College, City University of New York, New York, USA ³国立医薬品食品衛生研 ⁴豊橋技科大院工電子・情報工学 Environmental chemical exposures may act through pathologies, including the direct disruption of cells and structures of the nervous system, and epigenetic changes. Glyphosate, main compound of broad-applied herbicide, might affect developmental neural toxicity, however, it is Negative Control substance of Organisation for Economic Co-operation and Development test guideline. To mature animals, its neurotoxicity was associated with rigidity, slowness and resting tremor in all four limbs with no impairment of short-term memory. In addition, we had little knowledge about the developmental neurotoxicity of aminomethylphosphonic acid (AMPA), which was the only metabolite detected. The primary herbicidal function of glyphosate is to inhibit a key plant enzyme, namely 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). This enzyme participates in the biosynthesis of aromatic amino acids (phenylalanine, tyrosine and tryptophan) via the shikimate pathway in bacteria, fungi, and plants, whereas glyphosate could act as antibiotics in the mammalian gut microbiome, and may have the potential to modify the animal and human microbiota. In this study, we administrated glyphosate or AMPA to pregnancy rat at embryonic day 16 p.o. and observed the cerebellar development of offspring. Glyphosate-administrated rat showed decrease of Purkinje cells and activation of microglia on the Purkinje layer, while their lobule formation in the vermis was almost normal. We suggest that glyphosate and its derivatives might have some neurotoxicity in developing brain.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-018 新生仔ラット脳においてミクログリアはヘテロな細胞集団である Kohichi Kawahara（川原浩一）¹，Hitoshi Nakayama（中山仁）²，Takuya Hasegawa（長谷川拓也）¹，Takehiko Maeda（前田武彦）¹ ¹新潟薬科大学 ²熊本大学 To differentiate subtypes of microglia (MG), we developed a novel monoclonal antibody, 9F5, against one subtype (type 1) of rat primary MG. The 9F5 showed high selectivity for this cell type in Western blot and immunocytochemical analyses and no cross-reaction with rat peritoneal macrophages (Mφ). We identified the antigen molecule for 9F5: the 50- to 70-kDa fragments of rat glycoprotein nonmetastatic melanoma protein B (GPNMB)/osteoactivin, which started at Lys¹⁷⁰. In addition, 9F5 immunoreactivity with GPNMB depended on the activity of furin-like protease(s). More important, rat type 1 MG expressed the GPNMB fragments, but type 2 MG and Mφ did not, although all these cells expressed mRNA and the full-length protein for GPNMB. These results suggest that 9F5 reactivity with MG depends greatly on cleavage of GPNMB and that type 1 MG, in contrast to type 2 MG and Mφ, may have furin-like protease(s) for GPNMB cleavage. In neonatal rat brain, amoeboid 9F5+ MG were observed in specific brain areas including forebrain subventricular zone, corpus callosum, and retina. Double-immunostaining with 9F5 antibody and anti-Iba1 antibody, which reacts with MG throughout the CNS, revealed that 9F5+ MG were a portion of Iba1+ MG, suggesting that MG subtype(s) exist in vivo. We propose that 9F5 is a useful tool to discriminate between rat type 1 MG and other subtypes of MG/Mφ and to reveal the role of the GPNMB fragments during developing brain.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-020 マウス脳におけるグアニンヌクレオチド交換因子ARHGEF9の発現解析 Koh-ichi Nagata（永田浩一），Kyoko Ibaraki（茨木京子），Makoto Mizuno（水野誠），Ikuko Iwamoto（岩本郁子），Hidenori Tabata（田畑秀典），Hidenori Ito（伊東秀記）愛知県心身障害者コロニー発達障害研 ARHGEF9, also known as Collybistin, is a Dbl (diffuse B-cell lymphoma)-family guanine nucleotide exchange factor (GEF) for Rho small GTPases. ARHGEF9 has been reported to regulate the recruitment of gephyrin, a scaffold protein essential for clustering glycine and GABAA receptors at the cell surface. The critical role of ARHGEF9 in brain development is supposed by mutations associated with neurodevelopmental disorders such as epilepsy and mental retardation. However, precise expression analyses of ARHGEF9 have not so far been conducted. In this study, we prepared a specific polyclonal antibody against ARHGEF9, anti-ARHGEF9, and carried out expression analyses with mouse tissues especially brain. Western blotting analyses demonstrated tissue-dependent expression profiles of ARHGEF9 in the young adult mouse. In expression analyses with the whole brain extracts, ARHGEF9 was strongly detected at embryonic day (E)13.5 and then gradually decreased throughout the developmental process to postnatal day (P)30, suggesting a role during brain development. Immunohistochemical analyses revealed developmental stage-dependent expression profiles of ARHGEF9 in cerebral cortex, hippocampus and cerebellum. When mouse cultured primary hippocampal and cortical neurons were stained with anti-ARHGEF9 together with anti-Gephyrin, a maker for inhibitory postsynapses, their partial colocalization was observed in dendrites, suggesting that ARHGEF9 interacts with Gephyrin at inhibitory synapses. From the obtained results, anti-ARHGEF9 was found to be a useful tool for biochemical and pathophysiological analyses of ARHGEF9 in neurodevelopmental disorders such as epilepsy and intellectual disability		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-021 海馬ニューロン樹状突起形成過程におけるミトコンドリアの局在および動態解析 Akane Hatsuda（初田茜）¹，Mineko Kengaku（見学美根子）^1,2，Kazuto Fujishima（藤島和人）² ¹京都大院生命統合生命 ²京都大iCeMS Neurons consume a large amount of ATP for the construction and maintenance of their complex structures and for the regulation of neuronal activities. In order to meet these requirements, mitochondria, a source of ATP, need to be transported throughout neuronal processes. Previous papers have shown that defects in mitochondrial function or trafficking induce abnormal dendritic morphology and precocious neurodegeneration, indicating that mitochondrial trafficking is important for neuronal development and survival. In hippocampal neurons, dendrites dynamically change their shape during development (postnatal 1-2 weeks) before they construct complicated dendritic arbors which receive afferent neuronal signals on mature spines. We hypothesize that mitochondria may change their dynamics in response to the rapid increase in ATP demands during dendritic growth. In order to visualize mitochondrial dynamics during neuronal development, we transfected mitochondrial matrix-targeted GFP (mito-GFP) in CA1 pyramidal neurons and in primary hippocampal neural cultures. Immunofluorescence signals revealed that mitochondrial density in distal dendrites was very low in young neurons and gradually increased during dendritic growth until mitochondria were regularly arranged along the dendritic shaft in later stages. Time-lapse imaging showed that mobile mitochondria decreased as the dendrite matured. We currently seek for a possible feedback regulation of mitochondria mobility by local ATP concentrations.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-022 若齢マウスの脳の発達過程における交連下器官でのPax6の発現 Hitoshi Inada（稲田仁）¹，Kotaro Hiraoka（平岡宏太良）²，Shun Araki（荒木峻）¹，Noriko Osumi（大隅典子）¹ ¹東北大院医創生応用医セ発生発達神経 ²東北大サイクロ核医学研究部 The subcommissural organ (SCO) is one of the circumventricular organs located at the dorsal side of the boundary between the third ventricle and the cerebral aqueduct. The SCO secretes several proteins such as SCO-spondin that forms Reissner's fiber complex (Munoz et al., 2018) and is thought to be involved in the homeostasis of the cerebrospinal fluid. It has been reported that a transcription factor Pax6 is essential for the formation of the SCO during brain development in the embryonic stage (Estivill-Torru et al., 2001). However, function of Pax6 in the SCO in the postnatal stage remains unknown. To address the function of Pax6 in the SCO, we first examined Pax6 expression in the SCO at different postnatal stages. Apparent Pax6 expression in the SCO was observed from postnatal day (P) 0 to P28. At P0 mice, the Pax6 expression is observed in both the SCO and the circumventricular region, although the expression gradually became restricted in the SCO, suggesting that Pax6 might be required for maintenance of the SCO at postnatal stages. To characterize Pax6 expressing cells in the SCO, expression of several cell markers was investigated using antibodies against neurons (NeuN), astrocytes (anti-GFAP), oligodendrocytes (anti-Olig2), microglia (anti-Iba1), and neural stem/progenitor cells or ependymal cells (anti-Sox2) at P28 mice. The SCO showed a specific expression of Sox2, suggesting a possibility that the SCO may contain neural stem/progenitor cells. Considering the expression of Pax6 in the SCO, the organ could possibly be a niche for neural stem cells.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-023 VPAやMS-275によって誘導されるPC12細胞の分化初期段階に関与するnurr1遺伝子と下流遺伝子の発現上昇の分子機構 Hiroki Maruoka（丸岡弘規），Ryota Takahashi（高橋亮太），Ryosuke Yamazoe（山添亮輔），Koji Shimoke（下家浩二）関西大学化学生命工学部生命生物工学科 Since histone deacetylase (HDAC) inhibitors are known to induce related genes in both neuroprotection and neuronal differentiation, they are attracting an attention as effective therapeutic compounds for neurodegenerative diseases or nerve injuries. Recently, class I-selective HDAC (HDAC 1, 2, 3 and 8) inhibitors are applied for the treatment of not only cancer but also epilepsy or bipolar disorder. Moreover, it has been suggested that several class I-selective HDAC inhibitors have the effect of neuronal differentiation. However, detailed roles of these class I-selective HDAC in nervous system are largely unknown. In this study, we analyzed the role of HDAC isoform in neuronal differentiation using multiple class I-selective HDAC inhibitors or HDAC isoform specific siRNA. We revealed that class I-selective HDAC inhibitors, VPA (valploic acid) or MS-275, induced neurite outgrowth in PC12 cells, but HDAC1 and 2 specific inhibitor, SHI-1:2 did not, meaning that one of the HDAC isoforms, HDAC3, may contribute to neurite outgrowth. Thus, we demonstrated whether HDAC3 plays an important role in neurite outgrowth. Moreover, we confirmed the contribution of immediate early gene, nurr1, to extend neurites in the early stage of VPA or MS-275-induced differentiation. In addition, we analyzed what molecules are controlled by Nurr1 after the treatment with VPA or MS-275. The analysis using the neuronal marker proteins discovered that Nurr1 regulated the increased expression of Synapsin1, β-tubulin III and NeuroD. Then, immunoblotting analyses showed strong reduction of Synapsin1, β-tubulin III and NeuroD compared with cells treated with a negative control siRNA after the treatment with nurr1 siRNA for 24 h in the presence or absence of VPA or MS-275. These results indicate that the Nurr1 plays a pivotal role in the differentiation process by VPA or MS-275 via the upregulation of Synapsin1, β-tubulin III and NeuroD in PC12 cells.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-024 嗅球糸球体近傍ニューロンに高発現する α-シヌクレインの機能解析 Katsutoshi Taguchi（田口勝敏）¹，Yoshihisa Watanabe（渡邊義久）²，Atsushi Tsujimura（辻村敦）²，Masaki Tanaka（田中雅樹）¹ ¹京都府立医科大学　生体構造科学 ²京都府立医科大学　基礎老化学 α-Synuclein (αSyn), the major constituent of Lewy bodies and Lewy neurites, is generally expressed in presynapses and is involved in synaptic function. However, we previously demonstrated that some neurons, including those in the olfactory bulb, show high expression levels of αSyn in the cell body under normal conditions. αSyn is also known to be important for adult neurogenesis. In present study, we examined the role of αSyn in juxtaglomerular neurons (JGNs) with high αSyn expression in the mouse olfactory bulb. Most αSyn-enriched JGNs expressed Sox2, which functions to maintain neural immature identity. Interestingly, in αSyn homozygous (-/-) knockout (KO) mice, Sox2-positive JGNs were significantly increased compared with heterozygous (+/-) KO mice. Following global brain ischemia using wild-type mice, there was also a significant decrease in Sox2-positive JGNs and in the co-expression ratio of Sox2 in αSyn-enriched JGNs. By contrast, the co-expression ratio of NeuN, mature neuronal marker, was significantly increased in αSyn-enriched JGNs. However, this ischemia-induced decrease of Sox2-positive JGNs was not observed in αSyn homozygous KO mice. Overall, these data suggest that αSyn functions to promote the maturation of immature JGNs in the mouse olfactory bulb.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-025 神経回路形成因子LOTUSの記憶機能における役割 Ryohei Nishida（西田遼平）¹，Rie Ishikawa（石川理絵）²，Satoshi Kida（喜田聡）²，Kohtaro Takei（竹居光太郎）¹ ¹横浜市大院生命医生体機能医 ²東京農大院農バイオ Overcoming the higher brain dysfunction that exhibits memory disorders, such as dementia, is an important issue for mankind and many researches for the issue are currently conducted all over the world. Nogo binds to Nogo receptor 1 (NgR1) and the binding inhibits axon growth and synapse formation, thereby is associated with impaired nerve regeneration and memory function. We previously identified lateral olfactory guide substance (LOTUS) as an endogenous NgR1 antagonist. It was reported functional relationship between decrease of LOTUS expression in hippocampus in accordance with aging and memory function impairment. However, the role of LOTUS in memory function has remained to be elucidated. Therefore, we examined whether LOTUS is involved in memory function. We first performed social recognition test and Morris water maze test in wild type (WT), LOTUS-gene knocking-out (LOTUS- KO) and LOTUS gene overexpressing transgenic (LOTUS-Tg) mice. We found that memory function was impaired in LOTUS-KO, whereas the function was enhanced in LOTUS-Tg mice. Next, we examined roles of LOTUS in synapse formation in cultured hippocampal neurons, and found that decrease of synaptic density in LOTUS-KO mice in contrast to increase of the density in LOTUS-Tg mice. Furthermore, we also found that decrease of neurogenesis in the adult hippocampus of LOTUS-KO mice, while LOTUS-Tg mice showed increase the adult hippocampal neurogenesis. Since neurogenesis in the hippocampus is known to enhance memory function, increase of LOTUS expression level in the hippocampus may relate to enhancement of memory function through increase of neurogenesis. These findings suggest that LOTUS may involve in memory formation accompanied with synapse formation and neurogenesis in hippocampus.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-026 Sar1aのプルダウンアッセイは、細胞分化時におけるSar1aの活性化を明らかにする Yuri Urai（浦井悠里），Junji Yamauchi（山内淳司）東京薬科大院生命科学 The intracellular molecular transport system is a basic and general cellular mechanism that is regulated by an array of signaling molecules. Sar1 small GTPases are molecules that play a key role in controlling vehicle transport between the endoplasmic reticulum (ER) and Golgi bodies. Like other small GTPases, the activities of Sar1a depend on their guanine-nucleotide-binding states, which are regulated by guanine-nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs). Despite the well-known function of mammalian Sar1 in the intracellular transport system, little is known about when and how Sar1 is activated during cell morphological changes. Here we show that the C-terminal, but not the N-terminal, regions of Sec23A and Sec23B, the effector proteins of Sar1a, specifically bind to the active, GTP-bound form of Sar1a. An affinity precipitation (pull-down) assay using a recombinant C-terminal region of Sec23B reveals that Sar1a is activated following differentiation in neuronal cell lines. In neuronal N1E-115 cells, GTP-bound Sar1a is increased when cells elongate neuronal processes. Similar results are observed in morphological differentiation in oligodendroglial FBD-102b cells. Additionally, prolactin regulatory element binding (PREB), the GEF for Sar1 (Sar1 activator), increases the binding ability to the nucleotide-free form of Sar1a when morphological differentiation occurs. Nucleotide-free small GTPases preferentially interact with the cognate, active GEFs. These results provide evidence that using previously unreported pull down assays reveals that Sar1 and PREB are upregulated following the induction of morphological differentiation, suggesting the potential role of signaling through Sar1a during morphological differentiation.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-027 海馬歯状回顆粒細胞の発達におけるRacとCdc42の機能 Hidenori Ito（伊東秀記）¹，Rika Morishita（森下理香）¹，Makoto Mizuno（水野誠）¹，Hidenori Tabata（田畑秀典）¹，Koh-ichi Nagata（永田浩一）^1,2 ¹愛知県心身障害者コロニー発達障害研神経制御 ²名古屋大院医 Rho family small GTPases including Rho, Rac and Cdc42 are known to play pivotal roles in biological processes such as transcriptional activation, actin cytoskeletal reorganization and cellular morphology. Through the regulation of these processes, Rho GTPases and their related molecules control developmental processes such as neuronal migration, neurite elongation and branching, and synapse formation. In this study, we tried to clarify the physiological roles of Rho GTPases in the neonatal development of hippocampal dentate gyrus. When we determined the Rac1 expression in the neonatal mouse hippocampus by immunohistochemistry, Rac1 was detected in pyramidal cells in the cornus ammonis and in granule cells in the dentate gyrus. Rac1 expression was also observed in precursor cells for dentate granule neurons that resided underneath the CA3 region. Using the electroporation-mediated gene transfer method that we have established recently, we introduced knockdown vectors against Rac1 into precursors for dentate granule cells at postnatal day 0. After 21 days, Rac1-deficient cells were frequently mispositioned between the granule cell layer (GCL) and hilus. About 60% of these mislocalized cells expressed a dentate granule cell marker, Prox1. Both the dendritic spine density and the ratio of mature spine were reduced when Rac1 was silenced. Notably, the deficient cells have immature thin processes during the early neonatal migrating period. Knockdown of another Rac isoform, Rac3, also resulted in mislocalization of neonatally born dentate granule cells. In addition, knockdown of Cdc42, another Rho family protein, also caused mislocalization of the cell, although the effects were moderate compared to Rac1 and 3. Despite the ectopic localization, Rac3- or Cdc42-disrupted mispositioned cells expressed Prox1. These results indicate that Rho signaling pathways differentially regulate the proper localization and differentiation of dentate granule cells.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-028 マウス神経幹細胞に対するマイクロRNA(let7b-5p and miR409-3p)導入による効果について Ryuju Hashimoto（橋本龍樹）¹，Akihiro Matsumoto（松本暁洋）²，Hiroki Otani（大谷浩）² ¹島根大医・臨床看護 ²島根大医解剖・発生生物 We searched the microRNAs (miRNAs) which were related to histogenesis of the cerebrum in mice and analyzed their roles in the development of the cerebral cortex. We extracted total RNA from the cerebra of mouse embryos from embryonic day (E) 12 to E15 and analyzed the change in expression of miRNAs using RNA microarray assay. From these results, we chose let7b-5p, which showed the most increased miRNA level from E12 to E15 and miR409-3p, which showed the most decreased miRNA level in the same period. In this study, we analyzed functions of these miRNAs using cell culture system of mouse neural stem cells (MNSC), which were derived from the cerebrum of mouse E12 embryos, were transfected the single strand (ss) RNA of let7b-5p (ss-let 7 group), double strand (ds) RNA of let7b-5p (ds-let7 group), miR409-3p (409 group) which were labeled with fluorescein, and the ds RNA that has been confirmed not to affect the developmental process (C group). We extracted the total RNA from the MNSC of the four groups and we analyzed the change in expression of messenger (m) RNA using RNA microarray assay (RMA). We also quantified cell proliferation using measurement of DNA content (Thermo Fisher Science Inc.). By RMA, expression levels of 5 mRNAs in ss-let7 group increased more than 2-fold as compared with C group, and 13 mRNAs in ss-let7 group decreased by half or less as compared with C group. N-myc downstream regulated gene 1, which was one of the RNAs with reduced expression level after transfection in ss-let7 group, was involved in cell division. Expression level of 2 mRNAs in ds-let7 group increased more than 2-fold as compared with C group and 6 mRNAs in ds-let7 group decreased by half or less as compared with C group. Thrombospondin 1, which was one of the RNAs with reduced expression level after transfection in ds-let7 group, was involved in glial differentiation. Expression level of 2 mRNAs in 409 group increased more than 2-fold as compared with C group and 15 mRNAs in 409 group decreased by half or less as compared with C group. Thrombospondin 1 and c-fos, which were two of the RNAs with reduced expression level after transfection in 409 group, were involved in glial differentiation and neural migration. The cells in 409 group showed decreased activity of cell proliferation. These results suggested that let7b-5p and miR 409-3p are related to cell division and cell differentiation in cerebrum-derived MNSC.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-029 フィーダーフリー培養系iPS細胞由来中脳特異神経細胞の特性化 Risa Nonaka（野中里紗）^1,2，Kei-ichi Ishikawa（石川景一）^2,3，Takahiro Shiga（志賀孝宏）²，Ryota Nakamura（中村亮太）³，Genko Oyama（大山彦光）³，Nobutaka Hattori（服部信孝）³，Wado Akamatsu（赤松和土）² ¹順天堂大学院医学研究科　認知症診断・予防・治療学講座 ²順天堂大学医学部、ゲノム・再生医療センター ³順天堂大学医学部、神経学 Human iPSC-derived dopaminergic neurons are expected as one of the potential sources for cell replacement therapy in Parkinson's disease (PD). However, FACS sorting is necessary to induce iPSC-derived highly-enriched dopaminergic neurons suitable for regenerative medicine. Recently, we have developed a neurosphere-based culture system to control the regional identity of hiPSC-derived neural progenitors by using small molecules. This system facilitates effective induction of midbrain specific neural progenitors that are specified to dopaminergic neurons. In this study, we tried to fit this system into feeder-free cultured iPSCs (FF-iPS cells) to obtain midbrain specific dopaminergic neurons for regenerative medicine. We examined whether midbrain specificity of neurospheres was conserved in the neurospheres derived from feeder-free cultured iPSCs by detecting midbrain specific maker expressions. EN1( ENGRAILED1), which is expressed in the midbrain and anterior hindbrain, were highly expressed in the neurospheres treated with 3μM CHIR99021, a GSK β 3 inhibitor. In contrast, forebrain and hindbrain specific makers were hardly detected in these neurospheres. Approximately 60% of neuronal Crass III β-tubulin (Tuj-1) positive neurons induced from these neurospheres with midbrain identity were Tyrosine Hydroxylase (TH) positive, suggesting that highly enriched dopaminergic neurons could be induced from feeder-free iPSCs by optimized culture condition. We confirmed that midbrain identity of neurospheres by the expression of EN1were similar in both fibroblast and T-cell derived iPSCs. Further, frequencies of TH positive dopaminergic neurons were also similar, suggesting that our protocol shows robust neuronal differentiation efficiency regardless of the type of the original cell. We are trying to transplant these midbrain specific neurospheres into the striatum of mouse PD models to confirm safety and effectiveness of these cells for transplantation.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-030 小腸幹細胞の細胞周期における概日時計の役割 Ryuta Nakao（中尾龍太）^1,2，Yuki Maeda（前田有希）¹，Kojiro Tsujihana（辻花光次郎）^1,3，Iori Murai（村井伊織）¹，Keiko Tominaga（冨永恵子）⁴，Miho Yasuda（安田美穂）¹，Hideo Tanaka（田中秀央）²，Shinshichi Hamada（濱田新七）^2,5，Hitoshi Okamura（岡村均）¹ ¹京都大学大学院薬学研究科 ²京都府立医科大学大学院医学研究科 ³京都大学大学院医学研究科 ⁴大阪大学大学院生命機能研究科 ⁵大津市民病院病理診断科 A circadian rhythm is an endogenously generated oscillation for the daily metabolic, physiological and behavioral processes observed from unicellular animals to human. Cell reproduction by cell cycle is also a fundamental feature of all living things, and a typical proliferating human cell divides on average every 24 hours. Recent advances of circadian researches have revealed that the circadian clock controls the expression of cell cycle-related genes, and controls the timing of cell cycles. For example, we demonstrated that the timing of cell division of regenerating hepatocytes was controlled by clock-regulated Wee1 gene which changes the expression of active Cyclin B1-Cdc2 kinase, a key regulator of mitosis. However, the actual role of circadian clock in cell cycle machinery in normal tissue maintenance has not been addressed yet. Here, we analyzed the role of clock in the intestinal crypts in cell cycles in wild type (WT) and clock-gene deleted mice. Two types of cells undergo proliferative activity in intestinal epithelium; crypt base columnar (CBC) cells which are thought to be intestinal stem cells, and the transit amplifying (TA) cells which arise from the stem cells and divide a limited number of times until they produce mature cell lineages. We first examined the timing of S-phase and M-phase, and found that the timing of DNA synthesis and the mitosis occur in circadian fashion in CBC cells but not in TA cells in WT mice. In clock-gene deleted mice, no rhythms were detected in both CBC and TA cells. We next examined whether intestinal stem cells oscillate in cultured dissociated stem cells in Per2::Luc mice, and found that their luciferase activity and proliferation showed the circadian rhythms. Further, we measured the cell cycle time (Tc) by using sequential double-labelling with two thymidine analogues, BrdU and EdU, and found that Tc of CBC cells were shorter in clock-gene deleted mice than that in WT mice, although the Tc of TA cells was unaffected by the deletion of clock gene. These finding suggests that intestinal stem cells in vivo continuously self-renew and differentiate at slow speed under the circadian clock, but the highly proliferative transit amplifying cells perform their cell division without the effect of the clock.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-031 軸索誘導におけるリン脂質代謝産物の機能解析 Tomonobu Hida（肥田友伸），Tatsuya Mori（森達也），Hiroyuki Kamiguchi（上口裕之）理化学研究所脳神経科学研究センター神経細胞動態研究チーム Axon guidance is a crucial process for establishing neuronal circuits, and growth cones have pivotal roles in this process by responding directionally to attractive or repulsive guidance cues (ref. 1-3) The signaling mechanisms that control growth cone turning have been investigated over the past few decades, however, the involvement of intracellular phospholipid signaling is poorly understood. Phospholipase D (PLD) catalyzes the hydrolysis of phosphatidylcholine to generate a lipid second messenger, phosphatidic acid (PA) in plasma membrane. We found that a gradient of attractive guidance cue, such as nerve growth factor (NGF), activated PLD asymmetrically across the growth cone leading to polarized PA distribution in the growth cone plasma membrane. Furthermore, this lipid signaling is necessary for asymmetric exocytosis that drives attractive growth cone turning. To conclude, we identified PLD-PA as a novel intracellular signaling pathway important for axon guidance. Reference 1. Tessier-Lavigne M, Goodman CS. The molecular biology of axon guidance. Science. 1996;274 (5290):1123-33. 2. Tojima T, Hines JH, Henley JR, Kamiguchi H. Second messengers and membrane trafficking direct and organize growth cone steering. Nat Rev Neurosci. 2011;12(4):191-203. 3. Akiyama H, Matsu-ura T, Mikoshiba K, Kamiguchi H. Control of neuronal growth cone navigation by asymmetric inositol 1,4,5-trisphosphate signals. Sci Signal. 2009;2(79):ra34.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-032 イノシトール三リン酸受容体タイプ３による成長円錐のガイダンスシグナルに対する感受性の調節 Noriko Ooashi（大芦典子）¹，Carmen Chan（Chan Carmen）¹，Hiroki Akiyama（秋山博紀）²，Toru Matsu-ura（松浦徹）³，Tomomi Shimogori（下郡智美）⁴，Katsuhiko Mikoshiba（御子柴克彦）⁵，Hiroyuki Kamiguchi（上口裕之）¹ ¹理研CBS 神経細胞動態 ²早稲田大学人間科学学術院 ³関西医大医実験病理学 ⁴理研CBS 脳発達分子メカニズム ⁵理研CBS 発生神経生物 Neuronal wiring depends on axon path-finding, a process in which extracellular molecular cues guide a migrating growth cone toward its target. The growth cone is able to decipher directional information from guidance cues presented as shallow concentration gradients via amplification of signals. However, it remains unclear as to how the growth cone controls this amplification process during its journey through an environment in which basal concentrations of guidance cues vary widely. Here, we identified the inositol 1,4,5-trisphosphate (IP₃) receptor type 3 as a crucial regulator of axonal sensitivity to guidance cues in vitro and in vivo. Growth cones lacking IP₃ receptor type 3 are hypersensitive to nerve growth factor (NGF), an IP₃-dependent attractive cue, and incapable of turning toward normal concentration ranges of NGF to which wild-type growth cones respond. This failure in turning is due to globally, but not asymmetrically, activated Ca²⁺ signaling in the hypersensitive growth cones. Remarkably, lower concentration ranges of NGF can polarize growth cones for turning if IP₃ receptor type 3 is deficient. These data suggest a subtype-specific IP₃ receptor function in sensitivity adjustment during axon navigation. References: Akiyama, H., Matsu-ura, T., Mikoshiba, K., and Kamiguchi, H. (2009). Control of Neuronal Growth Cone Navigation by Asymmetric Inositol 1,4,5-Trisphosphate Signals. Sci. Signal. 2, ra34-ra34. Tojima, T., Hines, J.H., Henley, J.R., and Kamiguchi, H. (2011). Second messengers and membrane trafficking direct and organize growth cone steering. Nat. Rev. Neurosci. 12, 191-203.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-033 神経回路形成因子LOTUSは細胞接着分子との相互作用によって神経突起伸長を促進する Yuji Kurihara（栗原裕司）¹，Yuki Ohta（太田悠葵）²，Nana Kawasaki（川崎ナナ）²，Kohtaro Takei（竹居光太郎）¹ ¹横浜市大院　生命医　生体機能医 ²横浜市大院　生命医　創薬再生 Injured axons in the central nervous system (CNS) have a limited regenerative capacity due to axonal growth inhibitors such as Nogo protein that commonly bind to Nogo receptor-1 (NgR1). We previously reported that lateral olfactory tract usher substance (LOTUS) binds to NgR1 and suppresses these inhibitors-induced axonal growth inhibition, leading to an improvement in the regenerative capacity of injured CNS axons. Namely LOTUS functions in axon growth as an endogenous antagonist for NgR1. However, another cellular function of LOTUS remains unknown. In this study, we found that cultured retinal ganglion cell (RGC) neurons drastically extended their neurites on LOTUS substrate. This action was not attenuated in RGC neurons from Ngr1-deficient mice, showing that the promoting action of LOTUS on neurite outgrowth may be mediated by unidentified LOTUS-binding protein(s). We further found that LOTUS bound to Neuro2A cells and promoted neurite outgrowth in retinoic acid (RA)-treated Neuro2A cells, and thereby proteins expressed in Neuro2A cells were screened for LOTUS-binding protein(s) by using a liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). LC-MS/MS analysis showed that some membrane proteins such as cell adhesion molecules, transporters, epidermal growth factor-family proteins and proteases were identified as candidates for LOTUS-binding proteins. RNAi knockdown of a certain cell adhesion molecule inhibited LOTUS-promoting nerite outgrowth in RA-treated Neuro2A. These results suggest that LOTUS may promote neurite outgrowth by interacting with this cell adhesion molecule. These findings provide a novel mechanism underlying the promoting activity of LOTUS on axonal regeneration after CNS injury.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-034 神経回路形成因子LOTUSの発現制御機構 Junpei Matsubayashi（松林潤平）¹，Ryohei Nishida（西田遼平）¹，Keita Takahashi（高橋慶太）^1,2，Kohtaro Takei（竹居光太郎）¹ ¹横浜市大院生命医科学研究科・生体機能医科学研究室 ²横浜市大院医神経内科学・脳卒中医学 Neurons in the central nervous systems (CNS) undergo limited axonal regeneration after trauma and neurological disorder. Lateral olfactory tract usher substance (LOTUS) contributes to axonal tract formation as an endogenous Nogo receptor-1 antagonist. LOTUS is expected to be useful for future therapy for the CNS damage such as spinal cord injury and brain ischemia. However, the expression level of LOTUS is down-regulated and decreased by the CNS damage. How LOTUS expression is regulated remains to be clarified. In this study, we examined molecular mechanism of LOTUS expression regulation. Since immune responses induce inflammatory after trauma and neurological disorder in the CNS, we hypothesized that inflammatory substances produced by immune responses such as pro-inflammatory cytokines may down-regulate LOTUS expression level. Here, we established screening system to investigate effect of the inflammatory substances on LOTUS expression in cultured hippocampal neurons from embryonic day 18 rat fetuses. Addition of culture supernatant of spleen lymphocyte derived from experimental autoimmune encephalomyelitis (EAE) mouse to the culture media induced decrease of LOTUS expression in cultured hippocampal neurons, suggesting that the supernatant contained the factor decreasing LOTUS expression. We next examined various inflammatory substances as a candidate for a key regulator of LOTUS expression. We found that the substances A and B slightly decreased LOTUS expression level in cultured hippocampal neurons, but the substances C and D increased LOTUS expression level. Furthermore, the substance C increased LOTUS expression in mice. These findings suggest that the substances A&B and C&D may act as a negative and positive regulator of LOTUS expression, respectively. These substances may be involved in functional recovery from CNS damages through regulation of LOTUS expression level.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-035 運動誘発性成体神経新生に伴う神経回路形成因子LOTUSの発現変動 Reina Sugiyama（杉山礼奈），Ryohei Nishida（西田遼平），Kohtaro Takei（竹居光太郎）横浜市大院生命医科学 It has been well known that adult hippocampal neurogenesis (AHN) is derived from neuronal stem cell differentiation in hippocampal dentate gyrus. It has been also known that the increase of AHN in hippocampus promotes memory function. Nogo Receptor-1 (NgR1) is well known to be the common receptor of neuronal regeneration inhibitors including Nogo protein. We previously identified lateral olfactory tract usher substance (LOTUS) as an endogenous NgR1 antagonist. It is reported that genetic deletion of NgR1 inhibits AHN and impairs cognitive function. Therefore, there is a possibility that LOTUS may increase AHN and promote cognitive function. In fact, we found that LOTUS gene overexpressing transgenic (LOTUS-Tg) mice increase AHN and promote memory function, while LOTUS gene knocking out (LOTUS-KO) mice decrease AHN and impair memory function. These findings suggest that LOTUS is involved in neurogenesis and memory function. Since it has been reported that mild exercise enhances AHN, we investigated relationship between LOTUS expression level in hippocampus and exercise-induced AHN. Mice were housed in a standard cage (non-running) or in a cage with a running wheel (running) for 3 weeks. After exercise the mice were sacrificed and LOTUS expression level in hippocampus was determined by Western blotting and AHN was examined by 5-Bromo-2'-deoxyuridine (BrdU) incorporation. As expected, AHN was significantly increased in mice with exercise of 3-week-running (running mice). However, immunoblots revealed that the running mice showed a tendency of decrease of LOTUS expression in hippocampus when compared with non-running mice. Our data suggest that excise-induced AHN may be involved in regulation of LOTUS expression. It is speculated that excise-induced AHN may decrease LOTUS expression, thereby inhibition of AHN by increase of Nogo signaling as a negative feedback system. Further investigation is required for clarification of molecular mechanism regulating LOTUS expression in AHN.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-036 ニューロンIDの可視化：クラスター型プロトカドヘリンの発現解析 Ryosuke Kaneko（金子涼輔）¹，Manabu Abe（阿部学）²，Yusuke Takatsuru（高鶴裕介）³，Yukiko Ueno Inoue（井上上野由紀子）⁴，Masahiko Watanabe（渡辺雅彦）⁵，Kenji Sakimura（崎村建司）²，Yuchio Yanagawa（柳川右千夫）¹，Takeshi Yagi（八木健）⁶ ¹群馬大院医 ²新潟大学脳研究所 ³中沢会上毛病院 ⁴国立精神・神経医療研究センター　神経研究所 ⁵北海道大院医 ⁶大阪大院生命機能 Molecular tags, which organize complex networks in mammalian brain, could specify neuronal identity and should have diverse characteristics participating in discrimination between individual neurons. It has been speculated that clustered protocadherins (Pcdhs), which encode cadherin-related transmembrane proteins as gene clusters in vertebrate genome, could provide these kinds of molecular tags. The murine Pcdhs are further classified into three subfamilies: Pcdh-a (14 genes), Pcdh-b (22 genes), and Pcdh-g (22 genes). Their loss of function in mice revealed that the Pcdhs play important roles in neuronal survival, axonal projection, synaptic connectivity, and several brain functions including learning and memory. As revealed by histological examinations and single-cell RT-PCR, the Pcdhs show the scattered expression in each cerebellar Purkinje cell. The scattered expressions of the Pcdhs will provide a potential neuronal identity at the single-neuron level. The involvement of the scattered Pcdh expression in neural circuit formation has been inferred on the basis of several genetic analyses including loss of Pcdh-g and loss of gene regulators of the Pcdhs (CTCF and Dnmt3b). However, several key questions remain unanswered. For example, are the Pcdh expressions scattered in other cell types? How frequently does each cell type express one Pcdh isoform? Are the Pcdh expressions spatially different between individual animals? What condition alter Pcdh expression? In order to answer these questions, we generated knock-in mice that harbor cDNA encoding red fluorescent protein, tdTomato, under the control of endogenous Pcdh-b3 promoter. The mice showed scattered tdTomato fluorescence in various cell types, including cerebellar Purkinje cells, hippocampal CA1 pyramidal cells, dentate gyrus granule cells, cerebellar molecular layer interneurons, astrocyte etc. However, none of microglia expressed tdTomato. The percentage of cells expressing tdTomato at 4-week-old was 2% in cerebellar Purkinje cells and 0.2% in hippocampal CA3 pyramidal cells, suggesting differences in gene regulatory mechanism and/or expression level between these cell types. Interestingly, spatial distributions of tdTomato revealed inter-individual variation. We are currently addressing the key questions about the scattered Pcdh expression.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-037 ライブセルイメージングを用いた神経細胞におけるクラスター型プロトカドヘリンのホモフィリック相互作用の解明 Natsumi Hoshino（星野七海），Etsuko Tarusawa（足澤悦子），Hiroaki Kobayashi（小林裕明），Takeshi Yagi（八木健）大阪大院生命機能心生物学 Neural networks are precisely formed via specific synaptic connections. It is thought that cell surface recognition molecules play an essential role in forming the synaptic connections, and clustered protocadherins (cPcdhs) which have diverse isoforms are likely candidate molecules. In mice, cPcdhs consist of 58 isoforms and about 15 isoforms out of 58 are expressed in each individual neuron with different combinations. They form various dimers in each neuronal surface so that the variety of surface recognition is defined by the combination of their expression. It has been shown in the adhesion assay of non-neuronal cells that the trans homophilic interaction is strict and depends on the complete matching of isoform combination between adhering cells. This result suggests that cPcdhs generate neuronal individuality and function as recognition molecules between neurons to form proper neuronal networks through their homophilic interactions. However, their homophilic interactions between neurons have not been observed yet, because endogenous cPcdhs bind to exogenous cPcdh and inhibit its homophilic interaction. Here we aim to visualize the homophilic interaction of an exogenous cPcdh isoform between neurons. Multi-color time lapse imaging (different color sets for different neurons for cell morphology and fluorescent-tagged cPcdh proteins) in cultured cPcdh-deficient neurons allows us to analyze the dynamics and the localization of the cPcdh homophilic interaction at cell-cell contacts. We found that cPcdh proteins are highly expressed in extending axons and dendrites in immature neurons. The cPcdh proteins at the shaft of dendrites spontaneously enter into the filopodia. The co-localization of cPcdh proteins between different neurons is formed quickly after the touch of their neural fibers, and then the co-localization of cPcdh proteins is comparatively stable. Based on these findings, we will discuss roles of the homophilic interaction between neurons during neural network formation.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-038 神経の発生と可塑性に関与するリン酸化タンパク質GAP-43の解析 Masayasu Okada（岡田正康）^1,2,3,4，Asami Kawasaki（河嵜麻美）^3,4，Atsushi Tamada（玉田篤史）^3,4,5，Manabu Natsumeda（棗田学）²，Makoto Oishi（大石誠）²，Kazunobu Sawamoto（澤本和延）⁶，Kosei Takeuchi（武内恒成）^7,8，Yukihiko Fujii（藤井幸彦）²，Michihiro Igarashi（五十嵐道弘）^3,4 ¹新潟大医歯総合病院 ²新潟大脳研　脳神経外科分野 ³新潟大学研究推進機構超域学術院 ⁴新潟大院医歯学神経生化学分野 ⁵関西医大医 iPS・幹細胞応用医学講座 ⁶名古屋市大医再生医学 ⁷愛知医科大医学部細胞生物学 ⁸愛知医科大研究創出支援センター Elucidation of the mechanisms by which an axon grows at developmental stages, will contribute to the understanding of nerve regeneration as well as of the plasticity of neural circuits. Some researchers discovered GAP-43 as a phosphoprotein during long-term potentiation in the hippocampus, and axonal transport of this molecule is known to increase when an axon grows and regenerates. Thus, GAP-43 is thought to be important in the rearrangement of neuronal circuit and plasticity. Not only in the developing brain but also in the mature one, however, its exact roles are not clear. Here, we focused on the phosphorylation of GAP-43. As a result of phosphoproteomic analysis of the growth cone membranes, its serine (S) 96 and threonine (T) 172 were highly phosphorylated. We discovered that JNK is responsible for phosphorylation of these sites. So far, we reported that pS96 is a maker of axonal growth and regeneration after rodent nerve development and nerve injury (Kawasaki, Okada, Tamada et al.: iScience 2018). On the other hand, GAP-43 T172 is a more highly conserved sequence not only from rodents but also primates. Therefore, we started the study as a molecular marker involved in neural development and regeneration in primates. The antibody (Ab) against pT172 was confirmed its specificity in various regions of wild type fetal brain and those of the knock-in mice, in which pT172 was inactivated by amino acid substitution (T172A). The histological and the biochemical analyses using this pT172 Ab revealed that even the primate neurons, such as in marmoset brain and derived from human iPS cells, reacted with this Ab. In addition, histological analysis was also performed using tissue of hippocampal sclerosis as a disease model showing plasticity. In the hippocampal sclerosis tissue, pT172 was colocalized with synaptophysin, a presynaptic marker than GAP-43 itself. As a conclusion, pT172 is expressed in the elongating nerve of not only rodents but also primates. Taken together, our observations suggest that pT172 is expressed in the developing and the regenerating axons.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-039 発生脳におけるMAP1Bの高頻度リン酸化部位 Yuya Ishikawa（石川裕也）¹，Atsuko Honda（本多敦子）²，Yasuyuki Ito（伊藤泰行）²，Asami Kawasaki（河嵜麻実）²，Atsushi Tamada（玉田篤史）²，Naoto Endo（遠藤直人）¹，Michihiro Igarashi（五十嵐道弘）² ¹新潟大院医歯整形 ²新潟大院医歯分子細胞機能 The growth cone is an essential structure for formation and rearrangement of the neuronal network. Since phosphorylation is the most important signaling modification in all of the cells including neurons, we focused a new technique "phosphoprotemics", which can identify both the phosphorylation sites of proteins and their frequency simultaneously and help us understanding the comprehensive signaling pathways in a given system. To clarify the molecular mechanisms operating growth cone behaviors, we have performed phosphoproteomic analysis of the growth cone membrane (Kawasaki A, et al. iScience, 2018). This approach identified 4,600 phosphorylation sites corresponded to 1,200 proteins. Among these sites, we focused that two of the most abundant sites were Ser25 and Ser1201 of rat microtubule-associated protein 1B (MAP1B). MAP1B is a protein known to stabilize axonal microtubules and to be involved in axon formation. Although phosphorylation of MAP1B is reported by the distal portion of axons, its biological function is not well known. So far, the phosphorylation sites that we identified have been hardly elucidated. These two sites are highly conserved between rodents and human, suggesting that they have important functions. We produced the phospho-specific antibodies against these sites, and obtained a result that a developing axon is preferentially labelled by these antibodies in cultured mouse cortical neurons. In addition, we found that these antibodies successfully recognized the growing axons in embryonic mouse brain using immunohistochemistry. Western blotting analysis revealed that phosphorylation of these sites gradually decreased after development. These data suggest that these phosphorylation sites of MAP1B are spatiotemporally regulated in neuronal development.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-040 軸索誘導因子SLIT2は霊長類高次連合野に発現する -RBP4 ・PNMA5 との発現比較- Tetsuya Sasaki（佐々木哲也）^1,2,5，Yusuke Komatsu（小松勇介）^3,5，Tetsuo Yamamori（山森哲雄）^4,5 ¹筑波大・医　生命医科学・解剖学・神経科学 ²筑波大院　人間総合科学感性認知脳科学 ³(株)ACD ⁴理研BSI高次脳機能分子 ⁵基生研　脳生物 To elucidate the molecular basis for the specialization of cortical architectures, we searched for genes differentially expressed among neocortical areas of Old World monkeys. We previously reported that SLIT1, an axon guidance molecule, is abundant in the prefrontal cortex but with developmentally related changes. SLIT is a chemorepellent guidance molecule, which is well conserved in various species. The chemorepellent effect of SLIT is mediated by receptor, Roundabout (ROBO). In situ hybridization analysis revealed that SLIT1 mRNA was mainly distributed in the middle layers of most cortical areas, robustly in the prefrontal cortex and faintly in primary sensory areas (Sasaki et al., 2010). Our comprehensive expression analyses of other SLIT (SLIT2 and SLIT3) mRNAs showed enriched expression in the higher-order association areas with a distinct laminar pattern. Among them, the SLIT2 mRNA expression was high in layers II, III and V in the prefrontal association area. On the other hand, the excitatory population that expressed weak SLIT2 mRNA signals was restricted to the upper part of the supragranular layers of the primary visual area. These patterns are reminiscent of those of RBP4 and PNMA5 mRNAs, which were identified as an association area-enriched gene (Komatsu et al., 2005; Takaji et al., 2009; Yamamori 2011). Scattered signals of SLIT2 mRNA were identified in layers III-VI were in GAD67-mRNA-positive inhibitory neurons. Double ISH analysis showed that SLIT2 and RBP4 mRNAs were colocalized in the cortical neurons. Thus, SLIT2, RBP4 and PNMA5, whose expressions show the RBP4-like expression in the cortex, may all exert influence over a similar type of cortical neuron.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-041 Brainbow法と組織透明化によるニワトリ脳幹聴覚神経回路の3次元形態解析 Hiroshi Sekikawa（関川博）¹，Ryo Egawa（江川遼）²，Hiroshi Kuba（久場博司）² ¹名古屋大医 ²名古屋大院医細胞生理 Auditory brainstem circuit contributes to sound localization by detecting interaural time difference (ITD) on the order of microseconds at each sound frequency. In avian brainstem, the ITD detection is supported by the precise neuronal wiring from nucleus magnocellularis (NM) to nucleus laminaris (NL), which shows tonotopic organization and delay line pattern. However, it is not well understood how this characteristic neural circuit is constructed during embryonic development due to the labor-intensive process of single axon tracing. In this study, we established an experimental system which enables 3-D visualization of whole avian auditory brainstem circuits using Brainbow labeling and tissue clearing method. Plasmid vectors which randomly express three kinds of fluorescent proteins were introduced into NM progenitor cells at embryonic day 1.5 (E1.5) by in ovo electroporation method. At around E12, the 1.5-mm thick brainstem slices were cleared with CUBIC protocol and imaged under confocal microscopy. We found that the NM neurons were labeled with various colors and the individual axons were traceable using the color differences. Single axon tracing visualized the characteristic wiring and allowed the comparative analysis at each developmental stage. From the above, our system would facilitate further understanding of the developmental mechanisms of auditory brainstem circuits.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-042 フェレットとマウスにおける形成期大脳皮質の神経線維層の解析 Kengo Saito（齋藤健吾），Keishi Mizuguchi（水口敬司），Toshihide Horiike（堀池俊秀），Tung Anh Dinh Duong（Dinh Duong Tung Anh），Yohei Shinmyo（新明洋平），Hiroshi Kawasaki（河崎洋志）金沢大院医脳神経医学 Changes in the cerebral cortex of mammals during evolution have been of great interest. Ferrets, monkeys, and humans have more developed cerebral cortices compared with mice. Although the features of progenitors in the developing cortices of these animals have been intensively investigated, those of the fiber layers are still largely elusive. Interestingly, although the two fiber layers, the inner fiber layer (IFL) and the outer fiber layer (OFL), were identified in the cerebral cortex of monkeys and humans during development, corresponding structures in rodents and ferrets were unclear. By taking the advantage of our in utero electroporation technique for ferrets, we found that ferrets also have the two fiber layers in the developing cerebral cortex, as is the case in monkeys and humans. We therefore systematically investigated the cellular origins and projection patterns of axonal fibers in the IFL and the OFL in the developing ferret cortex. We found that axonal fibers in the IFL projected contralaterally and subcortically, whereas those in the OFL sent axons to neighboring cortical areas. Furthermore, we performed similar experiments using mice and found that fibers corresponding to the IFL and the OFL in ferrets were also present in mice. These findings raised the possibility that both IFL-like and OFL-like fibers commonly exist among various mammals. Our results shed light on the cellular origins, the projection patterns, the developmental processes, and the evolution of fiber layers in mammalian brains.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-043 PTPδノックアウトマウスにおける小脳・登上線維の表現型解析 Erika Shimizu（清水恵利香）^1,2，Aoi Jitsuki（實木葵）²，Fumio Nakamura（中村史雄）² ¹東京女子医大医学部 ²東京女子医大院医生化学 In mature cerebellar cortex, one Climbing fiber (CF) forms synapses on one Purkinje cell (PC). This one-to-one connection is important to proper cerebellar function. This projection is accomplished by synaptic pruning. During the first three postnatal weeks, multiple CFs make synapses on one PC. Afterward, only one CF is chosen and strengthened, the other redundant synapses are eliminated. Several molecules including Sema3A and Sema7A have been shown to be involved in this process, however, the molecular mechanism of CF-PC maturation is yet completely understood. We previously demonstrated that PTPδ, one of type IIa receptor protein tyrosine phosphatases, is involved in the development of cortical pyramidal dendrites. We also observed the expression of PTPδ in inferior cerebellar peduncle, which contains CFs from inferior olive nucleus. Therefore, we examined whether PTPδ is involved in the developing CF-PC synapse in vivo. The sections from wild-type and PTPδ knockout cerebella were double-immunostained with anti-VGlut2 (CF marker) and anti-Calbindin (PC marker) antibodies. We assessed them at several developmental stages, P17, P22, and P25. While PTPδ knockout cerebella from P17 did not show particular phenotype, the sections from P25 exhibited thinner molecular layer than wild type especially on anterior lobe, lobe1 to lobe5. The P25 sections displayed shallow CFs invasion into cerebellar molecular layer. The sections from P22 was intermediate. These results suggest that PTPδ may participate in the maturation of CF-PC synapses in developing cerebellum. The abnormalities of specific molecular layer and CFs may have an influence on functional impairment. We are designing further experiments by behavioral and electrophysiological analysis of PTPδ knockout mice.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-044 小脳プルキンエ細胞の樹状突起形成におけるphospholipase Cβの役割 Masahiko Tanaka（田中正彦），Miyu Naruse（成瀬美結），Naohide Hirashima（平嶋尚英）名市大院・薬・生体超分子システム解析学 Cerebellar Purkinje cells have the most elaborate dendritic trees among neurons in the brain. Dendritic trees of Purkinje cells receive numerous glutamatergic synaptic inputs from granule cells, forming a main component of cerebellar neuronal circuitry. Purkinje cell dendrites and granule-Purkinje synapses are formed during postnatal cerebellar development. Previous studies showed that glutamate and its downstream Ca²⁺ signaling are important for the differentiation of Purkinje cell dendrites. In the present study, we investigated the role of phospholipase Cβ (PLCβ), a key component in glutamate-Ca²⁺ signaling, in the dendritic differentiation of Purkinje cells, using an inhibitor and siRNA against PLCβ. When a PLC inhibitor U-73122 was added to the culture medium for 10 days, the elongation and branching of Purkinje cell dendrites were markedly inhibited. Unexpectedly, however, Purkinje cell death was induced when U-73343, an inactive analog of U-73122, was added to the culture medium. To precisely examine a role of PLCβ, we transferred siRNA against PLCβ3 and PLCβ4 into Purkinje cells in cerebellar cell cultures, using single-cell electroporation technique (J. Neurosci. Meth. 178: 80-86, 2009; Neuromethods 65: 129-139, 2012). Single-cell electroporation enables us to transfer siRNA into specific cells in the environment composed of different types of cells. In addition, we can easily and reliably transfer multiple siRNAs into a cell simply by loading them together in one micropipette. PLCβ3 and PLCβ4 are the two major isoforms of PLCβ expressed in Purkinje cells. Although PLCβ3 and PLCβ4 show complementary expression patterns in the cerebellum in vivo, it is reported that Purkinje cells lose such heterogeneity of PLCβ isoform expression and individual Purkinje cells express both PLCβ3 and PLCβ4 in culture (Kamikubo et al., 2007). Therefore, we transferred two types of siRNA, one against PLCβ3 and the other against PLCβ4, simultaneously. Alternatively, we transferred one type of siRNA against both PLCβ3 and PLCβ4. Ten days after both types of transfection, dendritic branching of the electroporated Purkinje cells was significantly inhibited. These results suggest that PLCβ3 and PLCβ4 promote dendritic differentiation of Purkinje cells.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-045 ノックアウトマウスを用いたSema7Aの発現解析 Yuka Ohyama（大山裕可）^1,2，Aoi Zitsuki（實木葵）²，Fumio Nakamura（中村史雄）² ¹東京女子医科大学医学部 ²東京女子医科大院医生化学 Axon guidance molecules regulate neural circuit formation during embryonic and early postnatal stages. Semapjorin-7A (Sema7A) regulates the projection of olfactory tract, however, its expression sustains in the nervous system of mature mice. In addition, the protein expression pattern of Sema7A has not been examined in detail. In order to investigate the postnatal function of Sema7A in the central nervous system, we examined the expression pattern of Sema7A in murine brains (2 Mo) by immunohistochemistry. To validate the specific expression of Sema7A, the sections from wild-type and Sema7A knockout mice were immunostained with anti-Sema7A antibody in the same condition. Sema7A expression was observed in olfactory bulb, neocortex, basal ganglia, thalamic nuclei, inferior colliculus, cerebellum, pons, and medulla oblongata. Strong immunoreactive signal was observed in glomerular and mitral cell layers of olfactory bulb, cortical II, III, V and VI layers, caudate putamen, pontine nuclei, and molecular layer of cerebellum. Hippocampal sections were darkly stained except pyramidal cell layer in CA1 to CA3 and granule cell layer in dentate gyrus. These results well correlate with previous observations. For example, Sema7A expression in cerebellar molecular layer has been shown to be involved in the pruning of climbing fibers. Sema7A expression in dentate gyrus regulates the number of dendrites from molecular cells. However, the postnatal function of Sema7A in cerebral cortex and striatum has not been addressed. We will investigate neuroanatomical and behavioral analyses of the knockout mice to reveal the neuronal function of Sema7A in neocortex and striatum.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-046 プロスタグランジンD2はDP1受容体を介して神経細胞の発達や形態に影響を及ぼす Atsuko Hayata（早田敦子）^1,2，Ikuko Mohri（毛利育子）³，Kenta Okuda（奥田健太）²，Masaya Tachibana（橘雅弥）³，Taiga Kikuchi（菊池泰河）²，Yusuke Shintani（新谷勇介）²，Takanobu Nakazawa（中澤敬信）^2,4，Masako Taniike（谷池雅子）³，Hitoshi Hashimoto（橋本均）^1,2,5,6 ¹大阪大院連合小児発達分子統御機構研セ ²大阪大院薬　神経薬理 ³大阪大院連合小児 ⁴大阪大院歯薬理 ⁵大阪大データビリティフロンティア機構 ⁶大阪大　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 Prostaglandin D2 (PGD2) is an inflammatory mediator produced by the activities of cyclooxygenase (COX) and either hematopoietic PGD synthase (HPGDS) or lipocalin-type PGD synthase (L-PGDS). In the central nervous system (CNS), L-PGDS is mainly expressed in oligodendrocytes and meninges while HPGDS is expressed in microglia. In the CNS, PGD2 also plays important roles as a neuromodulator in the regulation of the sleep-wake cycle, body temperature, hormone release, and pain responses via the G protein-coupled receptors DP1 and DP2/CRTH2. Previously, we found that HPGDS is expressed in activated microglia during perinatal period when the neural circuits develop and synapses are actively formed and pruned in the CNS. This finding suggests that PGD2 might be involved in neuronal development. However, the role of PGD2 signaling in the neuronal development remains largely unknown. In this study, we investigated whether PGD2 is responsible for mediating neuronal morphological development via DP1 and/or DP2 receptors. First, we examined DP1 and CRTH2 expression levels in the primary cultured cortical neurons. RT-PCR revealed the expression of DP1, but the CRTH2 expression was lower than the detection limit. At days in vitro (DIV) 5, the DP1-selective agonist BW245C, but not the CRTH2-selective agonist 13,14-dihydro-15-keto prostaglandin D2, increased the primary neurite number and total length of dendrites in the primary cultured cortical neurons. BW245C increased the PSD-95-negative dendritic spines at DIV 21 in primary cultured cortical neurons. In addition, repeated administration of BW245C to the mouse during early postnatal period caused alterations in the dendritic morphology and the number of synapses in the prefrontal cortex. These results suggest that PGD2 is implicated in neuronal development via DP1 receptor and the over-activation of PGD2-DP1 signaling during early development could be a risk factor for neurodevelopmental disorders.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-047 神経突起とスパイン形成を制御する新規キナーゼLMTK1のキナーゼ活性について Anni Huo（カクアンニ），ran wei（wei ran），Toshiyuki Takasugi（高杉俊之），Koji Tsutsumi（堤弘次），Kanae Ando（安藤香奈絵），Shinichi Hisanaga（久永眞市）首都大学東京 Lemur kinase 1(LMTK1) is an uncharacterized serine/threonine kinase highly expressed in mammalian brain. LMTK1 was originally reported as a tyrosine kinase upregulated at the time of apoptotic cell death, thus, it was called previously AATYK (apoptosis-associated tyrosine kinase). As a matter of fact, however, LMTK1 is expressed abundantly in neurons and has a primary role in regulation of axon and dendrite outgrowth and spine formation through recycling endosomal trafficking in a Rab11 GAP TBC1D9B-depandent manner. However, it is not known how LMTK1 regulates TBC1D9B. This is because the kinase activity of LMTK1 has not been clearly demonstrated. We thought it important to detect the kinase activity of LMTK1 to understand its function convincingly. In this study, we tried to detect the kinase activity of LMTK1 by several assay methods with various LMTK1 mutants including the kinase domain fragments. Because LMTK2 is reported to display the activity against myelin basic protein (MBP), we tested phosphorylation of MBP by immunoprecipitated LMTK1 in the presence of γ -32p-ATP. But we could not detect 32p incorporation into MBP more than control lacking LMTK1 expression or kinase-deficient-mutant LMTK1 D205V. We also analyzed the autophosphorylation of immunoprecipitated LMTK1-myc because many protein kinases have the autophosphorylation activity. However we did not find any autophosphorylation signaling more than control. LMTK1 has a long C-terminal tail. We through that the C-terminal tail in an inhibitory domain against the kinase activity. Then, we constructed the N-terminal fragment composed of only the kinase domain, but we failed to detected the activity of those fragments. Previous report described the kinase activity of LMTK1 in cerebellar granule neurons undergoing cell death. Now, we are going to detect the kinase activity of LMTK1 by using neuronal cell lines treated with cAMP for differentiation or ionomycine for cell death induction. We would like to show the distinct kinase activity of LMTK1 at the meeting.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-048 B4galt5と6遺伝子によってコードされるラクトシルセラミド合成酵素は神経系構築とミエリン鞘形成に必須である Toru Yoshihara（吉原亨）^1,2，Hiroyuki Satake（佐武寛之）²，Toshikazu Nishie（西江敏和）²，Nozomu Okino（沖野望）³，Toshihisa Hatta（八田稔久）⁴，Hiroki Otani（大谷浩）⁵，Chie Naruse（成瀬智恵）^1,2，Hiroshi Suzuki（鈴木紘史）²，Kazushi Sugihara（杉原一司）^1,2，eikichi Kamimura（神村栄吉）²，Noriyo Tokuda（徳田典代）⁶，Keiko Furukawa（古川圭子）⁷，Koichi Furukawa（古川鋼一）^6,7，Makoto Ito（伊藤信）³，Masahide Asano（浅野雅秀）^1,2 ¹京都大学医学研究科附属動物実験施設 ²金沢大学学際科学実験センター ³九州大学農学研究院 ⁴金沢医科大学分子細胞形態学 ⁵島根大学医学部 ⁶名古屋大学医学部 ⁷中部大学生命健康科学部 It is not certain which β4-galactosyltransferase (β4GalT; gene name, B4galt), β4GalT-5 and/or β4GalT-6, is critical for the production of lactosylceramide (LacCer) synthase, which functions in the initial step of ganglioside biosynthesis. Here, we generated nervous system specific B4galt5 knockout (B4galt5 cKO) mice, using Nestin-Cre mice, and crossed these with B4galt6 KO mice to generate B4galt5 and 6 double KO (DKO) mice in the central nervous system (CNS). LacCer synthase activity and major brain gangliosides were completely disappeared in brain samples of the DKO mice, although LacCer synthase activity was approximately half its normal level in B4galt5 cKO mice and B4galt6 KO mice. The DKO mice were born normally but they showed severe growth retardation and motor deficits at 2 weeks and died by weaning period (4 weeks of age). In histological analyses, myelin-associated proteins (MBP, MAG, MOG and PLP) were rarely found localized in axons in the cerebral cortex, and axonal and myelin formation were remarkably disturbed in the spinal cords of the DKO mice. Neuronal cells, differentiated from neurospheres that were prepared from the DKO mice, showed impairments in neurite outgrowth and branching formation, which can be explained by the fact that neurospheres from DKO mice could weakly interact with laminin due to lack of gangliosides, such as GM1a. In addition, the neurons showed immature traits and perineuronal nets (PNNs) were poorly formed in DKO cerebral cortices. These results indicate that LacCer synthase is encoded by both B4galt5 and 6 genes in the CNS, and that gangliosides are indispensable for neuronal maturation, PNN formation, and axonal and myelin formation.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-049 PTP deltaノックアウトマウス脳におけるSIRP alphaの過剰リン酸化 Miyu Wakatsuki（若槻実祐）東京女子医科大学医学部生化学教室 Hyperphosphorylation of SIRP alpha in PTP delta knockout mouse brains Miyu Wakatsuki 1, 2), Fumio Nakamura 2, 3) 1) School of Medicine, Tokyo Women's medical university 2) Department of Biochemistry, School of Medicine, Tokyo Women's medical university 3) Department of Pharmacology, School of Medicine, Yokohama City University Protein tyrosine phosphatase delta (PTP delta) is one of vertebrate Leukocyte common antigen-related (LAR) class PTPs. We previously reported that PTP delta is involved in the development of cortical pyramidal neurons through the activation of Fyn tyrosine kinase. However, in vivo substrates of PTP delta are largely unknown. As we observed hyperphosphorylated proteins in PTP delta knockout mouse brains by immunoblotting with anti-phosphotyrosine (pY) antibody, we carried out phospho-proteomics analysis of the knockout and wild-type brains (postnatal day 25). Among the identified proteins, Signal Regulatory Protein alpha (SIRP alpha) gave the strongest phosphorylated ratio of knockout to wild-type. We then examined the phosphorylation of SIRP alpha in mouse brains (postnatal day 22) using antibodies against phosphotyrosine (pY), SIRP alpha, and phosphorylated Tyr501 in SIRP alpha (pY501). Combination of SIRP alpha-immunoprecipitation and pY-blotting as well as anti-pY501 immunoblotting revealed the hyperphosphorylation of SIRP alpha in the knockouts. We next examined immunohistochemistry. Almost entire CNS regions were immunostained with anti-pY antibody in both genotypes, however, the signal was stronger in the knockout sections than in wild-type. Cortical layers from II to V, reticular thalamic nucleus, and subthalamic nuclei were hyperphosphorylated in the knockouts. Phospho-Y501 SIRP alpha immunostaining revealed that in the knockout sections, olfactory bundle, cortical II and III layers, striate body, fimbria, corpus callosum, reticular thalamic nucleus, ethmoid thalamic nucleus, and pyramidal tract were hyperphosphorylated. In addition, rostral migratory stream (RMS) was strongly immunostained with pY501 SIRP alpha in both genotypes. SIRP alpha-hyperphosphorylated regions in the knockouts partially correlated with the regions expressing PTP delta in wild-type including neocortex and thalamic reticular nucleus. This suggests that SIRP alpha may be one of the endogenous substrates of PTP delta.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-050 トランスサイトーシスを介したTrkAシグナル複合体の軸索への局在化 Naoya Yamashita（山下直也），Takashi Sakurai（櫻井隆）順天大院　医細胞・分子薬理 In the nervous system, communication between neurons and their post-synaptic target cells is critical for the formation, refinement and maintenance of functional neuronal connections. Diffusible signals secreted by target tissues influence diverse neuronal developmental events. Neurotrophins are one of the best-known examples of target-derived instructive cues that regulate distinct aspects of neuronal development. Upon nerve growth factor (NGF), a prototypical neurotrophin, binds to its receptor TrkA at axon terminals, these complexes are internalized and then retrogradely transported back to cell bodies. We have previously found the new function of retrograde signaling by which target-derived NGF enhances soma-to-axon transcytosis of TrkA. However, it is unclear whether the transcytosis machinery is a general mechanism which NGF utilizes to recruit additional membrane proteins necessary for axon growth and synapse maturation. We show evidence that NGF enhances soma-to-axon transcytosis of amyloid-beta precursor protein (APP). APP interacts with TrkA at the extracellular domain just adjacent to the transmembrane domain. Ectopic expression of APP induces the formation of neurite-like processes in non-neuronal cells, which might be mediated by TrkA-interacting domain of APP. Moreover, NGF regulates proteolytic processing of APP, a crucial determinant of APP function. Since APP and its proteolytic products play an important role in pathogenesis of Alzheimer's disease (AD), our data suggests that the TrkA-APP anterograde transcytosis is involved in NGF functioning and its defects might be related to the onset of AD.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-051 セルタイプ特異的に発現するスプライシング因子SLM2がGABA作動性介在性ニューロンのNeurexin依存的なシナプス特性に果たす役割 Yuji Sato（佐藤悠司）¹，Satoko Suzuki（鈴木暁子）²，Yoko Iijima（飯島陽子）^1,2，Takatoshi Iijima（飯島崇利）^1,2 ¹東海大学医学部基礎医学系　分子生命科学 ²東海大学創造科学技術研究機構　　医学部門 GABAergic interneurons occur in many different subtypes, that play exquisitely precise functions in neural networks. Each type of interneuron is highly selective, making synapses with particular populations of target cells and only with specific subcellular compartments. Neurexins (NRXs) play important roles in the synapse specification with several trans-synaptic receptors, Neuroligins, LRRTMs and Cbln-GluD complexes in central nervous system. The alternatively spliced segment 4 (AS4) of NRX genes (Nrxn) is a key element in selective trans-synaptic interactions. The incorporation of exon 20 of neurexin genes (Nrxn) in alternatively spliced segment 4 (AS4) generates NRX4(+) protein variants containing a 30-amino acid insertion, whereas skipping exon 20 results in the production of the NRX4(-) variant. Importantly, this alternative splicing of Nrxn at AS4 underlies a synaptic adhesive code between glutamatergic and GABAergic synapses. However, the splice code on NRXs on synapse specification and function on the GABAergic interneuron diversity is still not fully understood. To examine the distinct role of NRX AS4 isoforms between GABAergic interneurons, we initially analyzed the expression of SLM2, a selective and dominant regulator of AS4 splicing, in the interneurons of adult brains. We previously showed that SLM2 expression was quite low in all the cerebellar GABAergic neurons. Similarly, we found that the majority of interneurons did not express SLM2 in cortical interneurons. However, subpopulation of interneuron subtypes expressed high level of SLM2. We then ectopically expressed SLM2 in the interneurons in cultures, and evaluated the synaptogenic receptor activity of Neuroligins in a neuron-fibroblast co-culture system. In cerebellar neurons, Neuroligins induce VGAT+ GABAergic contacts in the control culture, but not properly in the SLM2 E/E culture. Lentivirus-based expression of Nrxn3 containing exon 20 restores the reduced NL-induced GABAergic contacts in the SLM2 E/E co-culture. This isoform-selective effect indicates the potential role of AS4 insertion in presynaptic differentiation of GABAergic cerebellar neurons. We are further analyzing effect of ectopic SLM2 expression on synaptogenic receptor activity between cortical interneuron subtypes. Our findings may provide further insights into understanding synapse specificity and diversity of interneuron networks.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-052 膜電位の光学的測定法によるマウス舌咽神経回路網の機能発生過程の解析 Katsushige Sato（佐藤勝重）¹，Yoko Momose-Sato（佐藤容子）² ¹駒沢女子大人間健康健康栄養 ²関東学院大栄養管理栄養 The glossopharyngeal nerve (N.IX) transfers motor and sensory information related to visceral and somatic functions, such as salivary secretion, gustation and the control of blood pressure. N.IX-related neural circuits are indispensable for these essential functions. Compared with the strenuous analysis of morphogenesis, we are only just starting to elucidate the functiogenesis of these neural circuits during ontogenesis. In the present study, we applied voltage-sensitive dye recording to the embryonic mouse brainstem, and examined the functional development of the N.IX-related neural circuits. First, we optically identified the motor nucleus (the inferior salivatory nucleus (ISN)) and the first-order sensory nucleus (the nucleus of the tractus solitarius (NTS)). We also succeeded in recording optical responses in the second/higher-order sensory nuclei via the NTS, including the parabrachial nucleus. Second, we pursued neuronal excitability and the onset of synaptic function in the N.IX-related nuclei. The neurons in the ISN were excitable at least at E11, and functional synaptic transmission in the NTS was first expressed at E12. In the second/higher-order sensory nuclei, synaptic function emerged at around E12-13. Third, by mapping optical responses to N.IX and vagus nerve (N.X) stimulation, we showed that the distribution patterns of neural activity in the NTS were different between N.IX and N.X from the early stage of ontogenesis. We discuss N.IX-related neural circuit formation in the brainstem, in comparison with our previous results obtained from the chick and rat embryos.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-053 膜電位の光学的測定法によるマウス前庭神経核の機能発生過程の解析 Yoko Momose-Sato（佐藤容子）¹，Katsushige Sato（佐藤勝重）² ¹関東学院大栄養管理栄養 ²駒沢女子大人間健康健康栄養 One major challenge in developmental neurobiology is clarifying when and how the brain is functionally organized during embryogenesis. Although such investigations are significant, they have been hampered by the limited conventional electrophysiological means available for immature neurons and assessing spatio-temporal patterns of neural responses. In the present study, a multiple-site optical recording technique with a voltage-sensitive dye was applied to the mouse embryo, and functional organization of the vestibular nucleus was examined. Stimulation of the vestibular nerve in E12-13 mouse brainstems elicited fast and slow optical signals, which corresponded to the action potential and the excitatory postsynaptic potential (EPSP), respectively. The EPSP was mediated by glutamate and was sensitive to extracellular Mg²⁺, which suppresses the NMDA receptor. In the E13 embryo, the EPSP-related signals were detected from the region extending longitudinally to the levels rostral and caudal to the vestibular ganglion, with high signals concentrated in the caudal region. At E12, the EPSP was lower and generally restricted to the caudal region even when extracellular Mg²⁺ was removed to enhance the glutamate receptor function. These results suggest that the developmental sequence of functional synaptic expression is different between the vestibular subnuclei, and that the EPSP initially appears in the caudal vestibular nucleus in the mouse embryo.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-054 CDP-ジアシルグリセロール合成酵素であるCdsAはショウジョウバエにおいてPIP2依存的に神経細胞を変性から保護する Yohei Nitta（新田陽平），Atsushi Sugie（杉江淳）新潟大学研究推進機構超域学術院 Lipids are the major component of the cells and function not only as energy storage, but as membrane anchoring, protein trafficking, and signaling molecules. Recent studies suggest that lipid metabolism regulate multiple aspects of the neural development, such as dendritic expansion, synaptic growth and axonal bifurcation. In addition, the lipid metabolism abnormalities play an important role in many neurodegenerative disorders, such as Alzheimer's disease, Parkinson's disease and Huntington's disease. Despite the importance of lipid metabolism in neuronal health, the molecular mechanisms of how lipid metabolism regulates these processes are still unclear. Here we will show CdsA, which encodes the CDP-diacylglycerol (CDP-DAG) synthase, is essential for the development of mushroom body (MB), the olfactory memory center of Drosophila. Knockdown of CdsA induced the neurodegeneration in MB neurons. This degeneration progresses in a distal-to-proximal manner and is not due to the apoptosis and the Wallerian degeneration. Furthermore, the phenotype is not due to the dysfunction of the maintenance of mature MB neurons but the developmental defects. We assume that the alteration of the composition of specific phospholipids causes the degeneration in MB neurons because the reduction of CDP-DAG affects the amount of various phospholipids, such as phosphatidic acid, cardiolipin and phosphatidylinositol. To identify the phospholipids that are involved in this phenotype, we performed RNAi screening for genes that synthesize each phospholipid. As a result, we have identified Phosphatidylinositol synthetase (Pis) and Phosphatidylinositol 4-phosphate 5-kinase at 59B (PI4P5K), which synthesizes PI(4,5)P2, as a candidate. However, the loss of Phospholipase C (PLC), which catalyzes PI(4,5)P2 to inositol triphosphate (IP3) and diacylglycerol (DAG), did not cause any phenotypes. Taken together, the reduction of PI(4,5)P2 results in the neurodegeneration, but this degeneration would be IP3/DAG-independent mechanisms. In this poster, we will discuss how the reduction of CDP-DAG and/or phosphatidylinositol affects the development of the nervous system.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-055 ナノマテリアルによる酸化ストレスと神経細胞死 Yuji Kamikubo（上窪裕二），Tomohito Yamana（山名智人），Yuriko Inoue（井上由理子），Yoshie Hashimoto（橋本祥江），Takashi Sakurai（櫻井隆）順天堂大医薬理 With the development of nanoscience, nanomaterials with various sizes, chemical elements, shapes, modification, and surface functionalization have been produced for industries and consumers. Although bulk materials have certain characteristics almost irrespective of their size and shape, nanomaterials possess novel physical and chemical properties with a size and shape dependent manner, and have been used in a broad range of processes and products including paint, food, cosmetics, clothes, polish of fine structure, and pharmaceutical products. Nanomaterials including nanoparticles (NP) of titanium dioxide, zinc oxide, and silicon dioxide (silica) are well studied and already applied in various fields. Nanomaterials have a variety of unique physical and chemical properties, and are studied for biotechnological, pharmacological, and medical utilization. Because some nanoparticles including silica are inexpensive, innocuous, and easy to make and functionalize, they are applied as adsorbent, catalyst carrier, materials for bio-imaging and drug delivery systems (DDS). On the other hand, some reports raised a question of the biocompatibility and potential toxicity when they are in contact with tissues and cell surface. Recent studies showed that SiNP treatment induced cytotoxicity and inflammatory responses on lung epithelial and endothelial cell lines. In this study, we investigated the toxic effects of NPs on primary cultures of hippocampal cells. We showed that several types of NPs caused oxidative stress and cell death. Furthermore, we found that these cytotoxicity were dependent on the particle size, particle concentration, and as well as the treatment condition. The toxicity was reduced by SiNP surface functionalization or protein coating and by pretreating cells with an antioxidant, suggesting that contact-induced oxidative stress may be responsible for NP-induced cell death. These data will be valuable for utilizing NPs in biomedical applications.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-056 マウス神経幹細胞におけるカルパイン／カルパスタチン経路を介したアポトーシス制御 Hanako Yamamoto（山本華子），Masashi Kurachi（倉知正），Koji Shibasaki（柴崎貢志），Masae Naruse（成瀬雅衣），Yasuki Ishizaki（石崎泰樹）群馬大学大学院医学系研究科分子細胞生物学 We have reported that BMP4 promoted survival of NPCs (neuronal precursor cells) derived from ganglionic eminence by increasing their Id1 and Bcl-xL expression in the presence of FGF-2. Subsequent to a death signal, it is well known that pro-apoptotic protein Bax is freed from regulation of anti-apoptotic factor Bcl-2 or Bcl-xL and translocated to the outer mitochondrial membrane, where it promotes its permeabilization, favoring the release of different apoptogenic factors, including cytochrome c. To investigate whether NPCs show the tolerance to an extrinsic death signal in the presence of both FGF-2 and BMP4 (FGF-2/BMP4), we treated the cells with staurosporine (STS), a protein kinase inhibitor. FGF-2/BMP4 reduced Bax cleavage and suppressed cytochrome c release in NPCs, and promoted their survival after STS treatment. To examine which of FGF-2 or BMP4 regulates Bax cleavage, we cultured NPCs in various doses of them. Bax cleavage was suppressed by FGF-2, and its effect was stronger in presence of BMP4. Conversely, BMP4 by itself did not reduce Bax cleavage. It has been demonstrated that calpain is responsible for Bax cleavage. Because calpain activity is critically controlled by calpastatin, an endogenous proteinase inhibitor, we examined effects of calpastatin on survival of NPCs cultured in the medium containing BMP4 alone. We found that overexpression of calpastatin suppressed Bax cleavage and promoted NPC survival in the presence of BMP4 alone. When we treated NPCs with PD150606, a cell-permeable, noncompetitive calpain inhibitor, it also reduced Bax cleavage and promoted NPC survival in the presence of BMP4 alone. Furthermore, overexpression of Bax/p18, but not full-length Bax, negated the effects of FGF-2/BMP4 on NPCsurvival. These results suggest that FGF-2 suppresses Bax cleavage by inhibiting calpain activity. Taken together, it is likely that calpain/calpastatin system plays a role for control of NPC survival by FGF-2/BMP4 via cleavage of Bax.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-057 幼若ラットの顔面神経切断時に起こる運動ニューロン死へのミクログリアの応答性 Kazuyuki Nakajima（中嶋一行），Maasa Koshimoto（越本茉亜紗），Takashi Ishijima（石嶋貴志）創価大学、理工学部 Young motoneurons are highly vulnerable to insult. Transection of infant rat facial nerve leads to motoneuronal cell death in the ipsilateral nucleus. In this study, we investigated the age at which motoneurons are sensitive or robust to axotomy and determined the stage at which activated microglia emerge as dead-cell scavengers. Following nerve transection, living motoneurons were determined by Nissl staining. At 1 week after transection of the facial nerve in rats aged 2 days (2D), 1 week (1W), 2W, 4W, and 8W, the proportions of living motoneurons were approximately 10%, 25%, 75%, 100%, and 95%, respectively. We thus found that younger motoneurons are vulnerable to insult. A time-course experiment revealed that injured motoneurons of 2D rats were almost all alive at 1 day post-insult, but approx. 90% of motoneurons were dead within 3 days post-insult. These results indicated that almost all injured motoneurons die within 3 days post-insult. We next examined the microglial response to the process of motoneuronal cell death by using ionized Ca2+ binding adapter molecule-1 (Iba1) and cFms (cFms proto-oncogene) as microglial markers. Immunohistochemical analysis demonstrated that the number of anti-Iba1 antibody-staining cells increased in the axotomized facial nucleus during 3-5 days post-insult. Immunoblotting quantitatively indicated that the levels of Iba1 and cFms in the axotomized facial nucleus were enhanced during 3-5 days post-insult. These results demonstrated that microglia were activated/proliferated from 3 to 5 days post-insult. We further confirmed that the microglia that emerged in the axotomized facial nucleus were anti-CD68 antibody-positive, indicating that the microglia are phagocytic. Taken together, these findings indicate that younger motoneurons more easily die when injured, and that dead motoneurons are cleaned by phagocytic microglia.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-058 脳微小血管内皮細胞由来の細胞外小胞に含まれるPDGF-Bはオリゴデンドロサイト前駆細胞の増殖を促進する Masashi Kurachi（倉知正）¹，Bin Xu（徐彬）¹，Sho Osawa（大澤祥）²，Toshiyuki Matsuzaki（松崎利行）³，Yasuki Ishizaki（石崎泰樹）¹ ¹群馬大院医分子細胞生物 ²群馬大院医脳神経外科学 ³群馬大院医生体構造学 We previously showed that transplantation of brain microvascular endothelial cells (MVECs) greatly enhanced remyelination in the white matter infarct of the internal capsule of rats induced by endothelin-1 injection by increasing the number of oligodendrocyte precursor cells (OPCs). This transplantation also improved the animals' behavioral outcome, when assessed by footprint test. We further revealed extracellular vesicles (EVs) from MVECs promoted survival and proliferation of OPCs in vitro. In this study, we investigated the molecular mechanism how EVs from MVECs (MVEC-EVs) contribute to OPC survival and proliferation in vitro. As EVs from Rat fibroblast-like cell line (Rat-1) cells (Rat-1-EVs) promoted neither survival nor proliferation of OPCs, we compared the contents of MVEC-EVs and those of Rat-1-EVs. Protein mass spectrometry revealed 1,388 proteins in MVEC-EVs and 695 proteins in Rat-1-EVs, and 869 proteins were found only in MVEC-EVs. Among these 869 proteins, we focused on PDGF-B, which is well known to promote OPC survival and proliferation. Enzyme-linked immunosorbent assay (ELISA) revealed approximately 10 pg PDGF-B in 1 μg of MVEC-EVs, while it was undetectable in Rat-1-EVs by ELISA. Western blot analysis using an anti-PDGF-B antibody revealed a clear band in the samples prepared from MVEC-EVs, while no clear band was seen in those from Rat-1-EVs. A neutralizing antibody against PDGF-B significantly reduced the promoting activity of MVEC-EVs on OPC proliferation, while it did not affect their activity on OPC survival. These lines of evidence suggest that PDGF-B in MVEC-EVs plays a role for their promoting activity on OPC proliferation. We are now trying to reveal the localization of PDGF-B in MVEC-EVs by immunoelectron microscopy.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-059 坐骨神経傷害後のシュワン細胞遊走におけるSlit-Roboシグナルの機能 Sachiko Saito（齋藤瑳智子）¹，Shiro Saito（齋藤志郎）^1,2，Hidetoshi Iwata（岩田英敏）^1,2，Hina Fukutomi（福冨妃菜）¹，Naoko Kaneko（金子奈穂子）¹，Kazunobu Sawamoto（澤本和延）^1,3 ¹名古屋市立大院医再生医学 ²名古屋市立大医整形外科 ³生理学研究所神経発達・再生機構研究部門 Following axonal degeneration induced by peripheral nerve injury, Schwann cells surrounding the damaged axons de-differentiate and migrate toward the damaged area. Previous studies suggest that the migrating Schwann cells are involved in axonal regeneration for functional rewiring. However, the molecular mechanisms of Schwann cell migration during nerve regeneration have been largely unknown. A diffusible protein Slit and its receptor Robo guide axonal extention and cell migration in the developing central nerve system. It has been reported that the expressions of Slit and Robo are increased in the injured peripheral nerve tissue. However, their precise localization and involvement in axonal regeneration have not been elucidated. Here, using Slit1-knockout (Slit1-KO) mice, we studied the role of Slit-Robo signaling in the axonal regeneration process after sciatic nerve transection. First, to study the expression patterns of Slit and Robo proteins, we performed immunostaining using dissociated Schwann cells derived from sciatic nerve and dorsal root ganglia (DRG) neurons. Consistent with a previous report, Slit1 and Robo1 were expressed in Schwann cells and DRG neurons. Signals for Slit1 and Slit2 were observed in extended neurites in cultured DRG neurons. We next analyzed neurite extension of DRG neurons and migration of Schwann cells. These cells were isolated from wild-type or Slit1-KO mice, and embedded in Matrigel. While there was no significant difference in the length of neurites extended from the DRG explants between control and Slit1-KO groups, migration distance of the Slit1-KO Schwann was significantly shorter than the control. Time-lapse imaging of living cells showed that the migration speed was significantly decreased in the Slit1-KO cultures. Furthermore, axonal extension in the injured tissue tended to be delayed in the Slit1-KO mice. These results suggest that Slit-Robo signaling is involved in the peripheral nerve regeneration processes.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-060 組織損傷に応答して発現誘導するプロテアーゼDINEは神経依存性に組織修復を促す Sumiko Kiryu-Seo（桐生寿美子）¹，Reika Matsushita（松下鈴佳）¹，Teppei Shimamura（島村徹平）²，Hiroshi Kiyama（木山博資）¹ ¹名古屋大院医機能組織学 ²名古屋大院医システム生物学 Damage-induced neuronal endopeptidase (DINE) is a type II membrane-bound metalloprotease which we originally identified as a nerve regeneration-associated gene. DINE has a unique feature; the expression is highly up-regulated in response to nerve injuries occurred in both peripheral and central nervous system. Due to structural similarity with neprilysin, the amyloid β degrading enzyme, DINE has been supposed to be associated with degradation and/or processing of peptides, which have an important role in injured neurons, although the substrate for DINE remains unclear. Genetic manipulations of DINE gene have recently provided new insights into DINE functions during development, regeneration and pathogenesis in the nervous system. DINE-deficient (KO) mice develop the corneal haze characterized by keratitis and ulcer along with aging. In addition, they delayed tissue regeneration after damage such as pinna damage. These phenotypes suggest that DINE is involved in tissue repair. Interestingly, the expression of DINE is not found in non-neuronal peripheral tissues such as cornea, skin, muscle and cartilage, nor in uninjured sensory neurons which abundantly innervate the peripheral tissues. Upon tissue damage, the innervated sensory nerve was damaged and the expression of DINE was specifically induced in the injured primary sensory neurons. It is, therefore, very likely that the lack of the DINE expression could lead to the delayed tissue regeneration in KO mouse. We further found the injured sensory nerve was more significantly degenerated in damaged tissue of KO mouse than in that of wild type (WT) mouse. The microarray analysis of damaged tissue between WT and KO mice showed that inflammatory signaling pathways were elevated in the damaged tissue of KO mouse. In consistent with this, co-culture experiment of nerve-injured dorsal root ganglion (DRG) neurons with immune cells showed the increased number of degenerating fiber in DINE-deficient DRG neurons. These findings suggest that injury-induced neuronal expression of DINE regulates microenvironment of damaged tissues in which the cleaved peptide by DINE functions as a mediator to reduce scar formation and promote tissue regeneration.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-061 線維芽細胞増殖因子2 処理歯髄細胞の性状と移植による脊髄損傷ラットの運動機能回復効果 Makoto Yamada（山田まこと）¹，Kosuke Nagashima（長島光介）¹，Yumi Sato（佐藤友美）¹，Mana Onoue（尾上愛）¹，Noria Okada（岡田のりあ）¹，Ryoji Ohnishi（大西諒司）¹，Erika Imoto（井本愛梨香）¹，Ken-ichi Tezuka（手塚健一）²，Hitomi Soumiya（宗宮仁美）¹，Shoei Furukawa（古川昭栄）¹，Hidefumi Fukumitsu（福光秀文）¹ ¹岐阜薬大　分子生物 ²岐阜大院医再生医・組織器官形成 Objective: The spinal cord is the major route for afferent/efferent axons, connecting brain and peripheral nervous system. Patients with spinal cord injury (SCI) usually have permanent neurological deficits and disabilities, as upper/lower-limb paralysis, respiration and lower urinary tract dysfunction. Unfortunately, there is no therapeutic method available, which recovers function of the injured spinal cord. In this study, we examined the effect of dental pulp cells (DPCs) transplantation on the locomotor function of SCI rat. Methods: (cell culture) DPCs were prepared as previously reported. The DPCs were sub-cultured in basal medium supplemented with or without 10 ng/ml of FGF2. (Animal Surgery and DPCs Transplantation) Seven weeks old female Wistar rats were anesthetized, and the spinal cord was completely transected by microsurgical scissors at the level of the 10th thoracic vertebra. Immediately after surgery, the DPCs were transplanted into the lesion epicenter. After the cell-transplantation, the locomotor function of the both hindlimbs of the SCI rats was assessed by the Basso, Beattie, and Bresnahan (BBB) locomotor scale in open field. (Tissue processing) Seven weeks after the surgery and cell transplantation, the animals were deeply anesthetized by i.p. injection of sodium pentobarbital and then cardio-perfused with 4% PFA. Frozen sections were prepared with a cryostat and used for histological analysis. Results: FGF2 priming facilitated the DPCs to promote axonal regeneration and to improve locomotor function in the rat with spinal cord injury. To investigate mechanism underlying the effect of FGF2-priming, we selected and analyzed seven DPC lines based on a screening for gene expression, cell proliferation, major neurotrophic factors production, and resistant properties against hydrogen-peroxide-induced cell death, in addition to their functionality in a rat SCI model. The data suggest that FGF2 priming might protect DPCs from the post-trauma microenvironment in which infiltrates and resident immune cells generate cytotoxic reactive oxygen species. Surviving DPCs could increase the availability of neurotrophic factors in the lesion site, thereby promoting axonal regeneration and locomotor function recovery. Conclusion: The dental pulp cells transplantation pre-treated with FGF-2 is a novel and promising cell therapy for the treatment of spinal cord injury

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-062 神経再生阻害因子に対するドラッグスクリーニング　－脊髄損傷治療への展開ー Kosei Takeuchi（武内恒成）^1,2，Hasashi Ikeno（池野正史）¹，Hiroyuki Sasakura（笹倉寛之）¹ ¹愛知医大医細胞生物 ²愛知医大研究創出支援センター Injured adult neurons in the mammalian central nervous system (CNS) rarely regenerate, because some of the intracellular and cell-surface environmental factors inhibit axon regrowth. Chondroitin sulfate (CS) is the most abundant and potent exogenous inhibitor of axonal regeneration. We generated null (KO) mice of CSGalNAcT1/T2 and GalNac4S-6ST, a key enzyme in CS biosynthesis. We have showed that T1KO mice recovered much faster and more completely from induced SCI than do wild-type mice and even ChondroitinaseABC treatment mice (Takeuchi et al., Nature Commun.). Our results show that reduction of CS synthesis by the controlling the CSGalNAcT1-expression is a best strategy for spinal cord injury and stroke treatment. We try to establish the accurate inhibition systems of CS-expressions in vivo from the drug screening system (small molecule compound and natural organic compound), to regulate CS-expressions and modifications in the injury area. We selected the drugs to down-regulate the CS-expressions. The sponge forms biomaterials impregnated with a mixture containing small compounds was placed on the lesion area in mice suffered neural injuries. The recovery of these mice which treated with drug delivery systems reached the levels of satisfactory amelioration comparable to those of KO mice. Taken together, our results indicated that our screened compounds and drug and delivery system is a promising therapeutic target for treatment of the spinal cord injury and brain infarction. In our drug screening procedures, we got the several up-regulated natural organic compound. We have already showed CSPG is involved in the regulation of adult hippocampal neurogenesis and suggest that increased synthesis of CSPG by CSGalNacT1 may mediate promotion of granule cell production and improvement of cognitive memory (Yamada et al, J Neuroscience). We try to promote the expression of CS in the normal and developmental brains. We would like to report these trials to regulate up-and down- expression of CS in vivo nervous systems.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-063 Age-related changes in whole-brain connectivity across the adult lifespan: The BMRC aging cohort study Epifanio Jr. T Bagarinao（Bagarinao Epifanio Jr. T）¹，Hirohisa Watanabe（Watanabe Hirohisa）¹，Satoshi Maesawa（Maesawa Satoshi）¹，Daisuke Mori（Mori Daisuke）¹，Kazuya Kawabata（Kawabata Kazuya）²，Kazuhiro Hara（Hara Kazuhiro）²，Shuji Koyama（Koyama Shuji）¹，Minoru Hoshiyama（Hoshiyama Minoru）¹，Haruo Isoda（Isoda Haruo）¹，Shinji Naganawa（Naganawa Shinji）^1,3，Gen Sobue（Sobue Gen）¹ ¹Brain and Mind Research Center, Nagoya University, Nagoya, Japan ²Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan ³Department of Radiology, Nagoya University Graduate School of Medicine, Nagoya, Japan Using resting state functional magnetic resonance imaging (rsfMRI), several studies have shown that the human brain is functionally organized into large-scale brain networks consisting of regions that are spatially distributed but functionally linked. Characterizing the brain's normal connectivity pattern throughout development is important as recent studies have indicated that disruptions in connectivity in critical brain regions could be associated with psychiatric as well as neurodegenerative disorders. In this study, we investigated age-related changes in whole-brain functional connectivity across the adult lifespan. We used rsfMRI data from 129 healthy volunteers ranging in age from 21 - 86 years old and carefully selected from participants of our ongoing aging cohort study. Each rsfMRI data was preprocessed by first discarding the initial 5 images, then performing slice-timing correction, realignment, co-registration to the anatomical image, normalization to MNI space, resampling to 2 x 2 x 2 mm³ voxel resolution, and smoothing using an 8-mm FWHM 3D Gaussian filter. Additional preprocessing steps were also performed to remove the effect of head motion and other physiological noise. Using a whole-brain parcellation consisting of 499 regions-of-interest (ROI), we then computed the connectivity matrix using the preprocessed rsfMRI data. For each ROI, we computed the mean functional connectivity (mFC) and correlated the mFC values from all participants with the participants' age, sex, and ACE-R total score. Our results showed that ROIs located in the bilateral temporal lobe, cerebellum, and orbital frontal regions showed significant decrease in mFC values with age. These ROIs were associated with the default mode, language, and executive control networks. On the other hand, ROIs associated with the sensorimotor and visual networks showed significant increase in mFC values with age. Some ROIs in the temporal cortex, medial prefrontal cortex, and dorsolateral prefrontal cortex had also mFC values that were positively correlated with ACE-R total score, whereas a number of ROIs in the visual cortex had mFC values that showed negative correlation. Overall, our findings suggest that regions in the temporal lobe are increasingly disconnected during healthy aging, while primary processing regions (sensorimotor, visual) tend to be increasingly connected. These changes may also be associated with general cognitive performance.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-064 肥満と加齢に伴う脳の構造変化の関係：Human Connectome Project-style paradigmを用いた脳マルチモーダルMRI研究 Toru Ishihara（石原暢）¹，Atsushi Miyazaki（宮崎淳）¹，Hiroki Tanaka（田中大貴）¹，Takayuki Fujii（藤井貴之）¹，Muneyoshi Takahashi（高橋宗良）¹，Yoshie Matsumoto（松本良恵）¹，Toko Kiyonari（清成透子）²，Haruto Takagishi（高岸治人）¹，Tetsuya Matsuda（松田哲也）¹ ¹玉川大脳研 ²青山学院大 Although mechanisms common to aging and obesity are hypothesized to increase susceptibility to neurodegeneration, evidence to support this is still scarce. Here, we studied the cross-sectional relationships of body mass index (BMI) and body composition (i.e., body fat percentage and visceral fat level) with multimodal brain structure acquired using the Human Connectome Project-style paradigm. The body weight and body composition data were obtained for 230 individuals (females = 112, males = 118; 27 to 69 years old) using a Tanita DC-320 bio-impedance analysis device (Tanita Corp., Tokyo, Japan). The height was assessed with a height scale. The BMI was calculated as body weight (kg)/height (m²). All MRI data (T1, T2, and diffusion image) were collected using a 3T Trio Tim scanner (Siemens, Erlangen, Germany). After controlling for sex, we analyzed the relationships of age, BMI, and body composition with cortical thickness, myelin content (estimated by T1w/T2w ratio), neurite density index (NDI), and orientation dispersion index (ODI) in 360 parcels of brain regions. All statistical analyses were conducted with Bonferroni corrected α = 0.05 (α = 0.00014). The cortical thickness decreased, and myelin content, NDI, and ODI increased with age. The BMI and body composition showed no correlation with cortical thickness. The BMI, body fat percentage, and visceral fat level were positively associated with myelin content (141 parcels, including the following areas of the cortex: dorsal and ventral prefrontal, posterior cingulate, inferior parietal, medial and lateral temporal, insular, and frontal opercular; partial r = .25 to .39; p < .00014), ODI (6 parcels, including medial temporal and medial prefrontal cortexes; partial r = .25 to .32; p < .00014), and NDI (54 parcels, including dorsal, ventral, and medial prefrontal cortex; medial and lateral temporal cortex; insular cortex; and frontal opercular cortex; partial r = .25 to .40; p < .00014). Further, our results showed that the brain structure indices in overweight and obese individuals (≧ 25 kg/m²) were associated with maximal effects with an estimated increase in brain age of 15 to 20 years (myelin content and ODI: 20 years, NDI: 15 years). Overall, our results suggest that overweight and obesity are associated with neurodegeneration, particularly increasing myelin content, neurite density, and orientation dispersion.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-065 認知機能維持メカニズムの解明 Tomoko Tanaka（田中智子），Haruo Okado（岡戸晴生）東京都医学総合研 Dementia, one of the major causes of cognitive decline mostly found in the elderly patients, has becomes a serious social problem. RP58, a zinc-finger transcriptional repressor, plays a critical role in the formation of cerebral cortex. Recently, it has been reported that the expression level of RP58 decreases in aged human cortex. Furthermore, it has been shown that the level of RP58 is declined in hippocampal CA1 region of aged rat. Therefore, RP58 may play a crucial factor in aging. In the present study, we investigated the involvement of RP58 in age-related cognitive decline. We used RP58 hetero knockout mice and wild-type mice from 1.5 to 17 months. We performed an object location test and a fear conditioning test to measure cognitive function. After behavioral test, we performed immunohistochemistry to determine whether microglia, single stranded DNA and gamma-H2AX respond to the decline in RP58 level. We indicated that, with advancing age, the wild-type mice showed impairment of place memory for object and increase of reactive microglia in hippocampus. Furthermore, wild-type mice also showed increase of single stranded DNA foci and gamma-H2AX in response to exogenous DNA damage, which indicate the increase of DNA damage or the dysfunction of DNA repair in hippocampus as age-related changes. On the other hand, RP58 hetero knockout mice showed these symptoms faster than wild-type mice. Therefore, these results suggest that the decline of RP58 may lead to progression of aging.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-066 加齢におけるBDNFプロペプチド Toshiyuki Mizui（水井利幸），Konomi Matsui（松井このみ），Masami Kojima（小島正己）産総研バイオメディカル Aging is a normal physiological process accompanied by cognitive deterioration. This aging process has been the primary risk factor for development of aging-related diseases such as Alzheimer's disease. The cognitive deficit is related to alterations of neurotrophic factors level such as brain-derived neurotrophic factor (BDNF). Yet the synaptic and molecular mechanisms responsible for cognitive aging are not well understood. Recently, we identified a new subtype of BDNF, BDNF pro-peptide. It is produced together with BDNF by proteolytic processing of precursor BDNF. Interestingly, however, while BDNF elicited long-term potentiation (LTP) and promoted dendritic spines, the BDNF pro-peptide facilitated long-term depression (LTD) and exerted spine shrinkage, suggesting that the BDNF pro-peptide is a new facilitator of synaptic depression and synapse loss. In the present study, we investigated how the protein levels of BDNF, its receptor TrkB, BDNF pro-peptide and its receptor component p75 changed in the mouse brain with age. Very interestingly, while a synaptic facilitator BDNF significantly declined the levels with age, the pro-peptide and its receptor p75 pronouncedly raised with age, showing that the biological action of BDNF and its pro-peptide are attenuated and enhanced, respectively, during aging. More interestingly, we found that, in line with the increased amount of the BDNF pro-peptide and p75, tau protein was phosphorylated with age, raising a new possibility that the BDNF pro-peptide/p75 signaling elicits the progression of the tau-phosphorylation-dependent Alzheimer disease.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-067 線条体におけるAQP4依存的なATP/アデノシンの増加によるドパミン神経伝達の調節 Satoshi Okada（岡田聡史），Masato Kobayashi（小林昌人），Mitsuhiro Morita（森田光洋）神戸大学大学院理学研究科 Accumulating evidence indicates astrocytic ATP release for intercellular communication in the brain. Astrocytic ATP triggers responses of glial cells, because ATP receptors are mostly expressed in astrocytes and microglia. Subsequent conversion of ATP to adenosine by extracellular enzymes modulates functions of neurons, vasculature and immune cells, which abundantly express adenosine receptors. Our previous study indicated AQP4-dependent ATP release after hypotonic treatment and subsequent adenosine increase in hippocampal slices by using an originally-developed biosensor for adenosine (Yamashiro et al, J Neurochem 2017). In order to investigate the functional implication of the AQP4-dependent ATP/adenosine increases, this study analyzed adenosine and dopamine in the striatum of AQP4 knockout mice by using biosensors for adenosine or dopamine. Adenosine or dopamine were measured by calcium imaging of CHO cells expressing adenosine A1 or dopamine D2 receptor together with Gqi5. An A1 receptor antagonist, DPCPX increased evoked dopamine release after repetitive electrical stimulation to wild type striatum, indicating tonic suppression of dopamine release by ambient adenosine activating A1 receptor. Meanwhile, DPCPX did not increase evoked dopamine release in AQP4 knockout mice, suggesting that the ambient adenosine is derived from AQP4-dependent ATP release from astrocytes. Since higher basal dopamine level and lack of motivation were reported in AQP4 knockout mice, the crosstalk between AQP4-dependent increase of astrocyte-derived ATP/adenosine and dopaminergic neurotransmission is very likely participated in mental processes.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-068 ショウジョウバエ脳においてドパミン放出を誘導するグアニル酸シクラーゼの同定 Kohei Ueno（上野耕平），Minoru Saitoe（齊藤実）（公財）東京都医学総合研究所・学習記憶プロジェクト Dopamine is a key neurotransmitter for neural plasticity in both mammal and Drosophila. However, its releasing mechanisms are still not clear. Previously, we have demonstrated that the synaptic transmission between antennal lobe (AL), primary olfactory center, and mushroom body (MB), center of olfactory memory, is enhanced for long-period (< 2 hours) after simultaneous stimulation of AL and ascending fibers of ventral nerve code (AFV), which send somatosensory information to brain (Ueno et al., 2013). This enhancement requires D1 dopamine receptor activity on the MB. Imaging analysis of exocytosis from the dopaminergic neurons on the MB demonstrated that the dopamine release is occurred upon simultaneous stimulation of AL and AFV, but not by each AL and AFV stimulation alone. Moreover, the exocytosis is localized on the MB neurons which had been coincidentally activated by AL and AFV inputs. Thus, these results suggest that coincidentally activated MB neurons generate a retrograde signal that gates dopamine release (Ueno et al., 2017). In previous meeting, we demonstrated carbon monoxide (CO) as a candidate of the retrograde signals. Thus, suppression of CO synthesis in the MB impaired dopamine release and application of CO induces exocytosis from dopaminergic neurons. However, there is still a large gap between CO and synaptic vesicle release. It is known that CO activates soluble guanylyl cyclases (GCs). We also found that CO stimulation increases cGMP level in the dopaminergic neurons, and application of the cGMP analogue, 8-Br-cGMP, induces dopamine release. We knock down various GC genes in the dopaminergic neurons, and found an atypical GC gene, Gyc-88E, in the dopaminergic neurons is required for olfactory memory. We also confirmed that this GC is also required for dopamine release in our ex vivo brain system. Taken together, we suggest that CO activates Gyc-88E in the dopaminergic neurons to induce exocytosis of synaptic vesicles.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-069 線条体のD2ドーパミン受容体発現間接路神経におけるNMDA受容体のマグネシウムブロック低下による運動異常 Toshikuni Sasaoka（笹岡俊邦）¹，Nae Saito（齊藤奈英）¹，Manabu Abe（阿部学）¹，Meiko Kawamura（川村名子）¹，Yoko Nabeshima（鍋島曜子）²，Yo-ichi Nabeshima（鍋島陽一）²，Kazuki Tainaka（田井中一貴）¹，Kenji Sakimura（﨑村建司）¹ ¹新潟大脳研 ²先端医療研究センター The striatum, an input station of the basal ganglia, receives glutamatergic inputs from the cerebral cortex and dopaminergic inputs from the substantia nigra. The striatum plays a pivotal role in motor learning. The efficacy of cortico-striatal neuronal transmission is modulated by dopaminergic inputs, which convey motor error signals. This synaptic plasticity is believed to be the basic mechanism of motor learning. In Parkinson's disease (PD), dopaminergic projections to the striatum are lost, and failure of cortico-striatal neurotransmission is supposed to cause parkinsonian symptoms. Although L-dopa treatment is one of standard therapy for PD, long term treatment of L-dopa for PD often induces dyskinesia. Thus, plastic changes may occur in the cortico-striatal neurotransmission in dyskinesia. Therefore, to understand the functions of the striatum in motor control, it is a key to analyze the interaction between the glutamatergic and dopaminergic inputs to the striatum and its effect on the synaptic plasticity. Striatal projection neurons are classified into dopamine D1 receptor expressing direct pathway neurons and dopamine D2 receptor expressing indirect pathway neurons. NMDA receptors are known to play an important role in synaptic plasticity. While NMDA receptors are inactive at rest by Mg2+ blockade, depolarization relives Mg2+ blockade, causes Ca2+ influx through the NMDA receptors, and induces synaptic plasticity. In order to analyze the effect of dopaminergic inputs on the plasticity of cortico-striatal neurotransmission, we developed genetically modified mice expressing activated NMDA receptors specifically in the indirect pathway. A single asparagine residue in the second membrane-associated segment of GluN2 subunits is critical for Mg2+ blockade. We have developed a new method for introducing a mutation into the gene of interest in a spatially restricted manner. Using this technique, we selectively introduced an amino acid substitution (N595Q) into the GluN2 subunits by Cre-loxP recombination using the Cre-expressing mice under control of D2R gene. Only when the wild-type exon sequences between two loxP sites were deleted by the action of Cre recombinase, the mutant exon would be spliced into mRNA. This mutation reduced the Mg2+ block property of the NMDA receptors. We performed analysis of Cre-loxP recombination by whole brain imaging technique and several behavior tests regarding spontaneous motor activity and learning and memory.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-070 シグマ1受容体アンタゴニストBD1047は海馬ニューロンのIhチャネルを抑制する Takako Ohno-Shosaku（少作隆子），Satoko Gamo（蒲生智子），Haruka Aoki（青木晴香）金沢大学保健学リハビリテーション科学 Sigma-1 receptors are a promising target for treatment of various diseases, including neurodegenerative diseases, neuropsychiatric disorders, and neuropathic pain. Sigma-1 receptor agonists enhance brain plasticity, and are effective in the treatment of major depression. Sigma-1 antagonists attenuate neuropathic pain, such as allodynia. However, precise mechanisms by which the sigma-1 agonists or antagonists exert their therapeutic effects are poorly understood. In the present study, we examined effects of the sigma-1 receptor agonist PRE-084 and antagonist BD1047 on synaptic currents and Ih channels in rat cultured hippocampal neurons. Application of PRE-084 did not influence the frequency of spontaneous postsynaptic currents, but induced an increase in the frequency of miniature excitatory postsynaptic current (mEPSC) without changing its amplitude. The frequency of mEPSC was decreased after PRE-084 was washed out, indicating that the stimulating effect of PRE-084 is reversible. Application of BD1047 did not influence the frequency of mEPSC. Unexpectedly, we found that application of BD1047 reversibly decreased the inward holding current at -80 mV. This effect was completely suppressed by the Ih channel blocker ZD7288, indicating that the BD1047-sensitive channel should be Ih channels. In some experiments, we observed a slight recovery of the inward holding current by BD1047 in the presence of ZD7288, suggesting that BD1047 binds to Ih channels directly, and competes with ZD7288 for a common binding site. Our results concerning the effect of PRE-084 on mEPSCs are consistent with a previous report indicating that the excitatory neurosteroid pregnenolone sulfate (PREGS) increases the frequency, but not the amplitude, of mEPSCs in hippocampal neurons, and the effect of PREGS on mEPSCs is blocked by sigma receptor antagonists. Our results concerning the effect of BD1047 on Ih channels are rather surprising. As far as we know, there are no studies reporting effects of BD1047 on Ih channels. Ih channels have been suggested to be involved in induction of allodynia, and Ih channel blockers including ZD7288 are effective for preventing allodynia. Therefore, it is likely that BD1047 attenuates neuropathic pain through two separate pathways; through antagonizing sigma-1 receptors and through suppressing Ih channels.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-071 ヒト遺伝子由来ペプチドエンドキニンC/Dは痛みと炎症の抑制効果を有する Rumi Naono（直野ー中山留美）¹，Hideki Funahashi（船橋英樹）²，Yu Miyahara（宮原裕）²，Toshikazu Nishimori（西森利數）²，Yusuke Ishida（石田雄介）¹，Keijyu Kamijyo（上条桂樹）¹ ¹東北医科薬科大学　医学部　解剖学教室 ²宮崎大学医学部　臨床神経科学講座　精神医学分野 Endokinin peptides (EKs) are encoded on the human TAC4 and consist of endokinin A (EKA), B (EKB), C (EKC) and D (EKD), respectively. It is known that EKs are a member of tachykinin peptide family, which belonging to substance P (SP). When intrathecal administration of endokinin A and B (EKA/B, the common C-terminal decapeptide in EKA and EKB) elicits nociceptive behavior like as SP-induced behavior; however, pretreated with endokinin C and D (EKC/D, the common C-terminal decapeptide in EKC and EKD) is attenuated SP-induced nociceptive behavior and inflammatory pain. These physiological differences may contribute to the 12th amino acid at the carboxyl terminus of EKA/B and EKC/D, and they are leucine and methionine, respectively. Furthermore, in order to the lasting anti-nociceptive and anti-inflammatory effect of EKC/D, it may be useful for the change some amino acids with D-type amino acid. Indeed, to press for the improvement in the anti-nociceptive effect of EKC/D, we synthesized EKC/D-derived peptides (D-EKC/Ds) having some of D-type amino acids. These peptides were estimated by SP-induced nociceptive behavior and inflammation induced by chemical compound, carrageenan and formalin, respectively. Pretreatment with D-EKC/Ds were long-lasting attenuated on SP-induced nociceptive behavior and inflammation compared with the only EKC/D. These results indicate that, D-EKC/Ds were not only anti-nociceptive effect but also anti-inflammatory effect and some of D-type amino acids may have a crucial role of these effects.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-072 Evaluation of synaptotagmin7 function in the unique properties of somatodendritic dopamine release Takuya Hikima（Hikima Takuya）¹，Paul Witkovsky（Witkovsky Paul）¹，Konstantin Ichichenko（Ichichenko Konstantin）²，Margaret E Rice（Rice Margaret E）^1,3 ¹Dept. Neurosurgery, New York Univ School of Medicine, New York, USA ²Dept. Biochemistry & Molecular Pharmacology, New York Univ School of Medicine, New York, USA ³Dept. Neuroscience and Physiology, New York Univ School of Medicine, New York, USA Midbrain dopamine (DA) neurons in the substantia nigra pars compacta (SNc) exhibit somatodendritic release of DA. Released DA acts at D2 autoreceptors to regulate neuronal firing patterns that regulate DA release throughout the brain. We have reported previously that: 1) the Ca²⁺ dependence of somatodendritic DA release differs from that of axonal release (Chen et al., 2011); 2) aspects of molecular organization for somatodendritic DA release are distinct from those for conventional synaptic release (Witkovsky et al., 2009). Additional evidence from our group implies that D2 receptors on a given DA neuron are activated primary by DA released from that same cell, literally autoregulation, via exocytosis. We focused on the function of synaptotagmin 7 (syt7), a high affinity Ca²⁺ sensor required for neurotransmitter release, and evaluated whether syt7 contributes to the unique Ca²⁺ dependence of somatodendritic DA release in the SNc. We tested this using voltage-clamp recording of SNc DA neurons in acute midbrain slices. Locally evoked D2 DA-dependent inhibitory currents (D2ICs) (5 pulses, 40 Hz), mediated by D2 receptor G-protein coupled K⁺ (GIRK) channels, were used as an index of somatodendritic DA release. Immunohistochemical studies indicated that syt7 is present in somata and dendrites of DA neurons. Evoked D2ICs were attenuated when antibody against syt7 was included in the recording pipette. This attenuation of D2IC amplitude was attenuated when a syt7 blocking peptide was included with syt7 in the pipette. Moreover, consistent with our previous findings, D2ICs persisted in submillimolar extracellular Ca²⁺ concentrations that do not support axonal DA release. However, this increased Ca²⁺ sensitivity of evoked release was lost when the antibody of syt7 was applied in the recording pipette. Together, these data support a key role for syt7 in SNc DA neurons in the unique Ca²⁺ dependence of somatodendritic DA release.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-073 ストレスによるてんかん発作モデルにおけるノルアドレナリン神経伝達の役割 Hitoki Sasase（笹瀨人暉），Shiho Ito（伊藤志穂），Ryuma Yaguchi（矢口立真），Shoma Izumi（泉翔馬），Kazuhei Niitani（二井谷和平），Satoshi Deyama（出山諭司），Eiichi Hinoi（檜井栄一），Katsuyuki Kaneda（金田勝幸）金沢大学大学院医薬保健学総合研究科薬理学研究室 Stress is one of the most frequently self-reported precipitants for seizure induction in epilepsy patients, but how stress triggers seizures remains unknown. In the medial prefrontal cortex (mPFC), stress has been known to enhance the release of noradrenaline (NA), which excites mPFC layer 5 (L5) pyramidal cells. Here, we investigated the possible contribution of NA in the mPFC to stress-induced epileptic seizures. Intra-mPFC infusion of picrotoxin (0.1 nmol/side) and NA (10 nmol/side) induced seizures with shorter latency than that of picrotoxin alone in C57BL/6J mice. In vitro whole-cell patch-clamp recordings from mPFC L5 pyramidal cells revealed that, in the presence of picrotoxin (30 μM), bath-application of NA (10 μM) induced rhythmic and frequent epileptiform activities (EAs) consisting of prolonged depolarization with burst firings in short latency. On the other hand, picrotoxin alone induced sporadic and long-latency EAs. Simultaneous whole-cell and field potential recordings revealed that EAs in individual cells were time-locked with population spikes. The NA-induced EAs were inhibited by α1 adrenoceptor antagonist terazosin (5 μM), but not α2 adrenoceptor antagonist atipamezole (3 μM) or β adrenoceptor antagonist timolol (10 μM). Additionally, in the presence of picrotoxin, bath-application of α1 adrenoceptor agonist phenylephrine (100 μM) induced rhythmic and frequent EAs. These results suggest that NA released in the mPFC might contribute to the expression of stress-induced seizures in epilepsy patients via the activation of α1 adrenoceptors.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-074 カリオフェリンαノックアウトマウスの行動解析と神経化学的分析 Koki Sakurai（櫻井航輝）^1,2，Yoichi Miyamoto（宮本洋一）³，Taichi Ito（伊藤大一）¹，Makiko Morita（森田真規子）¹，Yoshihiro Yoneda（米田悦啓）⁴，Masahiro Oka（岡正啓）³，Takatoshi Hikida（疋田貴俊）^1,2 ¹大阪大蛋白質研・高次脳機能学 ²大阪大院理生物科学 ³医薬健栄研・基盤研・細胞核輸送 ⁴医薬健栄研 Karyopherins (KPNs) are a family of proteins that assist newly synthesized polypeptide molecules to be transported from the cytoplasm into the nucleus. KPNαs (KPNAs), otherwise known as Importin-αs, are a type of KPN that bind to the classical nuclear localization signal contained in the sequence of cargo protein polypeptides and allow them to move across the nuclear membrane by passing through nuclear pore complexes. Their molecular characteristics related to nucleocytoplasmic transport of polypeptides have been extensively studied, but its other functions are still under investigation. To investigate the role of KPNAs related to brain functions, we developed kpna knockout (KO) mouse lines. The behavioral phenotypes of these mice were examined by conducting a battery of behavioral tasks. A KO mice line lacking a specific subtype of KPNA showed behavioral abnormalities including increased anxiety, indicating that KPNAs are important in brain function. To further characterize the molecular and biochemical states occurring in these KO mice, we performed further molecular/biochemical characterization. HPLC quantification of monoamine neurotransmitters and its metabolites in the KO mice brains showed that the levels of neurotransmitter metabolites were significantly reduced the brains of the KO mice. We aim to elucidate further details in the molecular and biochemical mechanisms underlying these these behavioral abnormalities. Further insight into the mechanisms may provide new insights on the brain functions and pathology for neuropsychiatric disorders.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-075 ストレス時の母体の咀嚼運動は、仔のストレス誘発性の肥満および糖尿病を改善する Sakurako Hayashi（林櫻子）¹，Masahisa Katano（片野雅久）¹，Kyoko Kajimoto（梶本京子）¹，Suzuko Ochi（越智鈴子）¹，Chie Hisada（久田智詠）¹，Eri Yoshikawa（吉川英里）¹，Hiroko Tsugane（津金裕子）¹，Kumiko Yamada（山田久美子）²，Mitsuo Iinuma（飯沼光生）¹，Kagaku Azuma（東華岳）³，Kin-ya Kubo（久保金弥）⁴ ¹朝日大学歯学部小児歯科 ²名女大　家政 ³産業医科　第一解剖 ⁴名女大　院　生活 Maternal stress in mice increases the risk of obesity and diabetes in the adult offspring. Increased levels of several peptides and hormones correlate with obesity and diabetes in mice. Adiponectin is secreted by adipose tissue and has an important role in the development of insulin resistance. Neuropeptide Y (NPY), a peptide hormone widely expressed in the neural system, is increased by food intake and decreased by physical activity. Ghrelin, released mainly by the stomach, stimulates appetite, increases food intake and promotes fat storage. We previously reported that prenatal stress-induced learning disabilities, anxiety-like behavior, and vulnerability to stress in mouse offspring can be ameliorated by allowing the pregnant dams to chew on a stick during periods of stress. Here we examined the effects of chewing during maternal restraint stress on changes in weight, insulin resistance, and metabolites of energy metabolism-related peptide and hormones in the adult offspring. Pregnant mice were exposed to restraint stress beginning on day 15 of pregnancy and continuing until delivery. Half of the dams were given a wooden stick to chew on during the restraint stress. The body weight of the offspring was measured weekly after birth and the amount of food ingested was measured weekly after weaning. Blood glucose, adiponectin, and ghrelin levels, and insulin resistance were measured in the offspring at 4 months of age. In addition, NPY mRNA expression levels in the arcuate nucleus of the hypothalamus were analyzed by in situ hybridization. Prenatal stress significantly increased body weight, amount of food ingested, blood glucose levels, insulin resistance, and NPY mRNA expression, and decreased blood adiponectin and ghrelin levels in the offspring. Allowing dams to chew on a stick during prenatal stress attenuated both the prenatal stress-induced increases in body weight, amount of food ingested, blood glucose levels, insulin resistance, and NPY mRNA expression, and decreases in blood adiponectin and ghrelin levels in the offspring. These findings suggest that maternal chewing during periods of stress improves energy metabolism and suppresses maternal stress-induced diabetes in the adult offspring.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-076 アストロサイト由来シナプス形成分子再活性化は注意欠陥・多動性を引き起こす Jun Nagai（長井淳）¹，Abha K. Rajbhandari（Rajbhandari K. Abha）²，Mohitkumar R. Gangwani（Gangwani R. Mohitkumar）¹，Ayaka Hachisuka（Hachisuka Ayaka）⁴，Giovanni Coppola（Coppola Giovanni）^3,4,5，Sotiris C. Masmanidis（Masmanidis C. Sotiris）⁴，Michael S. Fanselow（Fanselow S. Michael）^2,3，Baljit S. Khakh（Khakh S. Baljit）^1,4 ¹Dept Physiol, Univ of California, Los Angeles, USA ²Dept Psychol, Univ of California, Los Angeles, USA ³Dept Psychiatry, Univ of California, Los Angeles, USA ⁴Dept Neurobiol, Univ of California, Los Angeles, USA ⁵Semel Institute of Neuroscience and Human Behavior, Univ of California, Los Angeles, USA In order to understand how the CNS encodes, modifies, stores and retrieves information it is necessary to explore the diverse cell populations that comprise the CNS. One significant missing aspect to comprehensively understand the CNS is the largely unmet need to understand additional cell types such as astrocytes. Astrocytes represent around 40% of all CNS cells. An open question in neuroscience concerns exactly how neurons and astrocytes communicate with each other and modulate microcircuit activity and animal behavior. Here, we used multiple integrated approaches, including [Ca²⁺]i imaging, optogenetics, pharmacogenetics, animal behavioral tests, electrophysiology in slice and in vivo, and RNA-seq to address this question for the dorsal striatum of adult mice, where the underlying mechanisms of neuron-astrocyte interplay are incompletely understood. We found that upstate-like excitability of striatal medium spiny neurons (MSNs) triggered astrocyte calcium signaling via GABA_B1 receptor, a Gi GPCR, in slice and in vivo. Selective activation of this pathway in striatal astrocytes in vivo by a Gi-DREADD hM4Di, resulted in acute behavioral hyperactivity and disrupted attention. RNA-seq revealed that such responses resulted in up-regulation of the synaptogenic cue thrombospondin-1 (TSP1) in astrocytes, which has roles in developmental synapse formation but is down-regulated in adults. Astrocyte Gi activation also increased excitatory synapses, enhanced corticostriatal synaptic transmission, and increased MSN action potential firing in vivo. All of these changes were reversed by in vivo Gabapentin administration that antagonizes the TSP1 neuronal receptor α2δ-1. Our findings show not only that physiological activity of neurons triggers astrocyte signaling, but that signaling from astrocytes to neurons is also sufficient per se to alter synapses, circuits and behavior in adults by reactivating a single, latent astrocyte synaptogenic cue. The findings also suggest that behavioral phenotypes accompanying diverse brain disorders in adults may have an astrocytic component and that identifying and exploiting astrocyte based neuromodulation (e.g. via TSP1) affords new therapeutic opportunities for attention deficit hyperactivity disorder (ADHD)-like and possibly other psychiatric diseases.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-077 アストロタクチン2（ASTN2）はセロトニンおよびドーパミン作動性神経系に影響を及ぼすことで情動および認知機能を調節する Takahiro Ito（伊藤貴博）¹，Yuka Hiramatsu（平松愉加）¹，Mizuki Uchida（内田美月）¹，Yuki Imami（今見由貴）¹，Hirotake Hida（肥田裕丈）¹，Akira Yoshimi（吉見陽）¹，Itaru Kushima（久島周）²，Norio Ozaki（尾崎紀夫）²，Tomomi Aida（相田知海）³，Kohichi Tanaka（田中光一）³，Yukihiro Noda（野田幸裕）^1,2 ¹名城大学薬学部・大学院薬学研究科病態解析学I ²名古屋大学大学院医学系研究科精神医学 ³東京医科歯科大学難治疾患研究所先端分子医学研究部門分子神経科学 Astrotactin (ASTN)2, a glial-guided neuronal migration-related molecule, regulates the trafficking of ASTN1, a surface membrane protein, during glial-guided neuronal migration. Copy number variants (CNVs) of the ASTN2 locus are risk factors for autism spectrum disorder (ASD), attention deficit hyperactivity disorder (ADHD), schizophrenia (SCZ), and other neurodevelopmental disorders. We identified deletion of ASTN2 in three SCZ patients and one ASD patient in CNV analysis. However, it is unclear about physiological functions of ASTN2 in pathophysiology of neuropsychiatric disorders. In the present study, we investigated whether ASTN2 is involved in the emotional and cognitive functions using SCZ-like animal model (phencyclidine-administered mice) and Astn2 homozygous (-/-) mice. Phencyclidine-administered mice showed some behavioral abnormalities (e.g., the impairments of social and cognitive behaviors) related to dysfunctional serotoninergic and/or dopaminergic systems. The expression levels of Astn2 mRNA of phencyclidine-administered mice were increased in the prefrontal cortex/striatum and decreased in the olfactory bulb/amygdala, compared to those of saline-administered mice. While, Astn2 (-/-) mice showed the increased locomotor activity to the novel environment, increased stereotypic and social behaviors, or increased impulsivity, and the decreased anxiety-like behaviors, or decreased exploratory preference, compared to Astn2 (+/+) mice. In Astn2 (-/-) mice compared with Astn2 (+/+) mice, serotonin contents were decreased in the amygdala, and serotonin and dopamine turnovers were increased in the amygdala and the striatum, respectively. These results demonstrate that ASTN2 regulates emotional and cognitive functions via serotonergic and/or dopaminergic systems. ASTN2 is suggested to play an important role in the development of neuropsychiatric disorders.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-078 運動中の海馬で生じるグリコーゲン分解を介した活動依存性アストロサイトーニューロン乳酸シャトル：ドーパミンの役割 Takashi Matsui（松井崇）¹，Ignacio A Torres（Torres A Ignacio）²，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 ²Cajal Institute, CSIC, Madrid, Spain Astrocytic glycogenolysis-produced lactate is transported into neurons as a critical energy source and/or a neuromodulator, which is called "astrocyte-neuron lactate shuttle (ANLS) hypothesis". In the hippocampus, ANLS is required for memory functions. Although exercise, even in mild intensity, activates hippocampal neurons depending on running speed and enhances memory functions, hippocampal glycometabolism during exercise is less clear. Noradrenaline (NA) and serotonin (5-HT) are activators of astrocytic glycogenolysis, but they do not increase in the hippocampus during running exercise. Meanwhile, dopamine (DA), another monoamine, increases in the running hippocampus, and the agonism of the DAD2 receptor (DAD2R) elicits Ca2+ influx in astrocytes, a trigger for glycogenolysis. We thus hypothesized that running exercise induces DAD2R-mediated hippocampal ANLS through glycogenolysis in a speed-dependent manner. To test this hypothesis, we here employed a rat model of treadmill running and high-power (10 kW) microwave irradiation for accurate detection of glycogen, lactate, and monoamine levels in the hippocampus. Immediately after 30 min of exercise at different speeds (mild: 10 m/min, moderate: 20 m/min, and hard: 30 m/min), hippocampal glycogen decreased, while its lactate levels increased in a speed-dependent manner associated with DAergic, but not with NAergic or 5-HTergic activation. In vivo microdialysis during exercise revealed that hippocampal extracellular DA, but not NA or 5-HT, increased depending on running speed. Furthermore, in primary cultured hippocampal astrocytes, DA and a DAD2R agonist, but not a DAD1R agonist, induced dose-dependent glycogen decreases via Ca2+/cAMP/PKA signaling. Finally, a local injection of a DAD2R antagonist, but not a DAD1R antagonist, into the hippocampus prevented hippocampal glycogen decrease during mild exercise. These results clearly support our hypothesis for DAergic activity-dependent ANLS through glycogenolysis in the exercising hippocampus, suggesting a novel underlying mechanism in exercise-enhanced memory functions.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-079 線条体中型有棘神経からコリン作動性介在ニューロンへのGABA遊離はM1アセチルコリン受容体によりシナプス前性に抑制される Etsuko Suzuki（鈴木江津子），Toshihiko Momiyama（籾山俊彦）東京慈恵医大薬理 We have reported M1 muscarinic receptor-mediated inhibition of GABAergic transmission onto striatal cholinergic interneurons. This inhibition does not require intracellular G-protein cascade in the postsynaptic cholinergic interneurons. In the present study, the site of cholinergic modulation was examined. We used transgenic mice with restricted expression of channelrhodopsin-2 (ChR2) in the striatal medium spiny neurons (MSNs). Whole-cell patch-clamp recordings were made from striatal cholinergic interneurons in P10-17 mice brain slices. Neurons were voltage clamped at -60 mV. Light stimulation (470 nm, 5 ms duration) evoked postsynaptic currents in the presence of glutamate and glycine receptor antagonists. These postsynaptic currents were blocked by GABA_A receptor antagonist, bicuculline, suggesting they were GABA_A receptor-mediated inhibitory postsynaptic currents (IPSCs). A muscarinic acetylcholine receptor agonist, carbachol (1 μM), suppressed IPSCs by 49.5 ± 7.8%, (n = 5). To examine the changes in GABA release probability, we calculated coefficient of variation (CV) at baseline, after application of bicuculline , after washout of bicuculline and after application of carbachol. CV at baseline and after application of bicuculline were 0.2 ± 0.02 and 0.2 ± 0.03, respectively. The CV was not increased after application of bicuculline, suggesting the action site of bicuculline was postsynaptic GABA_A receptors. On the other hand, CV after application of carbachol was significantly increased (0.5 ± 0.06, p = 0.004), suggesting that GABA release probability was changed by carbachol. These results suggest that M1 muscarinic receptors inhibit GABA release from MSNs onto cholinergic interneurons.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-080 Morphological evidence for top-down innervation from prefrontal cortex to Locus coeruleus in mice Yu-Shan Kuo（Kuo Yu-Shan）^1,2，Ming-Yuan Min（Min Ming-Yuan）¹，Chao-Cheng Kuo（Kuo Chao-Cheng）¹，Jung-Chien Hsieh（Hsieh Jung-Chien）¹，Hsiu-Wen Yang（Yang Hsiu-Wen）² ¹Department of Life Science National Taiwan University, Taipei, Taiwan ²Department of Biomedical Sciences Chung-Shan Medical University Locus coeruleus (LC) contains high density of noradrenergic (NA) neurons that are the major supply of norepinephrine to the forebrain through global axonal projections. The LC-NA system plays important roles in modulating cognitive shift for behavioral adaption through adjusting the gain of neuronal networks. Accordingly, LC is expected to receive top-down afferent inputs from prefrontal cortex (PFC), which is involved in high cognitive functions, such as value evaluation of sensory stimuli and decision-making. Indeed, direct axonal projections from the PFC to the LC has been found in primates. Nevertheless, whether the top-down inputs from the PFC to the LC synapse directly on NA neurons or on local interneurons that integrate inputs from PFC with those from other cortical regions to LC NA neurons remains unknown. In this study, we aim to address this question. We first produced an adeno-associated virus (AAV) carrying an open reading frame of floxed wheat germ agglutinin (WGA) and injected the AAV into LC of tyrosine-hydorxylase cre (TH-cre) mouse. By using Immunohistochemistry, we not only observed numerous neurons showing both TH- and WGA-immunoreactive (ir) within LC as expected, but also a few neurons that do not show TH-ir but WGA-ir in LC and the surrounding areas. Since there are no other catecholamine neurons than LC NA neurons in dorsal potine tegmental area, and leak-out expression of cre in other cells than in LC NA neurons is very limited in the TH-cre mouse, we reason that these non-TH-ir but WGA-ir neurons are resulted from transneuronal transport of WGA from LC neurons and are the local interneurons making synaptic contacts with LC NA neurons. These interneurons are located predominantly in the medial aspect of peri-LC region, the gap between LC proper and the adjacent Barrington nucleus. Among the interneurons, 15% are found to express glutamate acid decarboxylase (GAD) and 40 % express FoxP2. To confirm whether these interneurons receive top-down cortical inputs, we injected AAV carrying reading frame of ChR2-mCherry into the PFC for anterograde tracing and the AAV-DIO-WGA into the LC of TH-cre mouse. We found that PFC fibers not only contact on LC NA neurons but also on the interneurons (non TH-ir but WGA-ir) expressing GAD and FoxP2. These results do support our arguments that there are local excitatory and inhibitory interneurons that have functional connection with LC NA neurons and can integrate PFC inputs onto LC NA neurons.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-081 ドコサヘキサエン酸によるグルタミン酸トランスポーター調節に関する研究 Kanako Takahashi（高橋華奈子）¹，Tomohiko Irie（入江智彦）²，Yasunari Kanda（諫田泰成）²，Kaoru Sato（佐藤薫）¹ ¹国立医薬品食品衛生研薬理・神経薬理 ²国立医薬品食品衛生研薬理 In the CNS, L-glutamate (L-Glu) transporter EAAT2 removes excitatory neurotransmitter L-Glu in synaptic cleft, thereby maintaining an efficient neurotransmission and preventing excitotoxicity. Docosahexaenoic acid (DHA; C22:6), one of polyunsaturated fatty acid (PUFA), is a constituent of astrocyte membrane phospholipids and is released after L-Glu stimulation. In recent years, DHA is sold as a brain function improving supplement. However, the precise effects and mechanisms of DHA are still unknown. To this aim, we applied two-electrode voltage clamp method to Xenopus oocytes overexpressing EAAT2. Exogenously-applied DHA (30-300 μM) reversibly increased the amplitude of L-Glu-induced EAAT2 currents, but had no effects on EAAT1. Transient application of Triton X-100 (200 μM), which increases membrane fluidity, had little effects on EAAT2 currents, suggesting that the DHA-induced enhancement of EAAT2 currents is independent of the membrane elasticity. The enhancement of EAAT2 was not prevented by the inhibitors of cyclooxygenase and lipoxygenase, indicating that DHA itself revealed the effects but not through the metabolites. DHA has a carboxyl group of which protonation is dependent on pH. When the extracellular pH was decreased from 7.5 to 5.5, i.e., in the deprotonated situation, the enhancement of EAAT2 by DHA was disappeared. In addition, DHA-methyl-ester (200 μM), non-chargeable DHA analogue, had no effects on EAAT2 current. Taken together, it is suggested that the negative charge is important. Next, we attempted to identify EAAT2 region important for the effects on DHA using EAAT1/2 chimeras. By substituting the transport domain of EAAT1 by that of EAAT2, the effects of DHA on EAAT1 turned out to be enhancement, suggesting that this region is important for the augmentation of EAAT2 current by DHA. Currently, we are further identifying the region in the transport domain essential to the effects of DHA.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-082 中枢性Gタンパク質共役型受容体を介する一次繊毛縮退ーRNAseqによる調節分子の同定 Yumiko Saito（斎藤祐見子）¹，Sakura Tomoshige（友重桜子）¹，Tatsuo Miyamoto（宮本達雄）²，Yuki Kobayashi（小林勇喜）¹ ¹広島大学総合科学研究科 ²広島大学原医研 The primary cilium is a plasma membrane-protruding sensory organelle supported by a 9 + 0 microtubule axoneme that extends from the basal body. Primary cilium coordinates multiple cellular signaling pathways through ion channels and conventional G-protein-coupled receptors (GPCRs) harbored in the cilium membrane. During postnatal development, the cilium membrane becomes equipped with a limited set of GPCR such as melanin-concentrating hormone (MCH) receptor 1 (MCHR1), which is involved in feeding and mood. The length of the cilium is a characteristic parameter that has implications for cilia function. We have previously found a new biological phenomenon, the MCH-mediated cilia shortening with an EC50 value of 0.54 nM in human telomerase-immortalized retinal pigmented epithelial (RPE1) cells. We further showed the MCH-MCHR1-Gi/o-Akt pathway causes a series of signal cascade that both depolymerize cytoplasmic tubulin and increase actin filaments. However, the precise mechanisms of this process remain unclear. In this study, we investigated changes in the transcriptome profile in MCHR1-expressing RPE1 cells with RNA-seq technology. There were 536 genes, which were significantly changed more than two-fold in response to MCH treatment. After performing qPCR to validate the results of RNA-seq, several candidates in the list showed great potential for further study as reflected by the fold change in mRNA expression along time-course. Finally, functional analysis by using siRNA knockdown and CRISPR-ObLiGaRe method revealed one molecule as the most significant regulator for MCHR1-mediated cilia shortening. We believe that our study will lead to a better understanding of and insight into the molecular mechanisms underlying ciliary GPCR-mediated cilia length control.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-083 ADAM22のリン酸化は脳内の抗てんかん蛋白質複合体、LGI1-ADAM22の量を制御する Norihiko Yokoi（横井紀彦）^1,2，Yuko Fukata（深田優子）^1,2，Makoto Sanbo（三宝誠）³，Teppei Goto（後藤哲平）³，Masumi Hirabayashi（平林真澄）³，Masaki Fukata（深田正紀）^1,2 ¹生理研分子細胞生理生体膜 ²総研大院生命科学生理 ³生理研行動・代謝分子解析センター遺伝子改変動物作製室 Epilepsy is one of the most common neurological disorders, and featured by recurrent, unprovoked seizures. Mutations of LGI1, a neuronal secreted protein, and its receptor, ADAM22, are reported in patients with seizures and intellectual disability. We previously reported that LGI1 knock-out mouse shows severe epileptic seizures and dies within three weeks after birth. LGI1 and ADAM22 form a heterotetrameric assembly that bridges the synaptic cleft and regulates AMPA receptor-mediated synaptic transmission. An epilepsy-causative mutation of LGI1 severely reduces the amount of the LGI1-ADAM22 complex in the mutant mouse brain, and its increase by a chemical treatment ameliorates the seizure phenotype of the mouse. Here, we report that phosphorylation of ADAM22 regulates the amount of the LGI1-ADAM22 complex in the brain. We found that most of ADAM22 is phosphorylated in the mouse brain and we determined the ADAM22 phosphorylation sites in vitro. To define the physiological role of phosphorylation of ADAM22, we generated the serine-alanine (SA) mutation knock-in (KI) mouse, in which phosphorylation of ADAM22 was deficient. In the KI mouse brain, the amount of the ADAM22 SA protein was robustly decreased to about 30% of the ADAM22 wild-type (WT) protein in the WT mouse brain. The amount of LGI1 in the KI mouse brain was also decreased to the same degree as ADAM22 SA. Consistently, ADAM22 SA was degraded much faster than the WT protein in cultured neurons. This is not due to the misfolding of ADAM22 SA, as ADAM22 SA in the brain was resistant to Endoglycosidase H. Furthermore, immunocytochemical analysis showed that the axo-dendric staining of ADAM22 SA was decreased. These results suggest that ADAM22 phosphorylation regulates the stability of the LGI1-ADAM22 complex at the synapse. We will also discuss the molecular mechanism to regulate the stability by ADAM22 phosphorylation.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-084 Atp1a3のハプロ不全は小脳プルキンエ細胞にグルタミン酸取り込み能の減弱を引き起こす：グリア細胞による補償とその影響 Shin’Ichiro Satake（佐竹伸一郎）^1,2，Kiyoshi Kawakami（川上潔）³，Keiko Ikeda（池田啓子）⁴ ¹自然科学研究機構・生理研 ²総研大・生命科学 ³自治医大・分子病態治療研究センター ⁴国際医福大・医 Sodium pump (Na,K-ATPase) plays a critical role in the maintenance of the electrochemical Na⁺/K⁺ gradients across the cell membrane. Utilizing this gradient, excitatory amino acid transporters (EAATs) are responsible for cellular uptake of the neurotransmitter glutamate. Mutations in the human Na pump α3 subunit gene, ATP1A3, have been identified as the cause for rapid-onset dystonia with Parkinsonism (RDP), alternating hemiplegia of childhood (AHC), and CAPOS syndrome (cerebellar ataxia, areflexia, pes cavus, optic atrophy, and sensorineural hearing loss). However, it is unknown whether EAATs contribute to the neurological disorders originated from the Na pump dysfunction. Considerable evidence suggests a possible involvement of the cerebellum in dystonia, although the basal ganglia have been proposed as the primarily responsible region. Therefore, we examined the EAAT activity in the cerebellum in Atp1a3 heterozygous knockout mice (Atp1a3^+/-). We found a remarkable reduction of the glutamate uptake-coupled currents mediated by the EAAT4 subtype in cerebellar Purkinje cells (PCs) in Atp1a3^+/- compared with those of wild type littermates. To our surprise, the amplitude of EAAT currents in the astrocyte Bergmann glia (BG) was profoundly higher in Atp1a3^+/-. Consistently, the protein levels of the glia-specific subtype EAAT1 increased in the cerebellum of Atp1a3^+/-. The glutamate uptake capacity will be augmented in BG to compensate the weakened neuronal EAAT activity. Furthermore in Atp1a3^+/-, long-term depression (LTD, postsynaptic origin and mGluR1-dependent) was diminished at the excitatory synapses from parallel fibers (PFs) to PCs. The impaired LTD was rescued by application of the EAAT1 inhibitor UCPH102 and the mGluR1 agonist DHPG. The result suggests that in Atp1a3^+/-, the enhanced glial EAAT activity attenuates the extracellular diffusion of the glutamate spilled out of the PF-PC synaptic clefts and thereby reduces the activation of the perisynaptically distributed mGluR1s on PC dendritic spines. It is quite likely that these events interfere with the LTD. Our findings would provide a clue for further understanding of pathophysiological mechanisms underlying RDP, AHC, and CAPOS syndrome.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-085 マウス脊髄後角におけるA線維性およびC線維性EPSCsに対するGABAトランスポーター阻害薬の効果 Misa Oyama（尾山実砂），Shun Wasatabe（渡辺俊），Takashi Iwai（岩井孝志），Mitsuo Tanabe（田辺光男）北里大・薬・薬理 Based on the evidence that the excessive excitation in the spinal dorsal horn is one of the crucial factors for central sensitization leading to chronic pain, we have focused on the importance of enhancing inhibitory influences to normalize the balance between excitation and inhibition in the superficial dorsal horn. GABA transporter (GAT1 and GAT3) inhibitors were reported to have analgesic effects in animal models of neuropathic pain. However, the influence of GAT inhibitors on excitatory and inhibitory synaptic transmission in the superficial dorsal horn remains unknown. We previously evaluated the effects of GAT inhibitors on miniature excitatory and GABAergic inhibitory postsynaptic currents (mEPSCs and mIPSCs) and evoked EPSCs and IPSCs (eEPSCs and eIPSCs) in the dorsal horn of spinal slices prepared from adult mice using whole-cell voltage-clamp technique. The GAT1 inhibitor NNC-711 largely attenuated the frequency of mEPSCs rather than mIPSCs, and decreased the amplitude of eIPSCs but not eEPSCs. By contrast, the GAT3 inhibitor SNAP-5114 reduced the frequency of mIPSCs and the amplitude of eEPSCs without affecting mEPSCs and eIPSCs. In the present study, we investigated the effects of GAT inhibitors on monosynaptic A- or C- fiber-evoked EPSCs derived from primary afferent fibers stimulated through a suction electrode in the spinal slices with an attached L4, L5 or L6 dorsal root. NNC-711 reduced C-fiber-evoked EPSCs, and SNAP-5114 decreased both A- and C-fiber-evoked EPSCs, all of which were not observed in the presence of the GABA_B receptor antagonist CGP55845. Consistently, intrathecally administered GAT inhibitors failed to produce the anti-allodynic effects in mice developing neuropathic pain after partial sciatic nerve ligation when mice were pretreated with CGP55845. NNC-711 generated the analgesic effects without affecting locomotor activity, while SNAP-5114 showed motor disturbance at the dose for pain relief. Therefore, endogenously increased GABA under the blockade of both GAT1 and GAT3 decreases excitatory neurotransmission via the activation of presynaptic GABA_B receptors, resulting in pain relief. NNC-711 is more favorable than SNAP-5114 to use as a novel analgesic agent because of less side effects.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-086 細胞表面上P2X4受容体を認識する抗P2X4受容体Fabの大腸菌発現系の確立 Masashi Otomoda（御供田将士）¹，Yoshito Abe（阿部義人）¹，Tatsuhiro Igawa（井川達弘）¹，Syuhei Kishikawa（岸川秀平）¹，Caaveiro MM Jose（Jose MM Caaveiro）²，Tadashi Ueda（植田正）¹ ¹九大院薬　蛋白創薬 ²九大院薬　グローバルヘルス Recently, we prepared the monoclonal antibody against rat P2X4 receptor, which is an ATP-activated ion channel and is involved in neuropathic pain. This antibody recognized native structure of the head domain of rat P2X4 receptor with high affinity (~20 nM) but did not inhibit the calcium influx function of P2X4 receptor (Igawa, Kishikawa and Abe et al. Purinergic Signal. 2019). Moreover, since its epitope is located near the ATP-binding region, we will be able to engineer and chemically modify the Fab of the antibody to additional functions. Since we reported the stable refolding system of humanized Fab fragments in vitro, in which H chain and L chain were expressed as inclusion bodies, respectively (Fujii and Ohkuri et al. J. Biochem 2007), we attempted to prepare of mouse Fab fragment of anti-P2X4 antibody using the refolding system. However, the method could not apply to the refolding of mouse Fab fragment of anti-P2X4 antibody, resulting in little recovery of the folded rat Fab. In this study, we cloned the co-expression vector with cDNAs of H chain and L chain and directly expressed the Fab fragment as a soluble protein. After purification using ion-exchange and affinity chromatography, we showed that the Fab fragment possessed the same activity as that of the antibody expressed by hybridoma cells and the refolded Fab prepared previously. Therefore, we established a simple and non time-consuming preparation method of recombinant Fab fragment against the P2X4 receptor.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-087 ラットP2X4受容体の天然構造を認識するモノクローナル抗体の作成 Yoshito Abe（阿部義人）¹，Tatsuhiro Igawa（井川達弘）¹，Shuhei Kishikawa（岸川秀平）¹，Tomohiro Yamashita（山下智大）²，Hiroyuki Tanaka（田中宏幸）³，Hidetoshi Tozaki-Saitoh（齊藤秀俊）⁴，Makoto Tsuda（津田誠）⁴，Kazuhide Inoue（井上和秀）⁵，Tadashi Ueda（植田正）¹ ¹九大院薬蛋白創薬 ²九大院薬グローバルヘルス ³九大院薬生薬 ⁴九大院薬ライフイノベーション ⁵九大院薬薬理 P2X purinergic receptors are ATP-driven ionic channels expressed as trimers and showing various functions. A subtype, P2X4 receptor is involved in neuropathic pain, which is occurred by activation of microglial cell after influx of calcium ion. Therefore, preparation of anti-P2X4 receptor monoclonal antibody will be available for the research tool for the functional P2X4 receptor and the imaging tool for the expression level of P2X4 receptor, which is strongly correlated with the quantitative level of neuropathic pain. In this study, in order to prepare monoclonal antibodies recognizing the native structure of rat P2X4 (rP2X4) receptor on the cell, we immunized mice with rP2X4's head domain (rHD, Gln111-Val167), which possesses an intact structure stabilized by three S-S bond formation (Igawa and Abe et al. FEBS Lett. 2015), as an antigen. The monoclonal antibody obtained here recognized rHD in native P2X4 receptor with the high affinity (about 20 nM) and could detect the rat P2X4 receptor expressed in 1321N1 human astrocytoma cells. Moreover, in order to characterize the monoclonal antibody, we examined the epitope on the P2X4 receptor using site-directed mutagenesis. As a result, we determined that Asn127 and Asp131 in turn structure of head domain of rat P2X4 receptor, in which combination of these amino acid residues are only conserved in P2X4 receptor among P2X family, are closely involved in the interaction with antibodies by using ELISA and surface plasmon resonance.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-088 Visualization of ionotropic glutamate receptor subunits by novel chemical labeling for electron microscopy Marijo Jevtic（Jevtic Marijo）¹，Shigekazu Tabata（Tabata Shigekazu）¹，Manabu Abe（Abe Manabu）²，Akio Ojida（Ojida Akio）³，Kenji Sakimura（Sakimura Kenji）²，Ryuichi Shigemoto（Shigemoto Ryuichi）¹ ¹Institute of Science and Technology Austria ²Dept. of Cellular Neurobiology, Brain Research Inst., Niigata Univ. ³Faculty of Pharmaceutical Sciences, Kyushu Univ. Ionotropic glutamate receptors (iGluRs) mediate fast excitatory neurotransmission and their role is implicated in complex processes such as learning and memory and various neurological diseases. These receptors are composed of different subunits and the subunit composition can affect channel properties, receptor trafficking and interaction with other associated proteins. However, no tools to visualize and study the subunit composition of single channels in situ have been available. Electron microscopy allows us to investigate protein localization with a high resolution, but conventional immunogold labeling approaches have limitations in the single channel analysis because of the large size of antibodies and steric hindrance hampering multiple labeling of single channels. Here, we apply chemical labeling using a short peptide tag and small synthetic probes, which form specific covalent bond with a cysteine residue in the tag (reactive tag system) to iGluRs. This system allows us to detect subunit of interest with a few nanometer resolution without interfering with detection of other subunits. Tagged subunits of iGluRs are expressed on the neuronal membrane, subsequently labeled with chemical probes conjugated with ultra-small gold particles and analyzed by SDS-digested freeze-fracture replica method for electron microscopy. The specificity, sensitivity and resolution of the chemical labeling are compared with conventional SDS-digested freeze-fracture replica immunogold labeling.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-089 トリ蝸牛神経核におけるカルシウムチャネル発現の発達変化 Kensuke Muto（武藤健右），Rei Yamada（山田玲），Hiroshi Kuba（久場博司）名古屋大院医細胞生理 In developing neurons, calcium ion plays significant roles in maturation of various cellular functions. In avian cochlear nucleus, the expression of voltage-gated potassium channels increases around hatch, which is prerequisite for the precise temporal coding in the mature neurons. We previously showed that activity-dependent calcium influx through voltage-gated calcium channels is critical for the upregulation of potassium channels. However, it is not well understood how the expression of calcium channels is regulated during development. In this study, we addressed this issue by recording calcium currents with patch-clamp technique in brainstem slices of chickens at different ages. We classified calcium currents into two groups according to their activation voltages, low-voltage-activated (LVA) and high-voltage-activated (HVA) currents, and analyzed them separately. During embryonic days, both LVA and HVA currents increased gradually and reached their peak just before hatch. LVA current decreased and almost disappeared after hatch, whereas HVA current did not change during the period. Moreover, deprivation of auditory input via cochlea ablation precluded the decrease of LVA current. These observations would support the idea that the developmental increase of calcium channels augments calcium influx and contributes to acquire the mature pattern of potassium channel expression in the auditory neurons. The results also suggested that the expression of LVA channels was regulated by auditory input.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-090 線虫C. elegansの酸素情報と温度情報の統合に関わる神経回路の解析 Misaki Okahata（岡畑美咲）^1,2，Aguan D Wei（Wei D Aguan）³，Akane Ohta（太田茜）^1,2，Atsushi Kuhara（久原篤）^1,2,4 ¹甲南大学自然科学 ²甲南大　統合ニューロバイオロジー研究所 ³Seattle Children's Research Institute, Seattle, USA ⁴国立研究開発法人日本医療研究開発機構 Temperature is one of the most important environmental factors for creatures. We are studying about cold acclimation in C. elegans to reveal the mechanisms for temperature response. We used previous DNA micro array analysis to identify novel gene involved in cold acclimation. kqt-2 mutant defective in KCNQ-type potassium channel showed abnormal cold acclimation. Interestingly, this abnormality was stronger when kqt-2 mutant cultivated at agar medium plate of 6 cm in diameter than cultivated at 3.5 cm in diameter. The expression of KQT-2 was observed in ASK and ADL head chemosensory neurons and intestine. To identify the cells where kqt-2 are involved in cultivation-space dependent-cold acclimation, cell specific rescue experiments were performed. The abnormal cold acclimation of kqt-2 mutant was fully rescued by expressing kqt-2cDNA in ADL chemosensory neuron. Ca2+ imaging suggested that ADL is responsive to temperature changes. Previously reported, neuronal activity of ADL for ascaroside was changed by oxygen concentration, which is sensed by URX sensory neuron connecting to ADL through RMG interneurons. To examine relationship between ADL and URX in cold acclimation, we used the mutation of GCY-35, an oxygen receptor in URX coelomic sensory neurons. We found that abnormal temperature response and cold acclimation of kqt-2 mutant were suppressed by gcy-35 mutation, implying that ADL works in downstream of URX oxygen sensory neuron, and temperature signaling in ADL is affected by oxygen signaling received by URX. We hypothesized that abnormal cold acclimation of kqt-2 depending on diameter of agar medium plate causes oxygen concentration in the agar plate. The oxygen concentrations of 3.5cm-diameter plate was 5% lower than that of 6cm-diameter plate. We also found that abnormally cold acclimation of the kqt-2 mutant was recovered to normal levels after cultivation with a lower O2 concentration in the incubator. Our genetic results elucidated that a simple neuronal circuit integrating two different sensory modalities, temperature and oxygen, that determines cold acclimation (Okahata et al., Science advances, 2019).		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-091 背側蝸牛神経核Cartwheel細胞におけるSKチャネルの細胞内Ca²⁺ソース Tomohiko Irie（入江智彦）国立医薬品食品衛生研薬理 The intracellular Ca²⁺ is a crucial second messenger which modulates directly or indirectly ion channels in excitable cells, including neurons. Some of the modulations are mediated by activation of Ca²⁺-activated K⁺ channels, resulting in the change of firing properties. We previously reported that activation of large-conductance, voltage- and Ca²⁺-activated K⁺ channels by Ca²⁺-induced Ca²⁺-release (CICR) through ryanodine receptors can control the generation of burst firings in cartwheel inhibitory interneuron of the mouse dorsal cochlear nucleus (Irie and Trussell, 2017). In this paper, we also found unexpectedly that the CICR does not activate small-conductance, Ca²⁺-activated K⁺ (SK) channels of cartwheel cells. This observation contrasts with the Ca²⁺ signaling seen in the SK channel activation of other types of cells, raising the question how intracellular Ca²⁺ is supplied in order to activate SK channels in cartwheel neurons. To answer the question, I recorded SK currents from mice cartwheel cells using in vitro patch-clamp recording, and explored the Ca²⁺ source pharmacologically. I will present data on what channels/receptors are involved in the gating of the SK currents, and on how the activation contributes to the firing mode in cartwheel cells.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-092 脳神経細胞における伸展感受性G-タンパク質共役受容体の存在 Nanae Takahasi（Takahashi Nanae），Zen'ichiro Wajima（輪嶋善一郎），Yoshihisa Kudo（工藤佳久）東京医科大学八王子医療センター　麻酔科 Aim: In the present study, we pharmacologically analyzed the mechanism behind the increase in intracellular Ca²⁺ caused by the brain cell swelling and regulatory volume change that occurs in mouse brain slices when they are exposed to hypotonic artificial cerebrospinal fluid (hypo-ACSF). Method: Horizontal slices of mouse (C57BL) were loaded with the fluorescent Ca²⁺ indicator Fura-2. Measure fluorescence when excited at 360 nm and 380 nm (F360 and F380) and exposed to hypo-ACSF (3/4 dilution of normal ACSF). Although F360 was not affected by Ca²⁺ concentration, it did respond to the increase in intracellular water resulting from cell swelling. As an increase in intracellular Ca²⁺ reduces F380, the intracellular calcium concentration can be determined by measuring the F360/F380 values. Results: Exposure to hypo-ACSF quickly causes brain cells to swell, and in approximately five minutes, shrinkage is induced by a homeostatic process called regulatory volume change (RVC). We observed that there was only a small change in fluorescence in preparations stained with sulforhodamine 101 (1μM), which is specifically taken up by astrocytes, and that neurons accounted for most of the observed swelling. The concentration of intracellular Ca²⁺ began to rise when RVC was induced. Using the voltage-sensitive dye JPW 1114, we observed that the brain cells depolarized after exposure to hypo-ACSF. Significant regional differences in the rise in Ca²⁺ concentration was seen in the cerebral cortex and the hippocampus. Although LaCl₃ (0.1mM) and GaCl₃ (0.1mM), which inhibit stretch-activated Ca²⁺ channels, tended to inhibit the increase in Ca²⁺ concentration, almost no inhibitory effect was observed with CNQX (0.1mM) or APV (0.1mM), and even a voltage-dependent Ca²⁺ channel inhibitor (nicardipine) (0.1mM) had no inhibitory effect. However, a significant inhibitory effect was observed when specimens were treated with the ER Ca²⁺ATPase blocker thapsigargin (1μM) 30 minutes before measurement, or with the specific IP₃ receptor blocker xestospongin C (10μM). Conclusion: In the present study, we observed that the increase in intra-neuronal Ca²⁺ accompanying RVC involves not only stretch-activated Ca²⁺ channels, but also G proteins (Gq) coupled process.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-093 皮質錐体細胞サブタイプに依存したオシレーション活動の運動学習における役割 Takeshi Ohtsuka（大塚岳），Yasuo Kawaguchi（川口泰雄）生理学研究所大脳神経回路論研究部門 The cortex outputs the information to several brain areas through different sets of pyramidal cells (PC). It has been shown that cortical PCs form intra- and inter-laminar subnetworks, depending on pyramidal projection subtypes. However, it remains unknown how individual PC subnetworks concern in cortical information processing and the high order brain functions. In the present study, we investigated the effects of optogenetic manipulations of each PC subtype activities on motor pattern learning. Channel rhodopsin (ChR2) or archaerhodops in (eArch) was selectively expressed in L2/3 PCs or L5 PC subtypes of rat motor cortical areas by in utero electroporation. Pattern learning task was examined using forced wheel running system, which animals learn irregularly-patterned intervals between step bars to drink water from the supply port, while M1 area was stimulated during running. To evaluate motor learning, we used the falling rate from step bars of a rotating wheel. Time courses of pattern learning formation were affected by optogenetic manipulations of specific PC subtypes. To examine cortical network activities during a task, in vivo recordings were obtained. We found that oscillatory activities in local field potentials during learning were generated and enhanced by light stimulations. Moreover, in slice preparations, oscillatory activities were observed in FS cells as well as in the L5 PC subtype by light stimulations to the specific PC subtype. Our results suggest that oscillatory activities induced by specific L5 PC subtype contribute to formation of the motor learning.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-094 ラット前頭皮質５層サブレイヤーに依存した興奮性-抑制性回路について Mieko Morishima（森島美絵子）^1,2，Yasuo Kawaguchi（川口泰雄）^1,2 ¹生理研基盤神経科学大脳神経回路論 ²総研大院生命科学生理 In the rat frontal cortex, layer 5 (L5) is divided into upper (L5a) and lower (L5b) sublayers on the basis of thalamic inputs. One of the major pyramidal subtypes in L5 is cortico-pontine (CPn) cells, projecting to the pontine nuclei. L5a and L5b CPn cells are morphologically different: L5a cells are longer in apical oblique branches and more extended in basal dendrites than L5b cells; L5a cells send their axons to the thalamus, whereas L5b cells project down to the spinal cord. However, it is still unknown that each pyramidal cell type forms a distinct local circuit including GABAergic cells in the sublayer. Low threshold spike (LTS) cells, a subtype of somatostatin (SOM) positive cells, more preferentially connected to CPn cells than other pyramidal cell types. Therefore, we compared the connections of CPn/LTS pair between L5a and L5b, using paired whole-cell recordings and morphological reconstruction of the recorded cells. The excitatory input from CPn cells to LTS cells was similar between the L5a and L5b CPn/LTS pairs. However, the excitatory connection patterns were different between them. Meanwhile, LTS cells mostly ascend their axons to layer 1 (L1) and innervate the dendrites of pyramidal cells, which are called Martinotti cell. The axonal arborizations in L1 were significantly longer in L5a LTS cells than in L5b LTS cells. The spatial overlap of LTS cell axons and CPn cell dendrites is also different between the L5a and L5b LTS/CPn pairs. However, the IPSC amplitudes recorded from L5a LTS/CPn pairs were similar to those of L5b LTS/CPn pairs. These results suggest that CPn cells and LTS cells make distinct local circuits in the sublayers, and perform information processing specific for the long-range subcortical outputs.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-095 小脳苔状線維シナプスにおけるシナプス小胞動態の解析 Takafumi Miki（三木崇史），Takeshi Sakaba（坂場武史）同志社大院脳科学シナプス分子機能 During neuronal activity, continuous synaptic vesicle release is required at presynaptic terminals firing at high frequency. As limited size of active zone (AZ) in presynaptic terminals and limited number of release site in AZ, vesicle recruitment is though to be crucial for sustained vesicle release. However, it has been little known how fast the recruitment occur in such synapses. In this study, we investigated synaptic vesicle dynamics near presynaptic membrane in cerebellar mossy fiber (cMF) terminals, which can fire up to 1- kHz. We labeled synaptic vesicle by loading FM-dye in dissociated cMF terminals, and observed individual synaptic vesicle movement upon electrical stimulations using total internal reflection fluorescence (TIRF) imaging with simultaneous electrophysiological recording. FM dye-labeled vesicles appeared rapidly near plasma membrane upon 100-ms depolarization pulse with a time constant of 100-200 ms, showing that vesicle recruitment occur as fast as vesicle release represented by disappeared events in TIRF image. The rapid vesicle recruitment was abolished by applying actin polymerization inhibitor, LatrunculinA. Electrophysiological recording revealed that LatrunculinA also suppressed sustained release upon a depolarization pulse. These results suggest that fast recruitment occur synchronously with vesicle release, and contribute on sustained release at cMF terminals.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-096 海馬苔状線維シナプス前終末におけるエキソサイトーシスおよびエンドサイトーシスのカルシウム依存性 Rinako Miyano（宮野里菜子），Takafumi Miki（三木崇史），Takeshi Sakaba（坂場武史）同志社大学大学院脳科学研究科 Hippocampal mossy fibers form large presynaptic terminals and make contacts with CA3 pyramidal cells. Owing to large sizes, we could use patch clamp technique. By applying membrane capacitance measurements to mossy fiber terminals, we examined the Ca²⁺ dependence of exocytosis and endocytosis, which remains unknown at cortical terminals. We used acute hippocampal slices from rats. Recordings were performed at room temperature. First, we measured the size and the release kinetics of the readily releasable pool (RRP) of synaptic vesicles. Capacitance changes increased with longer stimulation and saturated at a 100-ms depolarizing pulse. To examine the Ca²⁺ dependence of the release kinetics, we added various concentration of exogeneous Ca²⁺ buffers (EGTA and BAPTA) via the patch pipette. High concentration of EGTA had an effect on the time course of the RRP depletion. Next, we applied a pair of 100-ms depolarizing pulses at different interpulse intervals to examine the rate of vesicle replenishment to the RRP. The time constant of recovery from the RRP depletion was about 1 s and was insensitive to the concentration of Ca²⁺ buffers. We also applied a train of 100-ms depolarizing pulses with an interval of 250 ms and monitored the rate of vesicle replenishment. The rate constant of vesicle recruitment was similar to the recovery time constant of the RRP estimated from dual pulse experiments. In order to measure the kinetics of endocytosis, we analyzed capacitance decreases following a 100-ms depolarizing pulse. The capacitance decay was slow and was insensitive to the concentration of Ca²⁺ buffers. These results indicate that the kinetics of exocytosis is sensitive to Ca²⁺ buffers (EGTA and BAPTA). In contrast, vesicle replenishment to the RRP and endocytosis are insensitive to Ca²⁺ buffers.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-097 Electrophysiological, morphological, and connection characteristics of layer 2/3 somatostatin cells in mouse frontal cortex Fransiscus Adrian Agahari（Agahari Fransiscus Adrian）¹，Yasuo Kawaguchi（Kawaguchi Yasuo）^1,2 ¹National Institute for Physiological Sciences, Okazaki, Japan ²SOKENDAI, Okazaki, Japan Cortical somatostatin (SOM)-expressing cell is a major subtype of GABAergic interneuron which primarily innervates pyramidal cell dendrites. In layer 2/3 (L2/3), however, it remains to be investigated whether the SOM cells are differentiated to inhibit pyramidal cells of different projection types (e.g. contralateral, bottom up, and top down projections). We started our investigation by characterizing L2/3 SOM cells in mouse secondary motor (M2) cortex. These cells were identified with tdTomato red fluorescent protein in the progeny of SOM-IRES-Cre and Ai14 mice cross-breeding. Via immunohistochemistry, we indeed confirmed that most of the tdTomato-expressing cells were positive for SOM (92%), but negative for parvalbumin or vasoactive intestinal peptide. Electrophysiologically, based on the slope of action potential frequency-current injection (F-I) curve and coefficient of variation for inter-spike intervals, the SOM cells could be divided into two separated clusters. The first subgroup exhibited regular spiking pattern with weaker adaptation, whereas the second irregular spiking pattern with lower frequencies to the same current stimuli. Whilst the regular spiking cells comprised 74% (80/108 cells) of total population, the remainder 26% (28/108 cells) was the irregular spiking cells. The regular spiking cell had higher input resistance, slower time constant, and larger sag amplitude. Morphologically, the irregular spiking cell extended its dendrites vertically longer down to upper layer 5, whilst those of the regular spiking cell were limited within layer 1-3. The axonal arborization in layer 1A was longer for the regular than the irregular spiking cells. Furthermore, we found that the two subgroups of SOM had different connection patterns with pyramidal cells projecting to the contralateral cortex. These observations suggested that L2/3 has two SOM cell subgroups with different electrophysiological, morphological, and connection characteristics. Consequently, they exert different effects on the corticocortical output from L2/3.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-098 海馬神経細胞樹状突起におけるクラスター型プロトカドヘリンγの局在解析 Saki Hasegawa（長谷川沙紀），Etsuko Tarusawa（足澤悦子），Natsumi Hoshino（星野七海），Hiroaki Kobayashi（小林裕明），Takeshi Yagi（八木健）大阪大院生命機能 The clustered protocadherins(cPcdhs)belong to cadherin superfamily, that play a role in cell adhesion. In mice, cPcdhs consist of three gene clusters of Pcdh-α, Pcdh-β and Pcdh-γ on the same chromosome (ch18) with 58 isoforms, and show the transcellular homophilic interactions. cPcdhs are widely expressed in the central nervous system including hippocampus, and each neuron expresses about 15 isoforms in different combinations determining the neural individuality. Previously we found that deletion of cPcdhs caused the abnormality of hippocampal neural activity in the dissociated cell culture, indicating that cPcdhs are involved in the molecular mechanisms of formation of neural networks. However, it is still unclear how cPcdhs are involved in the process of the formation of synaptic connections. To reveal the functional role of cPcdhs in synaptogenesis, we examined the localization of cPcdh-γ proteins in the dissociated hippocampal neurons of 6-7 days in vitro (DIV6-7), DIV12 and DIV19, and also their colocalization with synaptic proteins during synaptogenesis by immunocytochemistry. Using an anti-cPcdh-γ antibody which recognizes all isoform of cPcdh-γ, we found that cPcdh-γ is localized in dendritic shaft, filopodia and spine in DIV12 cultured neurons. In addition, we investigated colocalization of cPcdh-γ with PSD95 and a subunit of NMDA glutamate receptor (NR1) localized in the postsynaptic sites. Double immunostaining revealed that cPcdh-γ colocalized with PSD95 or NR1 heterogeneously. The proportion of cPcdh-γ colocalized with PSD95 or NR1 increased from DIV6-7 to DIV12. We also observed cPcdh-γ expression in the spines without PSD95, indicating that cPcdh-γ expresses in immature synapses initially before the expression of synapse-related proteins. In conclusion, we found that cPcdh-γ colocalized with postsynaptic proteins in synaptogenesis. We would like to discuss functional roles of cPcdh-γ during synaptogenesis from the results of colocalization of cPcdh-γ and postsynaptic proteins. Also we try to examine the developmental change of localization of cPcdh-γ in the brain slices.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-099 神経回路形成におけるSmc3の新規役割の解明 Ayaka Yamaguchi（山口絢加）¹，Yuki Fujita（藤田幸）^2,3，Toshihide Yamashita（山下俊英）^1,2,3 ¹大阪大院生命機能分子神経科学 ²大阪大学免疫学フロンティア研究センター ³大阪大院医分子神経科学 Pathogenic mechanisms of neurodevelopmental disorders still remain to be fully understood. The symptoms of neurodevelopmental disorders depend on patients. However, abnormal synapse formation is frequently observed in patients with neurodevelopmental disorders such as autism. Recently, it has been shown that cohesin complex plays a significant role in formation of the central nervous system (CNS) and is involved in regulation of transcription through controlling higher order structure of the DNA. Cohesin complex is a ring-like structure consisting of four subunits (Smc1, Smc3, Scc3 and Scc1) and is well-known as an essential factor for chromosome segregation in cell division. Preceding studies have demonstrated that mutations that perturb the function of cohesion complex cause Cornelia de Lange syndrome (CdLS) with intellectual disability, limb abnormalities and characteristic facial features. Previous study showed Smc3-deficient mice exhibited anxiety-related behavior, which is similar to CdLS symptom, due to inhibition of synapse maturation that is involved in signal transduction between neurons. Moreover, it is reported that proteoglycans in which sugar chains are linked core protein are also involved in synapse formation. Interestingly, SMC3 protein was originally identified as the proteoglycan Bamacan. However, SMC3 protein has been mostly investigated as the subunit of the cohesin complex. Therefore, its function as a proteoglycan remains unclear. The purpose of this study is to reveal the Smc3/Bamacan roles for CNS formation. Through immunostaitning, it was revealed that SMC3 protein showed different subcellular localization depending on developmental stages. In addition, it was observed that overexpression of Smc3 with mutations in glycosylation sites caused subcellular localization changes in the neuroblast cell line N1E-115.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-100 マウス大脳皮質における自閉症関連遺伝子Kirrel3の発現 Tomoko Hisaoka（久岡朋子）¹，Tadasuke Komori（小森忠祐）¹，Toshio Kitamura（北村俊雄）²，Yoshihiro Morikawa（森川吉博）¹ ¹和歌山県立医大医解剖二 ²東京大医科研・先端医療研究センター・細胞療法分野 Kirrel3, a member of the immunoglobulin superfamily, is involved in the development of neural circuit, including axonal fasciculation and synapse formation. Mutations of KIRREL3 gene have been implicated in autism spectrum disorder (ASD). Previously, we showed that Kirrel3-deficient mice exhibited ASD-like behaviors, such as social and communication deficits, repetitive behaviors, and sensory abnormalities in addition to hyperactivity. Several reports suggest that the alterations of neuronal connectivity in excitatory and inhibitory cortical circuits disrupt the balance between excitation and inhibition in the neurons, which may induce ASD-like behaviors. However, it remains unknown how neuronal connectivity is altered in the cortex of Kirrel3-deficient mice. To get insights into the roles of Kirrel3 in the cortical circuits, we investigated the expression patterns of Kirrel3 in the neocortex of mice carrying the lacZ reporter gene with a nuclear localization signal under the control of Kirrel3 promoter. In the mice, lacZ activities were detected in the nucleus of Kirrel3-expressing cells. The expression of Kirrel3 was predominantly observed in layers II and V of the motor, visual, and primary somatosensory areas, whereas Kirrel3 was broadly expressed throughout layers II-V of the secondary somatosensory, auditory, limbic, insular, and rhinal areas. All Kirrel3-expressing cells were positive for NeuN, a marker of neurons. Majority of Kirrel3-expressing neurons in the primary and secondary somatosensory areas were positive for GluR2/3, indicating that Kirrel3 was expressed in the excitatory projection neurons. Double-immunofluorescence staining for Kirrel3 with Satb2 and Foxp2, markers of corticocortical and corticothalamic projection neurons, respectively, revealed that Kirrel3 was colocalized with Satb2, but not with Foxp2. In addition, a few Kirrel3-expressing neurons were positive for GABA, a marker of inhibitory interneurons. Therefore, Kirrel3 was expressed in corticocortical projection neurons and small subpopulations of cortical interneurons. These results suggest that Kirrel3 may be involved in the formation and maintenance of corticocortical circuits and specific inhibitory microcircuits in the neocortex. Altered neuronal connectivity in these circuits may contribute to ASD-like behaviors in Kirrel3-deficient mice.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-101 Effect of inhibitory interneuron activation in locus coeruleus on startle and pre-pulse inhibition behavior in mouse Hsing-Chun Tsai（Tsai Hsing-Chun）¹，Ming-Yuan Min（Min Ming-Yuan）¹，Chao-Cheng Kuo（Kuo Chao-Cheng）¹，Hau-Jie Yau（Yau Hau-Jie）³，Hsiu-Wen Yang（Yang Hsiu-Wen）² ¹Department of Life Science, National Taiwan University ²Department of Biomedical Sciences, Chung-Shan Medical University ³Graduate Institute of Brain and Mind Sciences, College of Medicine, National Taiwan University The locus coeruleus (LC) consists of noradrenergic (NA) neurons that have global axonal projection to the forebrain. LC neurons exhibit two spiking patterns, the tonic and phasic, which are known to regulate cognitive shift for behavioral adaptation through gain modulation of the neuronal networks. However, the cell mechanism underlying the regulation of LC tonic and phasic activations remains largely unknown. The pre-pulse inhibition (PPI) is a commonly used operational measurement of sensorimotor gating, which is a fundamental and preattentive form of information- processing allowing organisms to filter information from external and internal domains. Previous study has shown that pharmacological activation of LC tonic activity decreases the PPI. In this study, we report an enhancement of the PPI by inhibiting local inhibitory interneurons in the LC with method of designer-receptor -exclusively-activated-by designer-drug (DREADD). We injected adeno-associated virus (AAV) carrying an open reading frame of floxed hM4Di-mCherry into the LC of vesicular GABA transporter (Vgat)-Cre mice and found that the mice receiving intraperitoneal injection of clozapine-N-oxide (CNO) displays a significant increase in the PPI. On the contrary, the effect on the PPI was observed in mice receiving neither the AAV injected to other pontine areas nor control AAV carrying floxed mCherry only into the LC. Furthermore, the linear regression analysis of startle response and PPI shows no significant relation. The results demonstrate that LC of the mice show more attentive when the inhibitory interneurons in the LC are inactivated. Moreover, the increase in PPI is not due to the virus infection or leakage in other regions, and is independent from the slight increase of startle response. Thus, the effect of inhibitory interneuron in LC might lead to a tendency of increasing LC phasic activation.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-102 自閉症モデルマーモセット大脳皮質樹状突起のin vivo２光子観察 Jun Noguchi（野口潤）¹，Satoshi Watanabe（渡辺恵）¹，Akira Mishima（三嶋晶）¹，Keiko Nakagaki（中垣慶子）¹，Kazuhisa Sakai（境和久）¹，Eriko Sugano（菅野江里子）²，Hiroshi Tomita（冨田浩史）²，Akiya Watakabe（渡我部昭哉）³，Tetsuo Yamamori（山森哲雄）³，Hiroaki Mizukami（水上浩明）⁴，Noritaka Ichinohe（一戸紀孝）^1,5 ¹国立精神・神経セ神経研微細構造 ²岩手大理工生命コース視覚神経科学 ³理研CBS 高次脳機能分子解析 ⁴自治医大分子病態治療研究セ遺伝子治療 ⁵理研CBS 高次脳機能分子解析一戸G Autism spectrum disorder (ASD) is a developmental disorder characterized by several traits such as social communication impairments, perseverative behaviors, and sensory abnormalities. To understand the mechanisms causing the core symptoms of ASD, it is necessary to clarify how neuronal circuits in the higher cognitive centers of patients are altered. Marmosets present a credible animal model for this purpose, since they have high cognitive ability and similar neuronal circuits to humans. A marmoset model of ASD can be created using the anti-epileptic drug valproate (VPA), which has known teratogenic effects, and increases the probability of offspring developing ASD upon fetal exposure. We raised ASD model marmosets by 7-day oral administration of VPA to pregnant mother marmosets from ~60 days post-fertilization. Our group have previously used this model to show increases in dendritic spine density of neocortex layer 2/3 pyramidal neurons in several cortical areas during development, revealing a possible mechanism of how neuronal circuits could be altered (1). The increase in spines could increase competition between candidate neuronal circuits, resulting in less stable circuitry. In the present study, we attempted to directly observe the cerebral cortex dendritic spine stability by using two-photon microscopy to evaluate the efficacy of the neuronal circuits in living marmosets. Adeno-associated virus (AAV) vectors which express fluorescent proteins were injected into the cortex of marmosets to visualize dendrites and axons of pyramidal neurons. Approximately 3 weeks after the AAV injection, the skull and dura were carefully removed and replaced with a cover slip to build an observation window using a similar method described in a previous report (2). The observation window often remained clear for more than 60 days and was suitable for two-photon fluorescence imaging of the same dendrites during the period. Using this method, we evaluate the stability of synapses of neocortex pyramidal neurons in ASD model marmosets. 1. Sasaki T. et al. (2016) Annual Meeting Japan Neuroscience Society, P3-281. 2. Sadakane O. et al. (2015) Cell Reports. 13(9):1989-1999.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-103 シンタキシン１Bの熱性けいれんへの関与:シンタキシン1B遺伝子欠損マウスの神経機能解析 Tatsuya Mishima（三嶋竜弥）¹，Tomonori Fujiwara（藤原智徳）^1,2，Takefumi Kofuji（小藤剛史）^1,3，Ayako Saito（齋藤綾子）¹，Yasuo Terao（寺尾安生）¹，Kimio Akagawa（赤川公朗）¹ ¹杏林大医細胞生理 ²埼玉医科大学保健医療学部 ³杏林大医共研ＲＩ部門 Two syntaxin 1 (STX1) isoforms, HPC-1/STX1A and STX1B, are coexpressed in neurons and function as neuronal target membrane (t)-SNAREs. However, little is known about their functional differences in synaptic transmission. STX1A null mutant mice develop normally and do not show abnormalities in fast synaptic transmission, but monoaminergic transmissions are impaired (Fujiwara et al. J Neurosci, 2006). We previously reported that STX1B is primarily involved in the regulation of different types of fast synaptic vesicle exocytosis, including spontaneous and evoked release of glutamatergic and GABAergic synaptic transmission (Mishima et al. PloS one, 2014). Recently, STX1B haploinsufficiency have been shown to provoke a broad spectrum of fever-associated epilepsy syndromes. In the present study, in order to examine involvement of STX1B in pathogenesis of fever-associated epilepsy syndromes, we assessed for susceptibility to seizures induced by hyperthermic stimulation or systemic administration of PTZ in STX1B gene-ablated mice. We found that STX1B heterozygote mice showed increased susceptibility to hyperthermia-induced and PTZ-induced seizures. PTZ-induced seizures were blocked by anti-convulsant drug VPA and Phenytoin. We also examined the effect of acute high temperature to tonic GABA(A) current and burst activity of cultured neuronal networks which resembling epileptiform seizures. High temperature increased tonic GABA(A) current and decreased burst frequency in wild-type neurons but not in STX1B heterozygote neurons. To clarify the influence of temperature on synaptic functions of STX1B, we examined presynaptic properties of neurotransmitter release and transporter kinetics in glutamatergic and GABAergic synapses of cultured hippocampal neurons. Implication of STX1B in fever-associated epilepsy syndromes will be discussed.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-104 タンパク質変性剤TNBSのシナプス可塑性に対する作用はボツリヌストキシンの効果と類似している。 Yasuhiro Imafuku（今福泰浩）¹，Naoya Suzuki（鈴木直哉）²，Koh-ichi Enomoto（榎本浩一）³，Hiroko Kataoka（片岡裕子）³，Isao Ito（伊藤功）¹，Takashi Maeno（前野巍）⁴ ¹九州大理生物 ²名古屋大院理物理 ³島根大医 ⁴島根医大 Short-term synaptic plasticity (SSP) is composed of several synaptic processes that operate on millisecond-to-minute timescales and modulate synaptic efficacy in response to increases or decreases in presynaptic activity. SSP contains three components: short-lasting facilitation (τ_F < 1 s), intermediate augmentation (1 s < τ_A < 20 s) and slow potentiation (20 s < τ_P). Botulinum neurotoxins A and C (BoNT-A and BoNT-C), which were found to modify components of SSP (Lupa and Tabti, Pflugers Arch 1986; Maeno et al, Jpn J Physiol 1994 and 1998), cleave synaptosomal-associated protein of 25 kDa (SNAP-25) and syntaxin (Schiavo et al, FEBS Lett 1993 and J Biol Chem 1995). According to the current theory, a synaptic vesicle docks with its release site when synaptobrevin forms the SNARE complex with SNAP-25 and syntaxin distributed in the plasmalemma. Then, the docked synaptic vesicles undergo priming, thereby become readily available for release. We have already investigated SSP of frog neuromuscular transmission to clarify the roles of SNAP-25 and syntaxin by using BoNT-A and/or -C. We found that BoNT-A treatment eliminated slow potentiation by modifying SNAP-25, and BoNT-C poisoning modified syntaxin to abolish intermediate augmentation. Only fast facilitation survived after double poisoning with BoNT-A and -C, but the postsynaptic response became biphasic by an emergence of the novel depression termed repression. A novel depression, termed repression, emerged by double poisoning (Imafuku et al, 2P-049, The 41st Annual Meeting of the Japan Neuroscience Society, 2018). Here we investigated SSP by using 2,4,6-Trinitrobenzenesulfonic acid (TNBS), a membrane impermeable protein denaturing reagent, which modifies the transmembrane domain of SNAP-25 and syntaxin. We found that TNBS selectively inhibits potentiation component at low concentration as does BoNT-A. In addition TNBS almost eliminates augmentation at intermediate concentration. And finally TNBS mimics the effect of double ontoxication by BoNT-A and -C at high concentration. Thus, TNBS is eligible for a convenient alternative to BoNTs for the SSP study.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-105 Arf GEF, GAPはそれらのmRNAが樹状突起へ局在化し、スパイン形成・成熟に関与する Rie Ohashi（大橋りえ）^1,2，Yoshitaka Kimori（木森義隆）³，Nobuyuki Shiina（椎名伸之）^1,2,4 ¹基生研神経細胞生物 ²総研大院基礎生物 ³福井工大環境情報経営情報 ⁴自然科学研究機構・生命創成探究センター Local protein synthesis plays an important role in synaptic plasticity and memory formation. To achieve local translation in dendrites, translational machinery is assembled in a large complex called RNA granules and transported to dendrites. RNG105/Caprin1 is a major RNA-binding protein localized to RNA granules. RNG105 deficiency in mice leads to impairment of higher-order brain functions such as long-term memory and sociality. We previously found that many dendritic mRNAs are reduced in the dendritic layer of hippocampal neurons in RNG105-deficient mice, particularly mRNAs for Arf (ADP-ribosylation factor) regulators (GEFs and GAPs). The goal of this study is to reveal whether local translation of Arf regulators is involved in synaptic plasticity. First, we visualized and quantified the dendritic localization of 8 kinds of mRNAs for the Arf regulators using the MS2 system in mouse primary cultured cerebral neurons. We found that mRNAs for Arf GEFs were localized to dendrites independently on KCl stimulation (neuronal depolarization), whereas mRNAs for Arf GAPs were localized to dendrites in a KCl stimulation-dependent manner. These results suggested that Arf GEF and GAP mRNAs were transported to dendrites in response to different neural activity. Furthermore, we analyzed the knockdown effect of the Arf regulators on the spine formation/maturation and AMPAR (AMPA receptor) surface expression in dendrites of cultured neurons. The knockdown experiments suggested that Arf GEFs and GAPs were classified into two groups: one group positively regulated spine formation/maturation and AMPAR surface expression in dendrites, but the other group negatively regulated immature spine formation. This study suggested that mRNAs for Arf GAPs and GEFs are transported to dendrites dependently and independently on neuronal activity, respectively. Although we have not clarified whether the Arf regulator mRNAs are locally translated in dendrites, the Arf regulators are suggested to be important players in the tuning of spine formation/maturation and AMPAR surface expression in dendrites.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-106 RhoキナーゼはSynGAP1のリン酸化を介してRasの活性化を制御する Mengya Wu（呉夢雅）¹，Yasuhiro Funahashi（船橋靖弘）¹，Tetsuya Takano（髙野哲也）²，Daisuke Tsuboi（坪井大輔）¹，Rijwan Uddin Ahammad（Ahammad Uddin Rijwan）¹，Mutsuki Amano（天野睦紀）¹，Kozo Kaibuchi（貝淵弘三）¹ ¹名古屋大院医神経情報薬理 ²Department of Cell Biology, Duke University Medical School, USA The small GTPase RhoA and its downstream effector Rho-kinase/ROCK are considered as one of the key regulators in dendritic spine formation and synaptic plasticity. However, how RhoA/Rho-kinase signaling involved in modulating synaptic plasticity still remains largely unknown. We have recently developed a phosphoproteomic analysis method that uses affinity beads coated with 14-3-3 proteins to enrich phosphorylated proteins and established the kinase-associated neural phosphosignaling (KANPHOS) database that provides the phosphorylated sites identified by our phosphoproteomic approaches. Using the KANPHOS database, we identified SynGAP1, which is a synaptic Ras-GTPase activating protein, as a novel Rho-kinase substrate. In this study, we found that phosphorylation of SynGAP1 by Rho-Kinase increased its interaction with 14-3-3 but decreased with PSD-95, which is a major scaffolding protein in the postsynaptic densities of dendritic spines. SynGAP1 was dispersed from spines upon long-term potentiation (LTP) induction in cultured neurons, and this dispersion depends on phosphorylation of SynGAP1 by Rho-kinase. Moreover, we found that Rho-kinase increased Ras and ERK activity through phosphorylation of SynGAP1. Thus, the synaptic dispersion of SynGAP1 during LTP may be a key signaling link element that transduces RhoA/Rho-kinase activity to Ras-ERK signaling.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-107 AiCEマウス：入力依存的なスパインの変化を検出する新規ツール Kazuya Kuboyama（久保山和哉）¹，Takafumi Inoue（井上貴文）²，Yuki Hashimotodani（橋本谷祐輝）³，Takuya Itoh（伊藤拓矢）¹，Tohsuke Suzuki（鈴木東介）¹，Yosuke Ohtsuka（大塚庸介）¹，Ryo Kinoshita（木下諒）¹，Pooja Gusain（Gusain Pooja）⁴，Masanobu Kano（狩野方伸）³，Shigeo Okabe（岡部繁男）⁵，Maki K. Yamada（山田麻紀）^1,5,6 ¹徳島文理大香川薬薬理 ²早稲田大院先進理工生命医 ³東京大院医神経生理 ⁴徳島文理大香川薬神経科学研神経回路システム ⁵東京大院医神経細胞生物 ⁶JSTさきがけ Synaptic plasticity provides a basis for most models of learning and memory. One of the keys is the experimentally-induced long-term synaptic potentiation (LTP), which is thought to share the fundamental basis of an increase in synaptic weights. However, the real nature of the potentiation in synapses in the live brain has long been elusive, at least partially because of their sparsity. In the search for a marker for the synaptic change, we focused on CapZ, an F-actin capping protein, which accumulated and was maintained in dendritic spines (postsynaptic varicosities) in the region where an LTP-inducing stimulus was applied (Genes to Cells 15: 737-747, 2010). We herein observed a subtle change in green fluorescent signals in spines of a transgenic mouse line, named AiCE, expressing EGFP-tagged CapZ (EGFP-CapZ), in the visual cortex, twenty minutes after novel visual presentation, or in the sensory cortex after Rota-rod training. This change in fluorescence signal is likely to be derived from the recruitment of EGFP-CapZ into the dendritic spines and reflects molecular plasticity upon stimulation. This dynamic redistribution of EGFP-CapZ in AiCE-mouse was thought to detect a small subset of spines with synapses that underwent an LTP-like increase in synaptic efficacy in vivo and may provide a useful tool for a wider range of analyses in learning and memory.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-108 Modeling of AMPA receptor trafficking by the endosomal recycling system Sarah Yukie Nagasawa（Nagasawa 幸恵 Sarah）沖縄科学技術大学院大学 AMPA receptors (AMPARs) play a crucial role in synaptic transmission mechanisms throughout different regions of the brain. At the subcellular level AMPARs are transported between the synaptic surface, intracellular cytosol, and endosomal system. Currently, there are no realistic computational models which simulate such subcellular neuronal processes. Here we created a model of the vesicular trafficking system of endosomes in the STEPS software and combined it with simulation of stochastic reactions of molecules integrating diffusion rates and spatial effects enclosed in a neuronal mesh structure. STEPS incorporates vesicle objects, which collect and transport AMPARs throughout the tetrahedral mesh, and simulates lateral diffusion, endocytosis, exocytosis, and fusion and budding from the endosome. 1 In addition, biochemical interactions such as AMPARs binding to membrane docking proteins and endosomal sorting events of AMPARs in the endosome are included into the design of the model. To optimize the model, parametric tuning of diffusion rates, endocytosis and exocytosis rates of AMPARs were simulated. We believe by developing a model of the vesicular trafficking system of endosomes this will expand the capabilities of modeling research at the subcellular level in neuroscience and provide valuable insights into understanding the mechanisms of these neuronal processes important in learning and memory. 1. I. Hepburn, W. Chen, S. Wils, E. De Schutter. STEPS: efficient simulation of stochastic reaction-diffusion models in realistic morphologies. BMC Syst Biol. (2012)		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-109 分岐軸索枝間における枝の長さに依存した微小管の安定化は軸索輸送及び形態調節に寄与する Chiaki Imanaka（今中千秋）¹，Shino Ito（伊藤詩野）²，Marina Kassai（葛西真里菜）²，Satoshi Shimada（島田聡史）²，Yoshiyuki Konishi（小西慶幸）^2,3 ¹福井大院工 ²福井大学工学部 ³福井大学ライフサイエンスイノベーションセンター In the reconstruction of branched axonal pattern, competition between adjacent branches occurs via unknown mechanisms. We reported that retraction of branches was inhibited selectively in longer branches compared with adjacent shorter branches because of higher efficiency of axonal transport (Seno et al., 2016). Although the difference in microtubule stability between branches is supposed to contribute to branch-dependent axonal transport, it is unknown what kind of mechanism causes this difference. We carried out the simulation using a model in which the stability (lifetime) of the microtubule simply depends on the protrusion length. We found that the lifetime difference of the microtubules depending on the branch length could occur even in the vicinity of the branch point as observed in the actual axon. Furthermore, it was predicted that destabilization of the whole microtubule decreases the difference of microtubule lifetime between branches. Consistently with this result, we observed that difference of the stability of microtubules between branches of cultured cerebellar neurons was increased by stabilizing microtubules with taxol. In addition, taxol treatment increased the length of longer branches, but not that of shorter branches. We also observed that taxol treatment modulates the distribution of kinesin-head in axonal arbor. These results suggest that the competition between branches may depend on the stability of microtubules throughout the axons. Collectively we propose that stability of branch-dependent microtubule regulate the axonal morphologically via positive feedback mechanism exist in the longer branch.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-110 Cdk5はankyrin-Gのリン酸化を介して軸索起始部の形成を制御する Takeshi Yoshimura（吉村武）¹，Matthew N. Rasband（Rasband N. Matthew）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 ²Dept of Neurosci, Baylor College of Medicine, Houston, TX, USA The axon initial segment (AIS) is a structurally and molecularly unique neuronal compartment of the proximal axon that functions as both a physiological and physical bridge between the somatodendritic and axonal domains. The AIS has two main functions: to initiate action potentials and to maintain neuronal polarity. The cytoskeletal scaffold protein ankyrin-G is highly enriched in neurons at the AIS. Voltage-gated Na⁺ and K⁺ channels are clustered at the AIS through interactions with ankyrin-G. Ankyrin-G links AIS membrane proteins to the βIV-spectrin and actin-based submembranous cytoskeleton. Although all AIS have ion channels, cell adhesion molecules, extracellular matrix molecules and cytoskeletal scaffolds, the defining molecular component of the AIS is ankyrin-G since it is responsible for clustering ion channels and maintaining neuronal polarity; loss of ankyrin-G blocks ion channel clustering and causes axons to acquire the characteristics of dendrites. The human mutations in ankyrin-G were reported and shown to be associated with severe intellectual disability, attention deficit hyperactivity disorder, and autism. While ankyrin-G's role as an `ion channel accumulator' is fairly well understood, how ankyrin-G regulates neuronal polarity remains unknown. The molecular mechanism by which ankyrin-G is regulated remains unclear. Here, we report that Cdk5 phosphorylates ankyrin-G. Inhibition of Cdk5 impaired AIS formation and neuronal polarity. These results suggest that Cdk5 regulates AIS formation through the phosphorylation of ankyrin-G.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-111 神経細胞内におけるCRY2-CIBN相互作用を利用した空間時間的な脱チロシン化のコントロール Takuma Inoue（井上拓馬）¹，Yoshiyuki Konishi（小西慶幸）^2,3 ¹福井大院工 ²福井大工生物応用化学 ³福井大学ライフサイエンスイノベーションセンター Although it is known that complex nerve network establishment is important for its proper function, the mechanism of how neurons accomplish their morphology via various proteins is still remained unclear. One important factor for this establishment is microtubule function. The function of microtubule is controlled by tubulin modifications. Detyrosination on microtubule is carried out by a protein named Vasohibin, and its activity is dependent on its binding partner, Small Vasohibin Binding Protein, SVBP. In this study, we aimed to control detyrosination on microtubule at specific time and place by CRY2 and CIBN interactions whose binding is induced by only blue light. Using CIBN-CAAX which localizes at plasma membrane, we can trap interest protein that fused to CRY2 corresponding to blue light exposure. Thus, this method allows us to control interest protein spatiotemporally in living cells. Here, we expressed SVBP-CRY2, CIBN-CAAX, VASH into HEK293T cells. In this condition, blue light exposure resulted in colocalization of those three proteins at plasma membrane where CIBN exists. This result shows that we can control VASH as well as SVBP by interaction of CRY2 and CIBN-CAAX. Further more, controlling of those proteins by CRY2-CIBN-CAAX interaction for 2 hours showed decreased detyrosination level in HEK293T cells. This method may enable us to investigate how spatiotemporal detyrosination on tubulin is relevant to neuronal morphology by controlling endogenous VASH as well as exogenously expressed SVBP-CRY2.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-112 キネシンによる微小管構造変化のクライオ電子顕微鏡解析 Manatsu Morikawa（森川真夏）¹，Tomohiro Shima（島知弘）^2,3，Junichi Kaneshiro（金城純一）⁴，Taketoshi Kambara（神原丈敏）³，Shinji Kamimura（上村慎治）⁵，Toshiki Yagi（八木俊樹）⁶，Hiroyuki Iwamoto（岩本裕之）⁷，Sotaro Uemura（上村想太郎）²，Hideki Shigematsu（重松秀樹）⁸，Mikako Shirouzu（白水美香子）⁸，Taro Ichimura（市村垂生）⁴，Tomonobu M Watanabe（渡邉朋信）⁴，Ryo Nitta（仁田亮）^8,9，Yasushi Okada（岡田康志）^3,10，Nobutaka Hirokawa（廣川信隆）¹ ¹東京大院医分子細胞生物 ²東京大院理生物科学 ³理研生命機能科学研究センター細胞極性統御研究チーム ⁴理研生命機能科学研究センター先端バイオイメージング研究チーム ⁵中央大院理工生命科学 ⁶県立広島大学生命環境学部 ⁷JASRI, SPring-8 ⁸理研生命機能科学研究センター構造生物学グループ ⁹神戸大院医生理学・細胞生物学生体構造解剖学 ¹⁰東京大院理 Kinesin-1 (KIF5s) transports various cargoes to their cognate destination along the cellular microtubules. The microtubules are suggested to provide the directional cues for the polarized transport, but the molecular basis has yet to be fully elucidated. We have proposed that the conformational switching between the GTP- and the GDP- forms of microtubules would be recognized by KIF5C (Nakata et al., 2011; Yajima et al., 2012), and have performed biophysical studies to investigate the interaction between KIF5C and GDP-microtubules. The findings supported the idea that there is a positive cooperativity in the binding of nucleotide-free KIF5C to GDP-microtubules. Here, we present the cryo-EM structures of GDP-microtubules complexed with KIF5C in the various nucleotide states. The binding of nucleotide-free KIF5C to the intra-tubulin-dimer interface in the GDP-microtubule lattice pushes and rotates β-tubulin towards the plus-end side, leading the elongation of the tubulin-dimer periodicity without nucleotide exchange on β-tubulin. KIF5C can recognize the conformational differences of the microtubules by the interaction between the L11-α4 junction of KIF5C and H3' helix of α-tubulin, and binds to the longer pitch microtubules preferentially as reported in the previous studies (Nakata et al., 2011). This positive feedback mechanism would enable the self-organization of the high-affinity subset of the microtubules, which serve as the guidance cue for the polarized transport of KIF5C.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-113 社会的敗北ストレスが慢性疼痛モデルに及ぼす影響の解析 Hidetoshi Saitoh（齊藤秀俊），Takuya Hatta（八田拓弥），Makoto Tsuda（津田誠）九州大院薬 There are sustained interests among scientists and clinicians in understanding the interactions between stress and nociception. Many experimental studies have focused on emotional aspect of pain and stress through observation of anxiety and depressive behavior in rodents. However, studies linking various type of stresses and nociception are not fully examined. Here we examined some combination of chronic pain models with repeated social defeat stress model to address whether social stress modulates nociceptive behavior of mice subjected to chronic pain models. C57BL/6Jcl male mice preconditioned by repeated social defeat stress and separated to susceptible, resilient to stress and cage transfer (control) groups by social interaction test. Peripheral nerve injury induced neuropathic pain model, complete Freund's adjuvant induced inflammatory pain model and reserpine induced pain models were applied to each groups. In the all pain models, decreases of paw withdrawal threshold measured with von Frey filament were almost identical among groups. Recovery rates of decreased pain threshold were also similar between groups. However, after a recovery of pain threshold, we found that loading single social defeat stress induced a decrease of paw withdrawal threshold. This effect similarly occurred in every chronic pain models and mimicked by intraperitoneal injection of LPS, but was not seen in naive mice. Immunohistochemical analysis of iba-1 represented a sustained microglial activation in the spinal dorsal horn of stress loaded neuropathic pain model mice. Our results demonstrated that there are no visible nociceptive modulation of repeated social defeat stress in multiple chronic pain models, while social defeat stress itself could induce recurrence of experienced chronic pain state in which reactivation of spinal microglia may correlated with the phenomenon.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-114 反復社会挫折ストレスによる内側前頭前皮質ミクログリアのエピゲノム制御 Masayuki Taniguchi（谷口将之）¹，Shiho Kitaoka（北岡志保）¹，Shigehiro Kuraku（工樂樹洋）²，Mitsutaka Kadota（門田満隆）²，Tomoyuki Furuyashiki（古屋敷智之）¹ ¹神戸大院医薬理学 ²理研BDR 分子配列比較解析ユニット Stress is caused by various adverse environments, and often causes emotional changes including depression and elevated anxiety. Using social defeat stress in mice, we previously reported that repeated social defeat stress activates microglia in the medial prefrontal cortex (mPFC), then decreases dendritic arborization of mPFC pyramidal neurons and leads to social avoidance. Recently, we found that microglial activation in the mPFC occurs more rapidly and strongly with repetition of social defeat stress. This finding led us to speculate that repetition of social defeat stress induced persistent epigenomic changes of microglia in the mPFC. However, due to the limited sensitivity of chromatin immune precipitation sequencing (ChIP-seq), a brain region- and cell type-specific epigenomic analysis has been challenging. Here we optimized the protocol of ChIP-seq for whole-genome epigenomic analyses of mPFC microglia isolated by fluorescence-activated cell sorting (FACS). This protocol allows us to detect enrichment of active histone marks near microglia-specific genes in mPFC microglia. Our preliminary findings reveal that whole-genome epigenomic profiles in the mPFC microglia reflected the experience of repeated social defeat and stress sensitivity. Interestingly, these epigenomic changes in mPFC microglia were different from the changes in the nucleus accumbens, indicating brain-region-specific epigenomic changes. We are currently investigating transcription factors and epigenetic regulators related to regulation of mPFC microglial epigenomic changes associated with emotional changes induced by repeated social defeat stress.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-115 CDC42EP4欠損誘導性緑内障発症機構の解明 Shiho Takeda（武田詩穂），Youichi Shinozaki（篠崎陽一），Schuichi Koizumi（小泉修一）山梨大院医薬理 Glaucoma is progressive optic neuropathy, characterized by degeneration of retinal ganglion cells (RGCs) and visual field deficiency. Although elevated intraocular pressure (IOP) has long been considered as a primary cause of the pathology, there is a subtype of glaucoma less sensitive to IOP termed normal tension glaucoma (NTG) with statistically normal IOP. Excitotoxicity by excessive glutamate is considered as one of the causes of NTG. Indeed, model mice lacking glutamate transporters exhibit NTG-like symptoms. However, since these model mice cause acute RGC injury and visual dysfunction at young age, these pathological processes are significantly different from those in glaucoma patients because its progress develops very gradual after middle age. Here, we show CDC42EP4 as a new target molecule for the development of NTG because its deficiency causes NTG-like pathology. First, we investigated expression patterns of CDC42EP4 in the various brain regions and eyes, and found that its expression was highest in the retina. Second, using CDC42EP4KO mice, we asked whether these mice reveal glaucoma-like phenotype. Although CDC42EP4KO mice did not show any sign of RGC injury at 3 months, they revealed significantly higher damage of RGCs at 12 months compared with WT mice. Third, no difference was observed in IOP between WT and CDC42EP4KO mice. Thus, CDC42EP4KO mice could show NTG-like pathology with slow-onset/progression. Fourth, we asked which type of cells express CDC42EP4 in the retina. CDC42EP4 is known to be expressed mainly in Bergmann glia, an astrocyte-like cell in the cerebellum, and controls the localization of glutamate transporters. Considering this, we expected that CDC42EP4 is expressed in Müller cells, an astrocyte-like cell in the retina, and controls clearance of glutamate via its transporters. However, contrary to expectation, CDC42EP4 was mainly expressed in bipolar cells but not in Müller cells. Despite of its absence in Müller cells, CDC42EP4KO showed activation of Mlüler cells as well as microglia at 3 months. It should be noted that such glial activation occurs in the retina before RGC injury. Taken together, we demonstrated that CDC42EP4KO mice showed NTG-like symptoms, in which preceding glial activation might be involved.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-116 マウス大脳穿刺損傷における2ccPAのアストロサイト活性抑制効果 Mari Nakashima（中島麻里）^1,2，Kei Hashimoto（橋本恵）^1,2,4，Ayana Hamano（濱野文菜）^1,2，Hiroko Ikeshima-Kataoka（池島(片岡）宏子）^5,6，Mari Gotoh（後藤真里）^2,3，Kimiko Murakami-Murofushi（室伏きみ子）³，Yasunori Miyamoto（宮本泰則）^1,2 ¹お茶の水女子大学大学院人間文化創成科学研究科ライフサイエンス専攻 ²お茶の水女子大学ヒューマンライフイノベーション研究所 ³お茶の水女子大学ヒューマンウェルフェアサイエンス研究教育寄付研究部門 ⁴日本学術振興会特別研究員 ⁵早稲田大学理工学術院創造理工学部 ⁶慶応義塾大学医学部薬理学教室 Traumatic brain injury (TBI) is caused by physical damage, and the brain structure is disrupted due to the biomechanical insult to the cranium. We previously revealed that 2-carba cyclic phosphatidic acid (2ccPA) significantly suppresses the hemorrhage level and inflammation via the regulation of microglial activation in the stab-wounded cerebral cortex as a TBI mouse model. 2ccPA is a metabolically stabilized derivative of cyclic phosphatidic acid (cPA), and with phosphate oxygen is replaced with a methylene group at the sn-2 position of cPA. The suppression of hemorrhage level by 2ccPA is related to the recovering from blood-brain barrier (BBB) breakdown. Astrocytes, one of the major components of BBB, take a critical role in central immune responses by becoming activated. However, the mechanism how astrocytes are involved in the suppressions of the inflammation by 2ccPA in the stab-wounded cortex remains unknown. In this study, to investigate the effect of 2ccPA on astrocyte activation after the brain injury, we analyzed the glial fibrillary acidic protein (GFAP) and tenascin-C (TN-C), one of extracellular matrix proteins, expression levels in the stab-wounded mouse cortex. The effects of 2ccPA on astrocyte in the stab-wounded regions were examined. The administration of 2ccPA to brain injured mice decreased an astrocyte marker, GFAP mRNA and protein expression level, suggesting that 2ccPA attenuates the inflammation after the stab wound via astrocyte. Moreover, it is known that the expression of TN-C, is upregulated in activated astrocytes after experimental subarachnoid hemorrhage and is related to recovery from BBB breakdown. In the present study, the administration of 2ccPA decreased the number of GFAP- and TN-C-double positive astrocytes, whereas, increased that of GFAP-positive and TN-C-negative astrocytes in the stab wounded cortex. These results demonstrate that 2ccPA contributes to the regulation of the astrocyte activation in the aspect of the TN-C expression. Therefore, we are now trying to explore whether 2ccPA is involved in neuroprotection via the regulation of astrocyte activity in the stab-wounded cortex.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-117 マウスにおいてP2Y1受容体の欠損は高眼圧緑内障様の表現型を示す Kentaro Hamada（濱田健太郎），Youichi Shinozaki（篠崎陽一），Schuichi Koizumi（小泉修一）山梨大院医工薬理 P2Y1 receptor deficiency induces hypertensive glaucoma-like phenotypes in mice Kentaro Hamada, Youichi Shinozaki and Schuichi Koizumi Dept. Neuropharmacol., Interdiscip. Grad. Sch. Med. Univ. Yamanashi, Yamanashi, JAPAN Glaucoma is second leading cause of blindness worldwide which is characterized by progressive degeneration of retinal ganglion cells (RGCs). Although elevated intraocular pressure (IOP) is recognized as one of the highest risk factor, molecular target for IOP regulation has not been fully understood. A growing body of evidences has emerged that purinergic signaling may participate in pathogenesis of glaucoma. For example, ATP and nucleotides are present in the aqueous humor, and IOP and ATP levels in the aqueous humor are positively correlated in glaucoma patients.These reports indicate the possibility that dysregulated P2 receptor signaling trigger pathogenesis of glaucoma. Here we report that P2Y1 receptor is essential for IOP reduction in normal condition and its dysregulation causes glaucoma-like phenotypes such as sustained elevation in IOP and degeneration of RGCs. First, we investigated the role of P2Y1 receptor in IOP regulation. Topical application of MRS2365, selective agonist for P2Y1 receptor, caused dramatic reduction in IOP of wild-type (WT) mice. The hypotensive effect of MRS2365 was not observed in P2Y1 receptor deficient (P2Y1KO) mice, showing the essential role of P2Y1 receptor in IOP reduction. We then measured basal IOP levels and found that P2Y1KO mice show significantly higher IOP level regardless of their age (i.e. 3, 6, 12 and 18 month). Because sustained IOP elevation is one feature of hypertensive glaucoma, we checked RCG damages. Immunohistochemical analysis showed that the number of Brn3a-positive RGCs was moderately reduced in middle-age (12 months old) P2Y1KO mice but not in young adult mice (3 months old). Supporting this observation, the number of apoptotic RGCs was significantly increased in the middle-age P2Y1KO mice. Furthermore, optical coherence tomography (OCT) imaging revealed that middle-age P2Y1KO mice show thinner ganglion cell and inner plexiform layers, general diagnostic feature of glaucoma. Taken together, our results demonstrated that (1) P2Y1 receptor activation reduces IOP; (2) loss-of-function of P2Y1 receptor causes sustained elevation in IOP and (3) hypertensive glaucoma-like phenotypes in mice.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-118 ニューロン―アストロサイト間情報伝達のダイナミクス Eiji Shigetomi（繁冨英治）¹，Yukiho J Hirayama（平山幸歩）¹，Kazuhiro Ikenaka（池中一裕）²，Kenji F Tanaka（田中謙二）³，Haruhiko Bito（尾藤晴彦）⁴，Schuichi Koizumi（小泉修一）¹ ¹山梨大院総合研究部薬理 ²生理学研究所分子神経生理 ³慶應義塾大学医学部精神神経科学 ⁴東京大院医学研究科神経生化学 Emerging evidence indicate that astrocytes play a pivotal role in brain function. Astrocytes receive information from synapses and also transmit their information to synapses by releasing gliotransmitter. Astrocytes express a plethora of Gq-protein coupled receptors (GpPCR) to receive information from synapses. Recent evidence suggest that selective activation of GqPCR signaling in astrocytes can cause synaptic plasticity and memory enhancement in mice, suggesting the relevance of GqPCR functions of astrocytes to brain function. However, due to lack of spatio-temporal information on the bidirectional communication, our understanding of synapse regulation by astrocytes remains limited. Among GpPCRs natively expressed in astrocytes, we focused on P2Y1 receptor which is a central for Ca²⁺ signaling in astrocytes. To understand when and how P2Y1 receptor in astrocytes contribute to synaptic functions, we used transgenic mice whose astrocytes overexpressed P2Y1 receptors specifically by Tet-off system and imaged activities of neurons and astrocytes simultaneously using dual color of genetically encoded Ca²⁺ indicators (GECI) in combination with two-photon microscopy. We introduced red GECI, R-CaMP2 or jRGECO1a, and green GECI, GCaMP6f or Lck-GCaMP6f, into neurons and astrocytes, respectively. In CA1 region of acute hippocampal slices, we successfully visualized Ca²⁺ signals from both cell types. In astrocyte P2Y1 receptor overexpression mice, electrical stimulation of the Schaffer collateral resulted in fast Ca²⁺ rise in dendrites which was followed by slow-onset Ca²⁺ rise in astrocytes, suggesting existence communication from neurons to astrocytes. Ca²⁺ rise in astrocytes were abolished by TTX but still observed in the presence of D-AP5/CNQX/nifedipine. Evoked Ca²⁺ rise in astrocytes remained in the presence of DCPIB, a VRAC blocker. These data suggest that the ligands (e.g. ATP) for activation of P2Y1 receptor in astrocytes were released from presynaptic sites. Furthermore, duration of dendritic Ca²⁺ signals were reduced by MRS 2179, a P2Y1 receptor antagonist, suggesting that existence of excitatory signals from astrocytes to neurons. Overall, our data indicate that astrocytes regulate dendritic activities and reveal dynamic feature of information flow between neurons and astrocytes.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-119 LPS誘発性ミクログリアにおける脂肪酸アミド加水分解酵素(FAAH)の役割 Kazuya Yagyu（柳生和耶）¹，Mikiei Tanaka（田中幹衛）²，Scott Sackett（Sackett Scott）²，Yumin Zhang（Zhang Yumin）²，Yoko Hirata（平田洋子）¹ ¹岐阜大学大学院　連合創薬医療情報研究科 ²Uniformed Services University, Maryland, USA Fatty acid amide hydrolase (FAAH) is an integral membrane enzyme that hydrolases the endocannabinoid anandamide (AEA). A recent study has shown that FAAH inhibitor PF-3845 is neuroprotective and anti-inflammatory in animal models of traumatic brain injury and causes a shift of microglia from M1 to M2 phenotype. However, the role of FAAH in the inflammatory response and the mechanisms by which PF-3845 suppresses inflammation have not yet been elucidated in reactive microglia. In this study, we analyzed the effect of FAAH inhibitors PF-3845 and URB597, and FAAH knockdown on lipopolysaccharide (LPS)-induced inflammation using a mouse microglial cell line BV2. PF-3845 and URB597 reduced the expression of inducible nitric oxide (iNOS) and cyclooxygenase-2 (COX-2), and PGE2 production in LPS-treated BV2 cells. Furthermore, FAAH inhibitors decreased the mRNA levels of the pro-inflammatory cytokines such as IL-6 and IL-1β. In order to confirm whether the anti-inflammatory effect of FAAH inhibitors is due to FAAH inhibition, we analyzed the effect of FAAH knockdown on LPS-induced inflammation using siRNA. FAAH knockdown also reduced the expression of iNOS and COX-2, and PGE2 production. It also decreased the mRNA levels of the pro-inflammatory cytokines, whereas increased the levels of the anti-inflammatory cytokines including IL4 and IL10. Finally, cannabinoid receptor antagonists did not affect the above mentioned phenomena, suggesting that the effects of the FAAH are independent of cannabinoid receptors. These results indicate that FAAH is involved in the anti-inflammatory response in microglial cells which is independent of AEA receptor-mediated signaling.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-120 Olig2アストロサイトの機能解析 Kouko Tatsumi（辰巳晃子），Kouko Tatsumi（辰巳晃子），Ayami Isonishi（石西綾美），Shoko Takemura（竹村晶子），Tatsuhiude Tanaka（田中達英），Akio Wanaka（和中明生）奈良医大医第二解剖 Our lineage tracing study using Olig2^CreER; Rosa-CAG-LSL-eNpHR3.0-EYFP (Olig2 ^CreER; YFP) transgenic mice revealed that a subpopulation of Olig2-lineage mature astrocytes (Olig2-astrocytes) distributed widely but unevenly in the adult brain; regions rich in Olig2-astrocytes tended to lack GFAP-positive astrocytes (GFAP-astrocytes). Even within a single brain nucleus, Olig2-astrocytes and GFAP-astrocytes occupied mutually exclusive territories. External globus pallidus (GPe) is one of the representative nuclei. Interestingly, brain nuclei rich in Olig2-astrocytes tended to strongly express GABA-transporter 3 (GAT-3) in astrocytes and vesicular GABA transporter (vGAT) in neurons, suggesting that Olig2-lineage astrocytes may be involved specifically in inhibitory neuronal transmission by forming tripartite synapses. To compare molecular signatures of the two kinds of astrocytes (i.e., Olig2- and GFAP-astrocytes), we applied the laser micro-dissection (LMD) methods to the GPe in combination with immunohistochemistry. The LMD enabled us to distinguish and to selectively isolate two types of astrocytes from a single section of the GPe, where the territories of Olig2- and GFAP-astrocytes were intermingled. Feeding tamoxifen-containing chow for 1 week to adult Olig2^CreER; YFP mice successfully maximized recombination and subsequent YFP fluorescence. The Mice were then sacrificed and fresh brain were immediately frozen in dry ice. Frozen sections were cut on a cryostat and mounted on PPS membrane slides, and then subjected to immunohistochemistry with anti-GFAP antibody (mouse monoclonal) and secondary Alexa 594-labeled anti-mouse IgG. We differentially dissected YFP-expressing cells with bushy morphologies (Olig2-astrocyte) and Alexa 594-labeled star-like cells (GFAP-astrocyte) from single sections. mRNAs from each type of astrocytes were isolated and subjected to molecular comparison using qPCR. Consistent with our previous report, Olig2-astrocytes expressed lower GFAP mRNA than GFAP-astrocytes. The RT-qPCR analyses further showed that Olig2-astrocytes expressed higher level of GAT-3 gene than GFAP-astrocytes. These results strongly suggested that Olig2-astrocytes constitute a distinct subpopulation of astrocytes subsidiary to inhibitory GABAergic transmission.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-121 自然免疫を介して海馬のケラタン硫酸陽性ミクログリアがてんかん原生に関与する可能性の検討 Tomohiro Ohgomori（大篭友博），Shozo Jinno（神野尚三）九州大院医神経解剖学 Microglia represent a population of resident immune cells in the brain, and play various roles to maintain neuronal homeostasis, such as, phagocytosis of tissue debris, neuroprotection, and synaptic pruning. We have found that keratan sulfate (KS) recognized by monoclonal antibody 5D4 is expressed in a unique population of activated microglia during the latent period after pilocarpine-induced status epilepticus (SE). In this study, we aimed to elucidate the potential role of 5D4-positive (5D4 + ) microglia in epileptogenesis via innate immune system using lipopolysaccharide (LPS), because recent studies have reported that the innate immune system might also be involved in the pathogenesis of epilepsy. Mice were pretreated with intraperitoneal (i.p.) injection of LPS. The next day, SE was induced in all mice by i.p. administration of pilocarpine. The densities of pyramidal neurons with pykinotic nuclei in the CA1 region of the hippocampus were lower in LPS-treated mice than in vehicle-treated mice at 5 days post-SE. Although the increase in the densities of ionized calcium-binding adapter molecule 1-positive (Iba1 + ) microglia by pilocarpine-induced SE was not affected by LPS pretreatment, the expression levels of genes-related to phagocytosis in Iba1 + microglia were higher in LPS-treated mice than in vehicle-treated mice. Interestingly, the proportions of 5D4 + /Iba1 + microglia in total microglia were higher in LPS-treated mice than in vehicle-treated mice at 5 days post-SE. Together with our ongoing experiments, we hypothesize that induction of 5D4 + /Iba1 + microglia may be involved in inhibition of epileptogenesis via activation of the innate immune system.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-122 バルプロ酸ナトリウム曝露アストロサイトによる神経細胞発達・機能変化 Kotomi Takeda（武田琴水），Takuya Watanabe（渡辺拓也），Yuki Oba（大場雄貴），Shuntaro Tsukamoto（塚本春太郎），Kaori Kubota（窪田香織），Shutaro Katsurabayashi（桂林秀太郎），Katsunori Iwasaki（岩崎克典）福岡大薬 Valproic acid (VPA) is widely prescribed for epilepsy therapy. Maternal usage is, however, clinically restricted due to the severe risk that VPA may occur neurodevelopmental disorders such as autism spectrum disorder and mental retardation upon a newborn child. It is well known that the VPA exposure to cultured neurons exhibits abnormalities of synaptic formation and transmission during early development in vitro. In light of firm evidence that astrocytes play a vital role in neurodevelopment and synaptic function, the VPA-exposed astrocytes also expect to affect neurodevelopment and synaptic function. In this study, we examined whether the exposure of VPA into cultured astrocytes alters the neuronal morphology and/or synaptic function. Cortical astrocytes were obtained from newborn mice (postnatal day 0-1) and were chronically treated with VPA under the certain periods. Cortical neurons were co-cultured 14 days in vitro after the removal of VPA from the cultured astrocytes (VPA-exposed group). Cortical neurons with VPA untreated astrocytes were prepared in sister-culture as a control group. Inhibitory and excitatory spontaneous synaptic transmission were pharmacologically separated and recorded under the voltage-clamp condition. Dendritic morphology was evaluated using MAP2 (Microtubule-associated protein 2) antibody. The VPA-exposed group showed a decrease in the frequency of the miniature Inhibitory Postsynaptic Current (mIPSC) compared with the control group, whereas the frequency of the miniature Excitatory Postsynaptic Current (mEPSC) was unchanged. The number of dendritic branches was increased without change in the entire dendritic length of the inhibitory neuron. Together, the mean length of dendritic branches (total dendritic length /the number of dendritic branches) was statistically shorter in VPA-exposed group. However, the dendritic morphology of excitatory neuron was unchanged. These results would imply that the shortened length of individual dendrite causes the limitation of synaptic projection, eventually resulting in the lowered inhibitory synaptic transmission. We speculate that these alterations may be caused by abnormal expression of adhesion molecules and/or neurotrophic factors specific for the inhibitory neuronal network.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-123 脳内グリア細胞の形態とマウスの行動に及ぼす甲状腺機能低下症の影響と性差 Tetsushi Niiyama（新山哲士），Kosuke Aoi（青井孝介），Mami Noda（野田百美）九州大学・大学院・薬学研究院・病態生理学分野 Thyroid hormones (THs) are essential for the development and function of the central nervous system (CNS). In adult CNS, both hypo- and hyper- thyroidism, the prevalence in female being >10 times higher than that in male, may affect psychological condition, for example depression, and potentially increase the risk of cognitive impairment and neurodegeneration including Alzheimer's disease (AD). Microglia, one of the glial cells, is involved in the removal of degenerating neurons and axons in the central nervous system and maintains homeostasis in the central nervous system (Tanaka et al., 2009). On the other hand, it is known that various central nervous system diseases and neurodegenerative diseases occur due to malfunction of microglia. Especially in AD, abnormal activation of microglia is thought to contribute to the pathological progress (Akiyama et al., 2000). We have reported non-genomic effects of tri-iodothyronine (T3) on microglial functions and its signaling in vitro (Mori et al., 2015). In addition, we have observed sex- and age-dependent differences in neurological and psychological symptoms in mouse model of hyperthyroidism (Noda, 2015; Noda et al., 2016). Here we analyzed the effects of opposite thyroid dysfunction, hypothyroidism, on glial cells in vivo using young and old male and female mice. We observed that hypothyroidism also induces sex-dependent changes in glial morphology and animal behavior. These results may help to understand physiological and/or pathophysiological functions of THs in the CNS and how hypothyroidism affects behavioral and psychological conditions in sex-dependent manner.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-124 うつ病モデルマウスにおけるオリゴデンドロサイト再構成の機能解析 Takeshi Shimizu（清水健史），Akimasa Ishida（石田章真），Naoki Tajiri（田尻直輝），Hideki Hida（飛田秀樹）名古屋市立大学　医学研究科　脳神経生理学 In order to understand higher brain function and neurological disorders, it is important to investigate neuron-glial interactions and glial cell property itself. Recent studies demonstrate that myelin remodeling by oligodendrocytes in postnatal mice is involved in motor learning and social interaction, in which myelination is dependent on neuronal activity. To assess whether psychosocial experience during the period of postnatal development influences oligodendrocyte genesis or its property, we applied chronic social defeat stress to adolescent mice, a model for the examination of stress-related disorders in rodents. The social-defeated mice showed more anxiety-like behavior in the open field. The social defeat stress to adolescent mice led to decrease in the number of newly-born oligodendrocytes in the prefrontal cortex and in the number of PLP+ mature oligodendrocytes in the corpus callosum. In both of these regions of social-defeated mice, the number of BrdU-incorporated CC1-positive mature oligodendrocytes was also decreased. To assess whether oligodendrocyte remodeling plays roles in the anxiety-like behavior, we applied drug modulating oligodendrocyte generation to social-defeated mice. We found that the drug treatment rescued the behavioral abnormalities, suggesting that the promotion of oligodendrogenesis can ameliorate depressive symptoms in the mice. Taken together, these findings suggest that oligodendrocyte remodeling in psychosocial environments plays important roles in mental disorder.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-125 全身炎症時におけるミクログリアによる血液脳関門の制御 Koichiro Haruwaka（春若航一路）^1,2，Hiroaki Wake（和氣弘明）¹，Junichi Nabekura（鍋倉淳一）² ¹神戸大院医 ²総研大院生命科学生理 Microglia are sole immune cells in the central nervous system (CNS). Previous study showed that microglia are activated in neurological diseases such as multiple sclerosis, Alzheimer's disease and epilepsy, releasing cytokines and neuro-trophic factors, which act on neurons and vessels, to affect on disease pathologies. In particular, it has been shown that inflammatory cytokines released from activated microglia enhance the permeability of the blood-brain barrier (BBB), which causes infiltration of immune cells and worsens intracerebral inflammation. On the other hand, it is suggested that microglia are activated even during systemic inflammation such as severe infections or auto-immune diseases (e.g., systemic lupus erythematosus: SLE) that make increased BBB permeability. However, it is not clear how systemic inflammation causes microglia activation and brain pathologies. Since microglia are highly motile cells and frequently contact with vessels, this study focused on microglia, to study their dynamics and change of BBB permeability during systemic inflammation using SLE model mice and systemic inflammation induced by lipopolysaccharide (LPS) intraperitoneal injection. Using two-photon in vivo imaging, we showed that microglia migrate to the vessel responding to systemic inflammation which associated with increased BBB permeability. Genetic ablation of microglia caused increased BBB permeability in the early phase and reduced them in the late phase of systemic inflammation. While those results indicate that microglia prevent BBB leakage in the early phase of inflammation, they also appeared to have reversed effect under prolonged inflammation. Further detail shown by immunostaining revealed that vessel-associated microglia expressed Cldn5, a tight junction-related molecule in the early phase but phagocytosis marker CD68 in late phase. Those CD68 co-localized with BBB component aquaporin-4, suggesting their phagocytic function for BBB component to impair their functions. In addition, inhibition of microglial activation with minocycline suppressed increases BBB permeability on late phase. The present study showed that microglia have a dual function onto the BBB, suggesting they can be the therapeutic target for the brain pathologies induced by systemic inflammation.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-127 シンタキシン１B遺伝子欠損マウスのけいれん症状とGABA系異常の関連 Takefumi Kofuji（小藤剛史）^1,2，Tatsuya Mishima（三嶋竜弥）²，Tomonori Fujiwara（藤原智徳）^2,3，Ayako Saito（齋藤綾子）²，Yasuo Terao（寺尾安生）²，Kimio Akagawa（赤川公朗）² ¹杏林大医RI ²杏林大医細胞生理 ³埼玉医大保健医療 Syntaxin1B (STX1B) is thought to be important for exocytosis of synaptic vesicles. We previously reported that STX1B null mutant mice died within 2 weeks after birth, and exhibited structural abnormality in CNS partly caused by low cell viability of STX1B null neurons. Survival of STX1B null neurons was supported by wild type (WT) glial cells, while STX1B null glial cells were less effective, because the secretion of BDNF from STX1B null glial cells was reduced. We found that STX1B was expressed in glial cells, and the expression of STX1B in glial cells was predominant compared to that of HPC-1/syntaxin1A (STX1A), while it was known that both STX1A and STX1B were abundantly expressed in neurons. Furthermore, glutamatergic or GABAergic synaptic transmission was impaired in STX1B null mutant mice. Therefore, STX1B is associated with vesicle exocytosis in both neurons and glial cells. We also reported that STX1B heterozygote mutant mice tended to be seizure phenotype, and exhibited high susceptibility to seizure induced by administration of pentylenetetrazol (PTZ) or kainic acid. However, little is known how STX1B gene is associated with its seizure phenotype. In this study, we examined if its seizure phenotype in STX1B heterozygote mutant mice was related to GABAergic system. We found that PTZ-induced seizure was blocked by anti-convulsant drug, such as valproic acid and phenytoin. We also found that GABA uptake through GABA transporter was reduced in STX1B null and heterozygote glial cells. Expression of GABA transporters, such as GAT-1 and GAT-3, was decreased in STX1B null glial cells compared to that of WT glial cells. On the other hand, glutamate uptake into STX1B null or heterozygote glial cells was normal. Thus, in STX1B heterozygote mutant mice seizure phenotype was caused by impaired GABAergic system, and an imbalance of excitation and inhibition in neuronal network occurred. Implication of STX1B roles in excitation/inhibition balance will be further discussed.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-128 Quercetin inhibits viral-induced inflammatory response in murine microglia cells via suppression of JAK-STAT pathway and activation of HO-1 Yuan-Yen Chang（Chang Yuan-Yen）¹，Hui-Wen Lin（Lin Hui-Wen）² ¹Department of Microbiology and Immunology, School of Medicine, Chung-Shan Medical University ²Department of Optometry, Asia University Quercetin is one of the most common flavonoids found in fruits and vegetables, and has potent anti-inflammatory, anti-tumor, antioxidant properties and reduces cardiovascular disease risks. In this study, the anti-virus inflammatory capacity of Quercetin and its molecular mechanisms of action were analyzed. The cytotoxic effects of Quercetin were assessed in the presence or absence of pseudorabies virus (PRV) via LDH and MTT assays. The results showed that Quercetin (< 5 μM) had no toxic effects. In PRV-infected murine microglia (BV-2) cells, Quercetin potently inhibited the production of NO, iNOS, COX-2 and inflammatory cytokine production. We found that PRV-induced NF-κB activation is regulated through inhibition of JAK, STAT1and STAT3 phosphorylation in response to Quercetin. Additionally, Quercetin caused the induction of HO-1. Taken together, our data indicate that Quercetin diminishes the proinflammatory mediators NO, inflammatory cytokines and the expression of their regulatory genes, iNOS and COX-2, in PRV-infected MH-S cells by inhibiting JAK-STAT pathway and activation of HO-1. Keywords: Quercetin, pseudorabies virus (PRV), anti-inflammatory, proinflammatory mediators

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-129 青色光刺激したグリア前駆細胞株OS3ChR2からの細胞外小胞の放出 Kenji Ono（小野健治），Yosuke Hashimoto（橋本洋佑），Hiromi Suzuki（鈴木弘美），Makoto Sawada（澤田誠）名古屋大環境医脳機能 We have previously reported that photo-activated OS3ChR2 cells, ChR2-expressing clonal glial progenitor cells, differentiated into oligodendrocytes through alteration of intracellular Na⁺ and Ca²⁺, and that they recovered motor function when injected into demyelinated spinal cord in mice. However, it remains unclear how photo-activated OS3ChR2 cells communicated with parenchymal cells. Some groups reported that increase in intracellular Ca²⁺ promoted release of extracellular vesicles (EVs), which facilitate intercellular communication processes. In this study, we examined properties of EVs from photo-activated OS3ChR2 cells. EVs were separated from the conditioned medium, which was derived from OS3ChR2 cells with or without blue light exposure (BL) for 3 days. After removal of live cells and apoptotic vesicles, large EVs (LEVs) were collected by centrifugation at 10,000 x g for 60 min. Small EVs (SEVs) were prepared from the resulting supernatant by centrifugation at 110,000 x g for 60 min. From nanoparticle tracking analysis, average particle size of SEVs was observed smaller than that of LEVs and there was no difference in average particle size between EVs from OS3ChR2 cells with and without BL. In addition, SEVs concentration per OS3ChR2 cell was higher than LEVs concentration and the release of LEVs and SEVs rose up after photo-activation. To clarify whether EVs from OS3ChR2 cells played regulatory roles in intercellular communication, alteration of inflammatory-related mRNA expression in primary microglia was examined after treatment with EVs for 24 h. IL-1β and iNOS mRNA expression was increased in microglia treated with EVs from OS3ChR2 without BL, but not with BL. TNFα mRNA expression was also increased in microglia treated with LEVs and SEVs from OS3ChR2 cells without BL, whereas it was increased in microglia treated with LEVs, but not SEVs from OS3ChR2 cells with BL. These showed that alteration of intracellular ions in glial progenitor cells resulted in release of EVs with distinct properties. In addition, these suggested that glial progenitor cells might communicate to microglia with EVs in demyelinated mice.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-130 線条体神経細胞への長期的な光刺激によって起こる有髄神経軸索の構造解析 Yoshifumi Abe（阿部欣史），Kenji Tanaka（田中謙二）慶應義塾大学　医学部　精神神経科学教室 The learning of a new skill relies on changes in brain function. This functional plasticity can be accompanied by structural changes in myelinated axons. Additionally, it is known that neural excitation by motor learning or optogenetics stimulation induced a plasticity change of myelination. However, the causal relationship between neural excitation and structural alterations of myelinated axons has not been fully understood. To address these fundamental questions, we conducted a proof-of-concept study using mice that express channelrhodopsin-2 (ChR2) only in the striatal medium spiny neurons (MSNs) which are related to motor function. Optogenetic MSN stimulation enables long-term upregulation of specific neuronal populations. We utilized diffusion tensor imaging (DTI) for comprehensive visualization of structural alterations which were induced by the long-term optogenetics stimulation to the MSNs, then determined how the structural changes which were described by DTI reflected histological changes. One-week optogenetic stimulation was conducted to the dorsal striatum (dStr) of mice which express a ChR2-YFP in MSNs. A rotation behavior of the mice which was induced by continuous optogenetics stimulation to the hemispheral dStr was impaired one week after stimulation. DTI images were acquired with their fixed brains after stimulation to calculate a fractional anisotropy (FA) value (n=8). The stimulation decreased FA in the ipsilateral dStr, motor cortex (M), and substantia nigra reticular (SNr), compared with the contralateral side. Next, we observed fine structural changes at the dStr, the M, and the SNr using a super-resolution microscopy (SRM). This microscopy observation showed reductions in diameters of myelin proteolipid protein (PLP) positive axons and PLP positive myelin thickness in the dStr, the M, and the SNr. These histological structural changes were approved by observation with an electron microscopy and were highly correlated with the DTI-FA change. Additionally, we found reductions in the diameters of YFP positive axons and dendrites of MSNs. These results indicated that the long-term activation of dStr MSNs, leading to motor dysfunction, altered the myelinated axons in the cortico-striatal-thalamo-cortical circuit.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-131 ニワトリ脳幹聴覚神経回路における領域依存的なミエリン形成の3次元形態解析 Ryo Egawa（江川遼），Hiroshi Kuba（久場博司）名古屋大院医細胞生理 Oligodendrocyte regulates conduction velocity by myelination in central nervous system. In chick auditory brainstem circuit, it supports the integration of binaural information and contributes to sound localization. Specifically, on the axons projecting from nucleus magnocellularis (NM) to nucleus laminaris (NL), oligodendrocytes compensate the difference in conduction distance of the bilateral projection with conduction velocity by forming long myelin for long contralateral projection and short myelin for short ipsilateral projection. What mechanism causes such region-dependent myelination? To address this question, we established an experimental system capable of genetically labeling and 3-D morphometry of individual oligodendrocytes which myelinate NM axons. Plasmid vectors which express fluorescent protein under oligodendrocyte-specific promoter were introduced into neural tube at embryonic day 1.5 (E1.5) by in ovo electroporation method. At E20, the 2-mm thick brainstem slices were cleared with CUBIC protocol and imaged under two-photon microscopy. As a result, the oligodendrocytes on the NM axons sparsely expressed fluorescent protein, thereby visualizing the intact 3-D structure of their protrusions and myelin. Morphometry showed that the oligodendrocytes in the contralateral projection area have longer and less myelin than the NL peripheral area. In addition, co-expression of neurotrophin receptors allowed for functional perturbation on the region-dependent myelination. From the above, our system will facilitate further understanding of the mechanism of precise neural circuit regulation by oligodendrocyte.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-132 膜骨格複合体4.1G-Lin7-MPP6-CADM4のマウス末梢神経系における相互関係 Nobuo Terada（寺田信生）¹，Yurika Saitoh（齊藤百合花）^1,2，Jyunji Yamauchi（山内淳司）³，Akio Kamijo（上條明生）¹，Takeharu Sakamoto（坂本毅治）⁴ ¹信州大院医保健医療生命科学 ²帝京科学大医学教育 ³東京薬科大院生命科学分子神経 ⁴東京大医科研分子発癌 A fine meshwork structure under erythrocyte membrane is termed "membrane skeleton", and it has been well known to function for the membrane stabillity. 4.1R-membrane palmitoylated protein 1 (MPP1)-glycophorin C is one of the basic molecular complexes of the membrane skeleton. An analogous molecular complex, 4.1G-MPP6-cell adhesion molecule 4 (CADM4), is incorporated into the Schmidt-Lanterman incisure (SLI), a truncated cone shape in the myelin internode that is a specific feature of myelinated nerve fibers formed in Schwann cells in the peripheral nervous system (PNS). Under the stretched condition, the structures of peripheral nerve fibers had a beaded appearance, and the heights of SLI circular-truncated cones increased at the narrow sites of nerve fibers. We evaluated myelin ultrastructures under 4.1G and MPP6 deficiency (-/-). Using electron microscopy, 4.1G-/- mice exhibited myelin abnormalities: myelin was thicker in internodes, and attachment of myelin tips was distorted in some paranodes. In MPP6-/-mice, the sciatic nerves had thicker myelin in internodes. Under 4.1G-/-, CADM4 and a scaffolding protein, Lin7, were disappeared. In SLIs in MPP6-/- nerves, Lin7 was rarely detected by immunohistochemistry and western blotting, but the localization and amount of CADM4 and 4.1G were not altered. These indicate that CADM4 transportation is dependent on the presence of 4.1G. Motor activity was impaired with a tail-suspension test in 4.1G-/- mice, but not significant in MPP6-/- mice. Thus, we propose 4.1G and MPP6 function for proper formation of myelin in PNS.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-133 グロベニンはミエリン崩壊物への直接結合によりマウス末梢神経リゾレシチン誘導脱髄を修飾する Akiko Hayashi（林明子），Yuki Setoguchi（瀬戸口潔），Shinnosuke Kaneko（金子真之亮），Naho Suzuki（鈴木尚穂），Hiromu Tanaka（田中大夢），Daigo Hanaoaka（柳岡大悟），Hiroko Baba（馬場広子）東京薬科大薬学部機能形態学 The Intravenous Immunoglobulin (IVIg) therapy is first choice of the treatment of immune-mediated neuropathy. Recently we have reported that IgG antibodies against large-myelin protein zero (L-MPZ) are included in human γ-globulin preparation glovenin®-I (Nihon Pharmaceutical Co., LTD.). IVIg treatment shows protective effect on chemical demyelinating condition and induction of M2 macrophages in acute phase in mouse experimental model. To clarify the role of glovenin in neuropathy treatment, we examined immunohistological changes of demyelinating regions in glovenin-treated mice. ICR mice were injected with 1% Lysolecithin directly into bilateral sciatic nerves, and either glovenin or control saline was injected intravenously 24 hours later. The sciatic nerves were examined on the day 7 and 14 after Lysolecithin injection. On the day 7, abundant debris was found in the demyelinated regions and inside the phagocytic macrophages. Most of these debris was stained by anti-myelin basic protein (MBP) antibody. Some MBP⁺/human IgG (hIgG)⁺ and relatively large-sized hIgG⁺ debris was also observed in the IVIg treated mice. While substantial number of MBP⁺ debris was still present on the 14th day, hIgG⁺ myelin debris was significantly decreased. Instead, CD68⁺ phagocytic macrophages were labeled by anti-hIgG antibody, suggesting that direct binding of hIgG to myelin debris via L-MPZ enhanced its clearance by macrophages. Thus, IVIg treatment of the Lysolechithin-induced demyelination may be effective partly through IgG antibody reaction against L-MPZ in glovenin.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-134 A novel Olig2-binding protein in oligodendrocytes is indispensable for oligodendrocyte differentiation and myelination in the CNS Anna Simankova（Simankova Anna）¹，Norihisa Bizen（Bizen Norihisa）¹，Manabu Abe（Abe Manabu）²，Kenji Sakimura（Sakimura Kenji）²，Hirohide Takebayashi（Takebayashi Hirohide）¹ ¹Div of Neurobiol Anat, Niigata Univ ²Dept of Anim Mod Dep, Brain Res Inst, Niigata Univ Oligodendrocytes are the myelin-forming glial cells of the CNS, which enwrap individual axonal segments resulting in myelination in the CNS. Myelination is critical for the rapid conduction of electrical signals and integrity of the neuronal network required for the normal functioning of the CNS. It is a well-coordinated differentiation program regulated by multiple transcriptions factors and extrinsic growth and differentiation signals. Omission of the myelination process leads to serious, difficult to treat neurological disorders, such as multiple sclerosis and Pelizaeus-Merzbacher disease. One of the factors involved in myelination process is Olig2-basic helix-loop-helix transcription factor. Olig2 creates a complex with newly discovered Olig2-binding protein (Obp2). The Obp2 participates in miRNA biogenesis and transcriptional regulation. For investigation role of the Obp2 in OLs development, maturation and in the myelination process we creates Obp2 conditional knockout mice in MBP in synthesizing cells MBP-Cre: Obp2 cKO. These mice had a shorter life span, about 6 weeks of ages. According to immunohistochemical and in situ hybridization data, mice showed a decrease in the synthesis of myelin proteins, a significant decrease in the number of mature oligodendrocytes and oligodendrocytes progenitor cells, as well as changes in the morphology of oligodendrocytes in the spinal cord. Thus, Obp2 is essential for the development and maturation of oligodendrocytes and for the ordinary myelination.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-135 マウス海馬支脚におけるフィブロネクチンの発現分布 Tetsuhiko Kashima（鹿島哲彦）¹，Asako Noguchi（野口朝子）¹，Yuji Ikegaya（池谷裕二）^1,2，Nobuyoshi Matsumoto（松本信圭）¹ ¹東京大院薬薬品作用 ²脳情報通信融合センター The subiculum is heterogeneous anatomically and physiologically as is the case with the hippocampus. Recent studies have revealed that the dorsal subiculum is diverse in terms of gene expression. However, few studies have investigated the heterogeneity of the entire subiculum along anatomical axes. To reveal this anatomical heterogeneity, we focused on fibronectin because its mRNA (FN1 mRNA) is known to be expressed in the subiculum. We immunohistochemically characterized the expression pattern of fibronectin protein in the entire subiculum along three axes (i.e., the dorsoventral, proximodistal, and superficial-deep axes). We first found that FN1 mRNA is translated into protein inside cells. We then defined fibronectin-positive area in the subicular pyramidal cell layer and calculated the ratio of the number of fibronectin-positive cells to that of total cells in this area. Unexpectedly, the fibronectin-positive neurons were widely distributed in the pyramidal cell layer of the dorsal subiculum, whereas in the ventral subicular pyramidal field, fibronectin was concentrated around more superficial and distal corner. Moreover, we found that both in the dorsal and ventral subiculum were almost equal in the ratio of fibronectin-positive cells. These results suggest that fibronectin-positive excitatory neurons exist more locally in the ventral subiculum than in the dorsal subiculum. Therefore, fibronectin may be useful as a marker protein for investigating the heterogeneity of principal cells in the entire subiculum.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-136 Effects of sodium arsenite on proliferation and cytoskeletal gene expressions of mouse cortical astrocytes Nang Thinn Thinn Htike（Htike Nang Thinn Thinn）^1,2，Fumihiko Maekawa（Maekawa Fumihiko）⁴，Satomi Kobayashi（Kobayashi Satomi）²，Sho Maejima（Maejima Sho）³，Shinji Tsukahara（Tsukahara Shinji）^2,3 ¹Department of Physiology, University of Medicine Magway, Magway, Myanmar ²Area of Regulatory Biology, Division of Life Science, Graduate School of Science and Engineering, Saitama University, Saitama, Japan ³Area of Life-NanoBio, Division of Strategy Research, Graduate School of Science and Engineering, Saitama University, Saitama, Japan ⁴Center for Health and Environmental Risk Research, National Institute for Environmental Studies, Tsukuba, Japan Sodium arsenite (NaAsO₂) is a neurotoxicant affecting the viability and morphogenesis of neuron, and altering the cell cycle of astrocytes by inducing unscheduled S-phase-coupled cell death. Astrocytes response to a variety of insults with transforming into reactive phenotype as a defensive reaction. Reactive astrogliosis is characterized by alteration in morphological features with upregulation of the expression of intermediate filament proteins, and increasing proliferation. Arsenic alters cellular process, such as cell cycle check point, oxidative stress and inflammatory responses. The cortical astrocytes respond to inflammatory challenges differently between sexes. However, it is still unknown whether arsenic affects the ability of astrocyte to transform into reactive astrocytes. In this study, we examined the effects of NaAsO₂ on the mRNA expression of intermediate filament proteins involved in reactive astrogliosis and the proliferation of astrocytes in both sexes. Astrocytes isolated from the cerebral cortex of male and female C57BL/6 mice on postnatal day 1-2 were primarily cultured for each sex. The mRNA levels of glial fibrillary acidic protein (GFAP), vimentin, nestin and synemin of astrocytes were measured at the time point 72 hours after exposure to NaAsO₂ (0, 4, 8, 12, 16 and 20 μM) by a quantitative PCR method. Proliferation of astrocytes after NaAsO₂ exposure was examined by quantitative PCR of genomic DNA of GFAP gene. NaAsO₂ exposure significantly suppressed the mRNA expression of GFAP at the concentration of 12 μM, vimentin at 4 μM, synemin at 8 μM and nestin at 20 μM with concentration-dependent manner in both sexes. On the other hand, the proliferation of astrocytes estimated by the amount of genomic DNA of GFAP gene was not affected by NaAsO₂ exposure. These findings suggest that arsenic exposure suppresses the ability of astrocytes to transform into reactive phenotype with decreasing the expression of intermediate filament proteins. Additionally, we found a sex difference in the amount of genomic DNA of GFAP gene and the mRNA expression of GFAP and synemin independently of NaAsO₂ treatment. NaAsO₂ exposure may not exhibit sex-specific effect on astrocytes, though cortical astrocytes themselves exhibit sex differences in the proliferation and the mRNA expression of GFAP and synemin.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-137 海馬台の投射先選択的な神経細胞の分布とシナプス後細胞の投射先 Ryoko Umaba（馬場良子）¹，Takuma Kitanishi（北西卓磨）^2,3,4，Kenji Mizuseki（水関健司）^2,3 ¹大阪市大院医脳神経外科 ²大阪市大院医神経生理 ³大阪市大院医脳科学研究センター ⁴JSTさきがけ The subiculum is a major output region of the hippocampal formation that relays hippocampal CA1 output to various regions of the brain. Proximal and distal subicular neurons are known to project to different regions of the brain, and thus, potentially transfer distinct information in a target area-specific manner. However, the existence of such uneven distributions of target area-specific projection neurons along the superficial-deep axis of the subiculum remains unclear. Here, we systematically investigate the projection pattern of the rat dorsal subiculum using anterograde/retrograde tracing with immunohistochemical identification of the subicular laminar structure (Experiment 1). Further, we performed anterograde transsynaptic tracing to identify the projection targets of neurons that directly receive subicular output (Experiment 2). In experiment 1, we first used anterograde tracing to verify that the subiculum densely innervates the following five areas: nucleus accumbens (NAc), anteroventral thalamic nucleus (AV), interanteromedial thalamic nucleus (ITN), retrosplenial granular cortex (RSC), and medial mammillary body (MMB). Next, using retrograde tracing, we found that the subicular projection neurons innervating these target areas have a unique anatomical distribution: NAc-projecting cells were located in the anterior-proximal part of the subiculum; AV-projecting cells were in the deep-distal part; ITN-projecting cells were in the deepest layer containing PCP4+ cells; and RSC- or MMB-projecting cells were located more superficially than AV- or ITN-projecting cells. These results indicate that the distributions of projection neurons targeting distinct brain regions depend on the proximal-distal, superficial-deep, and anterior-posterior axes of the subiculum. To identify the projection targets of MMB neurons that directly receive subicular output, in experiment 2, we injected AAV1-hSyn-Cre (Zingg et al., 2017) into the dorsal subiculum and AAV-EF1α-DIO-EYFP into the MMB. We observed Cre+/EYFP+ somata in the MMB and EYFP+ axons in the AV, indicating that MMB neurons receiving synaptic inputs from the subiculum send efferent fibers to AV via the mammillothalamic tract. Taken together, our results suggest that the location of soma within the dorsal subiculum predicts projection targets and that the deep and superficial subiculum projects to the AV directly and indirectly via the MMB, respectively.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-138 Olfactory marker proteinのcAMP緩衝作用が反復嗅覚応答を実現する Noriyuki Nakashima（中島則行）¹，Kie Nakashima（中島輝恵）²，Akiko Taura（田浦晶子）³，Akiko Nakashima（中島明子）^1,4，Harunori Ohmori（大森治紀）⁵，Makoto Takano（鷹野誠）¹ ¹久留米大学医学部生理学講座統合自律機能部門 ²京都大院医 ³藍野大医療保健臨床工 ⁴東大病院臨床研修 ⁵金沢医大院医生理 Sensory cells generally undergo adaptation, reducing sensitivity to constant stimuli. Olfaction starts with olfactory receptor neurons (ORNs), utilizing cAMP as a second messenger. Olfactory stimulation produces cAMP surge, which activates cyclic nucleotide-gated Ca²⁺-permeable (CNG) channels in limited ciliary space of ORNs. The resultant Ca²⁺-accumulation inhibits CNG channels to cause prominent feedback adaptation of ORNs. Simultaneously, odour-source searching requires sufficient sensitivity of ORNs to repetitive stimuli during sniffing. However, how ORNs accommodate these conflicting responses remains unknown. Here, we show that cAMP-buffering machinery swiftly bypasses surplus cAMP during signal transduction to maintain sensitivity of ORNs to repetitive stimuli. We discovered by bioluminescence resonance-energy transfer experiments that cytosolic olfactory marker protein (OMP) directly captured cAMP with an affinity of approximately -37 kJ/mol. Cyclic AMP-buffering by OMP sharpened CNG channel activity and prevented excessive Ca²⁺-influx. OMP^-/--ORNs were immediately silenced after burst firing under repetitive stimulation, while OMP^+/+-ORNs resiliently maintained responses independently of enzymatic cAMP-production/degradation. Consequently, OMP^-/--mice showed serious impairment in odour-source searching by sniffing. Therefore, cAMP-buffering fundamentally operates to maintain discrete olfactory responses to ensure the olfaction by sniffing for identifying the location and nature of odours.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-139 抑制性ニューロンの活動により血流は増大しうる Mitsuhiro Fukuda（福田光洋）¹，Alexander J Poplawsky（Poplawsky J Alexander）¹，Bistra Iordanova（Iordanova Bistra）²，Alberto L Vazquez（Vazquez L Alberto）^1,2，Seong-Gi Kim（Kim Seong-Gi）^3,4 ¹Dept of Radiology, Univ of Pittsburgh, PA, USA ²Dept of Bioengineering, Univ of Pittsburgh, Pittsburgh, PA, USA ³Center for Neuroscience Imaging Research, Institute for Basic Science, Suwon, Korea. ⁴Dept of Biomedical Engineering, Sungkyunkwan University, Suwon, Korea. Introduction Functional magnetic resonance imaging (fMRI) relies on the hemodynamic response to neuronal activity. Although an fMRI signal increase is implicitly considered as an increase in excitatory neuronal activity, it is not clear whether activity of inhibitory neurons changes fMRI signals. Here we studied whether synaptically activated GABAergic cells can evoke hemodynamic responses in reciprocal dendrodendritic synapses in the external plexiform layer (EPL) of the main olfactory bulb of isoflurane anesthetized rats. Methods A stimulation electrode was implanted in the lateral olfactory tract (LOT), which is formed by axons of mitral cells (MCs), to antidromically excite MCs. Trains of rectangular pulses were delivered to the electrode to activate the dendrodendritic synapses in EPL. Blood volume and blood flow responses were measured by a contrast agent-based fMRI and laser-Doppler flowmetry (LDF), respectively. Neuronal activity was also recorded during LDF measurement. LOT-stimulation evoked hemodynamic responses were compared before and after application of drugs on the bulb surface. Results Maximal increases in blood volume measured with fMRI were evoked in EPL during LOT stimulation. This fMRI response was significantly suppressed when an NMDA glutamate receptor antagonist, APV, was topically applied. Consistent results were obtained for neuronal activity and blood flow responses evoked by LOT stimulation. We then examined whether astrocyte activity driven by glutamate reuptake was responsible for the evoked hemodynamic responses, as reported in the glomerular layer of the bulb. Topical application of an astrocyte-specific glutamate transporter inhibitor, DHK, did not suppress the evoked neuronal activity and blood flow responses in EPL, but rather enhanced them. Additionally, we examined whether nitric oxide, the predominant vascular mediator in the cerebellum and cortical GABAergic cells, initiated a hemodynamic response. A nitric oxide synthase inhibitor, L-NNA, did not change the evoked neuronal activity and blood flow responses in EPL. Conclusion These results suggest that presynaptic activity and astrocyte activity driven by glutamate reuptake were not enough to evoke the LOT-stimulation evoked hemodynamic responses and that activity of GABAergic cells was required. We therefore conclude that excitation of postsynaptic GABAergic cells increases blood flow and volume and could increase fMRI signals.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-140 線虫感覚神経における興奮性膜電位変化の伝播 Takashi Murayama（村山孝）¹，Tomomi Shindou（新道智視）²，Mayumi Shindou（新道まゆみ）²，Yuto Momohara（桃原悠人）¹，Ei-ichiro Saita（税田英一郎）¹，Jeffery R Wickens（Wickens R Jeffery）²，Ichiro Maruyama（丸山一郎）¹ ¹沖縄科学技術大院神経生物学研究ユニット ²沖縄科学技術大院神経生物学研究ユニット High-impedance neurons with short processes such as the nematode C. elegans neurons are thought to transmit electrical signals by passive propagation. Contrary to this dogma, in this annual meeting we will report regenerative depolarization of a major gustatory sensory neuron, ASEL, in C. elegans. Whole-cell patch-clamp recordings in vivo showed supralinear depolarization of ASEL upon current injection. Furthermore, stimulation of animal's nose with NaCl evoked all-or-none membrane depolarization in ASEL. In the ASEL sensory cilium, a receptor-type guanylyl cyclase, GCY-14, and a cGMP-gated cation channel complex, TAX-2/TAX-4, are required to respond to NaCl in the environment. Activation of GCY-14 by NaCl increases cGMP levels, which in turn, activate TAX-2/TAX-4 channels in ASEL cilia. Mutant analysis showed that EGL-19, the α1 subunit of L-type voltage-gated Ca²⁺ channels, is essential for the regenerative depolarization of ASEL. ASEL-specific knock-down of EGL-19 by RNAi demonstrated that EGL-19 functions in C. elegans chemotaxis along an NaCl gradient. These results demonstrate that a natural substance induces regenerative all-or-none electrical signals in dendrites, and that these signals are essential for activation of sensory neurons for chemotaxis. As in other vertebrate and invertebrate nervous systems, active information processing in dendrites occurs in C. elegans, and is necessary for adaptive behavior.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-141 片側内耳破壊の皮質神経活動への影響 Ryota Kai（甲斐竜太）¹，Kazuki Tainaka（田井中一貴）²，Kuniyuki Takahashi（高橋邦行）¹，Nae Saito（齊藤奈英）³，Toshikuni Sasaoka（笹岡俊邦）³，Shun Yamaguchi（山口瞬）⁵，Hiroyuki Nawa（那波宏之）⁴，Arata Horii（堀井新）¹ ¹新潟大院医歯生体機能調節医学感覚統合医学耳鼻咽喉科・頭頸部外科 ²新潟大脳研リソース研究セシステム脳病態 ³新潟大脳研リソース研究セ動物資源開発研究 ⁴新潟大脳研基礎神経科学分子神経生物 ⁵岐阜大院医神経統御・高次神経形態 Vestibular organs composed of semicircular canals and otoliths are the peripheral sensory system that detects acceleration. The vestibular information goes up the brain stem via this vestibular organ and is transmitted to the cerebral cortex. However, it is poorly understood where and how its vestibular information is processed in the cerebral cortex. Unilateral vestibular dysfunction in human produces strong vertigo symptoms and balance deficits in the acute phase. However, with the lapse of time, a phenomenon in which the symptom becomes inconspicuous gradually called "vestibular compensation" occurs. This study aims to identify the cerebral cortex regions and circuits that are involved in the dizziness cognition and its compensatory mechanisms. By monitoring the expression of an activity-dependent gene Arc, we visualized the neural activity change of individual cortical regions to identify the cortical circuits involved. We administered Sodium Arsanil into the inside of the tympanic membrane of transgenic mice expressing Arc-dVenus and destroyed the vestibular function. The mouse brain was taken out and transformed into transparent tissues by the CUBIC method, and then was subjected to 3D imaging using a light-sheet fluorescence microscope. In the vestibular-lesioned mice, the enhanced expression of Arc was observed in several cortical areas, including prelimbic cortex. Currently, western blotting is performed to quantify the expression of Arc, c-Fos, EGR-1 in the cortical regions. These results suggest the contribution of medial prefrontal circuits to the acceleration or balance cognition in mice.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-142 Brief perturbation of amygdala-cortical projections alters palatability coding in gustatory cortex Jian-You Lin（Lin Jian-You），Narendra Mukherjee（Mukherjee Narendra），Donald Katz（Katz Donald） Brandeis University Taste stimuli contain both chemical (taste identity, for instance sweet and salty) and hedonic value (taste palatability) information. Reflecting the complexity of tastes, gustatory cortex (GC) single-unit taste responses convey a time series of information-first about taste identity (e.g., sweet, bitter; Epoch 1: ~0.2-0.7s post-delivery) and then taste palatability (Epoch 2: ~0.8-1.5s post-delivery). We have hypothesized that the dynamic nature of this coding reflects the fact that GC integrates information from multiple subcortical nuclei, and that in particular the emergence of palatability-related information in GC specifically requires temporally-restricted inputs from basolateral amygdala (BLA). Previous work (Piette, et al., 2012) broadly confirmed the importance of BLA in cortical palatability coding; to more rigorously test our hypothesis, we examined whether taste-related GC firing could be altered by brief, targeted optogenetic perturbation limited to BLA-> GC projections. We first infected BLA of adult female Long Evans rats with AAV-ArchT; four weeks later, we implanted 1) unilateral or bilateral multi-electrode bundles with optical fibers (`optotrodes') in the GC and 2) intra-oral cannula (IOC) for taste delivery. With this preparation, we were able to specifically silence BLA-> GC projections (sparing both BLA and GC somae) while presenting 0.1M NaCl, 0.3M Sucrose, 0.1M Citric Acid and 1mM Quinine-HCl. We found that optogenetic inhibition of BLA-> GC projections had bidirectional effects on GC taste responses. Single unit responses were dramatically altered, but the effect was specifically restricted to the epoch of palatability-related firing; taste identity firing remains unaffected by the inactivation. This pattern of results is consistent with the hypothesis that relative to other inputs, BLA preferentially provides palatability information to GC. Future directions will involve experiments that aim to examine the impact of this brief, targeted optogenetic inactivation on taste-guided behavior, such as conditioned taste aversion.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-143 聴覚皮質の要素的な音成分の結合における脳リズムの役割 Sohei Kosaka（小坂奏平），Yoshiki Kashimori（樫森与志喜）電通大院情報理工学研究科基盤理工学専攻 Animals utilize auditory information for survival and communications of conspecifics. A sequence of sound is analyzed in animals' brain as elementary components such as notes and syllables. It has been reported that auditory information is represented by spatiotemporal activity of primary auditory cortex. However, how the elementary components of sound are extracted from the spatiotemporal activity of neurons is poorly understood. We hypothesize that the brain oscillations may separate elementary sound components such as vowels and consonants in spatiotemporal activity of the auditory cortex. To test this hypothesis, we incorporate a brain rhythm into a neural network model of auditory cortex, previously proposed by us. The model consists of three networks. Auditory information is encoded with spatiotemporal pattern of neuronal activity in the A1 layer that models the primary auditory cortex. Then the spatiotemporal aspect of the neuronal activity is detected by the feature-detective (FD) neurons in the second layer. These FD neurons integrate the spatiotemporal pattern over a short time period, thereby enabling the second layer to represent the information about notes and about the correlation of notes. The information encoded by the FD neurons is then combined as a linkage of attractors in the feature binding (FB) layer, providing a semantic information such as word. The three layers are linked with membrane oscillations of a beta rhythm (20Hz). Using the computational model, we show that the beta oscillation enables the FD layer to extract elementary sounds components such as vowels and consonants from spatiotemporal activity of the A1 network. We also show that the beta-coupling of the FD and FB layer facilitates the learning in the FB layer, required for combing syllables encoded in the FB layer. The results provide a clue for understanding the mechanism of how the information of notes, syllables, and phrases are constructed from spatiotemporal activity of the primary auditory cortex.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-144 視床網様核における異なる周波数の音入力の干渉 Akihisa Kimura（木村晃久），Hiroki Imbe（井辺弘樹）和歌山県立医大医生理一 The thalamic reticular nucleus (TRN), receiving excitatory inputs from cortical areas and thalamic nuclei, subserves modulation of thalamic sensory processing through its inhibitory projections to thalamic nuclei. Auditory cortical areas and thalamic nuclei send topographically organized projections to the caudoventral part of the TRN (auditory sector) that in turn sends topographic projections to the ventral division of the medial geniculate nucleus (MGV). The TRN is likely to subserve tonotopy-based modulation of auditory processing in the MGV. It remains unclear how cortical and thalamic inputs interact in the TRN to modulate thalamic auditory processing. The present study focuses on alterations of auditory response by two tones of different frequencies given with various intervals in single TRN cells to obtain insights into TRN-mediated gain and/or gate control of auditory processing. Experiments were performed on anesthetized rats, using juxta-cellular recording and labeling techniques that reveal precise cell locations and terminal fields of axonal projections. After estimating best frequencies based on responses to white noise burst and 11 pure tone stimuli, we examined how subthreshold sound stimulation affect auditory response (unit discharges) evoked by sound of another frequency given at 50-600 ms later following the subthreshold stimulation. 57 out of 65 TRN cells (88 %) showed alterations of auditory response with regard to response magnitude, latency, and/or burst spiking. Suppression predominated in response magnitude modulation. Preceding subthreshold stimulation altered not only onset but also late responses repeatedly evoked after the end of sound stimulation. Subthreshold stimulation given after onset responses also affected late responses. The majority of intervals between subthreshold stimulation and altered response were less than 200 ms, but some exceeded 500 ms. Alterations tended to be intense when frequencies of preceding sound were close to that of sound eliciting responses, but in some cases sound of a frequency widely separated from the frequency of sound eliciting responses specifically altered responses. These robust and complex interactions of auditory inputs across sound frequencies suggest that single TRN cells incorporate various sound information to compose temporal frameworks of information processing in the loop circuits between the cortex and thalamus crucial for auditory attention and perception.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-145 赤外光レーザー人工内耳における言語情報・パラ言語情報の符号化方法の検討 Haruka Yamasato（山里遥香）¹，Yuta Tamai（玉井湧太）²，Kazuyuki Matsumoto（松本和之）²，Miki Yoshida（吉田美紀）¹，Shizuko Hiryu（飛龍志津子）²，Kohta I Kobayasi（小林耕太）² ¹同志社大生命医科学 ²同志社大院生命医科学研究科脳神経行動工学研究室 An action potential can be evoked by irradiating a neuron with infrared laser. The laser stimulation has been paid much attention as a substitution of electric stimulation because the laser can stimulate nerves without contacting tissues. Our group want to develop novel auditory prosthesis with infrared laser for sensorial hearing loss. The purpose of this study was to develop speech encoding scheme for replicating linguistic and paralinguistic information (i.e., sexuality and individuality) with laser auditory prosthesis. Our previous study revealed that trans-tympanic membrane laser stimulation could activate cochlear nerves from broad part of a cochlea; therefore, we synthesized click-modulated speech sound (CMS) as simulated sounds of a novel auditory prothesis with infrared laser. The CMS replicate amplitude of four frequency band of an original sound because we hypothesized that four lasers were irradiated on four different part of a cochlear. The repetition rate of click followed fundamental frequency of the original sounds. Four speaker's speech sounds (two males and females) were used as the original sound. Each stimulus was calibrated as 70 dB SPL. Seven native Japanese speakers with normal hearing participated in the experiment. Before test session, the subjects practiced linguistic transcription and speaker identification with original sounds. In the test session, subjects listened to CMS and wrote down what they perceived and choose speaker from four alternative choices. As results, high correctly perceived linguistic (vowel: 96.3 %, consonant: 86.6 %) and paralinguistic (sex: 95.8 %) rate were achieved. The result suggests that novel auditory prosthesis with infrared laser may create linguistic and paralinguistic information by irradiating four different area of a cochlea with a repetition rate following fundamental frequency of an original sound.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-146 In vivoマウス聴覚皮質における音誘発応答を模擬した電気的刺激法の開発 Takashi Tateno（舘野高），Shuto Muramatsu（村松修斗），Masato Toda（戸田聖人），Jun Nishikawa（西川淳）北海道大学 In the neocortex, local application of electrical currents inside cortical tissue is one of the most common techniques to activate neurons, and intracortical stimulation (ICS) has a potential for new types of a neuroprosthetic device directly applied for the cortex. To both natural sensory and artificial electric stimuli for in vivo recordings of the same neuronal population, the comparison of the ICS-driven response patterns will give a direct answer to the question that the activated circuits are naturally physiological vs. profoundly artificial. However, few studies extensively reported simultaneous electrophysiological recordings combined with ICS in rodents. Here, to advance the field of cortical neuroprosthetics, we evaluated the similarity between the response patterns of cortical activity driven by sound and ICS in different cortical layers. For the auditory cortex of C57BL/6J mice, we performed laminar recordings via 16-channel silicon electrodes and ICS using sharp glass-pipette electrodes containing a marker (biocytin) for the site identification with the in-between distance from 86.3 ± 17.4 μm. In different cortical depth, current pulses (biphasic, 100-μs duration, 10 to 150 μA) were delivered to the mice in vivo under urethane anesthesia. For the recorded data, the properties of local field potentials and the current source densities (CSDs) were mainly analyzed. After the recordings, to determine the stimulation sites, the coronal slices of the mice were stained with Neuro Trace 500/525 Green and then imaged digitally. In this study, we demonstrate that electrical stimulation evoked different excitation patterns according to the stimulated cortical layer. Next, we propose a CSD-based stimulation method to artificially synthesize sound-driven responses, utilizing an approximation method associated with the linear combination of the CSD patterns electrically stimulated in the different cortical layers. Using these approaches, we provide a new technique that natural sound-driven responses could be mimicked by computationally well-designed stimulation pattern sequences in a layer-dependent manner, expecting a contribution to electrical stimulation methodology in a cortical prosthesis.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-147 鳥類高次聴覚領域における音声刺激に対する神経応答特性 Ayano Yoshida（吉田彩乃），Masahiro Inda（院田雅裕），Kohji Hotta（堀田耕司），Kotaro Oka（岡浩太郎）慶應義塾大学理工学部生命情報学科 Zebra finches (Taeniopygia guttata) use songs to communicate with others. Songs enable the birds to discriminate species and individuals, and previous studies suggested that females have high selectivity to individual male songs (Terpstra et al., 2006). That is why we focused on the song discriminability of females. In the avian brain, caudomedial nidopallium (NCM), one of higher auditory regions, might play the roles for discrimination and memorization of songs. Furthermore, recent studies showed that anesthesia has a great effect on the neural activity (Ruijssevelt et al., 2017). In this study, we recorded neural activities to several song stimuli from single NCM neurons in awake birds to investigate the difference between responses to songs. We also recorded the neural activities of single NCM neurons in birds under urethane anesthesia and awake birds, and compared the responses. In our experiment, three songs were presented as sound stimuli: direct song (sing for courtship), undirect song (sing in daily life) and hetero-specific song (Bengalese finches). We detected the significant differences in awake birds between the responses to undirect song and that to hetero-specific song. To find the reason why there was no significant difference between response to direct song and that to hetero-specific song, we investigate the difference of the several acoustic factors of songs: amplitude, fundamental frequency, mean frequency, frequency modulation and entropy. The comparisons of these acoustic factors between songs indicated that neurons in NCM encode these song properties and this ability caused the difference of responses. Furthermore, in the comparison between awake birds and anesthetized ones, there was no significant difference in the responses to the whole songs. However, each response to song syllables indicated significant differences. These results suggested the possibility that responses of awake and non-awake birds are different.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-148 MMN出現時における聴覚皮質ニューロンの発火特性 Eiichi Jodo（浄土英一）¹，Tadahiro Katayam（片山規央）²，Yoshiaki Suzuki（鈴木喜明）¹，Ken-yo Hoshino（星野賢洋）³，Satoshi Eifuku（永福智志）¹ ¹福島県立医科大学医学部システム神経科学講座 ²福島大学保健管理センター ³一陽会病院 The mismatch negativity is a change-specific event-related potential that is usually invoked by auditory stimuli deviating from established patterns of regularity. The reduction of MMN is a reliable biological property of patients with schizophrenia. The main electrical source of MMN is now considered to be located in the auditory cortex (AuC). However, it remains to be elucidated how the unit activity of AuC neurons is during elicitation of the MMN. In this study we recorded the MMN from the surface of the frontal cortex (FC) and unit activity of AuC neurons simultaneously during auditory stimulation in freely-moving rats. Recording was executed under two different conditions. In one condition two auditory stimuli with different frequency (3000, 6000Hz; 85dB SPL) and different probability of occurrence (10, 90%) were sequentially presented with stimulus onset asynchrony of 0.3s (odd-ball condition). In the other condition (many-standard condition) ten frequency-different auditory stimuli including former two stimuli (3000, 6000 Hz) were sequentially presented with the equal probability of occurrence (10%). The duration of each stimulus is 0.1s in both conditions. The human MMN-like potential with a peak latency of about 50 ms was recorded from the surface of FC near the midline. This MMN-like potential to the deviant tone (10%) in the odd-ball condition was notably larger than the ones both to the standard tone (90%), and to the non-deviant tone that was presented in the many-standard condition with the same probability of occurrence and pitch as the deviant tone. The waveform of local field potential (LFP) recorded in the primary AuC (pAuC) had notable bimodal peaks that temporally correspond to the rise and decay of the tone, and exhibited no significant change between deviant- and non-deviant tones. The LFP in the secondary AuC (sAuC) ventral to the pAuC exhibited a MMN-like waveform, and its amplitude depended on whether the stimulus was deviant or non-deviant. The unit activity of sAuC neurons differentially responded between deviant- and non-deviant tones, while pAuC neurons did not exhibit such change-specific differential responses. However, most of sAuC neurons exhibited inhibitory responses during elicitation of the MMN.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-149 鳥類高聴覚領域における時間スケールに依存した神経活動の性質について Masahiro Inda（院田雅裕），Ayano Yoshida（吉田彩乃），Kohji Hotta（堀田耕司），Kotaro Oka（岡浩太郎）慶應大理工生命情報 Zebra finches (Taeniopygia guttata) use their own songs for courtship. Female birds select her mating partner based on male's songs that consist of temporal structures of acoustic factors. Therefore, encoding temporal structures of songs in the brain are fundamental function in songbirds. The avian higher auditory regions, caudomedial mesopallium (CMM) and caudomedial nidopallium (NCM), are considered to have functions for song discriminability and memorization of mate song. It is known that song of zebra finches contains temporal hierarchical structures. Song is constructed from motif structures, and motif structure constructed from syllables and notes. Thus, we hypothesized that auditory neurons have complex temporal song encoding system depended on time scale. In this study, we investigated the relationship between neural activities and temporal structures of acoustic factors. We detected 124 sound responsive neurons and applied some computational analysis to these data. First, Fano factor as the function of time window size in CMM and NCM were calculated to investigate the detail of fundamental firing patterns. Change of Fano factor indicated clear differences by increasing of time window size in both regions. Especially Fano factor in NCM indicated over 1. Thus, spike pattern of sound responsive neurons in CMM and NCM have difference about fundamental temporal firing system at long time scale. Second, to evaluate temporal neural coding of these acoustic factors, we calculated Time Series Correlation (TSC) as a novel index of relationship between neural activities and acoustic factors. TSC could simultaneously evaluate positive and negative correlation intermixing. Change of TSC value of seven acoustic factors depended on increasing of window size indicated monotonically decreasing in all seven acoustic factors. Thus, to visualize the relative relationship of change of TSC values across acoustic factors, we calculated relative contribution to TSC. The relative relationship across seven acoustic factors changed with time window size. Remarkably, TSC of amplitude and entropy were dramatically changed compared with the control data, which were calculated from neural activity during silence. On the other hands, overall trends of TSC were similar between CMM and NCM.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-150 コモンマーモセット聴覚野の神経結合マップと周波数選択性マップ Toshiki Tani（谷利樹）¹，Hiroshi Abe（阿部央）¹，Hiromi Mashiko（益子宏美）¹，Naohito Kitamuara（北村尚士）¹，Kazuhisa Sakai（境和久）²，Taku Hayami（速水琢）²，Satoshi Watanabe（渡邉惠）²，Wataru Suzuki（鈴木航）²，Hiroaki Mizukami（水上浩明）³，Akiya Watakabe（渡我部昭哉）¹，Tetsuo Yamamori（山森哲雄）¹，Noritaka Ichinohe（一戸紀孝）^1,2 ¹理化学研究所　脳神経科学研究センター　高次脳機能分子解析チーム ²国立精神神経医療研究センター　神経研究所　微細構造研究部 ³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 Natural sound (voice, environmental sound, etc.) is composed of various frequencies. It has been well known that frequencies in sounds are represented in a systematic manner, i.e., tonotopic organization, in the core of the marmoset auditory cortex. Recently, by using optical intrinsic signal imaging (OISI), we examined evoked responses to band-pass noise stimuli in a range of sound frequencies (0.5-16 kHz) in anesthetized common marmosets (Callithrix jacchus) and found that the preferred sound frequency map existed in not only core but also belt and parabelt. Previous studies show that axonal projections reciprocally connected among auditory cortices in the common marmoset. However, the spacial relationship between the axonal projection map and the preferred sound frequency map within the auditory cortex is elusive. To address this issue, we injected an AAV vector, which works as an anterograde tracer by expressing either green or red fluorescent protein in infected neurons, into higher and lower frequency preferred regions, identified by OISI, in each auditory area, identified histologically. After a three-week waiting period, the animals were perfused, and the post-mortem brain was sectioned in a thickness of 50 um. The sections were divided into three series, one for fluorescent image scan and two for staining for myelin and Nissl substrate in order to identify cortical areas. We found that higher and lower responsive regions in the core, belt and parabelt tended to be connected with, respectively, higher and lower responsive regions in other auditory cortices. However, small fraction of the axonal projections from the higher and lower responsive regions were overlapped. These results suggest that the connections binding the similar frequency might differently contribute to sound frequency analysis in functionally and hierarchically distinct auditory cortices embedded in the core, belt and parabelt.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-151 グレア錯視の "まぶしさ"：心理物理学的研究 Ryo Sato（佐藤良）¹，Tomoka Matsumura（松村朋花）¹，Tomoya Ueda（上田誠也）²，Takuya Miyagi（宮城拓弥）²，Daiki Yoshioka（吉岡大貴）²，Saki Akaike（赤池早紀）³，Hidenori Horita（堀田英則）³，Megumi Ishizawa（石澤恵）³，Yuki Yamada（山田祐樹）⁴，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 ³スズキ株式会社 ⁴九州大学基幹教育院 Perceived brightness is illusorily increased in a region surrounded by luminance, which is gradient like scattered light (glare illusion). Tamura et al. (2016) psychophysically quantified the glare illusion and demonstrated that its illusory effect increased subjective brightness about 40%. In their study, however, participants had judged the "brightness" of visual stimuli but not its "glare." Illuminating Engineering Society (IES) defines glare as "the sensation produced by luminances within the visual field that are sufficiently greater than the luminance to which the eyes are adapted to cause annoyance, discomfort, or loss in visual performance or visibility." Thus, not only does glare include subjective brightness, but also discomfort and loss of visibility. In this study, we investigated the subjective "glare" of the glare illusion. Participants received a visual stimulus (luminance: 281, 315, 355, 450, 680, 1170, or 2350 cd/m²; duration: 300 ms) and judged whether the stimulus included glare or not. We set two conditions for the visual stimuli: glare-illusion and control conditions. In the visual stimuli for the glare-illusion condition, the illuminant central circle was surrounded by a luminance, which is gradient like scattered light. In the visual stimuli for the control condition, the illuminant central circle was surrounded by a ring of uniform luminance. The average luminances were the same in visual stimuli for the glare-illusion and control conditions. We calculated the threshold and the just noticeable difference (JND) from the proportion of the trials judged as "glare," as a function of the luminance of the stimuli, for each condition. In Experiment 1 (n = 12), we explained to the participants the IES definition of "glare." In Experiment 2 (n = 12), we did not provide the definition to the participants. As a result, there was no difference in the threshold and JND between the glare-illusion and control conditions in Experiments 1 and 2. Our results revealed that the "glare" illusion has no effect on subjective glare. Renaming this illusion will resolve the discrepancy between the name and the actual effect. This finding also suggests that by using the illusion, we can provide observers with increased-brightness displays, without any discomfort or loss of visibility that accompanies the "glare.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-152 ネコV1野におけるコントラスト依存の空間周波数チューニングダイナミクス Hiroki Tanaka（田中宏喜）京都産業大学情報理工学部 It was reported that V1 neurons often increase spatial frequency (SF) tuning bandwidth to cover higher spatial frequency range as stimulus contrast becomes higher (Sceniak et al. 2002). However, it is less clear how this contrast-dependent change of SF tunings is generated. It is known that many V1 neurons increase their preferred SFs during response. Therefore, one possibility is that SF tuning dynamics depend on stimulus contrast so that contrast-dependence of SF tunings develops during response time. To examine this, we measured time course of SF tunings of cat V1 neurons under different contrast conditions (6.25% ~ 50%). We vertically penetrated cat V1 with a multi-site electrode array and recorded the activities of 143 V1 neurons in response to random sequences of flashed sinusoidal gratings of various orientations, SFs and phases. Stimulus contrast of the gratings in each sequence was constant and sequences of different contrasts were interleaved. Time courses of SF tunings for each contrast were computed by a subspace reverse correlation technique. We found that shifts of preferred SF during response time become larger as stimulus contrast increases. Mean SF shift values were 0.19, 0.26, and 0.34 octaves for contrast 6.25%, 12.5%, and 50%, respectively. The preferred SFs of neurons for different contrasts were relatively similar at response onset. However, at response peak and offset, the preferred SF of a neuron for contrast 50% was typically higher than that for contrast 6.25%. This difference was larger for neurons with greater SF dynamics. These results show that SF tuning dynamics highly depend on stimulus contrast and that with these dynamics, contrast-dependence of SF tunings develops during response time.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-153 マウス一次および高次視覚野に分布するLPN投射軸索の反応特性 Yuko Kiyohara（清原悠嵩）¹，Satoru Kondo（根東覚）²，Kenichi Ohki（大木研一）^1,2 ¹東京大院医統合生理 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 The neurons of primary (V1) and higher visual cortical area (HVA) in mice have the characteristic response selectivity to visual stimulus features. These cortical areas receive visual input directly or indirectly from the two thalamic nuclei, lateral geniculate nucleus (LGN) and lateral posterior nucleus (LPN). Visual information captured by retina is mainly transmitted via LGN to V1 and further to HVAs. Because LPN receives visual information indirectly from retina via superior colliculus (SC), LPN could be the information ascending route different from LGN (Tohmi et al., 2014; Beltramo and Scanziani, 2019). The axons from LGN to V1 and from V1 to HVAs were well examined previously (Glickfeld and Reid, 2013; Matsui and Ohki, 2013; Kondo and Ohki, 2016). However, little is known about the response properties of axons from LPN to V1 or HVAs. To examine the response selectivity of LPN projections to visual cortical areas, we examined activities of LPN axons that innervate V1 and some HVAs including dorsal and ventral visual pathways (PM, AL and LM). We locally expressed genetically encoded calcium indicator GCaMP6s in the LPN neurons by AAV-mediated method and recorded responses of their axons to the visual stimulation in V1 and HVAs by wide-field and two-photon calcium imaging. We examined the differences between response selectivity of LPN-V1 and LPN-HVAs obtained in this study and LGN-V1 and V1-HVAs data from the previous literatures. This work was supported by MEXT/JSPS Kakenhi 17H0303540 to SK and Brain/MINDS to KO.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-154 腹側と背側視覚伝導路を連結する鉛直束(VOF)の白質解剖と神経束構成 Atsushi Yamaguchi（山口淳）¹，Seiichiro Hirono（廣野誠一郎）²，Tatsuya Yamamoto（山本達也）¹，Keiko Kitajo（北城敬子）¹，Yasuo Iwadate（岩立康男）²，Tatsuya Jitsuishi（實石達也）¹ ¹千葉大学　大学院医学系研究科　機能形態学 ²千葉大学　大学院医学系研究科　脳神経外科 The vertical occipital fasciculus (VOF), originally described by Wernicke, is located at the posterolateral corner of the brain. The VOF was re-evaluated by the recent neuroimaging studies as the major fiber tract to connect the dorsal and ventral visual cortices. Tractography studies showed the major cortical endpoints of VOF fall in the dorsal (i.e. V3A/B, V3d, IPS-0) and in the ventral visual areas (i.e. hV4,VO-1,VO-2), respectively. To validate the fiber tracking data of VOF previously reported, we here performed the white matter dissection in the post-mortem human brain and showed the major fiber streams of VOF connecting V3A/B areas with the fusiform gyrus. Glasser et al.(2016) recently reported the HCP MMP1.0 (the Human Connectome Project's multi-modal parcellation version 1.0) that is an updated map of the human cerebral cortex integrating information from several MRI modalities on data from the HCP. The 180 areas were delineated and identified by a combination of multi-modal neuroimaging techniques based on the degree of myelination, cortical thickness, and functional connectivity derived from task-based and resting-state fMRI. We then conducted the quantitative analysis of the structural connectivity of VOF to link relevant cerebral parcellations on an updated map of the human cerebral cortex(HCP MMP1.0). We found the laterality and the variability of VOF's subcomponents connecting dual visual streams. These findings could suggest a critical role of VOF to connect vision-related cortices, providing one of the common frameworks for future studies.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-155 単眼遮蔽による視覚野のヒゲ刺激への応答変化のin vivoイメージング Akari Hashimoto（橋本明香里），Akiko Miyamoto（宮本愛喜子），Yoshihisa Tachibana（橘吉寿），Koichiro Haruwaka（春若航一路），Hiroaki Wake（和氣弘明）神戸大院医生理学・細胞生物学システム生理学 External stimulations trigger sensory responses to exert the higher order of brain functions, but a part of them is disturbed in the patients of blindness or deafness. The concept of cross-modal plasticity is to compensate the impaired sensory system by the others. The previous studies using positron emission tomography (PET) showed that activity of visual cortex occurred in the blind people with reading Braille. Whisker-dependent activation of visual cortex was demonstrated in the eye enucleated mice. However, the functional changes of the compensated sensory cortical area in vivo have not been shown yet. In this research, we try to unravel the effect of early monocular deprivation (MD) on the activation of the visual cortex with whisker stimulation. We first visualized the axonal projection from S1 (primary somatosensory cortex) to V2 (extrastriate cortex) by injecting adeno-associated virus coded eGFP with synapsin promotor (AAV1-syn-eGFP). Retrograde labelling of S1-V2 axons by Cholera Toxin Subunit B (Recombinant) Alexa Fluor^TM 488 Conjugate also showed the existence of the projection. Then we hypothesized the activation triggered by whisker stimulation in S1 can be transmitted and processed in V2 as well. To verify our hypothesis, we combined in vivo two-photon imaging with the AAV injection to visualize the neuron in V2 with genetic Ca²⁺ indicators (GCaMP6f) and analyzed the correlation between neural activity and whisker stimulation. Furthermore, we assessed synaptic pruning by microglia, which might be a driving force to modify the neuronal circuit with MD. This study will be an important clue to understand the compensating ability of the cortex for the future therapeutic target.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-156 Perception of visual ensemble statistics of faces in autism and typically-developing individuals Mrinmoy Chakrabarty（Chakrabarty Mrinmoy）¹，Dr. Makoto Wada（Wada Dr. Makoto）^1,2 ¹National Rehabilitation Center for Persons with Disabilities ²Faculty of Informatics, Shizuoka University Humans invariably depend on environmental information to interact with the visual world. Visual targets are often represented and perceived in the context of surrounding visual information in the brain. How summary statistics conveyed by ensembles of contextual visual information influence the perception of a given target are less explored. We studied how summary intensities of goal irrelevant, high and low level visual information (emotion and brightness related information from facial stimuli, respectively) extracted from periphery affect the perception of a central target facial stimulus (centre task) in 19 typically developing (TD) and 10 autism spectrum condition (ASC) individuals. We found that graded alterations in the summary intensities of the emotion (sad to happy axis) and brightness (dark to bright axis) of the irrelevant context modulated the net perceptions of the relevant target face. In another experiment, perceptions of task relevant global mean emotion and brightness (mean task) correlated with centre task performance in TD but not ASC implying differences in task based visual cognitive flexibility between the two groups. Individual ASC sensitivities when compared to TD derived norms, largely differed. Finally, a cluster analysis applied with the variables experimental performance metrics and autism quotient (AQ) scores across all participants, separated TD from ASC group with appreciable accuracy. Participants with higher AQ had larger task variability, and most of them were ASC. The results suggest that summary statistics of visual ensemble information are differentially perceived by TD and ASC which influence their target visual perceptions based on task demands

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-157 ショウジョウバエにおける視覚認知メカニズム解明のための装置開発 Akinori Kayanoki（栢木秋典）¹，Kentaro Sugimoto（杉本健太郎）²，Junji Yamauchi（山内淳司）¹，Shuto Yamasaki（山﨑秀斗）¹，Takako Morimoto（森本高子）¹ ¹東京薬科大学 ²東京工業大学 ^{The neural mechanisms of visual perception attract much interest of many researchers. A number of studies have performed using vertebrates including primates and mouse. However, it has been reported that invertebrates also have vision and can discriminate and remember visual landmarks to gather important information from environment. Further, even insects possibly experience visual illusion, such as a perception of subjective contours. We think that it will be a very promising tool to study the neural mechanism of visual recognition using insects, especially like fruit flies. Here, we have established a new device to study visual perception in flies. In the previous report, the fly was conditioned to avoid certain visual patterns and tested whether it can discriminate the remembered pattern within other patterns by using the device like a flight simulator in which a fly tethered to a torque meter and can control its orientation. However, their equipment is very special and difficult to establish in the other laboratory. Therefore, we develop a new device that can be easily replicated at a low cost. This equipment basically consists of a mouse sensor and a servo motor to control visual stimulus. The mouse sensor can detect the rotation of a small floating ball and quantify walking distance of a fly on the floating ball. Then, the fly movement detected by the sensor is feedbacked to the movement of servo motor. Thus, our newly developed device can make the virtual reality world around a tethered fly. All these components are controlled by microcomputers, Arduino. Arduino supports a high performance in confined spaces. By combining with an external sensor that can trigger a thermal stimulus signal as a punishment for the fly, the device can also be used for the aversive conditioning. Therefore, using this device, we can study the neural mechanism of visual learning as well as visual perception. Currently, we examine the effectiveness of our device and try to know whether flies can percept the subjective contours such as kanizsa triangles.}		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-158 眼球運動と脳波による視覚認知に関する運転熟練度の評価 Keiichiro Inagaki（稲垣圭一郎），Tatsuya Maruno（丸野達也），Yuya Gotoh（後藤祐哉）中部大学工学部ロボット理工学科 Intelligent Transport Systems is aiming reduction of traffic accidents by developing various types of driving support infrastructures. It is reported that traffic accidents are reduced with the growth of ITS technologies. In the field of ITS, much technologies are usually developed in terms of improvement of traffic environment and vehicle functions to reduce traffic jam and accidents using latest information, communication, sensor technologies. On the other hand, it is necessary to understand driver's conditions including brain activities during vehicle driving to further improvement of ITS technology. To support this, it was reported that several approaches to understanding brain signal processing regarding vehicle driving are proposed in previous study. In those study, the relation between brain activities and driver's attention and body response are evaluated. In the vehicle driving, drivers search traffic scenes using eye movement, and cognize and judge traffic situations for maneuvering vehicle. Here, the brain processes numerous information related to those. On the other hand, the strategy for cognition, judgment, and operation is individually different for each driver. Moreover, eye movements, scan path, related those are also different. Those differences are thought to be arising from level of driving skill. To support this, it is reported that occurrence of traffic accidents during driving is frequent in beginner drivers than more experts. It indicated that the level of driving skill is crucial factor in driving. In the present work, we measure and analyze human brain activity (EEG: Electroencephalogram) and eye movement during vehicle driving, and evaluate the relationship between those and driving skill level.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-159 ワーキングメモリによるカテゴリー化タスク実行の神経機構 Hikaru Tokuhara（徳原光），Yoshiki Kashimori（樫森与志喜）電気通信大学基盤理工学 The ability to categorize objects is a fundamental function of visual recognition. In daily life, we can rapidly and effortlessly categorize huge numbers of objects. Much effort has been devoted to the studies of how category information is processed in the brain circuits. Freedman et al. trained monkeys to categorize the computer-generated images of dogs and cats. Monkeys performed a delayed match-to-category task that required to judge whether a sample and test stimulus were from the same category. Freedman et al. demonstrated the involvement of the interaction between inferior temporal (IT) cortex and prefrontal cortex (PFC) in the categorization task. Moreover, they showed that neurons in the IT were more sensitive to object features, whereas neurons in the PFC exhibited a larger sensitivity to object categories. However, it still remains unclear how working memory of category information is formed in the PFC and how the decision of the dogs/cats-categorization task is made from working memory activity. To address these issues, we develop a model of visual system that consists of a PFC and an IT networks. Each network has two-dimensional array of neurons, and single neuron was modeled with the leaky-integrator model. The PFC network has two neuron groups; one neuron group maintains category information during delay period, while another group encodes information of the time course of working memory activity and the activity elicited by a test stimulus in order to make a decision in a match or non-match trial. We show that PFC neurons with lower firing thresholds play a dominant role in formation and retention of working memory whereas PFC neurons with higher firing thresholds are highly sensitive to category information of dogs/cats images. We also show that temporal relationship between working memory activity and the activity evoked by a test stimulus is detected by a neuron group of PFC to make decisions relevant to a match or non-match trial. These results suggest that the two different functions of PFC neurons may be involved in efficient performance of categorization task.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-160 代謝型グルタミン酸6型受容体の細胞膜発現におけるC末端領域の役割 Takumi Akagi（赤木巧），Dilip Rai（Rai Dilip），Atsushi Shimohata（下畑充志），Toshiyuki Ishii（石井俊行），Mie Gangi（雁木美衣），Takuma Maruyama（丸山拓真），Ikuo Ogiwara（荻原郁夫），Makoto Kaneda（金田誠）日本医科大医生理（システム生理） The metabotropic glutamate receptor type 6 (mGluR6) is a retina-specific G-protein coupled receptor locating at dendritic tips of ON bipolar cells. mGluR6 senses glutamate released from photoreceptors and hyperpolarize ON bipolar cells via intracellular signaling once activated. To function properly, a well-organized localization of mGluR6 is essential at the subcellular level. However, the portion required for the regulation of intracellular receptor trafficking remains elusive. We here address whether the C-terminal domain (CTD) of mGluR6 (residues 840-871) is involved in cell surface expression using heterologous expression systems. We found that cell surface expression was not affected by removing up to 14 amino acids from C-terminus (residues 858-871), while it disappeared by further truncations from 15 (residues 857-871) to 20 (residues 852-871) amino acids. However, removal from 21 (residues 851-871) to 31 (residues 841-871) amino acids from C-terminus reverted cell surface expression, while loss of entire CTD (residues 840-871) again abolished it. We also eliminated residues sequentially from the border of intra-membrane position (residue 840) in the C-terminal direction, and found that all mutants except for that lacking entire CTD displayed cell surface expression. We further observed that an mGluR6 mutant, lacking the N- and C-terminal regions but containing a 6-aa middle segment (residues 851-856) at CTD, was deficient in cell surface expression. These observations suggest that cell surface expression of mGluR6 requires the sole residue at either N- or C-terminal end of CTD, while the CTD may contain a negative regulatory subdomain.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-161 深層学習を用いた嗅内野の視覚情報表現の同定 Seito Fukagawa（深川晴登）¹，Jumpei Ukita（浮田純平）¹，Nana Nishio（西尾奈々）^1,2，Kenichi Ohki（大木研一）^1,2 ¹東京大院医統合生理 ²東京大学ニューロインテリジェンス国際研究機構 The mouse has several higher visual areas that can be segregated into dorsal and ventral areas. Ectorhinal cortex (ECT, area TEa/36a) in the temporal cortex receives projections from the ventral areas but its function has little been elucidated thus far, except for their responsiveness to the moving objects (Nishio et al., Sci Rep., 2018). Recently, deep learning has not only revolutionized the artificial intelligence researches such as computer vision and natural language processing, but also been an emerging tool in the analysis of visual cortex, such as nonlinear receptive field estimation (Ukita et al., bioRxiv, 2018). In this study, we extended such deep-learning-based receptive field estimation to deciphering the spatiotemporal visual features represented in the mouse ECT. We first performed chronic wide-field Ca²⁺ imaging of the visual cortex, including ECT, while presenting natural movies to the awake mouse. Then using deep learning, we fitted the ECT Ca²⁺ responses to the natural movies. In order to model the response of ECT, we combined convolutional neural network (CNN) and long short-term memory (LSTM) layers. We compared many architectures with different numbers of CNN and LSTM layers. We confirmed that the networks were able to generalize; the learning loss and validation loss decreased, and the correlation between the predicted and the actual ECT responses became high. We then generated the optimal movie stimulus for ECT such that the movie would predictively maximize the output of the trained network. The synthesized movie contained contrast-changing objects and moving objects in a specific retinotopic position. These results imply that mouse temporal cortex may encode these visual features and a combination of CNN and LSTM can model the complex visual processing in the temporal cortex.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-162 ネコとげっ歯類動物における視覚皮質ニューロンの応答特性差異のモデル研究 Masanobu Miyashita（宮下真信）¹，Shigeru Tanaka（田中繁）² ¹沼津高専 ²電通大 In the cat primary visual cortex, most of neurons selectively respond to specific orientations of local contours presented in the visual field, and are regularly arranged according to neurons' preferred orientations, forming orderly orientation maps. Numerous experimental studies have been reported on the dependence of neuronal activity upon the size or luminance contrast of visual stimuli. On the other hand, little is known about the response properties of neurons in the rodent visual cortex, where orientation-selective neurons are sparsely and randomly distributed in the salt-and-pepper-like fashion. Recently we have reproduced qualitatively different orientation representations based on the same self-organization model only by changing the value of one parameter: the probability of excitatory intracortical connections, p. The salt-and-pepper-like orientation representation emerged at p < pc, whereas orderly orientation maps appeared at p > pc. In this study, in order to elucidate how the structure of orientation representation affects neuronal response properties, we performed computer simulations of the dynamics of neural networks composed of leaky integrate-and-fire units that receive the self-organized thalamo-cortical inputs. In the simulations, we employed sinusoidal grating patches as visual stimuli, and changed the luminance contrast and length of the grating patches along the stimulus orientations. In the model visual cortex showing either orderly map or salt-and-pepper-like representation, the neuronal firing rate in response to longer grating patches was saturated or suppressed, although firing rate in the salt-and-pepper-like representation was generally much lower than that in the orderly maps. Particularly at higher stimulus contrasts, neurons in the salt-and-pepper-like representation elicited maximum responses when the size of grating patches was similar to the size of classical receptive fields (CRFs). When only CRFs were stimulated by grating patches, the firing rate of neurons in orderly maps monotonically increased as luminance contrast increased up to 100%, whereas that in the salt-and-pepper-like representation showed plateau at contrasts higher than 50% and no responses were observed at contrasts lower than 10%. The simulation results suggest that the regularity of orientation representation contributes to the emergence of orientation-selective neurons with a wide dynamic range of luminance contra.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-163 知覚時間と赤/青刺激により誘発される瞳孔反応の関係 Yuya Kinzuka（金塚裕也）¹，Fumiaki Sato（佐藤文昭）¹，Tetsuto Minami（南哲人）²，Shigeki Nakauchi（中内茂樹）¹ ¹豊橋技科大工情報・知能工 ²豊橋技科大エレクトロニクス先端融合研 The effect of colors to our behavior and emotions are well known. In addition, recent studies have shown that observing emotional stimuli affects subjective experiences of time of the perceiver. Although, the potential effects of color to temporal perception has not been sufficiently investigated, when the effect of color red and blue are still controversial. The perceived duration is reported to be overestimated induced by hue-arousing color such as red (e.g. Shibasaki and Masataka, 2014), however, few studies suggest the color blue was perceived longer (Thönes et al., 2018). Also, study involving monkeys has reported a close relation between perception of passing time and pupillary response (Suzuki et al., 2016). To this end, this study aims to investigate how our perception of time is associated with pupillary response including the pupillary light reflex (PLR), which is closely linked to internal factors and neuronal activity in the locus coeruleus. Experimental procedure was based on the latest temporal perception study (Thönes et al., 2018), combining with pupillometry to investigate the correlation between subjective time perception and the pupillary response. In a two-interval duration-discrimination task, two subjectively equiluminant color stimuli (red/blue) were presented continuously on the screen (Display++ LCD Monitor). On each trial, participant indicated which stimulus (former/latter) had been presented for a longer duration using a NumPad. Pupil diameter was recorded by EyeLink 1000PLUS while presentation of the two stimuli. The behavioral data suggest that the color red was significantly perceived longer compared to blue stimuli indicated by a shift of the PSE, even though the based study reported the opposite result. Moreover, sampling trial with the same color condition and same duration condition (which means the two continuous stimuli, former/latter is physically identical), constriction of the PLR was significantly larger for stimuli that were judged longer in both color conditions. These results suggest that subjective temporal perception and pupillary response are somehow directly or indirectly connected, additionally, might be one factor to explain the inconsistent effects of color on time perception.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-164 ECoG電極を用いた視覚性ミスマッチ陰性電位のマーモセット全大脳領域からの計測 Masatoshi Yoshida（吉田正俊）^1,2，Takaaki Kaneko（兼子峰明）^3,4，Misako Komatsu（小松三佐子）⁵，Hideyuki Okano（岡野栄之）^3,4，Noritaka Ichinohe（一戸紀孝）^5,6 ¹生理学研究所・認知行動発達機構 ²総合研究大学院大学・生命科学研究科・生理科学専攻 ³理化学研究所・脳神経科学研究センター・マーモセット神経構造チーム ⁴慶應義塾大学・医学部・生理学教室 ⁵理化学研究所脳神経科学研究センター高次脳機能分子解析チーム ⁶国立精神・神経医療研究センター微細構造研究部 [Introduction] Mismatch negativity (MMN) is a well-established brain marker of schizophrenia. To establish an animal model of schizophrenia in marmosets, we previously measured auditory MMN, visual MMN and eye movements in marmosets and examined the effects of an intramuscular injection of ketamine in sub-anesthetic doses. We found that ketamine affected both the component of deviance detection in MMN and saccadic amplitudes in eye movements (Yoshida 2018). The aim of the current study is to extend the previous findings by measuring local field potentials from entire cortical surfaces of a hemisphere of marmosets and by examining which parts of the cortical areas contain information about deviance detection during visual MMN. [Methods] We implanted a 96-channel ECoG electrode epidurally to one of their hemispheres in three marmosets (Komatsu et. al. 2019). As visual stimuli, sinusoidal gratings with different orientations were presented 300 times (in 5 min), each with a 500-ms on period and a 500-ms off period. Two experimental conditions were alternated. One is an odd-ball condition in which 37 deviant stimuli (e.g., 45 deg in orientation) are intermixed with 263 standard stimuli (e.g., 135 deg in orientation). Another is a many-standards condition in which orientation gratings with all eight orientations were presented with an equal probability. By comparing the response to the deviants in the oddball condition and the response to the deviants in the many-standards condition, we evaluated deviance detection, which reflects surprises by unexpected stimuli. [Results] Significantly larger responses corresponding to deviance detection were found from various brain areas including V1, V2, V4, MT, V3/PPC, TPO/FST, and FEF. We also calculated correlation coefficients between LFPs of different channels and compared between the correlation coefficients for responses to the deviants in the oddball condition and those for responses to the deviants in the many-standards condition. We found significant differences in various channel pairs such as V1-V3/PPC, V3/PPC-TPO/FST, TPO/FST-FEF. [Conclusion] We succeeded in detecting the components of deviance detection in marmosets from various brain areas. Since most of them are overlapped with brain areas which are involved in visual salience (Veale et al 2017), our results suggest that deviance detection and visual salience share common computational processes in the brain.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-165 サルV4野神経細胞集団が表現する情報は画像特徴量から心理評価値へと切り替わる Yoshiyuki Suzuki（鈴木愛之）¹，Rina Kotani（小谷梨奈）²，Hiroshi Tamura（田村弘）^1,2,3 ¹大阪大基礎工 ²大阪大院生命機能認知脳科学 ³脳情報通信融合研究センター（CiNet）、大阪、日本 Visual features such as pattern, color and texture on the surface of objects help us to recognize them. Here, we investigated how these visual features are represented by neurons in the ventral visual pathway. We recorded single-neuron spiking activity from the primary visual cortex (V1), V4 and the inferior temporal cortex (IT) of analgesized and paralyzed macaque monkeys (Macaca fuscata, two males and two females, 5.9-8.6 kg) while 64 images from natural object surfaces were presented for 200 ms. We obtained 1333, 903, and 665 neurons from V1, V4 and IT. Among these, 738, 704, and 305 visually responsive neurons were selected for the analyses from each visual area. We examined relationship between neural population responses and image-based visual features or psychophysical features of stimulus images to clarify how the neurons representing object surface images in relation to these two types of visual features. We calculated 32 image statistics as image-based visual features, and obtained 13 psychological features using a semantic-differential scale method. To compare population-neuron responses with the two types of features, we computed representational dissimilarity matrix (RDM). RDM based on population neuronal activity during stimulus presentation period (200 ms) were correlated with image RDM in all the three visual areas, and were correlated with psychological RDM more strongly in V4 than in other visual areas, indicating the importance of V4 for the representation of psychological features. In all the three visual areas, however, relationship between image RDM and neuronal RDM as well as that between psychological RDM and neuronal RDM changed with time. Correlation between neuronal RDM of V4 and image RDM reached the peak around 35 ms after response onset (104 ms after stimulus onset), then showed gradual decrease. Correlation between the psychological RDM and neuronal RDM of V4 increased gradually with time and reached the peak around 140 ms. Both in V1 and IT, correlation between image RDM and neuronal RDM as well as that between psychological RDM and neuronal RDM reached the peak around 100 ms and gradually decreased. These results suggest that the information represented by the V4-neuron population switched around 35 ms after response onset from image features to psychological features.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-166 サルFST野における複雑運動刺激に対する反応特性 Takahisa M Sanada（眞田尚久）¹，Takahiro Kawabe（河邉隆寛）²，Shin'ya Nishida（西田眞也）²，Hidehiko Komatsu（小松英彦）³ ¹関西医科大学医学部生理学講座 ²NTTコミュニケーション科学基礎研究所 ³玉川大学脳科学研究所 Spatial distribution of visual motion in natural scene is typically non-uniform. For example, in liquid flows, various directions/speeds of motion vectors are spatially distributed in a complex manner, while we can easily discriminate liquid flows from noise motion just by the spatial feature of the motion flow. Recently, it was reported that the spatial smoothness of local motion vectors, characterized using the mean discrete Laplacian of motion vectors, correlated with rated liquidness impression by human psychophysics (Kawabe et al., 2015). Motion information is encoded in dorsal visual areas such as MT. Since majority of MT neurons are robustly driven by uniform motion, in which the directions/speeds are uniform across space, spatial feature of motion might be represented elsewhere. Neurons in area FST are selective to spatial structure defined by motion (Mysore et al., 2010), and we hypothesized that FST neurons represent spatial features of complex motion by integrating multiple direction components. In the present study, we tested this possibility by recording responses of FST neurons to complex motion stimuli with various degrees of liquidness. Each motion stimulus consisted of 16 x 16 array of small random dot motion patches, in which location, direction and log-speed were distributed within an arbitrary range. To generate different levels of liquidness of motion stimuli, we manipulated spatial smoothness of motion stimuli that was defined as discrete Laplacian of motion vectors by iteratively swapping positions of motion patches. Sixty-four stimuli were prepared in combination of 8 liquidness levels and 8 mean directions of motion. If a neuron purely represents motion direction regardless of the spatial smoothness of motion vectors, the neuron would respond to its preferred direction regardless of the Laplacian levels. On the other hand, if a neuron is related to represent liquidness, it should respond at particular Laplacian levels. We found that subset of FST neurons which showed broad direction selectivity responded when the spatial smoothness was high, suggesting that spatial feature of complex motion might be represented in area FST.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-167 サル下側頭葉皮質における質感属性の時空間周波数表現 Harunori Miki（三木治憲）¹，Kenta Anzai（安斎健太）^2,3，Masataka Sawayama（澤山正貴）⁴，Takeshi Matsuo（松尾健）⁵，Takafumi Suzuki（鈴木隆文）⁶，Takayuki Okatani（岡谷貴之）⁷，Isao Hasegawa（長谷川功）³，Keisuke Kawasaki（川嵜圭祐）³ ¹新潟大　工　福祉人間工学 ²新潟大学　大学院　自然科学研究科 ³新潟大学医学部　神経生理学分野 ⁴NTT コミュニケーション科学基礎研究所 ⁵東京都立神経病院　脳外科 ⁶情報通信研究機構脳情報通信融合研究センター ⁷東北大学大学院情報科学研究科 An object can be ideally created from arbitrary materials. A cup can be made of plastic, glass or ice. By means of material or shitsukan perception we can settle to an appropriate choice. It has been shown that the inferior temporal cortex (ITC) plays a pivotal role in both object and material perception. Previous neurophysiological studies about material perception mainly focused on gloss processing in view of the distribution of specular reflection and suggest that gloss strength and sharpness are encoded by a subpopulation of IT neurons. Gloss is however modulated not only by specular reflection but also by other object properties including the material of which the object is made. To comprehensively understand the role of ITC for material perception, in the present study we conducted the electroencephalogram (ECoG) recording from ITC by using the object images with multidimensional material properties. Specifically, we rendered three-dimensional computer graphics objects with manipulating five material dimensions of gloss strength, gloss sharpness, translucency, change from metal to glass (metal/glass), change from metal to plastic (metal/plastic). A 128-channel ECoG electrode was placed covering whole ITC area including the lower bank of the superior temporal sulcus. Visual evoked response was recorded while the monkey engaged in a passive fixation task. Event related spectral perturbation was calculated and the response selectivity in six frequency bands, delta, theta, alpha, beta, low-gamma and high-gamma along the five dimensions was examined. Response selectivity for gloss strength was pronounced in all frequency bands except theta band. Selective electrodes appeared in the posterior part of the ITC for delta and beta bands and in the middle and the anterior part of the ITC for alpha, low-gamma and high-gamma bands. Selectivity for gloss sharpness was prominent in low-gamma band observed in the anterior portion of the ITC. Response selectivities for translucency, metal/glass and metal/plastic dimensions were commonly observed in the posterior part of the ITC but in frequency domain, the metal/plastic and the other two dimensions were separable. Selectivity in high-gamma band was pronounced for metal/plastic dimension and delta to low-gamma bands were prominent for the other two dimensions. These results indicate that multiple material properties are represented in the overlapped specific spatial-time-frequency domains in ITC.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-168 局所的 in vivo ゲノム編集とウイルストレーシング法を用いた細胞種特異的なマウス視覚情報処理の解明 Sayumi Okigawa（沖川沙佑美）¹，Nao Morimoto（森本菜央）^1,2，Fumitaka Osakada（小坂田文隆）^1,2,3,4 ¹名古屋大院創薬科学 ²名古屋大高等研究院 ³名古屋大未来社会創造機構 ⁴JSTさきがけ/CREST Deciphering the structure and function of neural circuits including feedforward and feedback pathways is crucial to understand information processing in the brain. Many lines of evidence indicate that different types of neurons play distinct roles in sensory processing, cognitive function and behavioral output. In the visual system, parallel processing streams originate from more than a dozen distinct types of retinal ganglion cells, each of which organizes a unique feedforward pathway to the brain. The dorsal lateral geniculate nucleus (dLGN) of the thalamus and the primary visual cortex (V1) are interconnected via feedforward and feedback projections. However, it remains unclear how distinct cell types organize the feedback pathways between dLGN and V1 and contributes to visual functions. Here we focus on cell-type-specific feedback pathways from layer 6 (L6) neurons of V1 to dLGN. Using the trans-synaptic viral tracing and CRISPR/Cas9-mediated Knock-In system, we successfully dissected and visualized the various types of L6 neurons projecting to distinct domains of dLGN and analyzed their corticothalamic feedback projections in mice. We performed the monosynaptic rabies virus tracing combined with EnvA/TVA systems to visualize individual inputs to either the core or shell region of the mouse dLGN. In addition, we identified marker genes that are specifically expressed in distinct L6 cell types, and took a knocked-in strategy using homology-independent-targeted-integration (HITI) devised from the CRISPR/Cas9 system to label each L6 cell type in mice. Injection of adeno-associated virus (AAV) carrying the HITI cassette into the adult mouse V1 allowed targeted knock-in of a reporter gene at a marker gene locus and labeling of specific L6 cell types in the mouse V1. We successfully identified at least four distinct L6 cell types sending feedback projections to dLGN. These results provide evidence that each cell type of L6 cells has its own gene expression pattern, morphology and connection specificity, suggesting the existence of the parallel processing streams of corticogeniculate feedback in the mouse visual system. Our viral and genetic approaches combined with in vivo functional imaging will facilitate understanding physiological roles of L6 cell types and their parallel processing streams in the feedback visual pathways.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-169 大脳皮質感覚野神経細胞のスパイク発火は感覚刺激の有無よりも周囲細胞の活動状態に依存する Ryutaro Ozaki（尾崎龍太郎）¹，Fumitaka Kimura（木村文隆）²，Hiroshi Tamura（田村弘）^1,3,4 ¹大阪大基礎工、大阪、日本 ²大阪大院医 ³大阪大院生命機能 ⁴脳情報通信融合研究センター（CiNet）、大阪、日本 Spiking activity of sensory cortical neurons affected by a variety of factors. Here we examined contributions of each factor using generalized linear model (GLM). Neurons in sensory cortex changes spiking activity in relation to the presence of appropriate sensory stimulus. Activity of a neuron was also affected by activity of adjacent neurons. Furthermore, the current spiking activity of a neuron is related to its preceding activity. Thus, at least three factors, i.e., (1) sensory stimulus, (2) activity of adjacent neurons and (3) preceding activity, affect spiking activity of neurons. To quantitatively evaluate relative contribution of the three factors, we simultaneously recoded spiking activity of multiple-adjacent neurons within a columnar region of visual (V1) or somatosensory cortex (S1) of anesthetized (Urethane) rats while presenting appropriate stimuli using a 32-channel linear array electrode. We obtained 875 neurons from 5 penetrations and selected 288 neurons that showed response to stimuli for further analyses. GLM is applied to single neuron's spiking activity and the three factors as objective variable and explanatory variables, respectively. We set a FULL model including all three explanatory variables and a model excluding one explanatory variable (1, excluding the contribution of stimulus; 2, excluding the contribution of the average firing rate of adjacent neurons; 3 excluding the effect of preceding firing rate) and calculated deviance for each model. Contributions of each explanatory variable was quantified by comparing the deviance between FULL model and a model excluding one explanatory variable. We found that the most effective explanatory variable, i.e., removal of the variable resulted in the largest changes in the deviance, is the average firing rate of adjacent neurons within a columnar region. The proportion of neurons that are influenced mostly by the average firing rate of adjacent neurons is 80% (143/179) in V1 and 89% (98/109) in the barrel cortex. The proportion of neurons that are influenced mostly by stimulus is 17% (31/179) in V1 and 10% (10/109) in the barrel cortex. The contribution of preceding firing rate is small and the proportion was 3% (5/179) in V1 and 1% (1/109) in the barrel cortex. The results suggested that most neurons in sensory cortical areas were strongly influenced by ensemble spiking activity of adjacent neurons rather than by sensory stimulus or by preceding activity.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-170 ネコ視覚皮質での多細胞活動を用いた刺激方位推定　ー発火数相関の効果ー Hiroyuki Ito（伊藤浩之）¹，Reo Kohno（幸野怜歩）¹，Yoshiko Maruyama（圓山由子）²，Yoshiya Mori（森理也）¹ ¹京都産業大学情報理工学部 ²函館高専 Even under repeated presentations of the same stimulus, the spike counts of a single sensory cortical neuron show a considerable amount of trial-to-trial variability. One of the challenges in Brain-Machine Interface is to decode the stimulus information from the single trial activities of simultaneously recorded multiple neurons. We recorded multiple single neurons in the visual cortex of anesthetized cats under the moving bar of 16 different orientations (40 trials for each stimulus). We have shown that those variabilities were correlated between the neuron pairs (spike count correlation) and some neuron pairs showed stimulus dependent spike count correlations (Maruyama & Ito, 2013). We applied the multi-dimensional Gaussian model to decode the stimulus orientations based on single trial spike counts of simultaneously recorded multiple neurons. We compared the decoding performance between the model with stimulus dependent covariance and that with no covariance (assuming independent variabilities) to examine the effect of spike count correlations and their stimulus dependence. The decoding performance was tested by the cross validation. We also examined the increase of the performance by increasing the number of neurons included to the model. The model worked well leading to the performances of 6.4~76.7% over 48 sessions exceeding the chance level (6.25%). However, contrary to our expectation, the model with no covariance showed higher performance than that with stimulus dependent covariance, and the difference became more dominant for the models with a larger number of neurons. Further analyses concluded that the number of parameters becomes very large in the model with stimulus dependent covariance and the performance of the cross validation decreased due to over-fitting. To overcome the over-fitting, we tried three variations of the original model. At first, we ignored the stimulus dependence of the spike count correlation to decrease the number of parameters of the model. We also constructed the model in a lower dimensional feature space by applying the principal component analysis (PCA) and the linear discriminant analysis (LDA). The model applying the LDA showed the best performance for large numbers of sessions. Due to limited number of trials in experimental recordings, a special care should be taken in the selection of the decoding model to balance between the merit by inclusion of additional feature parameters and the demerit by the over-fitting.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-171 NKCC1及びKCC2はマウス網膜双極細胞の周辺部応答を決定する Chengzhu Yin（尹成珠），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） Center-surround antagonistic receptive field organization serves the basic platform for spatial information processing in the retina. In the case of bipolar cells (BC), the contribution of GABAergic inputs from horizontal cells is hypothesized. However, since surround responses between ON-BCs and OFF-BCs show an opposite polarity to light stimuli, it is puzzling how two types of surround responses are formed from GABAergic inputs only. Based on the immunohistochemical and electrophysiological studies, the difference of surround responses between ON-BCs and OFF-BCs is, currently, explained by the hypothesis that GABA currents with different reversal potential are formed by the different distribution of NKCC1 and KCC2, chloride transporters, in the dendritic region. Therefore, in the present study, we examined this hypothesis by electrophysiological and biological techniques. Under the perforated whole cell clamp mode, bumetanide, a NKCC1 antagonist, significantly shifted the GABA mediated reversal potential to the hyperpolarization side in ON-BCs (34.1 ± 3.9 mV, n=9) but not in OFF-BCs (5.2 ± 2.3 mV, n=6). In contrast, application of VU0255011, a KCC2 antagonist, shifted the reversal potential to depolarization side in OFF-BCs (30.6 ± 4.9 mV, n=7) but not in ON-BCs (4.9 ± 4.3 mV, n=8). To confirm the difference of reversal potentials by NKCC1 and KCC2, we further examined the expression of NKCC1 and KCC2 mRNA by fluorescence in situ hybridization. Expression of KCC2 mRNA was equivalently found in the PKCα (rod bipolar cells maker) positive ON-BCs (93.3%) and NK3R (Type 1 and Type 2 OFF-BCs maker) positive OFF-bipolar cells (92.9%). On the other hand, for NKCC1, the expression in ON-BCs (45.5%) was twice than the expression in OFF-BCs (20.0%). These results suggest that the balance of the activities and expression level of NKCC1 and KCC2 would contribute to determine the polarity of surround responses, and support the notion that the surround responses of retinal bipolar cells are mediated by GABA current.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-172 マーモセット視覚大脳皮質における自発的神経活動の時空間構造の解析 Teppei Matsui（松井鉄平）¹，Takayuki Hashimoto（橋本昴之）¹，Masato Uemura（上村充人）^1,2，Tomonari Murakami（村上知成）¹，Kohei Kikuta（菊田浩平）¹，Toshiki Kato（加藤利樹）¹，Kenichi Ohki（大木研一）^1,2 ¹東京大院医統合生理 ²ニューロインテリジェンス国際研究機構 Spontaneous neuronal activity in the resting-state is robustly observed throughout the human brain using functional magnetic resonance imaging and has been used widely to assess functional network structures at the scale of the whole-brain (Fox and Raichle, 2007). At the mesoscale, previous optical imaging in the visual cortex of cats and ferrets reported that spatial patterns of spontaneous cortical activity closely resemble spatial patterns of iso-orientation columns (Kenet et al., 2003; Smith et al., 2018) suggesting that spontaneous activity contain mesoscale network information. However, due to the lack of imaging technique applicable to primates, precise mesoscale structures of spontaneous neuronal activity in primates still remain largely unknown. Recently, genetically encoded calcium indicator applicable to common marmosets has been developed (Sadakane et al., 2013; Uemura et al., Society for Neuroscience Abstract, 2018). In this study, we utilized this technique and conducted widefield and two-photon calcium imaging of spontaneous cortical activity in a large cranial window (6.5 x 6.5mm) spanning the primary (V1) and secondary visual cortex (V2) of the marmoset monkey. Wide-field imaging of spontaneous activity in the marmoset V1 and V2 indeed revealed rich spatiotemporal structures at various spatial scales. At a large spatial scale, wave-like spontaneous activity propagating across the cortex was readily observed. Importantly, at a smaller spatial scale, columnar patterns resembling orientation maps were frequently observed and were often embedded within the wave-like activity. Cellular scale imaging with two-photon microscopy confirmed that the columnar spontaneous activity reflected spontaneously active clusters of neurons. Spike triggered average of spontaneous activity (Tsodykes et al., 1999) further showed significant overlap between the column-like spontaneously activity and iso-orientation columns. These results suggest that the analysis of spatiotemporal patterns of spontaneous activity provide useful information about the modular organization of primate neocortex.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-173 超微細空間スケールでのヒト両眼固視眼球運動の検出 Hiroyuki Fujie（藤江博幸）^1,2，Yasuto Tanaka（田中靖人）² ¹三城HD R&D ²神経数理学研究所 Recent progress of our research achieved detection of human miniature eye movement monocularly. Here we developed the system that is able to determine sub-micrometer scale detection of both eye and head movement simultaneously. We devised the fast optical system with high spatial and temporal resolution (Video occulography: VOG) which is image processed to extract statistical information to further increase spatial resolution which is applied to the eye and head movement detection. As results the components of drifts and micro tremors were determined in a sub-micrometer spatial resolution together with independent head movement. We also found that cardiovascular information (heart-rate variability) constituted in the miniature component of the eye movement by means of ECG/SpO2 processing in relation to chaos analysis (Fujie and Tanaka, 2018 JNS). In the current study, the paradigm was extended to the binocular regime. We used three high resolution cameras (300~1000Hz) to detect left and right eyes as well as head movement simultaneously. Cardiovascular signals (ECG/SpO2) were detected as well. We were successful to determine simultaneously 1) right and left eye movement together with head movement, 2) pure head movement and 3) pure eye movement estimated from the measurements using machine learning method. We device the method using machine learning of conducting two affine transformations and combine them to extract pure eye and head movements which is mutually independent. The precision and resolution of analysis in terms of pure eye/head movement as well as cardiovascular components are to be compared between monocular and binocular analysis.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-174 網膜視細胞変性におけるアクティブゾーンタンパク質CAST／ELKSの機能解析 Akari Hagiwara（萩原明）¹，Manabu Abe（阿部学）²，Yamato Hida（飛田耶馬人）¹，Naoko Sugiyama（杉山直子）¹，Kenji Sakimura（﨑村建司）²，Toshihisa Ohtsuka（大塚稔久）¹ ¹山梨大院医工生化学一 ²新潟大脳研基礎神経科学 In the mammalian retinal signal processing, the first-order neuron, photoreceptor, converts light into electrical signals. Before the signals are further transmitted to the brain by the ganglion cells, bipolar cells mediate the signal transduction with modulation of inhibitory neurons. Therefore, photoreceptor ribbon synapse forms unique triad synapse with bipolar and horizontal cells. In previous report, we have shown that the presynaptic active zone cytomatrix proteins, CAST and ELKS, have both redundant and unique roles in coordinating development for the structure and localization of ribbon synapses as well as visual processing. Remarkably, AAV-mediated photoreceptor specific ELKS deletion after synapse maturation induced neurodegeneration and loss of ribbon synapses. However, the role of CAST for the maintenance of photoreceptor has yet to be determined. Here, we generated a mouse line for conditional knock out (KO) of CAST, in which CAST gene exon14 flanked by loxP sites. As similar to the acute deletion of ELKS, the photoreceptor specific AAV-mediated Cre induction to CAST floxed mouse caused reduction of ribbon synapse density and degeneration of photoreceptors. And the photoreceptor degeneration was more prominent in conditional KO of both CAST and ELKS floxed mouse. From these results, we speculate that both CAST and ELKS have redundant roles to maintain the photoreceptor as well as its development. While retinitis pigmentosa is the most common progressive retinal degenerations, and in advanced stage, the degeneration of photoreceptor causes serious loss of eyesight, the pathophysiology has not been well characterized. Therefore, further investigation of functional and molecular mechanism on the roles of CAST and ELKS should contribute to more precise understanding of the pathogenesis and then development of novel therapeutic strategies.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-175 複合持続ストレスによる脊髄後角ニューロンにおける侵害受容感作 Daisuke Uta（歌大介）¹，Koji Wakatsuki（若月康次）^2,5，Katsuyuki Tsuboshima（坪島功幸）³，Masaya Yasui（安井正佐也）^4,5，Yohei Hamaue（濱上陽平）⁷，Hiroshi Kiyama（木山博資）⁵，Hisao Nishijo（西条寿夫）³，Kazue Mizumura（水村和枝）⁶，Toru Taguchi（田口徹）⁷ ¹富山大院医薬応用薬理 ²名古屋大環境医神経系２ ³富山大医システム情動（生理１） ⁴愛知医大解剖 ⁵名古屋大院医機能組織 ⁶日大歯生理 ⁷新潟医療福祉大理学療法 Persistent stress is known to cause pain hypersensitivity, while acute stress relieves it. The former type is called as stress-induced pain or hyperalgesia, although the underlying mechanisms are poorly understood. Here we examined the peripheral and spinal mechanisms of nociception using a stress-induced pain model. The model was made under continuous stress by keeping rats in a cage filled with water (1.5 cm in depth) for 5-6 days. In vivo extracellular recordings of the superficial dorsal horn (SDH) neurons were made under urethane anesthesia. The SDH neurons were found to be sensitized to mechanical stimulation applied with calibrated von Frey filaments (vFFs). The mechanically sensitized neurons exhibited higher spontaneous firings. In vivo patch-clamp recordings revealed that frequency and amplitude of the spontaneous excitatory postsynaptic currents (sEPSCs) to the SDH neurons were increased in the model. Frequency and amplitude of the spontaneous inhibitory postsynaptic currents (sIPSCs) to the neurons, on the other hand, were decreased in the model. In rats exposed to continuous stress, c-Fos-immunoreactive nuclei were significantly increased in the SDH. In the periphery, activities of single C-fiber nociceptors were recorded and analyzed. Neither general characteristics (conduction velocity, spontaneous activity and distribution of the receptive fields) nor responsiveness to noxious mechanical, cold and heat stimulus was changed in the stress model. These results suggest that spinal, but not peripheral, mechanisms is involved in the nociceptive sensitization of the SDH neurons in response to continuous stress. The mechanisms may be relevant to patients with stress-induced pain.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-176 Impact of prescription opioids on the brain in chronic pain patients: A multivariate pattern analysis Behnaz Jarrahi（Jarrahi Behnaz），Sean Mackey（Mackey Sean） Stanford University School of Medicine As prescription opioids are increasingly used for treating chronic pain, there is a need to better characterize how opioid analgesics impact the brain. Neuroimaging studies of substance misuse in healthy subjects have identified abnormalities in brain regions involved in cognitive functions (e.g., dorsolateral prefrontal cortex). However, it is unclear how opioid status in chronic pain patients affects brain regions associated with cognitive function and pain processes. Using functional magnetic resonance imaging, we explored this question by examining the time-course of whole-brain functional connectivity during cognitive control of evoked thermal pain in 20 chronic low back pain patients (10 females, 10 males, mean age, 46.5 years, half on long-term opioid regimen). Multivariate pattern analysis of whole-brain connectome (connectome-MVPA) was used to assess differences in whole-brain connectivity associated with cognitive control of pain between chronic pain patients on opioids and those who were not. Seed-based functional connectivity analysis was also performed to further explore connections between regions identified in the whole-brain analysis. To get statistically significant effects with modest number of subjects, non-parametric statistics were performed with 1000 simulations at the voxel level of p < 0.001, uncorrected (two-tailed), and at the cluster level of p < 0.05, false discovery rate corrected. The between-group differences in whole-brain MVPA revealed distributed regions including core hubs of the salience network (insula and anterior cingulate cortex), central executive network (dorsolateral prefrontal and posterior parietal cortices), and fronto-temporal attention networks. The functional connectivity analysis further revealed decreased functional connectivity in opioid-dependent patients between the left dorsolateral prefrontal cortex and two voxel clusters: the right medial orbitofrontal cortex (a key neural substrate of decision-making), and the left supplementary motor area. The second functional connectivity analysis with left insula as a seed showed decreased coupling between the left insula and right middle frontal gyrus. The results from this preliminary study demonstrate that prescription opioids can cause distinct alterations in brain's connectivity patterns in chronic pain patients. Future studies will have to expand upon the current data with a larger sample size and a more comprehensive assessment of cognitive functions.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-177 慢性炎症性マウスの前帯状回はGABAシナプス伝達に可塑的な変化を示す Kohei Koga（古賀浩平）^1,2，Shuji Shimoyama（下山修司）¹，Akihiro Yamada（山田彬博）²，Hidemasa Furue（古江秀昌）²，Kazuhiko Nakamura（中村和彦）³，Shinya Ueno（上野伸哉）¹ ¹弘前大学医脳神経生理 ²兵庫医大神経生理 ³弘前大学医神経精神医学 Chronic inflammatory pain is a persistent unpleasant sensation that produces pathological synaptic plasticity in the central nervous system. Human imaging study and animal studies consistently demonstrate that the anterior cingulate cortex (ACC) plays as a critical cortical area for nociceptive and chronic pain processing as well as emotions. So far, the mechanisms of excitatory synaptic transmission have been well characterized in the rodent ACC. Importantly, various models of chronic pain have induced synaptic plasticity on excitatory transmissions in the ACC. For example, nerve injury produces long-term potentiation in the ACC in vivo. On the other hands, the potential contribution of inhibitory synaptic transmission in the ACC, in models of chronic pain, is still unclear. In this study, we used a mouse model of chronic inflammation induced by Complete Freund Adjuvant (CFA). The CFA-induced inflammation model produced mechanical hypersensitivity 2-3 days after the injection into the left hindpaw. Then, we performed in vitro whole-cell patch-clamp recordings from layer II/III pyramidal neurons in 2 to 3 days after the CFA injection, and examined if the model could cause plastic changes on GABAergic transmissions, including transient and tonic type A γ-aminobutyric acid (GABAA) receptor-mediated inhibitory synaptic transmission, in the ACC. We analyzed miniature and spontaneous inhibitory postsynaptic currents (IPSCs), GABAA receptor-mediated tonic currents and evoked IPSCs. Furthermore, we studied whether GABAergic transmission-related proteins such as vesicular GABA transporter and GABAA receptors subunits in the presynapse and postsynapse of the ACC were altered. The protein expressions of GABAA receptors subunits in the ACC did not change after the CFA. However, the protein level of vesicular GABA transporter in the ACC was reduced in the model. These results suggest that early stage of chronic inflammation could alter GABAergic transmission in the ACC through the reduction of vesicular GABA transporter.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-178 慢性疼痛モデルラット（SNIモデル）の吻側腹内側延髄の神経活動への、運動野刺激の効果 Hiromasa Kitazawa（北澤宏理）東京医科大学 Motor cortex stimuli lead to anti-nociception in chronic pain model rats, however, the precise mechanism remains to be unknown. The purpose of this study is to examine whether the changes of the activity of rostral ventromedial medulla (RVM) neurons are involved in cortex-stimuli-induced anti-nociception. We made surgically one type of the chronic pain model, spared nerve injury (SNI) model rats under general anesthesia. The signs of chronic pain is confirmed with von Frey test. Then we tested the effects of motor cortex tetanic stimuli (50 Hz for 30minute pulse train, each pulse is >50μA, 200μs) on the spontaneous (ongoing) single unit activity and the field potentials in RVM evoked by single pulse stimulus (150-250μA 20μms pulse) to the motor cortex, in SNI rats fixed on a stereotaxic apparatus under general anesthesia. Prior to the test of tetanic /single cortical stimuli, an localization of the motor cortex by a train of electrical stimuli inducing limb movements and a classification of the single-unit activity recorded in RVM into the on, off, or neutral cells, based on their transient change by the nociceptive pinch stimuli applied to the hind paw, were performed. We found that the spontaneous single-unit activity of the on cells in the SNI rats decreased in compared with those of normal control rats, and also the field potentials recorded in parallel with the spontaneous activity depressed for at least 30 minutes after the end of the motor cortex tetanic stimuli. These results suggest that the decrease of the on cell activity may be involved in motor cortex-stimuli induced pain-relief.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-179 前脳基底部コリン作動神経による体性感覚刺激誘発皮質神経応答の変調 Shinnosuke Dezawa（出澤真乃介）^1,2，Kazuaki Nagasaka（長坂和明）¹，Yumiko Watanabe（渡辺由美子）¹，Ichiro Takashima（高島一郎）^1,2 ¹産業技術総合研究所 ²筑波大院　人間総合科学 The basal forebrain (BF) provides cholinergic projection that extensively releases acetylcholine in the brain such as the cerebral cortex, hippocampus and others. Previous studies revealed that specific lesion of cholinergic basal forebrain (CBF) neurons induced decline of cognitive function such as attention, memory and learning in the animal. In addition, nucleus basalis of Meynert (NBM) neurons that is one of the CBF had decreased in postmortem brains of patients with Alzheimer's disease and Parkinson's disease dementia. In recent years, it has been suggesting that deep brain stimulation (DBS) therapy targeted to the NBM may improve cognitive function in patients with dementia. However, it is unclear that what is the neurophysiological mechanisms underlying the improvement of cognitive function by NBM-DBS. In order to address this issue, we previously reported that electrical stimulation of NBM induces short-latency excitatory neural responses in the cortex including sensorimotor areas (Nagasaka et al., 2017). This result suggests that NBM electrical stimulation may affect not only to cognitive function but to sensorimotor function. In the present study, we investigated whether the NBM cholinergic projection affects to somatosensory neural response by using local field potential (LFP) recording. LFP was recorded in the primary somatosensory cortex (AP -0 mm to -1.0 mm, ML -4.0 mm, from bregma). Somatosensory stimulus was presented by electrical stimulation to the forelimb contralateral to the LFP-recording hemisphere (0.1 - 0.6 mA). Then we applied acetylcholine receptor antagonist mecamylamine and scopolamine to the cortex (1 mM, 30 min), or injected cholinergic neuronal immunotoxin 192 IgG-saporin to the NBM (0.3 μg/μL, 0.2 μL in total). The results showed that the amplitude of somatosensory evoked potential was increased due to both treatments. These results suggested that somatosensory evoked response is inhibitory modulated by NBM cholinergic projection. The next important issue is to investigate how the somatosensory-evoked LFP might be changed following NBM activation such as DBS. These approaches would contribute to understanding of neural mechanisms underlying NBM-DBS therapy.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-180 脊髄後角長距離介在ニューロンの分子マーカーの同定 Kazuhiko Nishida（西田和彦）¹，Seiji Ito（伊藤誠二）^1,2 ¹関西医大医医化学 ²大阪医大医麻酔 Spinal dorsal horn comprises a highly heterogeneous population of interneurons each of which interconnects each other to form complex neuronal circuitry responsible for the processing of somatosensory information. Among these neurons, the function of local-circuit neurons which form neuronal circuits within the same spinal segment has been extensively studied. In contrast, the significance of "propriospinal neurons (PN)" which send long axons rostrocaudally over several spinal segments remains totally unknown. As a first step to examine their function, we analyzed the spatial distribution of PNs within the spinal dorsal horn in the mouse. PNs were distinguished by their long projection range using retrograde axonal tracer cholera toxin B (CTB). We focally injected CTB into the spinal dorsal horn at L1 level, and analyzed the localization of retrogradely labeled neuronal cell bodies throughout all levels of the spinal cord. This analysis demonstrated that PNs with very long axons (~2 mm in length) were localized in the most dorsal part of the dorsolateral funiculus (DLF), whereas PNs with mid-range axons (1~2 mm) were localized not only in the dorsal DLF but also in the ventral DLF, deep (lamina III-IV), and superficial (lamina I) dorsal horn. Most PNs were rarely localized in lamina II. In order to characterize the identity of these CTB-labeled PNs, we next performed double immunostaining with antibodies for CTB together with candidate marker molecules of this population. We analyzed the expression of Cholecystokinin (CCK) and Cerebellin-2 (Cbln2), which have been shown to be markers for deep dorsal horn neurons transmitting innocuous mechanical stimulation. This analysis demonstrated that CCK- and Cbln2-expressing neurons were over-represented among PNs, indicating that some PNs are involved in the sensory processing of innocuous mechanosensation. Moreover, the POU family transcription factor Brn3a, which is highly expressed in the long-range neurons in other parts of the nervous system, was also expressed in the PNs. We are currently analyzing the function and cell fate specification of PNs.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-181 繰り返しの侵害刺激による痛覚閾値への影響 Setsuko Nagahama（長濱節子）帝京平成大学ヒューマンケア Nociceptors in the skin are present in free nerve endings of Aδ-fibers and C-fibers. The Aδ-fibers develop picking and fast pain in response to mechanical stimulation. It is well known that pain receptor is non-adapting type. There is little report about pain sensory and adaptation mechanism of Aδ fiber, and it is not clear of pain sensory mechanism of Aδ fiber. The author reported that repeated mechanical stimuli to nociceptor increase pressure pain thresholds, PPTs (Neuroscience 2018). A total of 16 participants took part in the study (ranging from 20 to 22 years; 8 males and 8 females). In 7, 10, 20, 30-seconds intervals the PPTs were significantly increased. In contrast, there were increase tendency of the PPTs in 5-seconds intervals, but there were no significant changes of them. This study focused on the adaptation of pain sensory by measuring the influence of repeated mechanical pain stimuli at 5-seconds intervals on PPTs. A total of 24 males participants took part in this study. The algometer used in the experiments measured pressure in units of 1g and in the range of 1g-10g. All experiments were done at intervals more than one week. The participant was pricked with the pressure algometer at the same point of right forearm medial cutis. Control was the average of PPTs measured 3 times at the same point. After the measurement of control, the participant was stimulated at the same point 12 times every 5 seconds successively, and measured a change of PPTs. The significance of the difference among samples was determined by a one-way repeated measures ANOVA followed by a Bonferroni multiple test. Differences at P<0.05 were termed significant. In the first experiments at 5-second intervals, PPTs s of the the 12^th stimulus significantly increased. In addition, PPTs were further measured by the similar method 1 hour, 4 hours and 24 hours after the first experiments. Then the control was measured in all cases and PPTs were measured by the similar method at the same point of the participant. A tendency of increase was seen in PTTs of the 6^th and the 12^thstimulus 24 hours later.PPTs of control stimulus increased 4 hours later, compared with the first experiments. These results showed that repeated mechanical stimuli at 5-secomds intervals increased PPTs. It was suggested that Aδ fiber was adapted by the repeated mechanical stimuli at short time intervals.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-182 若年女性において月経周期はメントール塗布時の足趾の皮膚温に影響する Yuki Uchida（内田有希），Chinami Tsunekawa（恒川千菜美），Izumi Sato（佐藤泉水），Keiko Morimoto（森本恵子）奈良女子大学生活環境学部 INTRODUCTION Menstrual cycle affected skin blood flow in the great toe during mild cooling of the sole in young women (Uchida et al., 2018, J Physiol Sci). Menthol is an agonist of TRPM8 that is a peripheral cold receptor. The aim of the present study was to investigate the effect of menstrual cycle on skin temperature (T_sk) of the foot during menthol application in young women. METHODS T_sk and partial cutaneous blood flow in the foot, tympanic temperature, blood pressure, heart rate, thermal sensation and pleasantness during the follicular (F), luteal (L), and menstrual (M) phases during menthol application in young women using thermography, laser Doppler flowmetry, a digital blood pressure monitor, and VAS scale were examined at 25°C. After application of the 0.5‰ menthol solution to the right foot, the measurements were continued for 20 min. RESULTS The T_sk of the second and third right toes in the F phase were lower than that in the L phase. The T_sk of the little right toe in the F phase was lower than that in the L and M phases. No significant differences were observed in the T_sk of the dorsum of right foot, cutaneous Laser-Doppler flow in the right great toe, tympanic temperature, blood pressure, heart rate, thermal sensation and pleasantness among the phases. CONCLUSION The menstrual cycle phase did not affect T_sk in the sole, but it affected the partial toes during menthol application.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-183 繰り返し拘束ストレス後の機械的痛覚過敏に関連した RVM GABA ニューロンにおけるヒストンアセチル化の増加 Hiroki Imbe（井辺弘樹），Akihisa Kimura（木村晃久）和歌山県立医大・医・生理学１ Psychophysical stresses have a great impact on central and peripheral nervous systems. Especially a substantial shift is found in sensory function. The sensitivity and response to pain are frequently enhanced by psychophysical stresses, which is termed stress-induced hyperalgesia (SIH). However, the mechanism remains unknown. The rostral ventromedial medulla (RVM) and locus coeruleus (LC) are key elements of the descending pain modulatory system, which modulate nociceptive transmission in the spinal dorsal horn. In the present study we examined the acetylation of histone H3 in the RVM and LC after repeated restraint stress for 3 weeks to clarify changes in the descending pain modulatory system in the rat with SIH. The repeated restraint stress (daily 6h for 3weeks) induced mechanical hypersensitivity in the hindpaw and an increase in acetylation of histone H3 in the RVM but not the LC. The number of acetylated histone H3-IR cells in the RVM was significantly higher in the repeated restraint group (282.9 ± 43.1) than that in the control group (134.7 ± 15.6, p<0.05). The number of acetylated histone H3-IR cells in the RVM was negatively correlated with paw withdrawal threshold (p<0.01). Furthermore, the repeated restraint stress increased acetylation of histone H3 in the RVM GABAergic neurons but not the RVM serotonergic neurons. The GAD67 protein level in the RVM was significantly higher in repeated restraint group (144.9 ± 17.0%) than that in the control group (100.0 ± 8.9%, p<0.05). These findings suggest the possibility that the stress-induced neuroplasticity in the RVM GABAergic neurons is involved in the mechanical hypersensitivity due to the dysfunction of the descending pain modulatory system.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-184 Exercise-induced hypoalgesiaにおける扁桃体-側坐核経路の役割 Katsuya Kami（上勝也）¹，Emiko Senba（仙波恵美子）² ¹和歌山県立医大医リハビリテーション医学 ²大阪行岡医療大医療理学療法 We have previously shown that voluntary exercise (VE), a strong reward for animals, induced exercise-induced hypoalgesia (EIH), which may be due in part to the activation of dopamine neurons in the ventral tegmental area (Kami et al., Sci Rep, 8:11540, 2018). In this study, we examined whether another pathway including amygdala and nucleus accumbens (NAc) also contributes to the EIH. Neuropathic pain (NPP) model mice were prepared by partial sciatic nerve ligation (PSL). PSL-Runner mice performed VE for 2 weeks prior to the surgery. Immediately after the surgery, these mice were returned to the cages with running wheels and performed VWR for 2 weeks post-surgery. Mechanical allodynia and heat hyperalgesia were evaluated by von Frey and plantar tests, respectively. Serial brain cross-sections including the amygdala and NAc were processed for immunostaining. Furthermore, a retrograde tracing technique was used to examine whether amygdala neurons projecting to the NAc are activated by VE. Withdrawal thresholds and latencies in PSL-Runner mice were significantly higher than those in PSL-Sedentary mice. Significantly decreased or increased FosB+ neurons were observed in the lateral region or the medial region of the NAc in PSL-Sedentary mice, respectively. Conversely, VE induced significant increase or decrease in FosB+ neurons in the lateral region or the medial region of the NAc, respectively. Increased FosB+ EAAC1+ neurons were detected in the lateral region (latBA) of the basal amygdala (BA), but not in the medial region (medBA) of BA in PSL-Sedentary mice, while VE increased FosB+ EAAC1+ neurons in the medBA, but not in the latBA. Using a combination of retrograde tracer and FosB immunostaining, we found that approximately 60 % of the RBR-labeled BA neurons was detected in the medBA, and about 75 % in these neurons showed FosB immunoreactivities. Most of the CeA neurons were GAD67+, i.e. GABAergic neurons. Although significantly increased numbers of FosB+ GABAergic neurons were detected in PSL-Sedentary mice, VE induced remarkable decrease in such neurons in the CeA, especially in the CeC. The present study suggests that EIH is achieved, at least in part by activation of the medBA-NAc pathway, which is considered to mediate appetitive behaviors, following VE, and that VE also contributes to the elimination of unpleasant emotion associated with chronic pain by inactivating the spinal dorsal horn-parabrachial nucleus-CeC pathway.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-185 Cellular and circuit mechanisms underlying descending modulation of itch Zhengrun Gao（Gao Zhengrun）¹，Wenzhen Chen（Chen Wenzhen）^1,2,3，Mingzhe Liu（Liu Mingzhe）^1,2,3，Xiaojun Chen（Chen Xiaojun）^1,2,3，Xinyan Zhang（Zhang Xinyan）^1,2,3，Yangang Sun（Sun Yangang）^2,3 ¹University of Chinese Academy of Sciences ²Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science ³Intelligence Technology, Chinese Academy of Sciences Uncontrollable itch-scratching cycles lead to serious skin and deep tissue damage in patients with chronic itch. Itch signal processing is known to be under tonic inhibitory control at the spinal level, but the neural circuit mechanism promoting the itch-scratching cycle remains elusive. Here we report that a group of tachykinin 1 (Tac1)-expressing glutamatergic neurons in the lateral/ventrolateral periaqueductal gray (l/vlPAG) facilitate the itch-scratching cycle via descending modulation of spinal neurons expressing gastrin-releasing peptide receptor (GRPR), the key relay neurons for itch sensation. Using extracellular electrophysiological recording, we found that some l/vlPAG neurons exhibited scratching behavior-related neural activity. Furthermore, suppressing the activity of l/vlPAG glutamatergic neurons significantly decrease itch-evoked scratching behavior. The glutamatergic neurons in l/vlPAG include two subpopulations, Tac1-positive and somatostatin (SST)-positive glutamatergic neurons. Suppression the activity or ablation Tac1-positive neurons decreased itch-induced scratching behavior, while ablation SST-positive neuron did not. In addition, we found that the activity of Tac1-positive in the l/vlPAG was elevated during itch-induced scratching behavior. Importantly, activation of Tac1-expressing neurons induced robust spontaneous scratching and grooming behaviors. The scratching behavior evoked by Tac1-expressing neuron activation was suppressed by ablation of spinal GRPR positive neurons. In summary, we found that Tac1-positive neurons in l/vlPAG promote scratching behavior by regulating GRPR-positive neurons in the spinal cord via a desending pathway. This study reveals the cellular and circuit mechanisms underlying itch descending modulation, and shows that Tac1-positive neurons in l/vlPAG represent a potential central therapeutic target for breaking the vicious itch-scratching cycle associated with chronic itch.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-187 マウス大脳皮質から腰髄への軸索投射解析 Hiroshi Kameda（亀田浩司）¹，Naoyuki Murabe（村部直之）¹，Hiroaki Mizukami（水上浩明）²，Keiya Ozawa（小澤敬也）³，Masaki Sakurai（桜井正樹）¹ ¹帝京大医生理 ²自治医大分子病態治療研究センター遺伝子治療研究部 ³東京大医科研遺伝子治療開発分野 Although the corticospinal (CS) tract is the main pathway for control of voluntary movement, CS neurons are distributed not only in motor-related cortical areas, but also in somatosensory areas. We previously showed that, in mice, CS neurons sending their axons to the gray matter of the seventh cervical spinal cord segment (C7) were located at the primary (M1) and secondary (M2) motor areas, and primary (S1) and secondary (S2) somatosensory areas. We further revealed that axons from each cortical area were distributed on its specific division in the C7 gray matter: (1) M2 and M1 project mainly to intermediate and ventral areas, (2) the rostral part of S1 to the mediodorsal area, (3) the caudal part of S1 to the dorsolateral area, and (4) S2 to the dorsal area. These projection patterns may reflect diverse functions of CS neurons located at each cortical area. However, little is known about the organization of the cortical region innervating the lumbar cord. In rodents, the region is relatively small compared to that innervating the cervical cord and restricted to M1/S1. However, it is still unclear to what extent the motor and somatosensory areas overlap with each other. In the present study, therefore, we aimed to comprehensively investigate the axonal-distribution patterns of lumbar-cord-projecting cortical neurons. We previously examined the cortical region innervating the gray matter of the fourth lumbar spinal cord segment (L4), so we injected adeno-associated virus vectors expressing fluorescent proteins into cortical sites separately to cover the region, and observed the distribution of labeled axons in the L4 gray matter. We found that there was a relation between locations of CS neurons and distribution patterns of their axons, suggesting the existence of topographic connections which are basically similar to those observed in the analysis of C7-projecting neurons. We further labeled both the CS axons and primary afferents to investigate the relationship between the termination areas of these axons.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-188 全身麻酔下の手術侵襲に関連する神経活動の初期応答遺伝子マッピング Yukimura Oe（大江将軍）¹，Maiko Hasegawa-Moriyama（長谷川麻衣子）¹，Keika Mukaihara（向原桂香）¹，Mayumi Nakahara（中原真由美）¹，Yuichi Kanmura（上村裕一）¹，Hiroyuki Okuno（奥野浩行）² ¹鹿児島大　医　麻酔蘇生学 ²鹿児島大　院医歯学総合　生化学・分子生物学 Delirium is the most common postoperative neurologic complication associated with anesthesia and surgical stress including pain. However, it is not clear whether the intraoperative changes in neuronal activity are linked to postoperative pain, cognition, and memory. This study thus aimed to detect neuronal activity in the brain during surgery under general (i.e., whole-body) anesthesia, in order to correlate it with postoperative sensation and cognition. We first investigated expression of c-Fos and Arc in the mouse brains by immunohistochemistry. Fos and Arc are products of the immediate early genes (IEGs) c-fos and Arc, respectively, and have been utilized as the most reliable markers for neurons that are recently activated. Mice were subjected to a surgical incision on the right hind paw on under isoflurane anesthesia. Control groups included mice underwent anesthesia without surgery as well as home-caged animals. Preliminary results revealed that several brain areas including the hippocampus specifically upregulated these IEG expression 2 hours after operation . Ongoing analysis suggested differences in expression patterns between Fos and Arc in a brain-area specific manner. We also plan to test cognitive performance of these groups of mice in memory tasks. Our analysis will provide a novel clue to understand cellular mechanisms underlying delirium after recovery from general anesthesia.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-189 後根神経節（DRG）細胞への細胞種特異的遺伝子導入法: 齧歯類と霊長類における比較 Moeko Kudo（工藤もゑこ）¹，Sidikejiang Wupuer（Wupuer Sidikejiang）¹，Ken-ichi Inoue（井上謙一）²，Masahiko Takada（高田昌彦）²，Kazuhiko Seki（関和彦）¹ ¹国立精神・神経セ神経研 ²京都大　霊長研統合脳システム Background : Common marmoset is the new world monkey drawing considerable attention in the field of biomedical research. While the structure and function of their central nervous system have been elucidated well in the past decade, comparable knowledge about their peripheral nervous system is lacking. In this study, we compared the distribution of the size of DRG cells in the lumbar spinal segment between common marmosets and rats, and examined the cell-type specificity of two serotypes of AAV. Methods: We injected the AAV6 or AAV9 containing GFP into the left sciatic nerve of six (marmoset) or four (rat) animals per serotype. Four weeks after the injection, each animal was sacrificed and DRG (L4, 5) was extracted for histological analysis. Every tenth DRG section was selected from the consecutive serial sections (3 to 6 sections for each DRG), and the number of GFP labeled cells were counted, and transduction efficiency was expressed as the percentage of total neuronal profiles revealed by NeuN staining. Further, we examined the cell type preference and transduction efficiency of each serotype by counterstaining with NF200 or Peripherin antibodies for neuronal identification. Results: We found two striking differences in the distribution of cell size of DRGs between two species. First, while the DRG cell-size of rats was distributed in smaller range (<700 μm² in 64 %) (see also Harper et al., J.Physiol. 1985), it was distributed in a significantly larger scale in the Marmoset DRG (<700 μm² in 32 %, p<0.0001). This result indicates that the DRG cell of marmoset is larger than that of rats. Second, the target cell type of AAV6 and AAV9 was different between the two species. In rats, we found the AAV9 preferentially transduced in myelinated (NF200 positive) neurons while AAV6 transduced unmyelinated nociceptors (Peripherin positive) (see also Black et al., Mol. Pain 2012). However, there was no significant difference between the proportions of myelinated and unmyelinated neurons for both AAV9 and AAV6 serotypes in marmoset DRG. This result may suggest the lower cell-type selectivity to the DRG cells of common marmoset. Different distribution of size and type could be ascribed to the phylogenetical evolution, and the difference in the AAV serotype specificity should be considered for their application to human gene therapy of peripheral nervous system.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-190 Small-animal neuroimaging analysis of noxious colorectal distension evoked pain in IBD model rats Tianliang Huang（Huang Tianliang）^1,2,3，Takashi Okauchi（Okauchi Takashi）¹，Yuping Wu（Wu Yuping）¹，Mika Shigeta（Shigeta Mika）¹，Di Hu（Hu Di）¹，Koichi Noguchi（Noguchi Koichi）²，Yasuyoshi Watanabe（Watanabe Yasuyoshi）¹，Yi Dai（Dai Yi）³，Yilong Cui（Cui Yilong）¹ ¹Laboratory for Biofunction Dynamics Imaging, RIKEN Center for Biosystems Dynamics Research, Japan ²Dept. Anat & Neurosci., Hyogo College of Medicine, Japan ³Dept. Pharm. Sch. Pharm., Hyogo University of Health Sciences Japan Abdominal pain is a common symptom of inflammatory bowel disease (IBD: Crohn's disease, ulcerative colitis). A majority of patients suffering from acute flares of IBD experience abdominal pain, which severely impairs quality of life. Both peripheral and central mechanisms are thought to be involved in such abdominal pain in IBD. Since most of previous researches have focused on the peripheral mechanisms of abdominal pain in IBD pathophysiology, the involvement of higher order brain regions remained incompletely understood. In the present study, we tried to investigate the regional brain activity induced by noxious colorectal distension (CRD) in a 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced IBD model rats, using a small-animal neuroimaging method combining 2-deoxy-2-[18F] fluoro-D-glucose ([18F]FDG) PET imaging and voxel-based statistical analysis. We show that the brain activity was decreased in primary somatosensory cortex, amygdala and thalamus, while the brain activity was increased in hypothalamus in response to the noxious CRD in the IBD model rats. Furthermore, The rostral ventromedial medulla (RVM), a key node of descending pain inhibitory pathway, was also activated by the noxious CRD and the activation was negatively correlated with pain severity , indicating enhanced brain activity in RVM might be involved in counteraction of subsequent noxious input.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-191 排尿時下部尿路からの脊髄感覚情報伝達 Tatsuki Nakagawa（中川達貴）^1,2,3，Atsushi Hakozaki（箱崎敦志）³，Nozomi Akimoto（秋元望）³，Masahito Kawatani（河谷正仁）⁴，Noriyuki Ozaki（尾﨑紀之）²，Keiji Imoto（井本敬二）³，Hidemasa Furue（古江秀昌）^1,3 ¹兵庫医科大学神経生理 ²金沢大学院医機能解剖学 ³自然科学研究機構生理学研究所神経シグナル ⁴秋田大学院医器官・統合生理学 The lower urinary tract (LUT) is composed of the bladder and urethra, and the LUT functions during voiding are controlled by the spinal cord neurons such as parasympathetic preganglionic neurons and Onuf's nucleus neurons. Sensory information originating from the bladder via dorsal roots composed of myelinated Aδ-fibers and unmyelinated C-fibers conveys into the spinal dorsal horn (SDH) at spinal segments of L6-S1, and has an important role on the precise coordination of the voiding reflex. However, it is still unclear how spinal dorsal horn neurons receive sensory information from the LUT during voiding. In this study, we performed simultaneous recordings of action potentials of SDH and LUT activities and examined spinal sensory signal transduction during voiding. SDH neurons were classified into two types based on their firing patterns in response to LUT activities. Type 1 neurons elicited firings at the peak of intravesical pressure during voiding, and type 2 neurons characterized by a typical burst action potentials with were correlated with a rise in the intravesical pressure for micturition. When capsaicin was perfused into the LUT, type1 neurons increased their firing frequency at the peak pressure and followed by a sustained firing even after voiding. However, Type2 neurons did not change their responses during voiding by intravesical capsaicin. These results indicate that sensory processing from LUT was separately conveyed to the different two types of dorsal horn neurons.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-192 脳幹で甘味を伝達する神経の特定 Ken-ichiro Nakajima（中島健一朗）¹，Ou Fu（傅欧）¹，Yuu Iwai（岩井優）²，Kunio Kondoh（近藤邦生）¹，Takumi Misaka（三坂巧）²，Yasuhiko Minokoshi（箕越靖彦）¹ ¹生理学研究所 ²東京大院農学生命科学 The gustatory system plays an important role in sensing appetitive and aversive tastes for evaluating food quality. In mice, taste signals are relayed by multiple brain regions, including the parabrachial nucleus (PBN) of the pons, before reaching the gustatory cortex via the gustatory thalamus. Recent studies show that taste information at the periphery and in the gustatory cortex is encoded in a labeled-line manner, such that each taste modality has its own neuronal pathway. Although the PBN consisting of multiple subnuclei has also been known to function as a hub to receive and transmit diverse information including signals of pain, hunger, and satiety, the molecular identity of gustatory neurons in the PBN is still unknown due to lack of molecular markers. Here we show that SatB2-expressing neurons, which are localized in the waist area in the PBN are essential for sweet taste transduction. Using SatB2-Cre mice, we found that ablation of SatB2^PBN neurons lead to large loss of sweet taste recognition, but no effects on other tastes. Importantly, in vivo calcium imaging experiments show that the majority of these neurons are activated when tasting sweet substances. Optogenetic activation of gustatory thalamus-projecting SatB2^PBN neurons induces appetitive licking behavior. Our data strongly suggest that SatB2^PBN neurons are the key gustatory neurons selectively transmitting sweet taste signals in the mouse brain. To our knowledge, SatB2^PBN neurons are the first molecularly-defined gustatory neurons not only in the PBN but also in the mammalian brain.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-193 エンジンの音と振動によるバイクシミュレータにおける動揺病の低減効果の検証 Masami Hayashi（林真光）¹，Kohsuke Aigou（相合皓介）¹，Yuki Sawada（澤田悠伎）¹，Takuya Miyagi（宮城拓弥）²，Tomoya Ueda（上田誠也）²，Masayuki Miki（三木将行）³，Tetsuya Kimura（木村哲也）³，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 ³ヤマハ発動機株式会社 A motorcycle simulator is an effective tool to safely evaluate driver behavior. However, the use of simulators is accompanied by the problem of motion sickness (i.e., simulator sickness). Developing techniques to reduce simulator sickness is necessary for the effective use of simulators. Based on the sensory conflict theory, we hypothesized that simulator sickness could be reduced by effectively adding the sounds and vibrations produced by a motorcycle engine. Forty participants (8 females) took part in our experiment. The participants wore a head-mounted display (HMD) and earphones and sat on the seat of a motorcycle-type chassis. The HMD presented a virtual reality scene of motorcycling along a winding road from a first-person perspective. The earphones presented the engine sounds and an actuator under the seat produced the vibrations. The sounds and vibrations were synchronized with the motorcycling scene. The participants were separated into two groups: the with-sound-vibration and without-sound-vibration groups (for each group: n = 20, 4 females). There was no difference in the scores on a motion sickness susceptibility questionnaire (MSSQ) between the two groups. In the experiments, the motorcycling scene was presented for 5 minutes; the participants responded to the fast motion sickness scale (FMS) at 1-minute intervals by verbally rating their current level of sickness from zero (no sickness at all) to 20 (severe sickness). Before and after the motorcycling scene presentation, they answered the simulation sickness questionnaire (SSQ) consisting of 16 items. Overall, the FMS scores increased over time, indicating that the motorcycling scene induced motion sickness. For male participants, however, FMS scores were significantly lower in the with-sound-vibration group than in the without-sound-vibration group. Moreover, for male participants, the SSQ scores after the motorcycling scene were also significantly lower in the with-sound-vibration group. Together, these results indicate that the engine sounds and vibrations reduced motion sickness, both during the motorcycling scene and after the experience. However, no reduction of the effects of motion sickness was found in female participants. Our results suggest that adding engine sounds and vibrations is effective for reducing simulator sickness but that this effect is limited to male simulator users.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-194 視覚および聴覚刺激に誘発されたサッカード眼球運動における刺激の顕著性・空間的一致性の影響 Shimpei Yamagishi（山岸慎平），Makoto Yoneya（米家惇），Shigeto Furukawa（古川茂人）日本電信電話株式会社コミュニケーション科学基礎研究所 The ability to direct the attention towards the interest object is an essential function of human and animals. The quick movement of the eye towards stimulus, namely saccade, has been used to estimate how the presented stimuli draw our attention. It is unclear, however, how the properties of eye movement towards the visual and auditory targets are affected by the saliency of stimulus and congruency of the position of visual and auditory stimuli. We hypothesized that the saccadic reaction time would be facilitated as increasing the stimulus saliency and the spatial congruency of visual and auditory stimuli. To test this we designed the experiment as follows: at the beginning of the experiment, the participant was asked to fixate on the target which was presented at the center of the display. The target disappeared for a period of 200 ms, then the target reappeared at the left or right side of the display (10 degrees from the center of the display in viewing angle). In visual-cue or auditory-cue conditions, the participant was asked to make a saccade towards visual or auditory targets, respectively. In the congruent condition, visual and auditory stimuli appeared at the same side in the saccade period, whereas in the incongruent condition, visual and auditory stimuli appeared opposite side of the display. We used two visual stimuli with different luminance (2.58 or 93.58 cd/m2) and two auditory stimuli with different intensity (A-weighted sound pressure level was 40 or 60 dB) to examine the effect of stimulus saliency. Results showed that the saccadic reaction time was shorter for the congruent condition than for the incongruent condition except for the salient visual-cue condition, implying that the salient visual flash has a strong influence on the saccade system. Although we expected that the saccadic reaction time would be shorter for strong stimulus than for weak stimulus for both modalities, the effect of stimulus saliency was observed for the visual-cue condition, not for the auditory-cue condition. A further examination is needed to design the effective salient auditory stimuli.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-195 場所と速度に関する情報は歯状回の神経細胞の集団活動にコードされている Tomoyuki Murano（村野友幸）¹，Nao Hirata（平田直）¹，Satoko Amemori（雨森智子）¹，Akito Nakao（中尾章人）²，Jun Yamamoto（山本純）³，Akira Murakami（村上晃）⁴，Yasuyuki Kamitani（神谷之康）⁴，Tsuyoshi Miyakawa（宮川剛）¹ ¹藤田医科大学　総合医科学研究所　システム医科学部門 ²京都大学大学院工学研究科　合成・生物化学専攻 ³Department of Psychiatry, Neuroscience Division The University of Texas Southwestern Medical Center, Dallas, USA ⁴京都大学情報学研究科 The hippocampal dentate gyrus (DG) is a region in which neural responses are highly sparse, and of diverse nature. Multiple types of information are presumably processed in the DG. However, it remains unknown how neural codes are distributed in the DG. Here, we examined neural representations of position and velocity in the population activity of DG neurons by performing in vivo Ca2+ imaging in freely exploring mice. Although spatial and velocity information encoded at the single cell level is not very informative, position and velocity of mice were accurately decoded when analyzing population activity of DG neurons by machine learning methods. We measured decoding accuracy of position or velocity while excluding neurons in the order of spatial and velocity information. Neurons with high spatial or velocity information were essential for position or velocity decoding. Furthermore, we performed position and velocity decoding in alpha-calcium-calmodulin kinase II mutant mice (αCaMKII+/-), an animal model with altered behavior that is relevant to psychiatric disorders. Position decoding accuracy of αCaMKII+/- mice was significantly lower than that of wild-type mice, whereas there was no significant difference in velocity decoding accuracy between wild-type and mutant mice. Additionally, spatial information during movement is reduced than those of resting state in αCaMKII+/- mice, whereas they are not significantly different in wild-type mice. These results indicate that spatial and velocity information in single neurons of the DG is not informative but population activity of DG neurons carries information sufficient to decode position/velocity information. We further propose that spatial and velocity information are distributed in different populations of DG neurons with an overlap.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-196 前障による大脳皮質抑制性ニューロンの活動制御 Ayako Ajima（安島綾子），Momoko Shiozaki（塩崎桃子），Yoshihiro Yoshihara（吉原良浩）理研CBS システム分子行動学 The claustrum is a thin sheet-like structure located between the insular cortex and the striatum, which has reciprocal connections with almost all the neocortical areas. Although it has been hypothesized that the claustrum may play roles in higher brain functions such as multisensory integration, saliency detection and consciousness, its physiological function is still enigmatic. Recently, we established a unique transgenic mouse line in which Cre recombinase is specifically expressed by a subpopulation of neurons in the claustrum of adult brain (Cla-Cre mice) and found that the insular cortex receives the densest inputs from the Cre-positive claustral neurons. In this study, we focused on the claustro-cortical connection and investigated electrophysiological responses of the insular neurons upon optogenetic stimulation of the claustrum in acute slice preparations. Cre-dependent, channelrhodopsin2 (ChR2)-expressing adeno-associated viral vector was injected into the claustrum of Cla-Cre mice. After three weeks, we prepared horizontal brain slices containing both the claustrum and the insular cortex and performed whole-cell patch-clamp recordings from neurons in the insular cortex in combination with optogenetic stimulation of the claustral neurons. The recorded insular neurons were classified into either regular-spiking or fast-spiking neurons according to their firing patterns generated upon current pulse injection. Photostimulation of the ChR2-expressing claustral neurons evoked EPSP in almost all the recorded neurons in both the regular-spiking and fast-spiking neurons in all the layers of the insular cortex. However, action potentials were observed only in the fast-spiking interneurons (64% in layer 2/3; 37% in layer 5/6), but not in the regular-spiking neurons. These results indicate that the Cre-positive claustral neurons selectively drive the inhibitory interneurons in the insular cortex. We also recorded neurons in the prefrontal cortex which also has dense connections with the claustrum and observed similar results. Thus, the present study raises a possible new concept that the claustrum modulates cortical activity by sculpting and/or synchronizing cortical activities through inhibitory interneurons.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-199 ラットの経血管灌流標本において内因性の硫化水素は正常呼吸に関与する Minako Okazaki（岡崎実那子）^1,2，Saori Uozu（魚津沙織）¹，Tadachika Koganezawa（小金澤禎史）¹ ¹筑波大　医学医療系生理 ²筑波大院　人間総合科学フロンティア医科学 Hydrogen sulfide (H₂S) is generally known as a toxic gas but constitutively generated in our body. Recently, it has been reported that endogenous H₂S has crucial roles in synaptic modulation as the third gasotransmitter after NO and CO. Although respiration is generated by neural network at respiratory center in the brainstem and the spinal cord, roles of endogenous H₂S to central respiratory pattern generation is not uncovered. In this study, we aimed to evaluate functional roles of endogenous H₂S in generating eupneic respiration at the respiratory center. We performed in situ arterially perfused preparations of decerebrated rats and recorded the activity of phrenic and vagus nerves to know central respiratory outputs. In order to block H₂S production, we pharmacologically inhibited H₂S producing enzymes, cystathionine β-synthase (CBS) and cystathionine γ-lyase (CSE) respectively. We also pharmacologically activated CBS. Further, we added a persistent sodium channel blocker to inhibit pacemaker property of neurons. And then, we compared the central respiratory pattern before and after administration of drugs. After inhibition of CBS, respiration was switched from three-phases eupneic pattern, which consists of inspiration, post-inspiration and expiration, to gasping-like pattern which is composed of only inspiration. Respiratory frequency and amplitude of phrenic and vagus nerves were also decreased compared with absence of CBS inhibitors. When we added persistent sodium channel blocker, gasping was disappeared or decreased in the presence of CBS inhibitors. On the other hand, after inhibition of H₂S synthesis via CSE and activation of synthesis via CBS, three phases respiratory patterns were maintained though respiratory frequency was increased and amplitude of phrenic nerve activities were decreased after inhibiting CSE. These results suggested that H₂S produced by CBS has crucial roles to maintain neuronal network to generate eupnea. Respiratory pattern generating mechanism might be switched from network-based system to pacemaker cells-based system under low H₂S condition.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-197 肘動作の筋活動を用いたロボットハンド錯覚における自己受容感覚ドリフト Toshihiro Kawase（川瀬利弘）^1,2,3，Soichiro Fujiki（藤木聡一朗）¹，Kenji Kansaku（神作憲司）^1,4 ¹獨協医大医生理（生体情報） ²東京医歯大生材研 ³東京工大科学技術創成研究院 ⁴電通大脳科学 Understanding how we consciously experience our bodies is a fundamental issue in cognitive neuroscience. The incorporation of non-body objects into the body representation has been investigated extensively using a rubber hand illusion task. In the former literatures, researchers have reported that the rubber hand illusion is accompanied by a change of perceived position of the hand towards the rubber hand, which is known as a proprioceptive drift. We formerly reported that the proprioceptive drift was observed in a robot hand illusion task using electromyography (EMG) activity on wrist movements (Sato et al., 2018). In the present study, we report that the same phenomena were observed in a robotic arm that moved synchronously with the participants' elbows. Three able-bodied participants were recruited. The robotic arm, using myoelectric control with one degree of freedom (elbow flexion and extension), consisted of a prosthetic glove and an actuator. The joint positions of the robotic arm were controlled continuously by means of the participant's muscular activity on the elbow flexor and extensor. The participants took part in the in-phase and out-of-phase movement conditions for 10 min each. Before and after each 10-min experiment, the participants pointed to indicate the sensed position of the right index finger with their left index finger with their eyes closed. The participants had to make the pointing movement by touching a panel placed vertically in front of them. We measured the position pointed to, and the proprioceptive drift in a vertical plane was then calculated by subtracting the two position measurements from each other. The median of the proprioceptive drifts in the in-phase movement condition (7.7 mm) were larger than mean in the out-of-phase movement condition (-9.2 mm), which is consistent with our results in wrist movements. The preliminary results suggest that our methods are applicable not only in wrist movements but also in elbow movements.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-198 延髄背側領域にあるPhox2B陽性ニューロンは吸啜および嚥下運動の発現に関与する Makito Iizuka（飯塚眞喜人）¹，Keiko Ikeda（池田啓子）²，Hiroyuki Igarashi（五十嵐敬幸）³，Kazuto Kobayashi（小林和人）⁴，Hiroshi Onimaru（鬼丸洋）¹，Masahiko Izumizaki（泉崎雅彦）¹ ¹昭和大・医・生体調節機能 ²国際医療福祉大・医・生理 ³ウエスタン大・生理薬理 ⁴福島医大・医・生体機能 Phox2B-positive interneurons in the ventrolateral part of the facial motor nucleus are essential to generate respiratory rhythm. Although there are other several regions where the Phox2B-positive neurons are clustered, the physiological roles are not fully elucidated. We developed a transgenic (Tg) rat in which Phox2b-positive neurons expressed one of channelrhodopsin variants; ChRFR(C167A), and found that the photo-stimulation of the Phox2B-positive neurons from the dorsal skull causes rhythmic opening/closing movement of the mouth in this Tg neonatal rat at free-moving conscious condition. To clarify the function of this rhythmic mouth movement, we recorded pressure in the oral cavity using anesthetized neonatal Tg rat. During this movement, the oral cavity showed rhythmic negative pressure at about 2 Hz. After euthanasia, the rat's stomach was filled with air that would come from the catheter to the pressure transducer in the oral cavity. Therefore, this movement should be both suckling and swallowing. In the next experiment, using isolated pons-spinal cord preparations, the area where photo-stimulation evoked rhythmic motor activity was explored. The motor activity was recorded from the motor branch of the trigeminal nerve, hypoglossal nerve, fourth cervical ventral root (C4VR) and phrenic nerve. When photo-stimulation was applied with a 0.2 mm optical fiber to the dorsal side of the brainstem, the area was restricted around the area postrema (AP). We also found that the rhythmic activity showed about 2 Hz, and these activities in trigeminal nerve, hypoglossal nerve, C4VR and phrenic nerve were synchronized. When the preparation was divided at about 0.4 mm caudal to the rostral end of AP, the rostral part of the preparation showed rhythmic activity in the hypoglossal nerve, and the caudal part of preparation also showed rhythmic activity in C4VR during photo-stimulation of the dorsal surface. These results showed the networks can be divided into at least two groups; the rostral one for sucking generator and the caudal one for swallowing generator, respectively.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-200 概日リズムの制御機構におけるドパミンD3受容体の機能解析 Naoya Matsunaga（松永直哉）^1,2，Yuya Yoshida（吉田優哉）²，Masaki Matsuda（松田将希）²，Chihiro Hayama（羽山千尋）²，Satoru Koyanagi（小柳悟）^1,2，Shigehiro Ohdo（大戸茂弘）² ¹九州大学大学院薬学研究院グローカルヘルスケア分野 ²九州大学大学院薬学研究院薬剤学分野 Many organisms have circadian clock mechanisms that cause periodic fluctuations in 24-h cycles and are involved in various physiological functions. The basis of circadian rhythm is a feedback loop mechanism of transcription and translation regulated by transcriptional factors encoded by clock genes.In addition, clock genes induce time-dependent oscillation in gene expression, including for enzymes and receptors important for biological functions.Therefore, it has been suggested that transcriptional alteration of clock genes is also related to the onset and pathology of various diseases. Dopamine-producing nerve cell is present in the mesenchyme of the hypothalamus and the area of the brain including the cerebral limbic system and the SCN, which is the center of circadian rhythm, and dopamine-producing nerve cell is also related to neurodegenerative diseases such as Parkinson's disease and Huntington's disease and to psychiatric conditions such as schizophrenia and depression. Therefore, dopamine receptor has been evaluated as a therapeutic target for these diseases. The expression of dopamine D3 receptor (DRD3) is regulated by clock genes and the relationship between functional abnormality of DRD3 and various neurological diseases is known. However, the relationship between DRD3 function and the circadian machinery is unclear. In this study, we investigated the influence of DRD3 on the circadian machinery. Locomotor activity was measured in wild-type (WT) and DRD3 knockout (Drd3-/-) C57BL/6J mice. The circadian expression of clock genes in the suprachiasmatic nucleus (SCN) of mice and primary astrocytes were also evaluated. The effect of Per1 expression change via DRD3 on the clock gene expression rhythm in mice and primary astrocytes was also determined. Drd3-/- mice demonstrated increased locomotor activity and period length compared with WT mice. Per1 expression increased in the SCN and cultured primary astrocytes of Drd3-/- mice compared with that in WT mice. The differences were caused by up-regulation of the extracellular signal-regulated kinase-cAMP response element binding protein (ERK-CREB) signaling pathway in Drd3-/- cells. The results of the study suggested that DRD3 is functionary for the circadian machinery and provide new insight into the pathology of neurological disorders related to DRD3 dysfunction.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-201 Phox2b陽性細胞にチャネルロドプシンを発現させたトランスジェニックラットを用いた呼吸中枢神経回路の光遺伝学的解析 Hiroshi Onimaru（鬼丸洋）¹，Keiko Ikeda（池田啓子）²，Hiroyuki Igarashi（五十嵐敬幸）³，Hiromu Yawo（八尾寛）⁴，Kazuto Kobayashi（小林和人）⁵，Satoru Arata（荒田悟）⁶，Kiyoshi Kawakami（川上潔）⁷，Masahiko Izumizaki（泉﨑雅彦）¹ ¹昭和大・医・生理 ²国際医療福祉大・医・生理 ³Dept Physiol and Pharmacol, Schulich Sch of Med and Dent, Western Univ, Ontario, Canada ⁴東北大学生命科学 ⁵福島医大・医・生体機能 ⁶昭和大・遺伝子組換え ⁷自治医科大細胞生物 The paired-like homeobox gene Phox2b is predominantly expressed in preinspiratory (Pre-I) neurons in the parafacial respiratory group (pFRG) in the rostral ventrolateral medulla of the newborn rat. To analyze detailed neuronal properties of respiratory rhythm generation using optogenetics, we used transgenic (Tg) newborn rats that expressed one of channelrhodopsin variants; ChRFR(C167A) in Phox2b positive cells. Effects of selective activation of Phox2b-positive neurons in the ventral medulla on respiratory rhythm generation were examined in brainstem-spinal cord preparations isolated from 0-2 day old Tg newborn rats. The rostral ventrolateral medulla corresponding to the pFRG was photo-stimulated by blue light-emitting diode via an optic fiber with 0.2 mm outer diameter up to 60 s (50 ms duration/ 150 ms interval). The photo-stimulation increased respiratory rate more than 200 % of control. The excitatory effects were maximum at immediately after start of the stimulation and gradually reduced during continuous stimulation. Short photo-stimulation (1.5 s) of the rostral pFRG reset the respiratory rhythm. We examined the effects of the photo-stimulation (up to 60 s) on membrane potentials of various types of respiratory-related neurons by whole cell patch clamp in the rostral ventrolateral medulla. The photo-stimulation depolarized Phox2b-positive (i.e. ChRFR-positive) Pre-I, inspiratory (Insp) and tonic (respiratory modulated) neurons as well as Phox2b-negative Pre-I neurons. In contrast, changes in the membrane potential of Phox2b-negative Insp and expiratory neurons are variable and dependent on types of the synaptic connections with other neurons activated directly or indirectly by the photo-stimulation. In the presence of TTX, the photo-stimulation depolarized Phox2b-positive cells, whereas it gave no significant changes in the membrane potential of Phox2b-negative cells. Depolarization of Phox2b-positive neurons in the absence of TTX was due to cell-autonomous photo-activation and summation of EPSPs. Membrane potential changes of Phox2b-negative neurons were dependent on properties of synaptic connections between respiratory-related neurons. Our findings further shed light on local networks among respiratory-related neurons in the rostral ventrolateral medulla and support the central role of Pre-I neurons in the respiratory rhythm generation in the neonatal rat.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-202 ゼブラフィッシュ幼魚の胸びれリズム運動を制御する神経回路の解析 Yuto Uemura（植村悠人）^1,2，Yukiko Kimura（木村有希子）²，Yoichi Oda（小田洋一）¹，Shin-ichi Higashijima（東島眞一）² ¹名古屋大院理生命 ²自然科学研究機構　基礎生物学研究所　神経行動学 Limbed vertebrates exhibit coordinated rhythmic movements of forelimbs and hindlimbs during locomotion. Neuronal circuits that control rhythmic limb movements in mammals have been investigated for decades, but our knowledge is limited because of the complexity of their limb. Rhythmic movement of pectoral fins during swimming in larval zebrafish is an attractive model. Pectoral fins of larval zebrafish show left and right alternated rhythmic movements, and they are actuated only by two types of muscles, i.e., abductor (Ab) and adductor (Ad). Due to the simplicity of pectoral fins, we expect that we will be able to characterize neuronal circuits that control rhythmic pectoral fin movements more deeply. We performed electrophysiological recordings of Ab motoneurons (MNs) and Ad MNs during fictive swimming. Both Ab MNs and Ad MNs showed spiking activities in particular phases of a swimming cycle. Activities of Ab MNs and Ad MNs on the same side essentially alternated. Both Ab MNs and Ad MNs received alternating excitatory and inhibitory inputs in a swimming cycle, and excitations mainly occurred in their preferential firing phase, and inhibitions mainly occurred for the rest of the period. To obtain insights into the sources of these inputs, we looked for synaptic connections between possible premotor interneurons and Ab or Ad MNs using optogenetics and electrophysiological recordings. The experiment reveled that neurons which express the dmrt3 gene made frequent synaptic connections onto Ab MNs, suggesting that dmrt3-positive neurons are one of the sources that provide inhibition to Ab MNs during rhythmic pectoral fin movements. This notions was further supported by analyzing firing activities of dmrt3-positive neurons during swimming: the timing of dmrt3-neuron firings coincided with the peak timing of inhibition that contralateral Ab MNs received. To reveal the functions of dmrt3 neurons, we generated transgenic zebrafish in which dmrt3 neurons near the pectoral fin MNs were ablated by the expression of diphtheria toxin. Electrophysiological recordings showed that the firing phases of Ab MNs in these fish shifted earlier than that of WT fish. Furthermore, behavioral analysis revealed that these fish were not able to close their pectoral fins sufficiently during swimming. These phenotypes are consistent with the idea that the inhibition provided by dmrt3-positive neurons on the contralateral side limit the firings of Ab MNs in a narrow time window.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-204 運動スキルトレーニングは糖尿病モデルラットの運動野の機能障害を改善する Ken Muramatsu（村松憲）¹，Masako Ikutomo（生友聖子）²，Toru Tamaki（玉木徹）³，Satoshi Shimo（志茂聡）⁴，Masatoshi Niwa（丹羽正利）⁵ ¹杏林大理学療法 ²東京医療学院大理学療法 ³健康科学大理学療法 ⁴健康科学大作業療法 ⁵杏林大作業療法 Recently, we found that diabetic rats exhibited reduction in the hindlimb and forelimb motor areas, with the hindlimb area reductions being the most severe. The effect of exercise on reversing motor cortex area loss is unknown. The purpose of this study was to investigate the effects of two types of exercise training - complex motor skills training (CMST) and aerobic training (AT) - on motor cortex area. Thirteen-week-old male rats were divided into a diabetic group (DM, n = 18), in which diabetes was induced by streptozotocin injections at 13 weeks, and a non-diabetic sedentary control group (CON, n = 6). Twenty-two weeks after the streptozotocin injections, movement representation in the sensorimotor cortex was assessed by intra-cortical microstimulation followed by 14 days of training for the DM-CMST group (n = 6) and DM-AT group (n = 6). As a control, we examined the age-matched diabetic sedentary group (DM-SED, n = 6) and CON group (n = 6). The area of hindlimb and forelimb motor cortex was smaller in the DM-SED and DM-AT groups than in the CON group (P < 0.01). Additionally, there was no significant difference between DM-AT and DM-SED. Interestingly, the area of the forelimb motor cortex of DM-CMST rats was almost equivalent to that of CON rats. Furthermore, the hindlimb motor area of DM-CMST rats was larger than that of DM-AT and DM-SED rats (P < 0.01), though this was smaller than the CON rats (P < 0.01). This study demonstrated that 14 days of AT did not prevent reductions of motor cortical area in diabetic rats. However, 14 days of CMST prevented reductions of motor cortical area. These findings suggest that CMST may be beneficial in prevention of reductions of motor cortical area in diabetic rats. Although it is known that both CMST and AT enhance functional and structural recovery after brain injury, it is unclear why only CMST demonstrated a therapeutic effect on motor cortex area recovery in the current study. It is recommended that future studies examine the mechanism of recovery.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-205 レム睡眠時筋脱力の神経回路 Shuntaro Uchida（内田俊太郎），Shingo Soya（征矢征矢），Takeshi Sakurai（櫻井武）筑波大　国際統合睡眠医科学研究機構 One of the hallmarks of REM sleep is bilateral muscle atonia. Recent studies in rodents showed that inhibitory neurons in the ventromedial medulla (VMM) are responsible for generating the muscle atonia. Consistently, we observed obvious increases in muscle tone and body movements during REM sleep in mice when we inhibited glycinergic neurons in the VMM (Gly^VMM neurons) by expression of tetanus toxin light chain. However, the precise neuronal circuit architecture that regulates the REM atonia has remained unknown. The purpose of this study to identify the upstream neuronal populations that make direct synaptic contact to Gly^VMM neuorns to delineate neuronal circuits that evoke the REM atonia. We injected Cre-activatable CAV2 vector carrying flippase (flp) into the anterior horn of the lumber spinal cord in GlyT2-iCre mice to express flp specifically in glycinergic neurons that send innervation to the anterior horn (AN). We next express TVA and rabies glycoprotein (RG) specifically in these neurons (Gly^VMM→AN neurons) by injecting flp-activatable contracts of AAV vectors carrying TVA and RG in the VMM. Then we performed retrograde tracing by injecting SADdG-GFP(EnvA) in the VMM. This study revealed that Gly^VMM→AN neurons receive direct synaptic inputs by neurons in the deep mesencephalic nucleus (DpMe), periaqueductal gray (PAG), superior colliculus (SC), inferior colliculus (IC), sublaterodorsal tegmental nucleus (SLD), and pontine reticular nucleus (Pn). This study suggests neuronal pathways that are implicated in the REM atonia. This information might be also important for understanding the pathophysiology of REM sleep behavior disorder, and cataplexy, which is sometimes accompanied by narcolepsy.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-206 発達期の皮質脊髄投射の退行と増生 Naoyuki Murabe（村部直之），Masaki Sakurai（桜井正樹）帝京大医生理 Studies in our laboratory showed that the corticospinal (CS) neurons extend the axons to the gray matter and form functional connections with the spinal neurons including interneurons and motoneurons in juvenile rodents. In the cervical cord, most of these connections are transient and later eliminated whereas distribution and density of the CS neurons projecting to the lumbar cord showed little changes during development. However, it is unclear when these connections were eliminated from the cervical cord in mice. To address this problem, we employed adeno-associated virus (AAV) that infects neurons from the C7 through axons. There are three major sensorimotor areas sending the axons to the cervical cord: M2, M1, S1 and S2. Injection of AAV2-retro encoding Cre recombinase into the C7 gray matter in Cre-inducible reporter mice on the postnatal day 7 (P7) showed that labeled neurons were distributed broader than these projecting to the C7 in the adult. Labeled areas expanded to the boundaries of adult distribution and even to the hindlimb areas, which is surrounded by M1 and S1 of the forelimb areas. The injection on P14 showed that the labeled areas started to change to the adult pattern such that labeled neurons in the boundaries and hindlimb area reduced in number. Therefore, period in which the maximum number of the cortical cells send axons to the C7 gray matter is P7 or before. Taking account of the previous findings that the CS axons projecting to the C7 reach a peak on P14 in ventrolateral region of the gray matter of the C7 and still increase beyond P14 in the intermediate zone, these results suggest that CS neurons eliminate exuberant axons while increase their arborizations in final target regions during development and highlight a dynamic property of arborization to the C7 in mice.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-207 外肛門括約筋の運動神経と筋にたいする高血糖の影響 Naomi Ohshiro（大城直美）¹，Ken Muramatsu（村松憲）²，Minori Fukuda（福田実乃里）¹，Masatoshi Niwa（丹羽正利）¹ ¹杏林大学保健学部作業療法学科 ²杏林大学保健学部理学療法学科 Diabetic neuropathy is caused by long-lasting hyperglycemic condition and mainly affects the sensory and autonomic nerves. Consequently, the normal functioning of the intestine is disturbed, and symptoms like repeated bad diarrhea and constipation occur. Recent studies have reported that diabetic neuropathy also affects the somatic motor nerves. The external anal sphincter (EAS) muscle is dominated by the pudendal nerve nucleus and has an important function in the suppression of incontinence and in control of the bowel movement. This study investigated the morphological alterations in EAS motoneurons and muscles of experimental type I diabetic rats. Type 1 diabetes was induced in 13-week-old female Wistar rats by streptozotocin administration (diabetic group). The age-matched control animals were injected with saline (control group). After 4-5 weeks, the EAS motoneurons of each animal were labeled. This was followed by administration of halothane aspiration anesthesia. The EAS muscle nerve was sectioned and dipped for 1 h into a small polyethylene tube containing 10% dextran-Texas red dissolved in saline. After 2 weeks survival period, the animals were re-anesthetized and perfused. The spinal cord was serially sectioned and observed with fluorescence microscopy; the number and size of cell bodies were examined. The EAS muscles were sectioned transversally and stained by hematoxylin-eosin (HE), to analyze the myocytes. There was no significant difference in the mean number and soma diameter of the labeled EAS motoneurons and the cross-sectional area of the EAS myocytes between the diabetic and control groups. However, the number and the soma diameter of the EAS motoneurons and the cross-sectional area of the EAS myocytes tended to decrease in the diabetic group. We suggest that the EAS is not affected much during the early stage of diabetes. However, there is a possibility that the EAS will be affected in prolonged diabetes. Therefore, we are currently investigating the effect of long-term hyperglycemia on the EAS. Furthermore, we plan to investigate the functional impairment of the EAS due to diabetes.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-208 糖尿病モデルラットでは速筋を支配する運動ニューロンと遅筋を支配する運動ニューロンで耐性が異なる Toru Tamaki（玉木徹）¹，Ken Muramatsu（村松憲）²，Masako Ikutomo（生友聖子）³，Masatoshi Niwa（丹羽正利）⁴ ¹健康科学大学　理学療法学科 ²杏林大学　保健学部　理学療法学科 ³東京医療学院大学　リハビリテーション学科　理学療法学専攻 ⁴杏林大学　保健学部　作業療法学科 Previous studies have suggested that the contractile strength of the medial gastrocnemius muscle (MG, a fast-twitching muscle) was reduced but that of the soleus muscle was preserved (SOL, a slow-twitching muscle) in streptozotocin-induced diabetic rats. Compared with disorders of the muscle itself, there is relatively scarce information on alterations to motor neurons (MNs). In the present study, we examined morphological alterations in the MNs of fast- and slow-twitch muscle, MG and SOL, respectively, in rats 12 and 22 weeks after diabetes onset. Diabetes was induced in 13-week-old male Wistar rats using streptozotocin (STZ) administration (n=10). Rats in the age-matched control group were injected with saline (n=10). The animals were anesthetized using halothane inhalation. Subsequently, the tibial nerve was cut near its MG insertion on one side, while the contralateral tibial nerve was sectioned near its SOL insertion. The central cut end was dipped into a small polyethylene tube containing 10 % 3-kDa dextran- Texas Red dissolved in saline for 1 h. After 14 days, the animals were re-anesthetized and perfused with 4% paraformaldehyde. After fixation, the L2-S3 portion of the spinal cord was removed and sectioned serially at 80 μm. The largest cross-sectional area of MN cell bodies and total number of MNs were recorded. The number of MNs in SOL muscle decreased to approximately 64% of the control value after 12 weeks of diabetes (P<0.01). On the other hand, the number of MNs in the MG muscle was unchanged at 12 weeks but decreased to approximately 78% of the control value after 22 weeks of diabetes (P<0.05). The mean cross-sectional area of the SOL MN cell bodies decreased to approximately 79% of the control value after 22 weeks of diabetes (P<0.05). On the other hand, the mean cross-sectional area of MG MNs showed a tendency to decrease, but this was not statistically significant at any of the evaluated time points. This study reports the difference in severity of MN loss between the MG and SOL muscles as determined using retrograde labeling. Notably, a higher rate of decrease in the number of SOL MNs was observed in the same animals. Given that previous studies have focused predominantly on diabetes-induced damage to fast-twitch muscles and their MNs, our findings reveal a novel aspect of diabetes pathology.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-211 ゼブラフィッシュ小脳発達におけるプルキンエ細胞集団の３次元形態学的解析 Narumi Fukuda（福田成美）¹，Kanae Hiyoshi（日吉加菜映）²，Kyo Yamasu（弥益恭）²，Sachiko Tsuda（津田佐知子）^2,3 ¹埼玉大理 ²埼玉大院理工生命科学 ³埼玉大学研究機構 The cerebellum is involved in various functions including motor control and learning, and has relatively simple and well-conserved neuronal circuits, providing an appropriate system for circuit analysis. A series of works have shown that the cerebellum is structurally and functionally compartmentalized, which are thought to function as functional modules in the cerebellum. However, it is not resolved how cerebellar functional circuits along with its compartments are formed during the development. To address this issue, we are using zebrafish, which has similar cerebellar circuits as mammals, and whose transparency and small size provide an ideal model system for studying neurogenesis and optical techniques. By high-speed and whole-cerebellar calcium imaging of Purkinje cell populations, we are identifying functional compartments in the developing cerebellum of zebrafish. However, we do not have a comprehensive map of cerebellar circuits in the developing cerebellum, which provides fundamental information for circuit analysis. Here, we sought to comprehensively characterize the anatomical properties of the developing cerebellum of zebrafish. First, to examine the 3D distribution of the entire Purkinje cell population in the cerebellum, we established an analysis platform by combining confocal imaging and 3D mapping of all Purkinje cells. At 3 to 14 day post fertilization, when the cerebellar circuits are being formed, the number of Purkinje cells was significantly increased and their distribution patterns were gradually changed. By detailed observation, we also found that the length, arborization and orientation of Purkinje cells were different depending on their positions in the cerebellum. In addition, Purkinje cell dendrites at the posterior region of the cerebellum showed band-like patterns. These results suggest the regionalization at cell population level in the developing cerebellum of zebrafish. We are now examining topographic maps of input and output neurons of the developing cerebellum. Combining the functional imaging data with our comprehensive and 3D morphological analysis of the Purkinje cells in the developing cerebellum would deepen our understanding of the structural and functional development of cerebellar circuits.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-209 脊髄損傷後の運動機能回復に関わる脳機能 Reona Yamaguchi（山口玲欧奈）¹，Toshinari Kawasaki（川崎敏生）^2,3，Chao C Zenas（Zenas C Chao）²，Masahiro Mitsuhashi（三橋賢大）^2,4，Satoko Ueno（上野里子）²，Tadashi Isa（伊佐正）^1,2 ¹京都大ASHBi ²京都大院医神経生物 ³京都大院医脳病態生理脳神経外科 ⁴京都大院医脳病態生理臨床神経学 Spinal cord injury (SCI) causes long-term devastating loss of physical functions in the patients. To elucidate the neuronal mechanism of recovery from SCI would contribute to development of better therapeutic strategies. We have previously shown that the activity of the ipsilesional motor cortices including the premotor (PM) and primary motor cortex (M1) is enhanced and contribute to the hand movement control during the recovery in a partial spinal cord injury model of macaque monkeys (Nishimura et al. Science, 2007). More recently, we have found that the motor-preparation-related interhemispheric interactions between the contralesional PM and ipsilesional PM in the α to low-β bands were enhanced after the lesion (Chao et al. Cereb Cortex, 2018). To further understand the contribution of ipsilesional PM, M1 and primary somatosensory cortex (S1) to the recovery, we chronically implanted the multi-channel ECoG electrodes on the bilateral PM/M1/S1 of the macaque monkey. We longitudinally monitored the cortical activity during a reach and grasp task, and used direct electrical stimulation to probe the cortical connectivity before and after the sub-hemisection of the mid-cervical spinal cord (C4-C5). In this lesion model, the impairment of hand movements was severer than the lateral funiculus-specific lesion model used in our previous studies. In the previous model, the precision grip mostly recovered in 1-3 months after the lesion. However, in the current model, the precision grip did not fully recover. For the brain activity, the high-gamma activity of the contralesional M1 during the reach and grasp movements was enhanced after the lesion. This enhancement continued to increase gradually after the lesion. To investigate the functional connectivities, we calculated the net information flows assessed by Granger causality analysis between the contralesional M1 and other brain areas. In the preoperative stage, the information flows to the contralesional M1 in the β band were mainly derived from the contralesional PM. However, the information flow from ipsilesional PM to contralesional M1 increased after the lesion. In addition, we found that the synaptic inputs from the ipsilesional PM to contralesional M1 were enhanced after the lesion as assessed by the direct electrical stimulation. These result suggested that the movement-related connectivity between ipsilesional PM and contralesional M1 contributes to the recovery after the lesion.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-210 衝動性眼球運動を用いた神経変性疾患における大脳基底核、小脳の機能異常の検出 Yasuo Terao（寺尾安生）¹，Shin-ichi Tokushige（徳重真一）²，Satomi Terada（寺田さとみ）¹，Akihiro Yugeta（弓削田晃弘）³，Yoshikazu Ugawa（宇川義一）⁴ ¹杏林大学医学部細胞生理学 ²杏林大学医学部付属病院神経内科 ³東京大学医学部附属病院神経内科 ⁴福島県立医科大学神経再生医療学 OBJECTIVE: Multiple system atrophy is a neurological disorder, featuring both cerebellar and basal ganglia pathology, and can theoretically present either with cerebellar ataxia, parkinsonism or both. Actually, it is difficult to distinguish MSA with predominant cerebellar ataxia (MSAC) from spinocerebellar ataxia with pure cerebellar presentation (SCD) at their early stages, solely on clinical grounds. On the other hand, MSAP with predominant Parkinsonism (MSAP) resembles Parkinson's disease (PD) at early stages, both presenting with parkinsonism. We studied whether these disorders showing cerebellar and/or basal ganglia dysfunction can be differentiated pathophysiologically by the changes in saccade parameters including the saccade velocity profiles METHODS: Visual guided (VGS) and memory guided (MGS) saccade tasks were performed by 16 patients with MSAP, 29 with MSAC, 63 with PD, 12 with SCD, as well as 36 age-matched normal subjects. Targets were presented 5, 10, 20, 30 degrees to the left and right of the fixation point. Saccade latency, amplitude, peak velocity, saccade duration (acceleration and deceleration periods) were compared among the subject groups, with reference to the clinical stage of the patients. RESULTS: Saccade parameters of both tasks showed similar changes with progressing disease in SCD and MSAC patients, as did those of MSAC and MSAP patients, although hypometria was slightly more pronounced in MSAP. In MSAC and MSAP, latency and success rate of MGS were stable throughout disease stages, whereas they deteriorated progressively with progressing disease in PD. Both MSAC and MSAP patients showed a decrease in peak velocity. In MSAP, saccade duration (both acceleration and deceleration periods) was prolonged, whereas MSAC patients showed shortening of the acceleration period. In SCD, the peak velocity decreased, the acceleration period was shortened whereas the deceleration period was extended. In PD, the peak velocity and the saccade acceleration period were normal, but the deceleration period was extended. CONCLUSIONS: The pathophysiological basis underlying these neurodegenerative disorders resulting from cerebellar and basal ganglia dysfunction can be differentiated by saccade parameters. The changes in saccade parameters may be used as a measure indexing the progression of cerebellar and basal ganglia dysfunction as well as for assessing the functional improvement through saccades when clinical treatment becomes available.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-213 眼球運動系神経積分器と前庭小脳との神経回路基盤の解明 Taketoshi Sugimura（杉村岳俊），Yasuhiko Saito（齋藤康彦）奈良医大医第一生理 Gaze holding is primarily controlled by the oculomotor neural integrators, which are separated into the prepositus hypoglossi nucleus (PHN) for horizontal gaze and the interstitial nucleus of Cajal (INC) for vertical gaze. Although it has been argued that the neural connections from the integrators to the vestibulo-cerebellar cortex are significant in gaze holding, the properties of the integrator neurons that project to the cortex have not been well defined. In the present study, we examined the types and distributions of PHN and INC neurons that were retrogradely labeled by injecting the dextran-conjugated Alexa 488 tracer into the cerebellar flocculus (FL) or uvula/nodulus (UN) using choline acetyltransferase (ChAT)-tdTomato transgenic rats. When the tracer was injected into the FL unilaterally or center of the UN, the retrogradely labeled neurons were observed in the PHN but not in the INC. The proportion of cholinergic PHN neurons that projected to the UN (21.7 ± 0.8 % in 6 rats) was significantly larger than the proportion of the neurons that projected to the FL (8.9 ± 1.5 % in 6 rats, P < 0.0001). The proportion of cholinergic PHN neurons that projected to the UN differed depending on the caudal (9.3 ± 1.4%), intermediate (23.3 ± 2.1%) and rostral (32.6 ± 2.0%) regions of PHN in 6 rats. To examine PHN neurons projecting to other cerebellar cortex than vestibulo-cerebellum, we investigated the neurons that projected to the spino-cerebellar cortex (centralis/culmen (CC)). The proportion of retrogradely labeled cholinergic PHN neurons by injecting the tracer into the CC (20.0 ± 4.8 % in 3 rats) were comparable to the proportion of the neurons by injecting the tracer into the UN. These results indicate that the projection to the vestibulo-cerebellum from the neural integrator is restricted to the PHN, in which the proportion of cholinergic projection neurons differs depending on the projection sites.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-214 マウス小脳細胞による歩行速度と加速度情報の表現様式 Koji Ikezoe（池添貢司），Kazuo Kitamura（喜多村和郎）山梨大医神経生理 Regulating speed of walking is necessary for animals to travel efficiently and cooperate with others. Monitoring speed and acceleration of their own walking are important to keep or change the speed as animal intends. Cerebellum is involved in control of locomotion. However, whether and how speed and acceleration are represented in neuronal activities in cerebellar cortex is poorly understood. Here, we sought to characterize speed and acceleration tunings of cerebellar neurons. We recorded activities of multiple single-units using silicon probes from lobule IV/V and VI of the cerebellum in a head-fixed mouse walking on a non-motorized treadmill in a virtual reality environment. The treadmill comprised a transparent circular disk and constrained a mouse to walk nearly straight forward. We recorded movements of mouse's paws from the side and the bottom using a CCD camera. Water-restricted mice were trained to keep still for one second and then walk for three seconds without stopping to obtain a water reward. Most of units changed their firing rate during walking. Preferred walking speed of individual units was uniformly distributed over all range of speeds. On the other hand, the distribution of preferred acceleration was biased to be positive. A joint tuning for speed and acceleration was well-fitted with a linear sum of tunings for individual parameters. From these results, we suggest that individual neurons in the cerebellum encode information of walking speed and acceleration, and thus these neurons and their downstream target can contribute to regulate walking speed properly. We are currently analyzing stride lengths and gait cycles of walking by offline video tracking of paws in relation to neuronal firing. We will also report cerebellar representation for these gait parameters in the presentation.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-215 複雑なステップパターンを走行中のマウスにおけるリズミカルな運動の制御 Kojiro Hirokane（広兼浩二朗）¹，Toru Nakamura（中村徹）¹，Yasuo Kubota（久保田康夫）²，Dan Hu（Hu Dan）²，Takeshi Yagi（八木健）¹，Ann M Graybiel（Graybiel Ann）²，Takashi Kitsukawa（木津川尚史）¹ ¹大阪大学生命機能研究科 ²Department of Brain and Cognitive Sciences and the McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, Massachusetts Behaviors of animals are often continuous and repetitive. Such behaviors like running and singing consist of many individual actions and often become rhythmic. In most cases, more than one body parts such as arms, legs, tongue, and throat are involved in continuous and repetitive activities. Notably, each of the body parts has its own timing and such timed actions are properly organized between the body parts. However, the mechanism for shaping such dynamical coordination between concurrent repetitive motions is not well understood. To understand how the dynamical coordination is organized in continuous movements, we analyzed the limb movement of mice running in complex stepping in the step-wheel. The step-wheel is a motor-driven running device with ladder-like pegs, the position of which can readily be changed. Mice drink water as a reward from the spout which intrudes into the wheel while running. Thus, mice run according to the rotation speed and the peg-pattern set by an experimenter. The timing of touches to each peg and spout was monitored by voltage touch sensors. We recorded the timing of touches and licks while mice were running on a complex peg-pattern in the step-wheel. We analyzed the relationship of right and left forelimbs and how this relationship changed in a course of stepping. The timing of a left-touch in one step of the right limb (two consecutive right touches), which is called the left-touch phase, was used as the index for the right-left relationship. The difference of phases in two consecutive steps, the phase shift, was calculated, and the average of phase shifts was used as the index for the changes in the right-left relationship. We found that there were several blocks of a few steps in a peg-pattern that the phase shift was small, which means that the relative position of the right and left limbs were conserved in the block. We also found that the variance of touch intervals was low inside such blocks compared to those of between-blocks, which indicates that running speed was controlled between blocks while it was kept relatively steady within blocks. These results suggest that mice adjusted their stepping so that they could hold constant phase and speed for a block of a few steps. This may be an easy and energy saving strategy for organizing concurrent movements of the right and left limbs in complex stepping.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-216 運足の感覚や位相をコードする線条体神経細胞 Takuma Terashita（寺下拓真）¹，Toru Nakamura（中村徹）¹，Yasuo Kubota（久保田康夫）²，Hu Dan（Dan Hu）²，Ann M Graybiel（Graybiel M Ann）²，Takeshi Yagi（八木健）¹，Tkashi Kitsukawa（木津川尚史）¹ ¹大阪大学大学院生命機能研究科 ²Department of Brain and Cognitive Sciences and the McGovern institute for Brain Research, Massachusetts Institute of Technology, Cambridge Our behavior encompasses many continuous movements, exemplified by those used in running, speech and playing musical instruments. Such behaviors consist of many individual actions of multiple body parts. When we play the piano, multiple fingers move sometimes simultaneously, or alternately, sequentially. The movement of fingers is often repetitive and rhythmic. In addition, each rhythm of fingers is coordinated temporally as if all of finger rhythms are connected each other. However, it is not clear how such coordination of movements is shaped in the brain. In our previous study, to investigate the coordination of limb movements, we developed a complex stepping task for mice, the step-wheel. The step-wheel is a motor-driven wheel in which mice run for water reward and can control the speed and the step-pattern of running mice. The step-pattern of mice can be controlled by the pattern of pegs which serve as footholds for mice. The step of mice can be detected by voltage sensors attached to each peg. In this study, we recorded spike activity from the dorsal striatum as mice performed the step-wheel task. We found neurons responding to events including the start or stop of drinking, licking of the spout, and touches of forelimb paws to pegs. Among them, we focused on neurons with phasic responses to touches. Even with complex peg-patterns, in which the arrangements of pegs were pseudo-random, the touch responding neurons showed phasic responses correlated with the touches of forelimb paws to pegs. However, the responses were not uniform to every touch; responses were high for some touches but low for others. By aligning the spikes to right or left touches, we found that the responses to touch-to-touch intervals and/or right-left timing of limbs were different neuron by neuron. To confirm such responsiveness of the neurons, we used two peg-patterns, the peg-pattern I and peg-pattern P. In the peg-pattern I, the space between two consecutive pegs (interval) were gradually changed from narrow to wide. In the peg-pattern P, the relative positions of right and left pegs (phase) were gradually changed. We found that considerable number of neurons responded to touches showed the modulation of firing rate by the interval in peg-pattern I and/or by phase in peg-pattern P. These results indicate that the striatal neurons may serve the coordination of body parts in repetitive movements using the parameters of rhythm, the phase and interval.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-217 Glutamatergic and GABAergic inputs to monkey subthalamic nucleus and its activity during motor task performance Zlata Polyakova（Polyakova Zlata）^1,2，Nobuhiko Hatanaka（Hatanaka Nobuhiko）^1,2，Satomi Chiken（Chiken Satomi）^1,2，Atsushi Nambu（Nambu Atsushi）^1,2 ¹Div System Neurophysiol, Natl Inst Physiol Sci, Okazaki, Aichi, Japan ²Dept Physiol Sci, SOKENDAI, Okazaki, Aichi, Japan The subthalamic nucleus (STN) receives inputs from the cerebral cortices through the hyperdirect and indirect pathways of basal ganglia (BG) and sends outputs to the internal segment of the globus pallidus (GPi), an output nucleus of the BG. STN neurons change their activity during motor execution and play an essential role in control of movements. Actually, abnormal activity in the STN has been reported in movement disorders, such as Parkinson's disease, and manipulation of its activity by deep brain stimulation or surgical lesion is an effective treatment for their symptoms. However, the control mechanism of STN activity is still not clear. In the present study, we have investigated how STN activity is controlled by glutamatergic inputs through the cortico-STN hyperdirect pathway and GABAergic inputs through the cortico-striato-external pallido (GPe)-STN indirect pathway. We recorded STN activity in awake Japanese monkeys. Electrical stimulation of the motor cortices induced early and following late excitations in STN neurons. Local injection of glutamatergic receptor antagonists into the STN reduced the early excitation. NMDA receptor antagonist (CPP) was more effective than AMPA-kainate receptor antagonist (NBQX). Injection of GABAergic agonist (muscimol) or antagonist (gabazine) into the GPe or striatum reduced late excitation. These results suggest that cortically induced early and late excitations are mediated by the hyperdirect and indirect pathways, respectively. Cortico-STN transmission is mainly mediated by NMDA glutamatergic receptors. Next, we recorded STN activity during task performance of goal-directed reaching task with delay, including go, no-go, and stop trials. In the control state, large portion of STN neurons demonstrated direction selectivity relative to left, center, or right target in go and/or stop trials. Most of STN neurons changed their activity in relation to go trials, and around 35% of them demonstrated stop-related activity, suggesting involvement of STN activity in both motor execution and cancellation. Then, we revealed the contribution of glutamatergic and GABAergic inputs to STN activity during voluntary movements by injection of their antagonists into the STN.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-218 直接路ニューロンから淡蒼球外節への投射は近接する間接路ニューロンの投射領域に包含される Shinichiro Okamoto（岡本慎一郎）^1,3，Jaerin Sohn（孫在隣）^2,3，Megumu Takahashi（高橋慧）³，Yoko Ishida（石田葉子）^1,3，Kenta Yamauchi（山内健太）¹，Masato Koike（小池正人）¹，Fumino Fujiyama（藤山文乃）⁴，Hiroyuki Hioki（日置寛之）^1,3 ¹順天大院　医神経生物・形態 ²生理研基盤神経科学研究領域大脳神経回路論 ³京都大院医高次脳形態 ⁴同志社大院脳科学研究科 The neostriatum (caudate-putamen, CPu), input nuclei of the basal ganglia, plays a key role in the motor control and cognitive function. Medium-sized spiny neurons (MSNs) constitute more than 90% of the neostriatal neurons and can be divided in two different subtypes, direct and indirect pathway neurons. The direct pathway neurons express dopamine D1 receptor, and project to the substantia nigra reticulata (SNr) and entopeduncular nucleus (EP). In contrast, indirect pathway neurons are characterized by the expression of D2 receptor and send axons only to the external globus pallidus (GPe). However, it was recently demonstrated that not only indirect but also direct pathway neurons innervate the GPe (Fujiyama et al., 2012). In the present study, we investigated whether or not direct and indirect pathway neurons located in vicinity send axon fibers to the same region(s) in the GPe. To visualize the direct and indirect pathway axons separately, we injected adeno-associated virus (AAV) vector into the CPu of D1-Cre transgenic mice. The virus vector expresses GFP or mRFP in the presence or absence of Cre, respectively, by using a Cre-dependent genetic switch (FLEx switch). Thus, D1-expressing direct pathway MSNs were labeled with GFP, whereas indirect pathway MSNs and some interneurons were visualized by mRFP. Since MSNs have numerous spines on the dendrites, we can easily distinguish indirect pathway MSNs from interneurons. After we observed the location of the direct and indirect pathway neurons in the CPu, we analyzed their axonal arborizations in the GPe. Both types of neurons often made two axonal bushes at the edge of the GPe. The axonal bushes of direct pathway neurons were sparse and narrow, and always distributed inside those of neighboring indirect pathway neurons. Since the cell bodies of direct and indirect pathway neurons examined in the present study were located nearby with each other, they may receive the common inputs from the brain regions such as the cerebral cortex and thalamus, and then transmit axon fibers to overlapping regions in the GPe.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-219 サルの運動タイミング制御におけるドーパミンの役割 Shigehiro Miyachi（宮地重弘）京都大霊長研高次脳 To clarify the roles of the dopamine in the motor timing control, we have trained macaques on repetitive button press tasks, in which the monkey presses a button in a rhythmically or in response to sensory cues. Previously, I have shown that the D2 antagonist halted the movement especially in the non-rhythmic task, whereas the D2 agonist caused timing deficits mainly in the rhythmic task. In the present study, effects of a dopamine D1 antagonist SCH39166 was examined using the modified versions of the same tasks. In the tasks, an LED button flashed, with a beep sound, 7 times/trial, and the monkey pressed the button in response to each flash (cue). In the rhythmic task, the button flashed rhythmically (stimulus onset asynchrony (SOA): 800 or 1000 ms). A liquid reward was given for each button press. Button press before the cue onset was allowed and the reward amount was maximized if the reaction time (RT) was 0 ms, to encourage the monkey to synchronize the press with the cue. In the ""random"" task, three SOAs (800, 1200, and 1600 ms) were randomized in each trial, and the presses before cue onset were not rewarded, to encourage the monkeys to press in response to each cue. Even though, after a long period of training, both monkeys frequently made anticipatory button presses after the longest (1600 ms) SOA. The monkeys well synchronized the button press with the cue in the rhythmic task, and also after the 1600-ms SOA in the random task. After systemic injection of the SCH39166 (0.03 mg/kg), motor timing was significantly deteriorated. The RTs of one monkey in the rhythmic task were 98±66 ms (median±QD) for 800 ms-SOA and 222±126 ms for 1000 ms-SOA, compared to 49±32 ms and -28±40 ms, respectively, in the control condition (p < 0.001, U-test). For the 1600 ms-SOA in the random task, the RT was 196±34 ms, compared to 66±43 ms in the control experiments (p < 0.001). RTs after the 800- and 1200-ms SOA in the random task were only slightly prolonged by the drug: 219±12 ms and 209±19 ms compared to 206±9 ms and 191±15 ms in control, respectively (p < 0.001). Essentially the same results were obtained for the other monkey. The results suggest that function of the dopamine D1 receptor is important for motor timing in the voluntary movement.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-220 ピアニストの指の運動機能 Tomoko Aoki（青木朋子）熊本県立大学 Motor function of individual fingers was compared between expert pianists (n = 11) and non-musician controls (n = 11) using fast tapping task and maximum voluntary isometric contraction task. In the tapping task, the subjects were instructed to hit the designated force sensor by designated finger as rapidly and as consistently as possible. The non-tapping fingers were required to maintain contact with the designated keys throughout the tasks. In the maximum voluntary isometric contraction task, the subjects were instructed to exert maxumum isometric force by the instructed digit to the instructed direction. In the tapping task, it was found that the tapping rate for each digit was significantly faster for the pianists than the controls. The group difference was largest in the ring finger and smallest in the index finger. In the maximum isometric contruction task, it was observed that maximum flexion and extension force of each digit was significantly stronger in the pianists than the controls. Maximum adduction forces of the thumb and the little finger were significantly larger for the pianists than the controls. Maximum abduction forces of the thumb and the index finger were significantly larger for the pianists than the controls. Thus, the present study demonstrated that dynamic and static motor function of individual fingers is strongly modulated by the long-term piano playing. These results suggest that long-term piano training can improve anatomical and neural factors determiming the dynamic and static motor function of individual fingers.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-221 Common neural network for different functions: an investigation of proactive and reactive inhibition Fan Zhang（Zhang Fan）¹，Sunao Iwaki（Iwaki Sunao）^1,2 ¹National Institute of Advanced Industrial Science and Technology (AIST) ²Graduate School of Comprehensive Human Sciences, University of Tsukuba Successful behavioral inhibition involves both proactive and reactive inhibition. Because simple stop cues cannot be used to distinguish between proactive and reactive inhibition processes, research in this area is relatively scarce. In the present study, we utilized the stop-signal paradigm to examine whole-brain contrasts and functional connectivity for proactive and reactive inhibition. The results of our fMRI data analysis show that the inferior frontal gyrus (IFG), the supplementary motor area (SMA), the subthalamic nucleus (STN), and the primary motor cortex (M1) were activated by both proactive and reactive inhibition. We then created 70 dynamic causal models (DCMs) representing the alternative hypotheses of modulatory effects from proactive and reactive inhibition in the IFG-SMA-STN-M1 network. Bayesian model selection (BMS) showed that causal connectivity from the IFG to the SMA was modulated by both proactive and reactive inhibition. To further investigate the possible brain circuits involved in behavioral control including proactive and reactive inhibitory processes, we compared 13 DCMs representing the alternative hypotheses of proactive modulation in the dorsolateral prefrontal cortex (DLPFC)-caudate-IFG-SMA neural circuits. BMS revealed that the effective connectivity from the caudate to the IFG is modulated only in the proactive inhibition condition but not in the reactive inhibition. Together, our results demonstrate how fronto-basal ganglia pathways are commonly involved in proactive and reactive inhibitory control, with a longer pathway (DLPFC-caudate-IFG-SMA-STN-M1) playing a modulatory role in proactive inhibitory control, and a shorter pathway (IFG-SMA-STN-M1) involved in reactive inhibition. These results provide causal evidence for the roles of indirect and hyperdirect pathways in mediating proactive and reactive inhibitory control.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-222 超低強度運動とカロテノイドの併用効果で高まる海馬認知機能と分子基盤 Hideaki Soya（征矢英昭），Jang Soo Yook（陸暲洙）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(ARIHHP) Regular exercise and dietary supplements with antioxidants each have the potential to improve cognitive function and attenuate cognitive decline. In some cases, they induce combination effects; however, additive and/or synergistic interactions and their molecular substrates remain unclear. Our current results reveal that mild exercise (ME) and astaxanthin (AX), a naturally occurring antioxidant carotenoid, have a neurogenic benefit on hippocampal neurogenesis and spatial memory function. Here we have found a strong synergistic effect of ME combined with AX potentiating hippocampus-based plasticity and cognition, mediated by acting leptin (LEP) in the hippocampus. In assessing the combined effects upon wild-type (WT; C57BL/6J) mice undergoing ME with or without AX for 4 weeks, we found that, when administrated alone, ME and AX each separately enhanced spatial memory and hippocampal neurogenesis, and their combination resulted in a substantial synergistic effect. Our subsequent omics analysis, including DNA microarray and IPA analysis, revealed that the up-regulation of LEP gene expression in the hippocampus of WT mice with ME alone was further increased when ME was combined with AX. Together, ME and AX have also increased hippocampal LEP protein levels and its downstream signaling (AKT/STAT3). In LEP-deficient (ob/ob) mice, synergistic enhancement by combined ME and AX was restricted, but chronic intracerebroventricular injections of LEP into the lateral ventricles restored the synergy. Collectively, our findings suggest that hippocampal LEP, but also exogenous LEP, mediates the effects of ME on hippocampal functions underlying spatial memory and neurogenesis, and that this effect is further enhanced by the antioxidant carotenoid, AX.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-223 体操選手と一般健常者の感覚運動野における運動イメージ中の事象関連脱同期の差異 Hirotaka Sugino（杉野広尭）¹，Junichi Ushiyama（牛山潤一）^1,2，Hisato Toriyama（鳥山央人）³ ¹慶應大環境情報 ²慶應大医リハビリ ³慶應大院政策・メディア Introduction: To clarify the neural basis of gymnasts' high motor imagery ability, this study examined difference in sensorimotor rhythm during motor imagery between gymnasts and non-gymnasts. Methods: 18 participants (9 gymnasts and 9 non-gymnasts) repeated motor execution and motor imagery, respectively, for isometric wrist dorsiflexion and shoulder abduction of dominant hand at 20% of maximal effort. The scalp electroencephalograms (EEG) over the sensorimotor cortex were recorded during the tasks. During motor execution and motor imagery, EEG power is known to decrease in alpha- (8-15 Hz) and beta-band (16-35 Hz) around the sensorimotor area, which is referred to as event-related desynchronization (ERD). We calculated the maximal peak of ERD (ERDmax) as a measure of corticospinal excitability both for sensorimotor areas contralateral and ipsilateral to the contracted/imagined arm. Results: Gymnasts showed significantly greater contralateral ERDmax than non-gymnasts in shoulder abduction imagery (p=0.005) but not in wrist-dorsiflexion imagery. Furthermore, ipsilateral ERDmax was significantly greater in gymnasts than in non-gymnasts during motor execution (p= 0.006), although its difference was slightly less than significant level during motor imagery (p=0.113). Discussion: These findings suggest that gymnasts have high motor imagery ability especially in the proximal muscle of upper limbs. We daily use distal muscles for accurate movements such as writing, tipping and so on. Thus, motor imagery ability of the distal muscle would be equivalent between two groups. However, gymnasts use proximal muscles uniquely, especially for postal control against gravity. Such specialized-use in the proximal muscle might lead to significantly higher ERD in gymnasts. Moreover, greater ipsilateral ERDmax in gymnasts suggests that gymnasts' cortex controls their proximal muscles bilaterally to accomplish their specialized performance. In conclusion, the present study suggests that gymnasts control their proximal muscles of dominant arm by bilateral cortex, which would be the basis of their high motor imagery ability.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-224 ニューロフィードバック訓練効果を決定するワーキングメモリモダリティ優位性の個人差 Mayuko Matsumoto（松本万由子）^1,2，Takeshi Sakurada（櫻田武）^3,4，Shin-ichiro Yamamoto（山本紳一郎）¹ ¹芝浦工大システム理工 ²自治医大脳機能研究部門 ³立命館大理工 ⁴自治医大脳外科 Neurofeedback (NF) is a useful neuromodulation approach to improve brain functions such as cognitive or motor performance. However, the effect of NF training widely varies among individuals. Our previous findings suggest that bilateral dorsolateral prefrontal cortex (DLPFC) reflects the individual modality dominance of working memory. In this study, based on these findings, we aimed to investigate the individual differences in the effect of NF training based on DLPFC activities. First, forty healthy adults performed a searching task where the participants had to find and memorize 6-target locations on a drawing tablet as quickly as possible by hand (Pre-sequential task). When their hand came close to a target location, either vibration stimuli on their hand (tactile condition) or visual stimuli on a monitor was presented (visual condition). To evaluate the individual modality dominance of working memory, we compared searching costs between both conditions. Subsequently, participants performed NF training task to increase the bilateral DLPFC activities. We randomly assigned participants into Real group (20 participants) and Sham group (20 participants). In the Real group, participants monitored visual feedback based on the own DLPFC activities. On the other hand, in the Sham group, participants monitored visual feedback based on the pre-recorded other's DLPFC activities. Finally, participants performed the searching task again (Post-sequential task). In the pre-sequential task, whereas 28 participants showed lower searching cost under the visual condition (visual-dominant), the other 12 participants showed the opposite trend (tactile-dominant). Tactile-dominant individuals in the Real group showed that strength of connectivity between bilateral DLPFC significantly increased through NF task. In the post-sequential task, we found that individual's working memory capacity in non-dominant modality improved, and this cognitive improvement was strongly observed in the Real group compared to the Sham group. These results suggest that the individual modality dominance of working memory characterized by the bilateral DLPFC activities is one of the influence factors to determine the NF training effect. The relationship between neural modulation in the bilateral DLPFC and motor improvement can contribute to developing a new tailor-made neurorehabilitation program.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-225 運動中における個人の最適注意戦略を決定づける低次感覚野応答特性 Takeshi Sakurada（櫻田武）^1,2，Masataka Yoshida（吉田将貴）³，Kiyoshi Nagai（永井清）¹ ¹立命館大理工 ²自治医大医脳外 ³立命館大院理工 Focus of attention is an influential factor for improving motor performance. We recently have found that directing attention to movement outcome (external focus; EF) did not always lead to a better motor performance compared to directing attention to body movement (internal focus; IF) in healthy and stroke populations. In a neuroimaging study, although we reported that the frontoparietal network is one of the neural basis of the individual optimal attentional strategy, it remains unclear whether sensory cortexes also reflect the individual differences. In this study, we aimed to explore the responses in the early sensory cortexes depended on the individual optimal attentional strategy by recording steady-state somatosensory/visual evoked potentials (SSSEP/SSVEP). Firstly, twenty-five participants performed a visuomotor learning task under the IF and EF conditions. Fourteen participants showed higher motor learning effect in the EF condition (EF-dominant), whereas the others showed the opposite trend (IF-dominant). Subsequently, we recorded the SSSEP from the somatosensory cortex during presenting vibration stimuli on the fingertips and the SSVEP from the visual cortex during presenting flickering visual stimuli. When the IF-dominant individuals actively directed their attention to vibration stimuli the amplitude of SSSEP got stronger. On the other hand, the amplitude modulation of SSSEP was not observed in the EF-dominant individuals. Regarding the SSVEP, there was no difference in amplitude modulation between IF- and EF-individuals. These results suggest that somatosensory cortex is one of the neural basis characterizing the individual optimal attentional strategy. Furthermore, the top-down modulation of the SSSEP amplitude implies that the neural activities associated with the individual attentional optimality reflect the cognitive ability of attention control rather than response characteristic of sensory processing to focused stimuli.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-226 Single unit activities in the marmoset parietal cortex during a saccadic task Liya Ma（Ma Liya），Janahan Selvanayagam（Selvanayagam Janahan），Lauren K.H. Schaeffer（Schaeffer Lauren K.H.），Kevin Johnston（Johnston Kevin），Stefan Everling（Everling Stefan） University of Western Ontario Abnormal saccadic eye movements are characteristic for patients with several psychiatric and neurological disorders, but are either difficult or impossible to study in non-primate animal models. The common marmoset (Callithrix jacchus) is a promising nonhuman primate model with a lissencephalic brain, which allows for easy and accurate targeting of brain regions that are hidden in sulci in the brains of macaque monkeys. Two trained marmosets demonstrated a gap effect commonly observed in humans, which is the reduction of saccadic reaction times (SRTs) conferred by the time `gap' between the disappearance of the fixation spot and the onset of a peripheral saccade target (Johnston et al., 2018). We implanted 32-channel microelectrode arrays (Utah array) in the posterior parietal cortex of both animals and recorded both spiking activities and local field potentials during this task. During this task, Gap trials (200ms between fixation spot offset and stimulus onset) and Step trials (no latency between the two events) were randomly interleaved. Among the 361 cells recorded from two marmosets, we found 56 gap-modulated cells (15.5%), the spiking activities of which changed significantly from the pre-gap fixation period (200 to 0ms before gap) to the gap period (-150ms to +50ms around stimulus onset). In trials where the stimulus was presented contralaterally to the neurons recorded, activities of gap-modulated cells were predictive of the subsequent SRTs, and whether the animals were about to make an express (SRT<=104ms) or regular (SRT>104ms) saccade. We also found 30% of all cells in Gap trials and 25% in Step trials that displayed a significant visual response 70-120ms after stimulus onset. When combined, these cells accounted for 40% of the population and their visual responses were predictive of the SRTs on all trial types.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-227 運動野から感覚野へのフィードバック信号による可塑性の向上 Ryuta Sakamoto（坂本隆太）¹，Rikiya Kameno（亀野力哉）²，Osamu Hoshino（星野修）^1,2 ¹茨城大院理工機械システム ²（財）脳神経疾患研究所付属総合南東北病院 Motor learning, which in general results from training, refers to the ability of animals to acquire neuronal representations that tailor their movements. The primary motor cortex (M1) that sends its projection to spinal motoneurons is known to control movements, and the M1 has considerable plasticity in neuronal circuitry. Motor learning modifies neuronal connections according to activity-dependent synaptic plasticity, which might be a candidate substrate for map reorganization in M1. This synaptic reorganization translates into a change in neural activity and results in refinement of movements. To elucidate how the motor cortical plasticity contributes to improving the motor function, we simulated a neural network model. In the network model, a sensory cortical network (NS) and a motor cortical network (NM) are reciprocally connected, and the NM sends projections to spinal motoneurons (Mns). During motor learning a sensory stimulus was presented to the NS and synaptic weights between NM pyramidal cells were modified according to a classic spike-timing-dependent plasticity (STDP) paradigm. The STDP strengthened recurrent excitation between NM pyramidal cells, which strongly activated Mns and thus shortened their reaction time to the sensory stimulus learned. The primary motor cortex is a dynamic modulator of the primary sensory cortex (somatosensation) during not only movement but also non-movement. Reciprocal connections between sensory and motor cortices infer intimate interactions between them. We implemented additional simulation in which the intensity of motor to sensory cortex signaling was changed by increasing or decreasing synaptic connection weights from NM to NS pyramidal cells. An increase in feedback signaling increased NS pyramidal cell activity, thereby strongly activating NM pyramidal cell activity and therefore effectively strengthening the synaptic connection weights. We suggest that motor to sensory cortex feedback signaling could be a key factor for enhancing motor cortical plasticity in motor learning.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-228 ラット大脳皮質における感覚・運動関連情報の分布 Masanori Kawabata（川端政則），Satoshi Nonomura（野々村聡），Tomohiko Yoshizawa（吉澤知彦），Alain Antonio Rios（リオスアントニオアライン），Tomomi Sakairi（坂入朋美），Yutaka Sakai（酒井裕），Yoshikazu Isomura（礒村宜和）玉川大脳研 The sensory cortex represents information coming from the sensory organs, and the motor cortex represents information to control muscle movements. When animals execute an action in response to a stimulus, sensory information must be converted to motor information in the brain. As a first step to elucidate where and how the sensori-motor conversion occurs, we investigated the distribution of sensory/motor-related information in the cerebral cortex. We performed a multi-channel recording from different cortical areas of head-fixed rats during task performance. In this task, when the rats pushed a right spout-lever and kept it for a varying hold time, a weak visual stimulus was presented on either right or left display in front of them. If the rats responded to the stimulus and pulled the lever quickly, they got a reward with a delay time. However, if they failed to respond, a second, strong stimulus was presented 1,200 milliseconds later. Then, they got a reward if they responded to the second stimulus by pulling it quickly. The weak stimulus intensity was adjusted around the perceptual threshold (50% in response rate) in each rat. The local field potential and spike activity were analyzed in relation with the stimulus intensity and behavioral reaction time. We found that motor-related information already appeared in higher-order (secondary) visual cortex (V2) as well as the posterior parietal cortex and motor cortex. Importantly, primary visual cortex (V1) neurons showed a visual response without behavioral motor response. These results suggest that motor information converted from sensory one can be processed in V2 but not V1 area.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-229 ショウジョウバエを用いた方向検知とそれに基づく行動における中心複合体の機能解析 Ryo Murakami（村上僚）¹，Masumi Akiba（秋葉真澄）¹，Kentaro Sugimoto（杉本健太郎）²，Taro Ueno（上野太郎）³，Junji Yamauchi（山内淳司）¹，Takako Morimoto（森本高子）¹ ¹東京薬科大学 ²東京工業大学院 ³東邦大学 Vision is extremely important for most organisms. Among visual parameters, motion is one of essential cues to identify the state of objects. Detection of motion direction is important to recognize whether the moving objects, for example, moving predators are approaching or going out. The neural mechanisms underlying motion detection and the behavior which is thereby induced have been extensively studied in mammals as well as insects although there are many unsolved questions. In our laboratory, we have been studying optomotor response (OMR) in Drosophila melanogaster. OMR is the turning response to the direction of moving objects, which is observed in most sight-reliant animals including mammals and insects. In the last Neuroscience meeting, we provided evidence that fly OMR can be modulated by dopamine and identified the central complex area in the fly brain as a target for dopaminergic modulation of OMR. Particularly, the dopamine receptor, D1R2, expressed in the special area named fan-shaped body (FB), in the central complex, was involved in the OMR. Here, we examined the necessity of the neural activity of this brain area in OMR. OMR was disappeared when the neural activity was inhibited in FB by using the region-specific and temperature-sensitive expression system (Gal4-Gal80^ts-UAS system) and the neural silencing protein (Kir potassium channel mutant). Moreover, when monitoring the walking behavior on a small floating ball of a fly suppressed with the neural activity of FB, walking distances were remarkably decreased and the time without the movement (silent time) was dramatically increased compared to that of the wild type. On the other hand, there seemed to be no problem in the speed during movement, suggesting that the fly can move normally if they start moving. Also their survival ability was normal since they can generate offspring normally. These results suggest the possibilities that flies suppressed with the neural activity of FB are defective in detecting the moving direction and/or they are defective in promoting moving, rather than moving itself. We further examine the function of FB in OMR and other behavior, which will reveal the neural mechanisms how OMR is promoted.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-230 音楽聴取、イメージ、演奏時の脳波：声楽とピアノ Shoji Tanaka（田中昌司）上智大学情報理工 Music performance requires highly integrated cognitive and emotional control. However, how the brain (regions and networks) contribute to such control is unknown. This study aims to characterize brain activity and the dynamics of brain networks during music performance as well as listening to music and music imagery. Electroencephalograms (EEG) were recorded during music performance, listening to music, and music imagery, using a 32-channel wireless EEG device (g.tec, Austria). For example, vocalists were singing an opera aria, such as Va! Laisse couler mes larmes from Opera Werther by Massenet. Pianists were playing a piece of classical piano music, such as Chopin Nocturne Op.62 No.1. The acquired data were analyzed using MATLAB, EEGLAB, and LORETA to estimate power spectral density, coherence, time-frequency characteristics, and source localization. To characterize EEG during singing, this study conducted EEG recording also during imagined singing, watching a video clip, listening to music, and resting. The power spectral density profile during singing is characterized to be lower theta/alpha powers, relatively higher beta/gamma powers, and highest delta-band power at the most of the electrodes. The source localization analysis showed higher activities in the prefrontal cortex and visual cortex during the most of the periods of singing. The coherence analysis showed higher coherence between electrode pairs in the central and parietal brain regions, suggesting that singing requires intimate network communication in the sensorimotor and parietal regions of the brain. The recorded EEG reflects cognitive and emotional control required for singing an aria. The power spectral density profile during piano performance differed depending on the degree of mastery of the music. In conclusion, several features of EEG characterize brain dynamics during music performance as well as listening to music and music imagery. The analysis enables the interpretation of neural processing of music.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-231 急峻な断続的等尺性随意筋収縮による初期運動誤差修正と皮質-筋コヒーレンスの関係性 Rina Suzuki（鈴木里奈）¹，Junichi Ushiyama（牛山潤一）^1,2 ¹慶應大環境情報 ²慶應大医リハビリ Introduction: Our motor performance is believed to be controlled under feedforward and feedback loop of sensorimotor system. Despite its popularity, its physiological mechanism is unclear. The present study evaluated corticomuscular coherence (CMC) to examine how the inaccuracy of feedforward control, such as initial movement error, is corrected by feedback control via sensorimotor integration. Regarding to signal-dependent noise theory, it was hypothesised that the magnitude of initial error associates with CMC. Methods: 17 healthy young adults joined in this study. We recorded electroencephalograms over the sensorimotor cortex and electromyogram of the tibialis anterior muscle. They performed two motor tasks by isometric ankle dorsiflexion at 10% or 15% of their maximal voluntary contraction (MVC) along the visual feedback. For sustained task, they kept static contraction for 70 s. For intermittent task, they performed 6 s contraction by ballistic torque development for 60 trials by randomised intensities. For analyses, maximal CMC magnitude (CMCmax) and torque coefficient of variation (CV) were calculated. Results: For sustained task, there was no significant difference for CMCmax (t = -0.961, p = 0.361) or torque CV (t = 1.144, p = 0.282) between intensities which supports the previous studies. For intermittent task, notably, significant differences for CMCmax (t = 2.934, p = 0.017) and torque CV (t = 4.117, p < 0.01) were revealed between intensities. Likewise, when the mean initial error relative to the target intensity was compared, 10% MVC showed more deviation (t = 4.092, p < 0.01) and variation (t = 3.588, p < 0.01) of the exerted torque. Discussion: Notably, different results between two tasks were found from the present findings. Indeed, significant difference in CMCmax was observed only in intermittent task between intensities. This could be due to the initial error emerged by ballistic contraction. Variation of initial error and torque CV could be taken as indicators for the necessity of feedback control. For 10% MVC of intermittent task, unstable feedforward control might affected enhanced CMC for sensorimotor integration. As a result of neural coupling between the motor cortex and the muscle, torque CV could be increased. Therefore, performance under unstable feedforward control may require more feedback control for error correction, resulting enhanced synchrony between the motor cortex and the muscle.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-232 サル一次体性感覚野における運動・感覚情報の混合表現. Kei Mochizuki（望月圭）¹，Katsumi Nakajima（中陦克己）²，Masahiko Inase（稲瀬正彦）¹，Akira Murata（村田哲）¹ ¹近畿大医生理 ²岩手医大医生理 Traditionally, neurons in primary somatosensory cortex (SI) have been thought to represent raw afferent signals from peripheral somatosensory neurons, thus acting as a primal entrance of somatosensory information to the cerebral cortex. However, it is also known that tactile response of SI neurons is modulated by motor execution, suggesting the influence of corollary discharge from motor cortices. In the present study, we recorded the activity of primate SI neurons during tactile self-stimulation. The monkey was required to push and pull a lever manipulandum while a brush moving synchronously to the lever stimulated its opposite hand. We compared tactile responses of SI neurons between two conditions of stimulation. In self-stimulation condition, the monkey stimulated its own hand by lever manipulation. In passive-stimulation condition, the same tactile stimulation recorded during self-stimulation condition was provided by automatic movement of the brush as a control. To examine hidden factors influencing the neuronal activity, we used a dimensionality reduction technique ("neuronal state space"). As a result, six principal components (PCs) could explain about 90% of the original variance of the SI neuron activity. Of those, the first three components were almost identical between self-stimulation and control passive-stimulation conditions, meaning that these components were mainly related to the approximately similar transient response to tactile stimulation, regardless of how the stimulus was provided. On the other hand, the 4th--6th PCs were considerably different between self- and passive-stimulation conditions. The 4th PC showed comparable transient rise of activation at tactile stimulus onset in both conditions, but then gradually started to diverge when that stimulus was resulted from animal's own lever manipulation. The 5th and 6th PCs showed difference between two conditions even before the stimulation onset, suggesting that they carry information about preparation and execution of lever manipulation before the brush stimulation. These results indicate that, although SI neurons strongly represent somatosensory input, it also conveys ongoing motor information in a high dimensional network dynamics.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-233 新奇環境におけるラット迷走神経の発火活動パターンの解析 Toya Okonogi（小此木闘也）¹，Yu Shikano（鹿野悠）¹，Yuya Nishimura（西村侑也）¹，Akihiro Yamanaka（山中章弘）²，Yuji Ikegaya（池谷裕二）¹，Takuya Sasaki（佐々木拓哉）^1,3 ¹東京大院薬薬品作用 ²名古屋大環医研　神経系２ ³JSTさきがけ The vagus nerve serves as a central pathway for communication between the central and peripheral organs. Despite traditional knowledge of vagus nerve functions, detailed neurophysiological dynamics of the vagus nerve in naive behavior remain to be understood. In this study, we developed a new method to record spiking patterns from the cervical vagus nerve while simultaneously monitoring central and peripheral organ bioelectrical signals in a freely moving rat. When the rats transiently elevated locomotor activity, the frequency of vagus nerve spikes was correspondingly increased, and this activity was retained for several seconds after the increase in running speed terminated. Spike patterns of the vagus nerve were not robustly associated with which arms the animals entered on an elevated plus maze. During sniffing behavior, vagus nerve spikes were nearly absent. During stopping, the vagus nerve spike patterns differed considerably depending on external contexts and peripheral activity states associated with cortical arousal levels. Stimulation of the vagus nerve altered rat's running speed and cortical arousal states depending on running speed at the instant of stimulation. These observations are a new step for uncovering the physiological dynamics of the vagus nerve modulating the visceral organs such as cardiovascular, respiratory, and gastrointestinal systems. In addition, we are now constructing a new experimental system to manipulate the vagus nerve using a transgenic mouse line in which channelrhodopsin2, a photosensitive protein, is specifically expressed in the vagus nerve. We present a preliminary electrophysiological data obtained from photostimulation to the nucleus ambiguous, the origin of vagal efferent fibers.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-234 後肢刺激時の動脈圧反応：反対側の外側腕傍核とKölliker-Fuse 核の関与 Hana Nozawa（野澤羽奈）^1,2，Rie Shimoju（下重里江）^1,3，Takamichi Taniguchi（谷口敬道）^1,2，Hideshi Shibata（柴田秀史）⁴，Mieko Kurosawa（黒澤美枝子）^1,5 ¹国際医療福祉大学大学院医療福祉学研究科保健医療学専攻 ²国際医療福祉大学保健医療学部作業療法学科 ³国際医療福祉大学保健医療学部理学療法学科 ⁴東京農工大学大学院農学研究院 ⁵国際医療福祉大学基礎医学研究センター Noxious mechanical stimulation (pinching) of the hindpaw reflexly increases arterial pressure in anesthetized rats. The pressor reflex is mediated via the supraspinal structure; however the precise brain mechanisms have not been elucidated. Since approximately 95% of the projection neurons from spinal lamina I terminate in the contralateral side of the lateral parabrachial nucleus (LPBN) (Todd, 2010), the present study firstly aimed to clarify the involvement of the LPBN in the pressor reflex responses to unilateral pinching of the hindpaw in anesthetized rats. Secondly, we also investigated the involvement of the Kölliker-Fuse nucleus (KF), a part of the parabrachial complex, since the KF provides descending projections to the rostral ventrolateral medulla and the intermediolateral cell column. Arterial pressure was recorded via a catheter inserted into the carotid artery. Muscimol, a widely used neuronal inhibitor, was nanoinjected into the unilateral LPBN or KF. Pinching was applied with a surgical clamp at a force of 3-5 kg to the hindpaw for 20 s. The administration of muscimol into the LPBN significantly attenuated the pressor responses to pinching of the hindpaw contralateral to the site of muscimol injection. Ipsilateral pinching also attenuated the pressor responses although the attenuation was significantly smaller, compared to the contralateral pinching. The administration of muscimol into the LPBN had no influence on the tonic arterial pressure. The effects of muscimol injected into the KF on the tonic pressure as well as the pressor responses to pinching of the hindpaw were similar to those observed by the injection into the LPBN. The present results demonstrate that the LPBN and the KF are both involved in the pressor reflex responses elicited by pinching of the hindpaw, and that the degree of their involvement was dependent on the laterality of stimulation.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-235 ラットの外側手綱核電気刺激に対する心臓血管応答の検討 Yuma Sato（佐藤優真）^1,2，Tri Huu Doan（ゾアンフュチー）^1,3，Masayuki Matsumoto（松本正幸）¹，Tadachika Koganezawa（小金澤禎史）¹ ¹筑波大　医学医療系　生理 ²筑波大院　人間総合科学　フロンティア医科学 ³筑波大院　人間総合科学　生命システム医学 The habenula is a pair of small nuclei which is located above the thalamus and divided into medial and lateral (LHb). It is known that an exogenous stress event activates neurons in the LHb. The stress event also induces various autonomic physiological changes such as changes of blood pressure and heart rate. However, it is little known the functional roles of neurons in the LHb into the cardiovascular responses to the stress event. To better understand the involvement of neurons in the LHb into the cardiovascular responses to stress events, here we tried to observe the autonomic cardiovascular responses induced by electrical stimulation of the LHb. In this study, Wistar male rats were used with introduction of anesthesia by urethane (1 g/kg, i.p.). Arterial pressure was recorded from the femoral artery via a catheter. Heart rate was analyzed from R-R intervals of the electrocardiograph. The LHb was electrically stimulated with a coaxial electrode (300 µ A, 0.5 ms duration, 100 Hz, for 10 s). As a result, electrical stimulation of the LHb significantly increased the mean arterial pressure by 27.9 ± 7.4 mmHg (n = 4, p < 0.05, 35.5 ± 9.2 % vs baseline). Electrical stimulation of the LHb also significantly decreased heart rate by 16.5 ± 2.4 bpm (n = 4, p < 0.05, 4.8 ± 0.7 % vs baseline). The onset latencies of the rise of blood pressure and the fall of heart rate were 4.5 ± 0.5 s and 0.7 ± 0.1 s, respectively. The onset latency of heart rate was significantly earlier than that of blood pressure by 3.6 ± 0.5 s (p < 0.05). These data suggested that the excitation of neurons in the LHb produced the pressor response and bradycardia with directly controlling the autonomic nervous system because the bradycardia was not induced by baroreflex which is triggered by the pressor response. The LHb stimulation-induced cardiovascular changes might be a mimic autonomic cardiovascular response under stress environments.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-236 5-HT1A受容体のCdk5によるリン酸化 Miyuki Takahashi（高橋美由紀）¹，Yuki Kobayashi（小林勇喜）²，Kanae Ando（安藤香奈絵）¹，Yumiko Saito（斎藤祐見子）²，Shin-ichi Hisanaga（久永眞市）¹ ¹首都大学東京　理学研究科 ²広島大学　大学院総合科学研究科 Mental disorders including depression are one of urgent issues to be addressed. To prevent the onset and develop the treatment, it is important to understand a mechanism of the diseases at molecular level. Serotonin (5HT) is a neurotransmitter involved in releasing anxiety and depression. Serotonin 1A receptor (5HT1AR) is a subtype of 5-HT receptors expressed highly in central nervous system and is thought to be involved in psychiatric activity. 5HT1AR is a seven transmembrane G-protein-coupled receptor, which binds to Gi or Go of trimeric G proteins to inhibit adenylyl cyclase or open K+ channels in neurons. Dysfunction of the serotonin signal is considered as a cause of many mental diseases. It is not fully known, however, how expression or activity of 5HTlAR is regulated. We found three possible Cyclin-dependent kinase 5 (Cdk5)-phosphorylation sites in 5HT1AR. Cdk5 is a neuron-specific membrane-bound Ser/Thr kinase. We examined phosphorylation of 5HT1AR by Cdk5-p35. 5HT1AR was indeed phosphorylated by Cdk5-p35. We constructed non-phosphorylatable Ala mutants at T149A, S245A, or T314A, and examined their phosphorylation. Thr314 was identified as a phosphorylation site in 5HT1AR. Expression level of 5HT1AR was decreased by cotransfection with Cdk5-p35, but not with kinase negative Cdk5-p35, in COS-7 cells. We investigated the effect of decreasing 5HT1AR expression by Cdk5 activity on serotonin signaling by measuring cAMP. Co-expression of Cdk5-p35 suppressed 5HT1AR's inhibitory action on cAMP synthesis. These results suggest that Cdk5-p35 modulates the serotonin signal through phosphorylation-dependent downregulation of 5HT1AR.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-237 脳内オートファジーは体温の恒常性維持に重要である Junnosuke Nakamura（中村準之助），Takuma Aihara（相原拓馬），Ban Sato（佐藤伴），Tomoki Chiba（千葉智樹），Fuminori Tsuruta（鶴田文憲）筑波大院　生命環境科学 The body temperature of mammals is controlled by the hypothalamus in the brain. Hypothalamus also maintains biogenic homeostasis. Interestingly, previous studies have reported that impairment of autophagy in hypothalamus causes abnormality of neuronal circuit formation involved in thermoregulation. Although autophagy is thought to be essential for the maintenance of brain homeostasis, the mechanisms that underlie autophagy-regulated thermoregulations is not well understood. Here, we report that autophagy plays important roles in maintaining body temperature, especially with cold exposure. The body temperature of ATG7^flox/flox: Nestin-Cre (Atg7 cKO) mice is slightly lower than of wild-type (WT) mice in room temperature. Moreover, we found that the body temperature of Atg7 cKO mice is rapidly decreased with chronic cold exposure (4 degrees). In addition, microarray analysis has revealed that the expression level of RNA binding motif protein 3 (RBM3) is increased in Atg7 cKO mice brain. Normally, RBM3 is upregulated in response to cold shock stimulation. This protein has been considered to protect neurons from their toxicity. We found that the expression of RBM3 is increased at hypothalamic arcuate nucleus (ARC) in Atg7 cKO mice. We also found that treatment with bafilomycin, which is an autophagy inhibitor, has little effect on an expression of RBM3 in neurons. On the other hand, RBM3 expression is significantly upregulated by treatment with astrocytes conditioned medium (ACM). Especially, it is likely that the ACM derived from autophagy-inhibiting astrocytes is more effective than the regular ACM. These results suggest that RBM3 expression in neuron could be regulated by the autophagic pathway in astrocytes. These findings suggest that impairment of autophagy in brain disrupts thermoregulation at cold temperature and increase of expression of RBM3 in ARC. Our study may shed light on the novel mechanisms by which RBM3 expression in ARC is a novel candidate linking autophagy defect to thermoregulation.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-238 esports選手の闘争-逃走反応による生理状態変化は対戦相手と強く相関する Ken Watanabe（渡辺謙），Naoki Saijo（西條直樹），Makio Kashino（柏野牧夫） NTTコミュニケーション科学基礎研 It is well known that a fight-or-flight response is generated when one has to fight enemies or quickly escape a threat. In our daily life, it may occur when one player tries to beat another as in fighting sports. However, the relationship between the responses of two opponents remains unclear. In this study, we observed the changes in the physiological states of two opponents who played an esports fighting game. Nine professional esports player participated in this study. In the fighting game, one player controls a game character and fights an opponent in a virtual world. We measured the electrocardiograms (ECGs) of the participants when they were playing the game. First, the participants were measured in a resting state. Next, they played against a computer for 5 minutes. After that, the participant competed with other participants until one player had achieved five wins. A group of three men of differing abilities engaged in a round-robin tournament, and a group of six men with comparable abilities engaged in a round-robin tournament. Heart rate time series were calculated from the ECGs as indices of autonomic nerve activity and the cross-correlation function (CCF) was calculated from two sets of time-series data obtained from a fighting pair. The results for a pair with different abilities showed that the CCF and the change in heart rate of a strong player was smaller than for other pairs, indicating that only the weaker player exhibited the fight-or-flight response. On the other hand, the results for a pair with comparable abilities exhibited a high CCF, indicating that they both induced the fight-or-flight response with a similar temporal pattern. These results suggest that the appearance of the fight-or-flight response is related to one's opponent's ability.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-239 Olanzapine Impairs Whole-Body Insulin Sensitivity via Suppression of Central K-ATP Channel-Mediated Glucose Uptake Chantel Kowalchuk（Kowalchuk Chantel）¹，Laura Castellani（Castellani Laura）¹，Pruntha Kanagasundaram（Kanagasundaram Pruntha）¹，Gary Remington（Remington Gary）^1,2,3，Margaret K Hahn（Hahn Margaret K）^1,2,3 ¹Centre for Addiction and Mental Health, Toronto, Canada ²Institute of Medical Science, University of Toronto, Toronto, Canada ³Department of Psychiatry, University of Toronto, Toronto, Canada Antipsychotics are the cornerstone of treatment for schizophrenia but are associated with weight gain and type 2 diabetes. Recent work suggests antipsychotics can induce insulin resistance independent of weight gain via the central nervous system (CNS). To this point, we published data that olanzapine abolishes the ability of a CNS insulin infusion to restrain hepatic glucose production in rodents, but the mechanism is unknown. The KATP channel is a key metabolic sensor downstream of CNS insulin signaling involved in the maintenance of glucose homeostasis. Thus, we investigated whether olanzapine inhibits CNS KATP channel activation to disrupt glucose metabolism. Pancreatic euglycemic clamps were performed in rats. During the clamp, a somatostatin infusion inhibits endogenous insulin secretion and insulin is replaced at basal levels. Glucose is infused at a variable rate to maintain euglycemia, and the glucose infusion rate represents whole body insulin sensitivity. A radioactive tracer infusion allows measurement of glucose uptake and production. Prior to the clamp, rats receive a single subcutaneous injection of olanzapine (OLA) or vehicle (VEH). An ICV infusion of the KATP channel activator Diazoxide (DIA) or VEH was administered throughout. Groups include (ICV-peripheral): VEH-VEH, VEH-OLA, DIA-VEH, DIA-OLA. The glucose infusion rate needed to maintain euglycemia during the clamp was higher in DIA-VEH rats compared to VEH-VEH/VEH-OLA. DIA-OLA rats had a decreased glucose infusion rate compared to DIA-VEH, indicative of impaired whole-body insulin sensitivity. DIA treatment suppressed glucose production, and this was undisturbed by OLA co-treatment (DIA-OLA). Glucose uptake was increased by DIA-VEH, and this effect was abolished by OLA co-administration (DIA-OLA). Thus, olanzapine inhibits CNS KATP channel activation to perturb whole body insulin sensitivity via inhibition of glucose uptake. Combining this with our previous findings that olanzapine impairs central insulin-mediated glucose production, this suggests olanzapine may act through divergent CNS pathways to separately regulate glucose production and uptake. Regardless, inactivation of KATP channels is a novel way that antipsychotics disrupt energy homeostasis. As KATP channels are also involved in CNS interactions of neurotransmitter systems (i.e. dopamine, glutamate), our findings may also have future implications beyond metabolic side-effects of these drugs to effects on psychopathology.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-240 脳海馬が合成する男性・女性ホルモンによる記憶シナプスの急性的制御 Mika Soma（相馬ミカ）^1,2，Mari Ikeda（池田真理）^1,2，Suguru Kawato（川戸佳）^1,2 ¹帝京大薬認知神経科学 ²順天堂大院医泌尿器外科 Sex-steroids are rapidly synthesized (within 20 min) in the hippocampal glutamatergic neurons (Kimoto et al., 2001; Kawato et al., 2002; Hojo et al., 2004, 2009). We investigated new rapid functions of hippocampus-synthesized androgen and estrogen. By using super-resolution confocal microscopy and Spiso-3D image analysis software (Mukai et al., 2011), we demonstrated that testosterone (T), dihydrotestosterone (DHT) , estradiol (E2) and progesterone (PROG) rapidly increased (within 2 h) the hippocampal dendritic spines (= postsynapses) in isolated hippocampal slices. Signaling pathways include synaptic AR, ER, PR receptors, kinase networks (LIMK, MAPK, PKA, PKC), actin-binding proteins (cofilin, cortactin), resulting in spine increase. Selective kinase inhibitors suppressed the spine increase by T, DHT, E2 and PROG. By using multi-electrode electrophysiology, we demonstrated that E2 induced the long-term potentiation (E2-LTP) of the hippocampal neurons. E2 rapidly drives PKA, PKC, and MAPK through synaptic ER. These kinases may phosphorylate NR2B subunit of NMDA receptor, resulting in induction of LTP. Surprisingly, we found that DHT suppressed E2-LTP via activation of phosphatase. Rapid kinase-dependent signaling is non-genomic, and is completely different from slow genomic signaling which include translocation of steroid receptors into nucleus and protein stnthesis. Detailed results are described in (Soma et al., 2018; Hatanaka et al., 2015; Hasegawa et al., 2015; Murakami et al., 2015; Kato et al., 2013, 2017).		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-241 分界条床核CRFニューロンの性差 Katsuya Uchida（内田克哉）¹，Hiroko Otsuka（大塚寛子）¹，Masahiro Morishita（森下雅大）²，Shinji Tsukahara（塚原伸治）²，Tatsuya Sato（佐藤達也）¹，Kenji Sakimura（崎村建司）^1,3，Keiichi Itoi（井樋慶一）¹ ¹東北大学　大学院情報科学研究科　情報生物学分野 ²埼玉大学　理学部　生体制御学科　調節生理学研究室 ³新潟大学　新潟大学脳研究所 The bed nucleus of the stria terminalis (BNST) contains the highest density of corticotropin-releasing factor (CRF)-producing neurons in the brain. CRF-immunoreactive neurons show a female-biased sexual dimorphism in the dorsolateral BNST in the rat. Since CRF neurons cannot be immunostained clearly with available CRF antibodies in the mouse, we used a mouse line, in which modified yellow fluorescent protein (Venus) was inserted to the CRF gene, and the Neo cassette was removed, to examine the morphological characteristics of CRF neurons in the dorsolateral BNST. Developmental changes of CRF neurons were examined from postnatal stages to adulthood. Gonadectomy (GDX) was carried out in adult male and female mice to examine the effects of sex steroids on the number of CRF neurons in the dorsolateral BNST. Most Venus-expressing neurons co-expressed Crf mRNA in the dorsolateral BNST. They constitute a group of neurons without calbindin immunoreactivity, which makes a contrast to the principal nucleus of the BNST that is characterized by calbindin immunostaining. In the dorsolateral BNST, the number of Venus-expressing neurons increased across developmental stages until adulthood. Sexual difference in the number of Venus-expressing neurons was not evident by postnatal day 5. In adulthood, however, there was a significant female predominance in the number of Venus expressing neurons in two subnuclei of the dorsolateral BNST, i.e., the oval nucleus of the BNST (ovBNST) and the anterolateral BNST (alBNST). The number of Venus-expressing neurons was smaller significantly in ovariectomized females compared with proestrous females in either ovBNST or alBNST, and greater significantly in orchiectomized males compared with gonadally intact males in ovBNST. The total number of neurons was also greater significantly in females than in males in ovBNST and alBNST, but it was not affected by GDX.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-242 ゼブラフィッシュ成魚の脳における神経ペプチド発現マップ Towako Kajiyama（梶山十和子），Yoshihiro Yoshihara（吉原良浩）理研CBS システム分子行動学 Various neuropeptides play important roles in modulation of neural activities in the brain. In addition, they are expressed in distinct types of neurons, allowing us to use the neuropeptide mRNAs and/or proteins as molecular markers to identify and distinguish different neuronal populations. In this study, we aimed to construct a comprehensive expression map of neuropeptide genes in the adult zebrafish brain. For the expression mapping, the adult zebrafish brains were sampled, embedded in paraffin, and cut into serial coronal sections of 10 μm thickness. The brain sections were subjected to in situ hybridization using cRNA probes for 36 neuropeptide genes listed below. The images of stained sections were captured by digital slide scanner to develop a comprehensive expression map of these genes. By utilizing this map, we searched for neuropeptide genes that are expressed in neurons along the olfactory pathway activated by sex pheromone stimulation. In the sex pheromone-stimulated adult male zebrafish, we observed the increase of c-Fos expression in aHc (caudal zone of periventricular hypothalamus, anterior part), where several neuropeptide genes were present in different types of neurons and one of them was identified as a gene expressed in the activated neurons. This comprehensive neuropeptide expression atlas will become a useful and fundamental basis for the future studies to dissect functional neural circuitry in the zebrafish brain. Mapped genes agrp, agrp2, cart1, cart2, cart3, cart4, ccka, cckb, crha, crhb, gala, galb, gnih, gnrh2, gnrh3, grp, it, kiss1, kiss2, npb, npy, orx, pdyn, penka, penkb, pmch1, pmch2, pomca, pomcb, sst2, tac1, tac3a, tac3b, vipa, vipb, vt		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-243 PDGF-AA 脳室投与は行動量を抑制することにより肥満を惹起し視床下部背内側核のMEF2C発現を抑制する Atsushi Fukushima（福島篤），Toshiya Funabashi（舩橋利也）聖マリアンナ医大医生理 Platelet-derived growth factor receptor α (PDGFRα) expressing cells in the brain are defined as oligodendrocyte progenitor cells (OPCs). It is suggested that OPCs are not only differentiated to cells formed myelin sheath but also developed to functional neurons. We previously found that intracerebroventricular (icv) infusion with one of endogenous ligands for PDGFRα, PDGF-AA (0.01 ng/0.11 μl/h, by osmotic pump), increased body weight by decreasing night time locomotor activity and O2 consumption. Since we also showed that PDGFRα protein in the hypothalamus was induced by fasting, we hypothesized that PDGF system via PDGFRα in the brain plays a role in metabolic control of the body. In the present study, we first examined what was a candidate gene for PDGF-AA-induced weight gain by the hypothalamus. PDGF-AA was started to icv infusion with osmotic pump at noon. The locomotor activity was significantly decreased at night. Next day, the brain was removed and RNAs were extracted. Expression Array showed this treatment significantly attenuated myocyte enhancer factor 2C (MEF2C) gene expression in the hypothalamus. Western blotting analysis showed that MEF2C was significantly decreased in the hypothalamus by PDGF-AA infusion. Immunohistochemical experiment showed that MEF2C expression in the dorsomedial hypothalamus seemed to be attenuated by PDGF-AA infusion. Semiquantitative analysis with Image-J revealed MEF2C expression in the dorsomedial hypothalamus but not in the lateral amygdala was significantly decreased. Since c-Fos expression in the dorsomedial hypothalamus was also attenuated by PDGF-AA ifusion, we suggested from the present study that PDGF system in the hypothalamus affects the body weight through the hypothalamus.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-244 レム睡眠制御における非選択的陽イオンチャネルNALCNの生理学的役割の解析 Tomoyuki Fujiyama（藤山知之）¹，Seiya Mizuno（水野聖哉）²，Manabu Abe（阿部学）³，Satomi Kanno（管野里美）¹，Miyo Kakizaki（柿崎美代）¹，Kanako Iwasaki（岩崎加奈子）¹，Aya Ikkyu（一久綾）¹，Noriko Hirashima Hotta（堀田平島範子）¹，Mana Yamada（山田麻奈）¹，Chika Miyoshi（三好千香）¹，Makito Sato（佐藤牧人）¹，Takeshi Kanda（上田壮志）¹，Kenji Sakimura（崎村建司）³，Satoru Takahashi（高橋智）²，Hiromasa Funato（船戸弘正）^1,4，Masashi Yanagisawa（柳沢正史）^1,5 ¹筑波大学　国際統合睡眠医科学研究機構　柳沢／船戸研究室 ²筑波大学　生命科学動物資源センター ³新潟大脳研基礎神経科学細胞神経生物 ⁴東邦大医解剖微細形態 ⁵テキサス大学サウスウェスタン医学センター Although sleep is a ubiquitous animal behavior, the molecular/neural basis mechanism of REM sleep (REMS) remains unknown. We performed high-throughput screening of ENU-mutagenized mice in order to identify genes regulating sleep/wake behavior, and established the Dreamless mutant pedigree shows about 50% reduction in 24-h REMS time. We identified a nucleotide change specific to Dreamless mutant mice within the exon9 in Nalcn gene. The single nucleotide substitution leads to a single amino acid substitution (N315K) of the gene product leak cation channel NALCN that we termed "Dreamless". We adopted CRISPR/Cas system to recapitulate Dreamless phenotype, then confirmed that the one base substitution was responsible for REMS abnormality, suggesting that the identified gene is related to the regulation of daily REMS time. To elucidate the molecular/neural basis of REMS regulation by NALCN, we utilized genome editing tools and generated genetically-modified Nalcn mutant bearing flox and FLEx (flip and excision) knock-in mice for loss-of and gain-of-function studies, respectively. In Nalcn-FLEx mice, we observed that the mice with a systemic Cre-expressing line Actb-iCre showed the Dreamless-like phenotype on the electroencephalogram and electromyogram (EEG/EMG) analysis. On the other hand, in Nalcn-flox mice, we confirmed the neuronal subtype-specific deletion of Nalcn mRNA expression in adult brain tissue. Now we are planning to analyze sleep and wakefulness by using these two Nalcn gene mutant mouse lines.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-245 摂食が誘発する睡眠における神経ペプチドの働き Mei Makino（牧野芽以）名古屋大院理生命理学 We spend almost one-third of our lives sleeping. Sleep deprivation contributes to decrease of productivity of an individual as well as of a society. Recent studies show that sleep is evolutionarily conserved from jelly fish, worms, flies to mammals. Yet, why we sleep or how sleep is regulated are still largely unknown. The recent study revealed that neuropeptide orexin regulates feeding and sleep in mice. This study suggests that feeding and sleep are closely linked through the neuropeptides. Therefore, investigation of neuropeptides clarifies the mechanism of feeding-induced sleep. A circadian clock is a main controller of sleep timing. Interestingly, the clock can be entrained by food; if food is given only during the hours when the animals are supposed to sleep, they re-adjust their clock and stay awake during the feeding hours. However, it is difficult to study how feeding regulates sleep in mammals because their brains are so complex. We use C. elegans, a powerful genetic model with a simple nervous system to investigate neuro-molecular mechanisms underlying sleep induced by feeding. The FMRFamid like peptides (FLP) is the neuropeptides that share the common RFamide sequence at their C-termini. Certain FLPs regulate complex behaviors such as locomotion and egg-lying in C. elegans and sleep in both C. elegans and zebrafish. Yet, the functions and the molecular mechanisms of FLPs remain unknown. In this study, an RNAi screen was performed to test if a certain group of neuropeptides FLP play a role in feeding-induced sleep. Among 26 tested flp genes, 10 candidates were found to regulate feeding-induced sleep. The mutant defective in each of the 10 flp genes also showed altered satiety quiescence, validating the results from the RNAi screen. Our results show that the specific FLP neuropeptides play significant roles in feeding-induced sleep in C. elegans.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-246 大脳皮質徐波活動における前障の関与 Kimiya Narikiyo（成清公弥），Momoko Shiozaki（塩﨑桃子），Yoshihiro Yoshihara（吉原良浩）理研CBS システム分子行動学 The claustrum is a subcortical neural structure which has reciprocal connections with wide areas of the neocortex but whose function is poorly understood. We studied physiological relationships between claustrum and neocortex using a transgenic mouse line expressing Cre recombinase specifically in the claustral neurons. We conducted in vivo electrophysiological recordings of the claustral unit activity with simultaneous recording of neocortical electro-encephalogram (EEG) in head-fixed condition and found that a majority of claustral neurons increased their firing activity when slow-wave activity (1-4Hz) appears in the neocortical EEG. To examine the causal relationships between claustral firing and the neocortical slow-wave activity, local field potentials (LFPs) and unit activities were recorded in the frontal cortex which receives massive inputs from the claustrum, while claustral neurons were optogenetically stimulated using Channelrhodopsin-2. The claustral photo-stimulation induced slow-wave activity in the LFP with long-lasting silencing (100 -150ms) of spike activities of frontal cortex neurons across all the layers. Single-unit analysis of cortical neurons revealed that the claustral photo-stimulation selectively drove spike activity in neocortical inhibitory interneurons. The claustral-induced slow-wave was distributed across entire frontal cortex and the distribution pattern resembled that of naturally occurring slow-waves. These results indicate a crucial role of the claustrum in the coordination of neocortical slow-wave activity. In the presentation, we will also discuss the results of ongoing loss-of-function experiments.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-247 ショウジョウバエの睡眠を制御する中心複合体の神経回路の解析 Yoshiaki Kato（加藤善章），Gosuke Ban（坂豪祐），Jun Tomita（冨田淳），Kazuhiko Kume（粂和彦）名古屋市大薬 Sleep is an essential physiological state, which is behaviorally defined, and is widely conserved across species from mammals to invertebrates such as insects. Sleep study using the genetically tractable model organism Drosophila melanogaster has revealed many molecular and neural bases of sleep conserved in both flies and mammals. In the sleep study of Drosophila, the central complex (CX) is one of the most well investigated region. The CX is a prominent brain structure, composed of four interconnected substructures, namely the protocerebral bridge (PB), the fan-shaped body (FB), the ellipsoid body (EB) and the noduli (NO), and plays important roles in various behaviors such as locomotion, flight, visual memory and sleep. We recently made the following two discoveries about sleep regulation by CX. First, a single dopamine neuron innervating the dorsal fan-shaped body of the CX in the Drosophila brain regulates sleep. Second, T1 dopamine neuron, PB inter neuron and PB-FB-NO neuron form a neural circuit and regulate sleep. Furthermore, we explored the output pathways from this PB circuit, and found the lateral accessory lobe (LAL) might be one of them. LAL is a brain neuropil that is anatomically and functionally connected to CX. By using over twenty Gal4 drivers that express in the LAL neurons, we found that several drivers were affected sleep amount. We are now examining the role of LAL neurons in sleep regulation and connectivity to the previously discovered PB circuit.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-248 Orexin and MCH neurons double ablated mice showed severe sleep attack and cataplexy Chi Jung Hung（洪啓栄），Daisuke Ono（小野大輔），Akihiro Yamanaka（山中章弘）名古屋大環境医神経系２ Orexin neurons and melanin-concentrating hormone (MCH) neurons are exclusively distributed in the lateral hypothalamus (LH) and projected to the whole brain to regulate sleep and wakefulness. Reciprocal interaction between orexin and MCH neurons has been reported. Orexin neurons innervate and suppressed MCH neurons. On the other hand, application of MCH peptides inhibited the firing rate in orexin neurons. Nevertheless, it is unclear how this reciprocal interaction functions to regulate sleep and wakefulness. To understand this, we generated new transgenic mice in which both orexin and MCH neurons are ablated by the Tet-off system. These mice express tetracycline-controlled translate activator (tTA) in both orexin and MCH neurons. The absence of doxycycline (DOX), tTA protein binds to tetracycline operator (TetO) and express diphtheria toxin A (DTA). DTA inhibits protein synthesis and induces cell death. Our result indicated that orexin and MCH neurons double ablation mice exacerbated cataplexy. These results suggested that MCH neurons play a suppressive role in cataplexy. In addition to this, these double ablation mice increased wakefulness and decreased non-rapid eye movement (NREM) sleep. Time in REM sleep also decreased in the dark period. Furthermore, a novel sleep episode, delta/theta (DT) sleep, was observed in the double ablated mice. DT sleep is defined as a high density of δ and θ wave in EEG with low EMG. Several behavior experiments suggested a difference between DT sleep and cataplexy. The administration of selective serotonin reuptake inhibitor decreased cataplexy but not DT sleep. The dopamine reuptake inhibitor decreased NREM sleep but not DT sleep. These results suggested a new role of MCH neurons in the regulation of sleep and wakefulness. Conflict of interest: The authors declare no competing financial interests.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-249 ラット海馬スライスにおけるカルバコール誘導振動の日内変化は恒常暗条件により抑制される Masaya Shigemoto（重本昌也），Kiyohisa Natsume（夏目季代久）九工大院生命体工学系 The cholinergic agonist, carbachol, can induce oscillations such as theta (4-13 Hz) and beta (13-30 Hz) oscillations in rat hippocampal slices. Diurnal rhythm under the light and dark cycle (LD cycle) modulated the frequencies of the oscillations. The frequency was significantly decreased at midnight compared with those at the other times. The decrease in frequency will be caused by the inhibition of GABAergic transmission. It is not clarified whether the modulation of oscillations is kept in intrinsic circadian rhythm observed under the constant dark (DD) condition. In this study, we investigated the relationship between carbachol-induced oscillations and circadian rhythm using hippocampal slices made from Wistar rats. We kept 13 rats under DD condition for at least a day after rat's being kept in LD cycle. The rats which was kept under DD condition for a few days were sacrificed. The amount of voluntary activity of rats was measured. Total amount of the activity was significantly increased after the onset of DD condition, especially during the daytime (*p < 0.05; Paired t-test). The peak time of the activity was shifted to later phase. The cosinor analysis showed the increase in the significant probability of the circadian cycle after DD condition. It is suggested that the circadian cycle of the activity can be transiently affected. The circadian cycle of the activity was disrupted in four rats. Then, we made hippocampal slices from 9 rats and applied carbachol to the slice. We measured the frequency of the carbachol-induced oscillations at the circadian time of the recording. As a result, the modulation of the frequency observed in the LD cycle was suppressed. These results suggest that DD condition transiently inhibits the diurnal rhythmic modulation of brain waves in rat hippocampus.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-250 レム睡眠関連脱力発作における扁桃体へのドーパミン放出の影響 Emi Hasegawa（長谷川恵美）¹，Takashi Maejima（前島隆司）²，Takayuki Yoshida（吉田隆行）³，Mitsuhiro Yoshioka（吉岡充弘）³，Masashi Yanagisawa（柳沢正史）^1,4，Michihiro Mieda（三枝理博）²，Takeshi Sakurai（櫻井武）^1,4 ¹筑波大学国際統合睡眠医科学研究機構 ²金沢大学大学医薬保健研究域医学系統合神経生理学 ³北海道大学大学院医学研究院薬理学分野神経薬理学 ⁴筑波大学生命領域学祭研究センター Although the lack of orexin signaling causes the sleep disorder narcolepsy, the precise neural mechanisms by which orexin neurons prevent narcolepsy remain unclear. Mice lacking orexin peptides, orexin neurons, or orexin receptors recapitulate human narcolepsy phenotypes, suggesting a critical role for orexin signaling in the maintenance of wakefulness. In a previous study, we found that targeted restoration of orexin receptor expression in the dorsal raphe nucleus (DR) and in the locus coeruleus (LC) of mice lacking both of orexin receptors inhibited cataplexy and pathological fragmentation of wakefulness (i.e., sleepiness), respectively. These results suggested that DR serotonergic and LC noradrenergic neurons play differential roles in orexin neuron-dependent regulation of sleep/wakefulness. As a next step, we used optogenetic and chemogenetic approaches to demonstrate that DR serotonin neurons suppress cataplexy by reducing the activity of the basolateral/lateral amygdala that plays an important role in emotional processing, as consistent with the fact that strong emotion often triggers cataplexy. Our results suggest that the orexin neuron-DR5HT-amygdala pathway is a critical circuit for preventing cataplexy. We also examined an effect of manipulation of dopaminergic (DA) neurons in the ventral tegmental area (VTA) in narcolepsy phenotype, because cataplexy is usually triggered by emotion with positive valence. Our results suggest a possible role of DA neurons in emergence of cataplexy.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-251 視床下部ニューロテンシン神経は覚醒の促進と体温上昇機能に重要な役割を持つ Fumito Naganuma（長沼史登）¹，Tadaho Nakamura（中村正帆）¹，Ramalingam Vetrivelan（ベトリベランラマリンガム）²，Nobuyuki Okamura（岡村信行）¹ ¹東北医科薬科大学医学部薬理学教室 ²ベスイスラエルディアコネスメディカルセンター、ハーバード大学医学部神経学部門、ボストン、アメリカ Sleep and wakefulness are greatly influenced by various physiological and psychological factors, but the neuronal elements responsible for organizing sleep-wake behavior in response to these factors are largely unknown. The lateral hypothalamus (LH) is considered to be an important brain region for sleep-wake regulation. Actually, LH neurons express several neuropeptides related to sleep-wake regulation, such as orexin and melanin-concentrating hormone. In addition to them, the LH comprises a large population of peptidergic neurons neurochemically defined by the presence of neurotensin (Nts). Previous studies reported that Nts neurons in LH heavily project to dopaminergic neurons in ventral tegmental area (VTA) and microinjection of Nts into the VTA increase locomotor activity. Thus, we hypothesized that Nts neurons may form another excitatory wake promoting cell group within the LH. To test this hypothesis, we studied the functional roles of Nts-LH neurons in sleep-wake, locomotor activity (LMA) and body temperature (Tb) regulations using chemogenetic and optogenetic tools. We found that selective activation of Nts-LH neurons with chemogenetic method produced sustained arousal, higher locomotor activity and hyperthermia. In addition, optogenetic activation of Nts-LH neurons elicited rapid transition from non-rapid eye movement (NREM) sleep to wakefulness. On the other hand, selective chemogenetic inhibition of Nts-LH neurons attenuates the arousal, LMA and Tb responses to a psychological stress (in a novel environment) and augments responses to a physiological stress (fasting condition). These results indicated that Nts-LH neurons are capable of initiating and sustaining wakefulness and increasing Tb and LMA. Considering the involvement of the LH in various physiological functions including sleep-wake, feeding and thermoregulation and close-interrelationship between these functions, our results suggest that Nts-LH neurons could play a crucial role in modulating sleep-wake states, LMA and Tb in response to a variety of physiologic and metabolic demands.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-252 前帯状皮質は学習意欲を司る Kenichi Makino（牧野健一），Yuji Ikegaya（池谷裕二）東京大院薬薬品作用 Willingness to perform a task fluctuates depending on outcome at each step in the task. Successes in the task may maintain the willingness, whereas failures may decrease it. However, it remains unclear which brain area regulates willingness. Inspired by previous human studies (Németh et al., 1988; Parvizi et al., 2013), we focused on the anterior cingulate cortex (ACC) as a candidate brain region that control willingness; we examined whether neuronal activity of the ACC alter willingness to perform the task in the learning process in rats. In this study, we designed a two-hole nose-poke test. In this test, rats were forced to choose one of the two nose-poke holes, which was randomly illuminated in each trial. When rats poked their noses into the non-illuminated hole, they were given a food pellet as a reward. In some trials, however, rats did not poke; we regarded these `omission' trials as an index of willingness, which reflects the demotivational state to perform the task. We then analyzed the transition probabilities between the three outcomes, correct (success) trials, incorrect (failure) trials, and omission trials in consecutive trials. When the ACC was specifically inactivated by local injection of the GABA_A receptor agonist muscimol, the transition probability from incorrect to omission increased, but the probability from correct to omission was not affected. Therefore, ACC activity is required to support the willingness, which otherwise may decrease following failure trials in the task. The ACC inactivation also increased the transition probability from omission to omission. Thus, ACC activity is also required to recover from the sustained demotivational state. Finally, we increased the ACC activity using the DREADD chemogenetic system. The ACC activation decreased the transition probabilities from incorrect to omission and from omission to omission. Thus, ACC activation enhances the willingness. These results suggest that ACC activity reduces the demotivation sensitivity to failure during learning.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-253 ドーパミンニューロンの持続的活動はダイナミックに変動する報酬価値をコードするＹａｗｅｉ Wang（王亜偉）¹，Osamu Toyoshima（豊島理）¹，Jun Kunimatsu（國松淳）²，Hiroshi Yamada（山田洋）²，Masayuki Matsumoto（松本正幸）² ¹筑波大院　人間総合科学研究科 ²筑波大　医学医療系 Imagine that a bartender is pouring a cocktail into your glass. The amount of the cocktail you will obtain is gradually increasing. How does our brain monitor such dynamically changing reward value? Although many cortical and subcortical structures have been identified to encode reward value under an ideal condition in which the reward value does not change with time, it is unclear whether these structures encode dynamically changing reward value that we experience in daily life. To answer this question, here we examined the activity of midbrain dopamine neurons, which have been known to encode stationary reward value by their phasic response, in a monkey during three types of classical conditioning. In one classical conditioning (increase condition), a bar stimulus was presented, and the length of the bar gradually increased (0.082 ml/s) during a random duration (0 - 2,450 ms). The final bar length was corresponding to the amount of a liquid reward that the monkey would obtain at the end of each trial. Thus, the predicted reward value gradually increased as the bar length increased with time. In another conditioning (decrease condition), the length of the bar gradually decreased during the random duration. Thus, the predicted reward value gradually decreased as the bar length decreased with time. In the other conditioning (control condition), a bar was presented with one of three fixed lengths (short, medium or long). Thus, the predicted reward value was fixed at small, medium or large. We recorded the activity of 45 dopamine neurons in the three conditions, and found that a subset of these neurons exhibited a tonic, gradual increase in their activity as the reward value predicted by the bar length gradually increased in the increase condition. To statistically identify this tonic, gradually increase, we calculated the correlation coefficient between the magnitude of activity and elapsed time during bar increase. Seven of the 45 dopamine neurons exhibited a significant positive correlation (P < 0.05). On the other hand, only one neuron exhibited a significant negative correlation between the magnitude of activity and elapsed time during bar decrease (P < 0.05). Our findings suggest that a subset of midbrain dopamine neurons encodes dynamically increasing reward value by a tonic, gradual increase in their activity but are unlikely to track decreasing reward value.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-254 遺伝子改変マウスを用いたマウスの自発行動に関わる遺伝子の制御機構の解析 Naoko Ueda（上田奈央子）^1,2，Ayako Ishii（石井亜矢子）¹，Yuji Imai（今井悠二）¹，Kazuto Yoshimi（吉見一人）¹，Tsuyoshi Koide（小出剛）^1,2 ¹国立遺伝研系統生物研究セマウス開発 ²総研大院生命科学遺伝学 Voluntary physical activity can keep our healthy body weight which prevents many kinds of chronic diseases, as well as keep our brain functions healthy which prevent cognitive decline and learning deficits. Tendency to engage in voluntary activity is heritable but its complex genetic mechanism is not well known. To reveal the genetic mechanism of voluntary activity, Quantitative trait loci (QTL) analysis is one of the most important approaches. In mice, home-cage activity test can be used for measuring voluntary physical activity. In this test, we found that a wild-derived mouse strain MSM/MS (MSM) showed higher activity than a laboratory strain C57BL/6J (B6). QTL analysis of consomic strains, which have one particular chromosome from MSM with B6 background, revealed that genes on several chromosomes are related to regulate home-cage activity. Next, we established congenic strains which have a part of chromosome 6 from MSM with B6 background. By behavioral and statistical analysis of congenic strains, we found three candidate genes: Arl8b and EdemI which may increase activity, and Lrrn1 which may inhibit the effect of the region that increase activity. First we focus on Arl8b which may increase home-cage activity. ARL8B protein is a small G protein which is essential for lysosome motility toward cell periphery. First, by qRT-PCR, we found the expression of Arl8b in cerebellum of MSM is higher than that of B6, suggesting that mouse showing higher activity may have higher expression of Arl8b. Next, we generated Arl8b mutant mice (Arl8b^Δ9), which have deletion of 9 bp in Arl8b coding sequence, and Arl8b knock out (KO) mice. Because mice homozygous for Arl8b mutant (Arl8b^Δ9/Δ9) and Arl8b-KO (Arl8b^-/-) are lethal, we conducted behavioral analysis of Arl8b^+/Δ9 and Arl8b^+/- mice. Although Arl8b^+/- mouse showed normal activity, Arl8b^+/Δ9 mouse showed higher activity than wild type. These results suggest that Arl8b can affect home-cage activity. Also, lethality of Arl8b^Δ9/Δ9 and Arl8b^-/- suggests that Arl8b might affect mice development, and the 9 bp which we deleted might affect the function of ARL8B. We are analyzing the function of Arl8b by focusing on G protein activity.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-255 報酬に基づく意思決定における腹側被蓋野ニューロンの情報処理 Hideyuki Matsumoto（松本英之），Kenji Mizuseki（水関健司）大阪市立大学大学院医学研究科神経生理学 Dopamine involves in various brain functions such as learning, motivation, movement, decision-making and regulation of sleep-wake states. Recent studies indicate that midbrain dopamine neurons convey specific information to distinct downstream brain targets. However, it is still under debate which dopaminergic projection pathways and what spike firing patterns (ex, phasic or tonic) of dopamine neurons are involved in individual brain functions. Furthermore, it remains elusive how spiking activities of dopamine neurons are incorporated into the local network mechanisms during freely-moving behavior. In this study, we recorded multiple single unit activities and local field potentials in the ventral tegmental area (VTA) while rats performed a self-paced, reward-based adaptive decision-making task that allows us to dissociate individual brain functions. Specifically, we manipulated the amount of reward following a left or right choice independently. Thus, the total amount of reward following their choice influences animals' response vigor (motivation to initiate trials) while relative rewards affect their choice. We labeled tyrosine hydroxylase-expressing dopamine neurons with a light sensitive protein channelrhodopsin-2 and identified them based on their responses to optical stimulation while recording. We implanted optical fibers into the VTA, and the ventral striatum and medial prefrontal cortex, the major projection targets of VTA dopamine neurons. This enabled us to optogenetically identify dopamine neurons among VTA neurons, as well as their target brain areas antidromically. We found that VTA neurons exhibited diverse response patterns to task-related parameters, such as not only reward amount, reward history and reward expectation, but also animals' choice and movement. We will discuss how these diverse responses of individual dopamine neurons are correlated with their target brain areas. We will also discuss how spiking activities of VTA neurons are incorporated into the VTA local network activities.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-256 消化管PPAR-alphaはドーパミンニューロンを活性化して運動モチベーションを高める Tetsuya Shiuchi（志内哲也）¹，Keisuke Hashimoto（橋本啓祐）^1,2，Ayaka Hashimoto（橋本采佳）^1,2，Sachiko Chikahisa（近久幸子）¹，Noriyuki Shimizu（清水紀之）¹，Hiroyoshi Sei（勢井宏義）¹ ¹徳島大学大学院医歯薬学研究部　統合生理学分野 ²徳島大学医学部　Student Lab Exercise habit has a benefit for the health of body and mind although it is hard to begin and/or keep up the exercise training for many people, who especially ought to exercise for their health. We found that transient intake of high fat diet increased spontaneous wheel running activity in rodents. In this study, we tried to clarify the mechanism which high fat diet-induced increase of wheel running activity in mice focusing on dopaminergic neuron and peroxisome proliferator-activated receptor (PPAR)-alpha in the gut. Wheel running activity for 2 hours after high fat diet feeding was higher in wild type mice than that of PPAR-alpha knock out (KO) mice. Oral administration of PPAR-alpha agonist (Wy-14643 or fenofibrate) increased wheel running activity in male C57BL/6J mice. This effect was reduced by intracerebroventricular injection of dopamine receptor antagonist and by capsaicin treatment. Moreover, oral PPAR-alpha agonist-induced enhancement of wheel running activity was not inhibited by the intracerebroventricular injection of PPAR-alpha antagonist, GW6471. Furthermore, rescue of PPAR-alpha expression in gut of PPAR-alpha KO mice recovered Wy-14643-induced enhancement in wheel running activity. These results suggest that PPAR-alpha in gut is an important molecule for the regulation of exercise motivation with activation of dopaminergic pathway via vagal nerve in mice.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-257 腹側被蓋野ドパミンニューロンの活動抑制が自然報酬によって強化されたラットの道具的行動の動機づけに与える効果 Yoshio Iguchi（井口善生），Shigeki Kato（加藤成樹），Kayo Nishizawa（西澤佳代），Kazuto Kobayashi（小林和人）福島県立医大医生体情報伝達研生体機能 Neuromodulation by ventral tegmental area (VTA) dopamine (DA) neurons in learning and cognition has been well documented. Previous lesion studies utilizing rodents' instrumental behavior reinforced on a progressive ratio schedule have revealed the role of VTA DA neurons in animals' effort-related motivation to gain access to their reward. However, it was not so easily expected that the combination of the chronic lesion method with the behavioral protocol whose quantitative performance is low is able to provide a measurable basis for comparison and translation between distinct experimental/preclinical settings employing different subject species and rewards. As a result, elucidation of pathophysiology of anhedonia and amotivation, which are shared by several psychiatric diseases as a key symptom and have been implicated in the mesolimbic DA systems, as well as physiology of VTA DA function in generation and regulation of motivated behavior, has been hindered. Present study sought to overcome the challenges by combining a reversible manipulation of DA neuronal activity (in the present report, inhibition) supported by the unique techniques of gene recombination, with estimation of the motivation-related parameters based on the application of an economic demand model to the appetitive instrumental behavior of rats in which "price of reward (required lever presses to gain access to a unit of food)" was systematically increased in a single session. Drd2-Cre rats uniquely developed in our group have got Cre recombinase in VTA DA neurons. We injected the AAV vector into VTA of the Drd2-Cre rats, which induces expression of the anion channel derived from Caenorhabditis elegans (GluCL) in a Cre-dependent manner, and sought to inhibit temporarily the DA neuronal activity by administration of ivermectin, a ligand of GluCL. This reversible inhibition of DA neuronal activity enhanced demand elasticity (α: inverse of motivation) estimated from the within-session analysis of the rats' lever presses, in a ligand dose-dependent manner; however, the second parameter Q0, free consumption, was not found to be affected by the DA neuron inhibition. Utilization of this experimental system with the chemogenetic/optogenetic DA neuronal excitation, or different subject animals, or different rewards in the future would provide the quantitative and flexibly-translatable bases to reveal detailed overview of the function of VTA DA neurons in motivation for reward.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-258 妊娠中の偏った必須脂肪酸摂取はドパミンニューロンの過剰産生を介して仔の嗜好食品摂取を促す Nobuyuki Sakayori（酒寄信幸）^1,2，Oki Higuchi（樋口央紀）^3,4，Ryoji Fukabori（深堀良二）¹，Yoshio Iguchi（井口善生）¹，Susumu Setogawa（瀬戸川将）¹，Takahiro Eitsuka（永塚貴弘）⁴，Teruo Miyazawa（宮澤陽夫）^4,5，Kazuto Kobayashi（小林和人）¹ ¹福島医大医生体機能 ²日本学術振興会 ³機能性植物研究所 ⁴東北大農機能分子 ⁵東北大未来科学技術共同研究センター Regardless of the fact that increasing prevalence of obesity and non-communicable diseases disrupt our civilized society, we remain ignorant of how greediness for food is escalated by recent environmental changes in the world. Omega-6 (n-6) and omega-3 (n-3) polyunsaturated fatty acids (PUFAs) are essential nutrients for normal brain development and function, and its dietary ratio (n-6:n-3) attracts sanitary attention due to a spreading meal style that consumes more seed oils (rich in n-6 PUFAs) and less fish (rich in n-3 PUFAs). Working in mice, we show that maternal consumption of a diet that is rich in n-6 PUFAs and poor in n-3 PUFAs induces consumption of palatable foods in the offspring through excess production of dopamine (DA) neurons in the ventral tegmental area (VTA), the brain core for palatable feeding. Lifelong exposure to the n-6 rich/n-3 poor diet induced the intake of sucrose and fat, despite no difference in the intake of water or standard diets. The pregressive ratio schedule-based operant conditioning analysis further confimed evoked motivation for sucrose reward intake in mice fed the n-6 rich/n-3 poor diet. In vivo microdialysis experiment showed that the DA release in the nucleus accumbens (NAc) of the n-6 rich/n-3 poor diet group was increased by sucrose reward exposure, which correlates with induced intake of sucrose. Consistent with these behavioral and neurochemical results, the number of DA neurons in the VTA innervating the NAc was increased in mice fed the n-6 rich/n-3 poor diet. Finally, such induced palatable feeding and excess DA neurogenesis were induced by exposure to the n-6 rich/n-3 poor diet before birth, but not induced by exposure after birth, indicating that imbalance of dietary PUFAs during pregnancy is critical to induce palatable feeding in offspring. Our findings reveal the novel environmental factor that has irreversible impacts on offspring's feeding behavior for palatable foods.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-259 学習者の興味の枠組に近付けて身心を一体化する科学の学習（3）：心理学/物理化学/認知行動哲学と創発細胞生物学の融合を意図した言葉による理解と「触覚情報」を用いた意識的体幹調整法から学ぶ Yoriko Atomi（跡見順子）¹，Yoshikazu Higashi（東芳一）¹，Miho Shimizu（清水美穂）¹，Kaori Uno（宇野カオリ）²，Hideo Kawamoto（河本英夫）³，Toshiyuki Watanabe（渡邊敏行）⁴，Aya Atomi（跡見綾）¹，Eri Fujita（藤田恵理）¹，Tomoaki Atomi（跡見友章）⁵ ¹東京農工大院工 ²筑波大人間系 ³東洋大文学部哲学科 ⁴東京農工大院工 ⁵杏林大保健学部理学療法 Although neuroscience and life science is progressing, the outcome is not enough contributing to contemporary society problems like the increase of psychiatric diseases. In 2016 Meeting we reported the intention and effects of the innovative trunk muscle contraction introduced by own hands in supine position together with lectures of brain science and cell biology. One-week practice showed a significant improvement of physical performance such as sit-up and standing posture and the will to carry out it. [Program] Expanding the attempt of last year (performed together with expert in psychology) three classes linked with three distinct areas of psychology / physics & chemistry / philosophy were carried out , keeping lectures of body-mind integrative science and core gymnastics are common. We investigated the degree of implementation and understanding. The class was 8 lectures in a row for 2 consecutive days and was the capacity of 43 students in each class. All curricula were composed of lectures with narrative explaining and showing dynamic images of self-emergent cell system using PPT. Trunk exercise was also explained for participants during lectures. Two reporting tasks were imposed for students. [Educational Effect] Students learned themselves as an emergent existence having a body composed of cells, evaluated that they were able to collaborate as entities by actually doing things and verbalization. In other words, by understanding the logical nature of statement memorization, showing the basis of understanding / essence of the mechanism of cooperation in physical relationship, conducting actions that must be connected to the mind as procedural memory. [Discussion] As Courtine reported gait learning is ineffective even in treadmill walking by electric stimulation, and it is inevitable to take a voluntary step with rat's will from standing in a towed state (2012). As rewards of this education program integrating body and mind five items were extracted as follows; (1) presentation of reasons convinced by themselves, (2) ease of task and time, (3) awareness of the effect, (4) himself customizing the correct exercise, activities and actions, and self-evaluate own state of physical condition and health, (5) to be able to convince both himself and his / her body of mind, and to logicize (verbalize) the subject. In this attempt, lectures by trustworthy instructors in different fields increased participant's reward, because it is placed in their interests.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-260 Locomotion changes in male mice with prenatal nicotine exposure Kun-Ruey Shieh（Shieh Kun-Ruey）¹，Shu-Chuan Yang（Yang Shu-Chuan）^2,3，Hsien-Yong Lai（Lai Hsien-Yong）⁴ ¹Department of Physiology, Tzu Chi University, Hualien, Taiwan ²Holistic Education Center, Tzu Chi University of Science and Technology, Hualien, Taiwan ³Institute of Radiological Sciences, Tzu Chi University of Science and Technology, Hualien, Taiwan ⁴Mennonite Christian Hospital, Hualien, Taiwan The nicotine exposure or cigarette smoking during pregnancy is associated with cognitive disabilities in the offspring. The prenatal nicotine exposure through cigarette smoking or even nicotine replacement therapy may directly and/or indirectly cause impairments in fetal and neonatal development. The direct effect of nicotine on the fetal brain is on the nicotinic acetylcholine receptors. Cholinergic signaling enable to regulate the release of dopamine neurotransmitters, and even those neurotransmitters which are involved in the regulation of neurogenesis, neuronal migration, and differentiation. Dopaminergic signaling plays key roles in the regulation of fetal brain development and locomotion. This study examined the locomotor activity in male mice with prenatal nicotine exposure to verify the effects of nicotine in brain development. Male mice with or without prenatal nicotine exposure were used. Using the behavioral indices of to the elevated plus maze and open field tests was done in 3, 6, 9 and 12 weeks old. The data showed that the experimental group compared to the control one had the higher locomotor activity in the open field test in 3, 6, 9 and 12 weeks old. Interestingly, no difference was found in the first trial of elevated plus maze, but significant difference was found in the second one. Behavioral changes were found in male mice when exposed to nicotine during prenatal and early postnatal life. These findings indicate that prenatal nicotine exposure may cause persistent locomotion changes when exposure occurs during a period of rapid brain growth.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-261 母親の接近-回避行動と乳児音声に含まれる感情の関係 Yuki Oishi（大石悠貴）¹，Daiki Hiraoka（平岡大樹）²，Michio Nomura（野村理朗）²，Ryoko Mugitani（麦谷綾子）¹ ¹NTTコミュニケーション科学基礎研 ²京都大 The emotional contents of infants affect communication between infants and mothers. For example, mothers' behavior alters when infants' emotion is changed from laughing to crying. Although many psychological and neural responses of mothers to infant emotion have been reported, few studies have examined maternal approach-avoidance behavior in response to infant emotion from the viewpoint of audition. In this study, we sought to determine how infant emotional vocalization affects maternal approach-avoidance behavior. Twenty mothers participated in this study, all of whom had infants of 24 months old or less. In the behavioral experiment, they stood on a Balance Board that collected real-time data regarding center of pressure (COP), while listening to a series of infant vocalizations including cry (unpleasant), laugh (pleasant), and babbling (neutral) via a speaker. In the self-report experiment, they then listened to the same vocalizations for a second time and rated their felt emotions in response to each vocalization. We found mothers' significant postural movements of approaching in response to infants' cry stimuli or to stimuli regarded as highly urgent. In contrast, they demonstrated postural movement of avoidance in response to infants' laugh vocalization. These findings suggest that maternal behavior in response to infant emotional vocalization is regulated not only by the category of the emotion embedded in infant vocalization but also by the urgency of the stimulus that mothers felt.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-262 Behavioral responses to open field test in male mice with prenatal nicotine exposure Shu-Chuan Yang（Yang Shu-Chuan）^1,2，Kun-Ruey Shieh（Shieh Kun-Ruey）³ ¹Holistic Education Center, Tzu Chi University of Science and Technology, Hualien, Taiwan ²Institute of Radiological Sciences, Tzu Chi University of Science and Technology, Hualien, Taiwan ³Department of Physiology, Tzu Chi University, Hualien, Taiwan The cigarette smoking during pregnancy is able to produce lifelong changes in the offspring brain. Recently cigarette smoking among pregnant women and women of childbearing age has dropped; however, exposure to nicotine during pregnancy via secondhand cigarette smoke, smokeless tobacco, and even nicotine replacement therapy has increased and it might be another serious issue in the world. The behavioral effects in cigarette smoke exposure are mainly from the nicotine on the developing and mature brain. The present study used animal models of prenatal nicotine exposure to support the hypothesis that nicotine may directly and/or indirectly cause impairments in fetal and neonatal development and induce the changes of behavioral responses. This study examined the behavioral responses to the open field test, a common anxious-like behavioral test, in male mice with prenatal nicotine exposure to verify the effects of nicotine in brain development. Male mice with or without prenatal nicotine exposure were used. Using the behavioral indices to the open field test was done in 3, 6, 9 and 12 weeks old. The data showed that the experimental group compared to the control one had the increase movement in the center and periphery arenas in the open field test. Behavioral changes were found at 6 and 9 weeks old, but not at 3 weeks old in male mice when exposed to nicotine during prenatal and early postnatal life. These findings indicate that prenatal nicotine exposure may cause persistent behavioral changes when exposure occurs during a period of rapid brain growth.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-263 前補足運動野へのtDCS刺激による笑顔認知の促進効果 Yuka Kondo（近藤友香）¹，Joy Hirsch（Hirsch Joy）^2,3,4,5，Yumie Ono（小野弓絵）⁶ ¹明治大院理工電気 ²Dept Psychiatry, Yale Sch of Med, New Haven, USA ³Dept Neurosci, Yale Sch of Med, New Haven, USA ⁴Dept Comp Med, Yale Sch of Med, New Haven, USA ⁵Dept Med Physics Biomed Eng, Univ Coll London, London, UK ⁶明治大理工電気電子生命 Recognizing a facial expression plays an important role in interpersonal communication. We investigated the role of the pre-supplementary motor area (preSMA) in recognizing the facial expression of smile. Although the involvement of preSMA in the recognition of smile expression has been reported in human brain imaging studies using implanted depth electrodes (Krolak-Salmon et al, 2005 ) and fMRI (Hennenlotter et al, 2005), a causal relationship of preSMA activity on the smile recognition ability has not yet well understood (Rochas et al, 2013). Eleven young adult participants (8 men, 3 women, mean and standard deviation 22.4 ± 1.3 years old ) judged the smile expression of randomly presented 169 Ekman face images that were morphed from full smile to no specific facial expressions with 13 different ratios. Two sessions of the smile recognition task were performed with anode (1.5 mA for 10 minutes) or sham transcranial direct current stimulation (tDCS) to the preSMA in between these sessions. The order of anode and sham conditions were randomized and counterbalanced across participants, and experiments of two conditions were separated at least one week to washout the effect of tDCS. We determined a smile recognition threshold for each session, a morph ratio ranged from no facial expression (0%) to full smile (100%) at which participants equally recognized smile or not. Anodal stimulation to preSMA but not sham stimulation decreased smile recognition threshold in more than 70% of the participants (n = 8; from 40.9 to 36.7% in anodal stimulation, from 41.4 to 41.2% in sham stimulation). These results suggest that anodal stimulation of the preSMA facilitates detection of smile expression, possibly through enhanced activity of the mirror neuron system.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-264 マウスにおける退屈様行動の発見 Yosuke Yawata（八幡洋輔），Yuji Ikegaya（池谷裕二）東京大院薬薬品作用 We are bored when we have nothing to do or when we continue to do a very simple task. The psychological definition of boredom is "an aversive state of wanting but being unable, to engage in satisfying activity" (Eastwood et al., 2012). To our knowledge, the neural mechanisms by which we become bored remain unknown. This may be because boredom has been investigated by human studies in which invasive methods can hardly be introduced, including single-neuron recordings and manipulation of neuronal activity. To solve this problem, we sought to establish a valid behavioral paradigm to quantify boredom of mice. In a human study, people do not enjoy spending several minutes in a room by themselves with nothing to do but think (Wilson et al., 2014). Moreover, in monotonous, boring conditions, people shock themselves more often and with a higher intensity, compared to the neutral condition (Havermans et al., 2015; Nederkoorn et al., 2016). We hypothesized that, like human, mice also choose giving themselves aversive stimuli rather than doing nothing. We put a male mouse for 15 min in an empty chamber that contained nothing but a nose-poke hole. When the mouse poked its nose into the hole, it received an air-puff stimulus that was naturally aversive for mice. We defined this nose-poke behavior as a "boredom-like" behavior and used its repeat number as an index of 'boredness'. For control conditions, we used an enriched chamber that was furnished with multiple toys, such as a ladder and a seesaw. We found that mice exhibited more nose-poke behaviors in the empty chamber than in the enriched chamber. In addition, mice in the empty chamber exhibited more jumping behaviors onto walls, which are considered escaping behaviors out of the chamber. These results suggest that mice exhibited more boredom-like behaviors in an impoverished environment and tried to avoid the aversive situation.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-265 暑熱・寒冷環境により活性化する神経回路に関する組織学的解析 Minami Kanai（金井みなみ），Ryota Kamiizawa（上伊澤涼太），Natsuko Hitora-Imamura（人羅 (今村) 菜津子），Hiroshi Nomura（野村洋），Masabumi Minami（南雅文）北海道大学薬学部薬学科　薬理学研究室学士課程5年 Seeking a more confortable ambient temperature is an important behavior conserved in many species to control body temperature. Preference/aversion to ambient temperature is based on pleasant/unpleasant emotion induced by environmental temperature. However, it remains unclear which neural pathways mediate unpleasant emotion induced by hot and cold environments. Using c-Fos mapping, we have revealed that the paraventricular thalamic nucleus (PVT) are activated by a hot environment. In this study, downstream regions of the PVT were examined with focusing on the nucleus accumbens (NAc), bed nucleus of the stria terminalis (BNST) and central nucleus of the amygdala (CeA), which are the brain regions implicated in unpleasant emotion. We injected fluorescent retrograde tracers (2% FluoroGold, 0.1 μL) into these brain regions of male Sprague-Dawley rats to label the neurons projecting to these brain regions. After exposure to a hot (38°C) or cold (8°C) environment or room temperature for 150 min, c-Fos expression was examined by immunohistochemical staining. In the animals exposed to a hot environment, 56.1%, 50.5% and 20.9% of c-Fos positive neurons projected to the NAc, BNST and CeA, respectively. In the animals exposed to cold environment, 40.0%, 48.1% and 24.0% of c-Fos positive neurons projected to the NAc, BNST and CeA, respectively. This study demonstrated that PVT neurons projecting to the NAc, BNST and CeA were activated by the exposure to hot and cold environments. These neural pathways may be involved in the induction of unpleasant emotion by hot and cold environments, and thereby play a role in the seeking behaviors to a more confortable ambient temperature.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-266 δオピオイド受容体を介した大脳皮質興奮性シナプス伝達の調節 Daisuke Yamada（山田大輔）¹，Jun-Ichiro Oka（岡淳一郎）¹，Hiroshi Nagase（長瀬博）²，Akiyoshi Saitoh（斎藤顕宜）¹ ¹東京理科大・薬・薬理 ²筑波大睡眠研究機構 It has been reported that δ-opioid receptor (δOR), one of the types of opioid receptors, regulates emotional behaviors in rodents. We previously reported that local perfusion of KNT-127, a selective agonist for δOR, in the prelimbic subregion of the medial prefrontal cortex (PL-PFC) decreased anxiety-like behavior induced by veratrine, a sodium channel activator, in mice, and simultaneously caused attenuation of veratrine induced increases in extracellular glutamate levels in PL-PFC using a in vivo reverse microdialysis study. These results suggested the possibility that KNT-127 exerted anxiolytic-like effect through modulation of glutamatergic excitatory synaptic transmission via δOR in the PL-PFC. To test this, we investigated in the present study the effect of KNT-127 on excitatory synaptic transmission in the PL-PFC. Male C57BL/6J mice (6-12 weeks old) was used for electrophysiological experiments. Brain slices (300 μm-thick) including PL-PFC were prepared using a vibratome. We performed whole-cell patch-clamp recordings from pyramidal shaped neurons in layer 2/3 of the PL-PFC. Spontaneous excitatory postsynaptic current (sEPSC) and evoked EPSC (eEPSC) induced by electrical stimulation in layer 5 of the PL-PFC were recorded. After baseline recordings of sEPSC and eEPSC in artificial cerebrospinal fluid (aCSF), KNT-127 was perfused for at least 15 min, and then sEPSC and eEPSC were recorded again. As a result, resting membrane potential and passive membrane properties were unchanged by KNT-127 perfusion. The frequency of sEPSC was significantly decreased by application of KNT-127. In addition, paired-pulse ratio of eEPSC was significantly higher in the recordings after KNT-127 application than that in baseline recordings, when the inter-stimulus intervals were set at 25, 50, and 100 ms. These results indicated that the reduction of glutamate release from presynaptic site by activation of the δOR in the PL-PFC in mice.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-267 2型糖尿病モデルラットにおける大脳辺縁系のコレシストキニン陽性ニューロンやパルブアルブミン陽性ニューロンと不安様行動 Ryosuke Ochi（越智亮介）¹，Naoto Fujita（藤田直人）¹，Natsuki Goto（後藤夏季）¹，Hisao Nishijo（西条寿夫）²，Susumu Urakawa（浦川将）¹ ¹広島大院医歯薬運動器機能医科学 ²富山大医システム情動（生理１） Otsuka Long-Evans Tokushima fatty (OLETF) rats exhibit hyperphagia, resulting in type 2 diabetes with obesity, hyperinsulinemia, and lipid metabolism disorders. It has been reported that OLETF rats show anxiety-like behavior in the open field and elevated plus maze test because of the spontaneous defect of the cholecystokinin (CCK)-1 receptor and/or type 2 diabetes. However, the neurophysiological mechanisms of anxiety-like behavior in OLETF rats is not fully understood. In this study, we investigated the anxiety-like behavior and CCK expressing (CCK+) and parvalbumin (PV+) neurons in the limbic system in OLETF rats. Twenty weeks old male OLETF rats and age-matched non-diabetic Long-Evans Tokushima Otsuka (LETO) rats were used. OLETF rats exhibit typical hyperglycemia indicating type 2 diabetes from eighteen weeks of age, so twenty weeks of age in OLETF rats is just after the onset of type 2 diabetes. The open field test was performed for assessment of anxiety-like behavior. Expressions of CCK and PV were assessed by immunohistochemistry in the basolateral amygdaloid complex, medial prefrontal cortex, and hippocampus. In the open field test, the time in the center zone was significantly less in OLETF rats than LETO rats. The densities of CCK+ neurons in the basolateral amygdaloid complex, prelimbic-infralimbic cortex, and hippocampal CA2 were significantly higher in OLETF rats than LETO rats. The density of PV+ neurons in the prelimbic-infralimbic cortex tended to be lower in OLETF rats than LETO rats. Time spent in the center zone in the open field test was negatively or positively correlated with the densities of CCK+ neurons in the basolateral amygdaloid complex and hippocampal CA2 or the density of PV+ neurons in the prelimbic-infralimbic cortex, respectively. We showed for the first time that OLETF rats exhibited alterations of the densities of CCK+ and PV+ neurons in the limbic system, while the mechanisms of these alterations were unclear. Previous studies reported that CCK+ and PV+ basket cells inhibited the activity of pyramidal cells, and the number of PV+ neurons and expression of PV mRNA in the limbic system correlated with anxiety-like behavior. CCK is also a neurotransmitter, which induces increased anxiety-like behavior via activation of the CCK-2 receptor. These results suggest that the alterations of CCK+ and PV+ neurons in the limbic system might involve in increased anxiety-like behavior in OLETF rats.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-268 相互に抑制する扁桃体インターカレーテッド細胞核による恐怖行動発現コントロール Kenta M Hagihara（萩原 M 賢太）¹，Olena Bukalo（ブカロオレーナ）²，Martin Zeller（ツェラーマーティン）³，Ayla Askoy-Aksel（アスコイアクセルアイラ）³，Ingrid Ehrlich（アーリッヒイングリッド）³，Andrew Holmes（ホームズアンドリュー）²，Andreas Luthi（ルティアンドレアス）¹ ¹フリードリッヒ　ミーシャー　インスティチュート ²えぬあいえっち ³ちゅーびんげん大 Excessive fear expression leads to loss of opportunities in many aspects and disadvantages animal survival. Thus, properly suppressing fear memory is as important for survival as the initial formation of a fear memory. Intercalated cells (ITCs) of the amygdala, clusters of small-sized unique inhibitory neurons encapsulating the lateral and basal amygdala, are anatomically well situated to provide inhibition on amygdala circuits underlying fear expression, and thus, have been implicated in fear suppression upon extinction. However, largely due to their small size and lack of genetic markers for targeting, it has been difficult to functionally dissect roles of individual ITC clusters. Here, to address this issue, we employed a multidisciplinary approach including in vivo calcium imaging from freely moving mice engaging a classical auditory fear conditioning and extinction paradigm, opto-/pharmaco-genetic activity manipulations, slice physiology, virus-based circuit tracings, and RNA-sequencing. We found that two transcriptionally distinguishable major clusters of ITCs, dmITC and vmITC oppositely represent fear-eliciting and extinguished auditory stimuli, and that manipulations of the two clusters changed the fear state of animals in opposite directions. We also found that dmITC and vmITC clusters directly and mutually inhibit each other while innervating to the basolateral amygdala (BLA), suggesting that they comprise antagonistic inhibitory control over BLA fear circuit. Furthermore, virus based projection mapping revealed that they target different brain regions. Overall, those results suggest that defined ITC clusters interact with each other to control the fear state of animals by influencing distributed brain-wide computation. The current study provides a new circuit motif where two mutually inhibiting nuclei perform "push-pull" calculation.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-269 音楽鑑賞による脳波および自律神経系への生体作用の電気生理学的研究 Mitsuyuki Ichinose（一ノ瀬充行），Satoshi Yoshikawa（吉川聡）岩手大理工学化学・生命理工学 To investigate physiological effects by listening several kinds of classical music, we measured EEG and the autonomic nervous system by measuring heart rate and heart rate variability. Psychological conditions before and after listening the music were monitored by describing two psychological tests, i.e. the multiple mood scale (MMS) and the General Arousal Checklist (GACL). Subjects, who were repeatedly examined up to 8 times, were young healthy 3 students. The subjects listened the classic music selected by commercial CD. By listening the music in subject #2, amplitude of δ band in frequency analysis (mFFT) of EEG was reduced in the central line (Fz, Cz, Pz) slightly and in lateral areas (C3,C4,P3,P4) significantly. Concerning α band, the amplitude was increased in Cz but in other positions little change was observed. The slope of power spectral density of EEG was reduced in Fz and O1, meaning that after listening the music arousal level was increased. On the other hand, in subject #3, amplitude of δ band was significantly increased in the central line (Fz, Cz, Pz) and in lateral areas (P3, P4, T5, T6). Concerning α band, the amplitude was decreased in the central line (Fz, Cz, Pz) and in lateral areas (C3, C4, P3, P4). The slope of power spectral density of EEG was almost no change in Fz and reduced in O1, meaning that after listening the music arousal level was increased in occipital regain but not in frontal area. These data suggested that the music stimulated nervous activities of subject #2 but reduced those of subject #3.In subject #2, heart rate was increased and the sympathetic nervous activity indicated by LF/HF was increased. On the other hand, in subject #3 heart rate was reduced and the para-sympathetic nervous activity indicated by HF was increased. The result of heart rate and its variability showed that by listening music subject #2 was stimulated the sympathetic nervous activity and subject #3 was stimulated the para-sympathetic nervous activity. In subject #2, liveliness and startle of mind in MMS was increased after listening the music. In subject #3, high activation of tense arousal in GACL was reduced and general deactivation was stimulated by the music. These data suggested that listening the classic music stimulated subject #2 but rested subject #3. By the results of the central and autonomic nervous activities and psychological effects, responses to the music were different depending on the subjects.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-270 自然風景の画像鑑賞によるリラックス効果の検討 Chong Chen（陳冲），Akiyo Takao（高尾晃世），Erika Nakagawa（中川恵里花），Kousuke Hagiwara（萩原康輔），Ayumi Kobayashi（古林亜由美），Yuko Fujii（藤井優子），Keiko Hirata（平田圭子），Toshio Matsubara（松原敏郎），Akiko Hashimoto（橋本亜希子），Hirotaka Yamagata（山形弘隆），Kenichiro Harada（原田健一郎），Jun Sasaki（佐々木順），Fumihiro Higuchi（樋口文宏），Shin Nakagawa（中川伸）山口大学大学院医学系研究科高次脳機能病態学講座 Due to increasing mental health issues and compromised efficacy of pharmacological treatment, the development of complementary and preventive strategies is becoming a focus of interest. One such strategy is nature therapy. Contact with nature has been reported to be healing. Yet, on one hand, whether nature therapy is effective for all people remains inconclusive, on the other hand, the underlying neurobiological mechanism of the therapeutic effect remains unclear. Therefore, in the present study, we tested the relaxing effect of viewing natural scenes with a randomized, crossover trial and explored its moderating factors as well as the underlying neural substrates using fNIRS. The study was approved by the IRB of Yamaguchi University Hospital and written informed consent was obtained from all subjects. 30 healthy volunteers were randomly assigned to view pictures of nature or cities on two different days in a counter-balanced order. While viewing the pictures, their brain activities were monitored by fNIRS. Before and after the intervention, they were asked to report their feelings of relaxation, calmness, and stress on a visual analog scale. Viewing pictures of nature but not cities had a significant relaxing and calming effect. Using repeated measures ANOVA controlling covariates of gender, age, working memory, mindfulness, etc., we found that the main effect of viewing pictures of nature on relaxation and calmness disappeared, while its interaction with mindfulness remained significant for relaxation. Further analysis confirmed that the relaxing effect of viewing pictures of nature was significant only in subjects with high than average level of mindfulness. Meanwhile, partial correlational analysis controlling covariates indicated a significant correlation between mindfulness and the increase in feelings of relaxation after viewing pictures of nature. Finally, we found that as subjects' level of mindfulness became higher, their brain activation in response to nature scenes became lower in the right dlPFC and lOFC. These results suggest that viewing pictures of natural scenes is relaxing only in mindful people and the dlPFC and lOFC may be involved in this moderating effect. As patients with depression typically show hyperactivation in dlPFC and lOFC at rest due to their ruminative tendency, nature therapy may be a promising treatment for depression. However, mindfulness training should be prescribed together in order to achieve therapeutic effects.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-271 情動の回路；幼少期ストレスの影響下における外側手綱核の構造と機能の可塑的変化 Tomoya Nakamura（中村友也），Kohei Kurosaki（黒嵜恒平），Munenori Kanemoto（兼本宗則），Takumi Maeda（前田拓海），Hiroyuki Ichijo（一條裕之）富山大医統合神経（生理２） It is reported epidemiologically excessive stress in early life such as neglect and abuse cause psychiatric disorder, but its neuronal circuit mechanism is largely unknown. Stress activates the lateral habenula (LHb) that projects to GABAergic neurons of downstream monoaminergic systems in the ventral tegmental area (VTA) and the dorsal raphe nucleus (DRN); thus, the LHb is involved in emotion and cognition through stress processing. Firstly, we investigated influence of early life stress on behavior in adult around P60. The pups were separated from the dam for 3 hours/day during postnatal (P) 10-20 (repeated maternal separation; RMS). The RMS group showed anxiety-like behavior on the light-dark box test in adults. Secondly, to know the cause of the anxiety behavior, structure of the LHb was investigated in the RMS group. The RMS group showed the larger number of immediately early gene product ZIF268/EGR1 positive cells after treatment 1-hour immobilization (IMO), the smaller number of PV positive cells than those of the control group in adults. Thirdly, to know the role of those neurons, normal maturation and charactrer were investigated. During the maturation of the LHb, the small number of PV positive cells gradually increased from P10 to P35 and reached a plateau. ZIF268/EGR1 positive cells were higher in P10-20 under the stress. Retrograde tracing showed that the ZIF268/EGR1 positive cells project to the VTA and the DRN. It was revealed that the PV positive cells in the LHb didn't express glutamic acid decarboxylase (GAD) 1 and GAD2 by using GAD1-GFP knock-in mice and GAD2-mCherry transgenic mice. Early life stress raised anxiety-like behavior, which is thought to result from the high activity of the LHb in adulthood that would inhibit monoaminergic neurons. Because the RMS group showed the similar concentration of plasma corticosterone to the control group, anxiety-like behavior is not due to activation of hypothalamic-pituitary-adrenal axis releasing corticosterone. The number of PV positive cells was fall in inverse proportion to that of ZIF268/EGR1 positive cells; thus, the PV positive cells may have some inhibitory function to ZIF268/EGR1 positive cells. Although the PV positive cells are known to be GABA inhibitory interneuron in general, almost all the PV positive cells have no enzyme to synthesize GABA; thus, the PV positive cells are thought to be involved in a novel manner in the LHb.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-272 不安様行動におけるカイニン酸受容体GluK3サブユニットの役割 Izumi Watanabe Iida（飯田渡辺和泉）¹，Kotaro Konno（今野幸太郎）³，Rie Natsume（夏目里恵）²，Manabu Abe（阿部学）²，Masahiko Watanabe（渡辺雅彦）³，Kenji Sakimura（﨑村建司）²，Miho Terunuma（照沼美穂）¹ ¹新潟大院医歯学口腔生化 ²新潟大脳研モデル動物開発 ³北海道大医解剖 Kainate receptor (KAR) is a member of the ionotropic glutamate receptor family which plays role in excitatory synaptic transmission in the central nervous system. KARs are tetramers composed of combinations of low-affinity GluK1-GluK3 and high-affinity GluK4-GluK5 subunits. Though large studies with AMPA and NMDA receptors, other types of glutamate receptors, have identified their important roles in long-term memory, a physiological character of KAR is still unclear. Here we generated GluK1-GluK5 single subunit knockout (KO) mouse and analyzed their phenotype using behavioral test battery. Surprisingly, only GluK3 KO mice showed reduction in anxiety-like behavior in both open field test and elevated plus maze test. GluK3 heterozygous mice showed mild anxiolytic behavior in open field test suggesting that the expression levels of GluK3 containing KAR is a determinant of anxiolytic behavior. Since fluorescent in-situ hybridization confirmed the expression of GluK3 in both excitatory and inhibitory neurons, we generated GluK3 conditional KO (GluK3 cKO) mice using Emx1-Cre mouse to knockdown GluK3 in excitatory neurons, and GAD67-Cre mouse to knockdown GluK3 in inhibitory neurons. Interestingly, the inhibitory neuron-specific GluK3 cKO mice showed anxiolytic behavior similar to GluK3 KO mice, but the converse was evident in excitatory neuron-specific GluK3 cKO mice. These results revealed that GluK3 containing KAR has a bidirectional role in anxiety behavior depending on the type of neuron it expresses. Together, our studies suggested the novel and yet specific role of KAR in anxiety-like behavior.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-273 高架式十字迷路上でのラット抗不安様行動に関連する下辺縁皮質および前辺縁皮質の神経活動 Tomoko Shimizu（清水朋子）¹，Chihiro Tsukagoshi（塚越千尋）²，Yilong Cui（崔翼龍）¹，Akira Mitani（三谷章）³ ¹理化学研究所生命機能科学研究センター生体機能動態イメージング研究ユニット ²藍野大学医療保健学部作業療法学科 ³京都大学院医学研究科人間健康科学系専攻 The medial prefrontal cortex (mPFC) is suggested to modulate the expression of anxiety, but the nature of this effect is somewhat controversial. Recently, some studies reported the anxiolytic effects of the mPFC, in contrast, others reported anxiogenic effects of the mPFC. One of the reasons of this disagreement is that the mPFC is a heterogenous structure including the prelimbic cortex (PL; the dorsal part of the mPFC) and the infralimbic cortex (IL; the ventral part of the mPFC). We previously reported that electrical stimulation of the IL, but not the PL, exerts anxiolytic-like effect. Based on this finding, we hypothesized that the activation of the IL neurons probably contribute to the anxiolytic-like behavior. To verify this hypothesis, we recorded neural activity from the PL and IL cortices in freely moving rats. Adult male SD rats aged 9-12 weeks were used for experiments. Rats were unilaterally inserted recording electrodes (two stainless steel wires of 50 μm in diameter) in either the PL or IL. Recording coordinates were 10.5-11.0 mm anterior to the interaural line, 0.7 mm lateral to the midline, and 2.5 mm and 4.2 mm ventral to the cortical surface for the PL and IL respectively. We assessed the anxiety-related behavior in the elevated plus-maze. The maze was made of two open and two closed arms extending from a central platform, and was elevated 50 cm above the floor. During the10-min elevated plus-maze test, behavioral date was recorded with a video camera fixed above the test apparatus and the neural activity was recorded using a wireless telemetry recording system. Behavioral and neural data was stored for further analyses. Neural activity was analyzed as multiunit activity and the mean firing rates in the open and closed arms and event-related firing rates were analyzed. IL neurons showed significantly higher firing rate prior to entries from a closed to an open arm than prior to entries from a closed to the other closed arm, whereas PL neurons didn't show significant differences among event related firing rates. These results suggest that the neural activity of the IL is associated with anxiolytic-like responses such as facilitation of open arm entry.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-274 腸内細菌叢の組成変化は早期離乳ストレスによるうつ様行動に関連する Itsuka Kamimura（上村いつか）¹，Eiji Miyauchi（宮内栄治）²，Noriaki Tsuchiya（土屋範晃）¹，Kanami Tamura（田村奏美）¹，Ayumi Uesugi（上杉愛海）¹，Takeuchi Tadashi（竹内直志）²，Kazutaka Mogi（茂木一孝）¹，Hiroshi Ohno（大野博司）²，Takefumi Kikusui（菊水健史）¹ ¹麻布大獣医 ²理研IMS 粘膜システム Early life stress increases the risk of psychiatric disorders. In rodents, early-weaning (EW), in which pups were weaned one week earlier than normal condition, led to increased anxiety and enhanced the hypothalamic-pituitary-adrenal (HPA) stress responses. On the other hand, recent studies reported that intestinal microbial community regulates the development of the HPA stress response and ingestion of probiotics reduces anxiety- and depression-related behavior via the vagus nerve. Thus, we hypothesized that heightened anxiety and the HPA stress response in early-weaned mice is due to changes in microbial community. In the present study, we conducted correlation analysis between the microbiota and behavior of EW and normal-weaned (NW) mice. Furthermore, we evaluated the impacts of the microbial community changed by early weaning by comparing germ free (GF) mice exposed to the fecal microbiota obtained from EW and NW mice on postnatal day 10 (GF-EW or GF-NW). There was no change in behavior in EW mice as compared to the NW mice in the open field test and the marble burying test. In the tail suspension test, the immobility time of the EW mice at 4 weeks of age was longer than that of NW mice, and this difference disappeared at 8 weeks of age. Similarly, GF-EW mice showed longer immobility time at 4 weeks of age in the tail suspension test, and this difference similarly disappeared at 8 weeks of age. There were differences in the intestinal microbiome composition between EW and NW mice, and similar difference was found in the GF-EW and GF-NW mice. These results suggest that early life stress leads higher depression-like behavior during the juvenile period and microbiota composition change is involved in this behavioral changes.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-275 MAPK-mediated phosphorylation of MKL2 regulates nuclear localization and transcriptional activity in striatal neurons Anthony Ariza（Ariza Anthony），Yasuhiro Funahashi（Funahashi Yasuhiro），Keiichiro Okuda（Okuda Keiichiro），Sachi Kozawa（Kozawa Sachi），Keisuke Kuroda（Kuroda Keisuke），Kozo Kaibuchi（Kaibuchi Kozo） Nagoya University Graduate School of Medicine Dopamine (DA) type 1 receptor (D1R) signaling activates cAMP/Protein kinase A (PKA), which then activates Mitogen-Activated Protein Kinase (MAPK) through Rap1 consequently activating transcription factors (TFs) in striatal medium spiny neurons (MSNs). These TFs play a pivotal role in reward-related behavior (Nagai T. et al. Neuron, 89, 1-16. 2016). However how D1R signaling regulates behavior through TFs is still largely unknown. CREB-binding protein (CBP) promotes transcription through hundreds of different TFs and cofactors, and is also important for reward-related behavior. To isolate TFs regulated by DA signaling in MSNs, we performed a Pull-down assay followed by a proteomic study for CBP-interacting proteins and identified more 300 novel candidates in the mouse striatum. We focused on Myocardin Like 2 (MKL2) protein, a transcriptional co-activator of Serum Response Factor (SRF) highly expressed in the brain and known to regulate dendritic complexity. We found that interaction of CBP and MKL2 increased by cocaine administration. Furthermore, MKL2 also formed a ternary complex with CBP and SRF in vivo. The C-terminal domain of MKL2, which can interact with KIX domain of CBP was phosphorylated by MAPK at Serine 913. MAPK signaling induced MKL2 nuclear localization and also increased SRF-response element promoter activity in striatal neurons. These results demonstrate that MAPK activation downstream DA signaling can phosphorylate MKL2 and promotes MKL2 nuclear import facilitating interaction with CBP and consequently regulating SRF-dependent gene expression in striatum.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-276 社会的伝染行動に対する社会性の影響 Keiko Takanami（高浪景子）¹，Olga Yakimenko（Yakimenko Olga）^1,2，Hiromichi Nagayama（永山博通）¹，Tsuyoshi Koide（小出剛）¹ ¹国立遺伝研　マウス開発研究室 ²Biomedicine, Karolinska Institutet, Stockholm, Sweden Socially contagious behaviors (e.g. scratching and yawning) are ubiquitous in human society. Though several reasons such as instinct, memory, sociality, emotion, empathy are conceivable, the mechanism is unclear. To determine whether sociality affects the social contagious behavior, we used tame mice and non-tame mice which we generated by behavior genetics. As a social contagious behavior, scratching behavior, indicative of itch, was used for analysis. At first, in a social exploration test, tame mice to human showed more contact behaviors for other mouse than non-tame mice. This result indicated that tame mice to human show high sociality for same species. Next, in social contagious itch behavior test, we presented the demonstrator mice which show scratching behavior with itch mediator to observer, tame mice or non-tame mice, which is not received itch stimulation. Tame mice showed more scratching behavior than non-tame mice. There was no significant different of grooming behavior. Finally, to investigate the general itch threshold of tame mice and non-tame mice, scratching behavior analysis was performed. Though, intradermal injection of peripheral itch mediator, histamine and chloroquine, induced the scratching behavior in both tame mice and non-tame mice, there was no significant difference of scratching bouts and scratching durations between 2 groups. In conclusions, our results showed that tame mice showed high sociality and high contagious itch behavior compared with non-tame mice, suggested that individual sociality is related to their social contagious behavior.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-277 長期的自発運動が向社会的行動に及ぼす影響 Yiran liu（劉伊然），Tatsuki Shiiba（椎葉竜生），Natsuko Kubota（久保田夏子），Maina Ishida（石田舞奈），Takeshi Nishijima（西島壮），Ichiro Kita（北一郎）首都大院人間健康行動生理 Prosocial behavior, which is defined as the voluntary behavior intended to be benefit for another, is often observed in animal, as well as human. Previously, several studies have shown that physical exercise may facilitate prosocial behavior, but the neural mechanisms underlying the facilitatory effect of exercise on prosocial action remains unclear. It has been suggested that empathy is one of the most essential factors for the prosocial behavior, and may be involved in oxytocin (OXT) neurons. Thus, it is possible that exercise may affect the prosocial behavior via activity of OXT neurons. However, few experimental studies for prosocial behavior have focused on the effect of exercise, and on its neural mechanisms. In the present study, we examined the effects of long-term voluntary exercise (4 weeks) on helping behavior for soaked conspecifics in male rats, and on neuronal activity of OXT neurons in the hypothalamic paraventricular nucleus (PVN) during helping situation using immunohistochemistry. We also measured neuronal activities in the central nucleus of the amygdala (CeA) and corticotropin-releasing factor (CRF) neurons in the PVN in both helping (observer) and soaked (demonstrator) rats, because these activities are suggested to related to empathetic concern in the prosocial behavior. Helping rats were assigned one of two groups to alter the exercise condition; housing in the cage with or without running wheel (EX or no-EX). All the helping rats were individually housed in each cage for 4 weeks, because the individual familiarity may affect ability of empathy even to prosocial behavior. Then, we observed a helping behavior that a helping rat opens the door to help a soaked rat in pool area containing water. The time to door-opened was recorded for 6 consecutive days (maximum for 5 min/day). In addition, the time spent in interaction zone that is near to door and the duration of grooming were also observed. There is no significant difference in the time to door-opened between EX and no-EX groups. Interestingly, the time spent in interaction zone and the duration of grooming in the EX group were longer than those in the no-EX group. These results suggest that long-term voluntary exercise with individually housing may not facilitate prosocial behavior, but may enhance an ability of empathy. The results of neuronal activity are under consideration.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-278 腹側被蓋野ドパミン神経の長期活動亢進による社会性行動異常の誘導 Hidekazu Sotoyama（外山英和），Hiroyuki Nawa（那波宏之）新潟大学脳研究所分子神経生物学分野 Dopamine neurons in the ventral tegmental area (VTA) regulate social interactions. Alterations in their activities are known to influence the suceptibility to social stress. However, it is still unknown how persistent dysregulations of VTA dopamine neurons provide consequences on social interactions. We investigated the role of activities of VTA dopamine neuron in putative schizophrenia animal models for social interactions. The animal model with schizophrenia-related endophenotypes, which was prepared by neonatal injections of EGF, showed higher firing rates of VTA dopamine neurons and larger basal dopamine release in the medial prefrontal cortex (mPFC). Chronic treatment with antipsychotic risperidone normalized these abnormalities as well as the social deficit, suggesting the role of dopamine firing. In agreement, chronic attenuation of VTA dopaminergic basal firing in this model rat by DREADD (hM4Di) manipulation ameliorated the abnormal social interaction. These results suggest that hyper-activities in the basal firing of the VTA dopamine neurons are responsible for social deficits in EGF model rats. Hyper-activities of the basal dopaminergic firing in the VTA were found in another schizophrenia animal model such as rats treated with MK801 as neonates. We tested effects of chronic stimulations of VTA dopamine neurons with DREADD (hM3Dq) on social interactions in normal rats. The chronic stimulations of the basal firing of VTA dopamine neurons leaded to social deficits. Therefore, it is suggested that the persistent activation of VTA dopamine neurons has a causal association to social deficits.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-279 マウスにおける巣作り行動の定量のための実験系の最適化 Natsuki Tagawa（田川菜月）¹，Keita Mori（森啓太）¹，Michinori Koebisu（古戎道典）²，Atsu Aiba（饗場篤）²，Yuichi Iino（飯野雄一）¹ ¹東京大院理生物科学 ²東京大院医疾患生命工学 One of the main undertakings of neuroscience is to describe the neural circuits and molecular mechanisms by which complex behaviors are generated. Nest building is one such behavior, and is widely preserved throughout the animal kingdom. Mice build nests for thermoregulation, housing of their pups, and self-defense. This behavior is composed of sequential stereotype behaviors and can be used as an indicator of well-being in mice. The purpose of this study is to clarify neural basis of nest building which, to date, has been only partly elucidated. To address this issue, we have developed a platform for assessing the nest-building behavior. In a conventional nest-building behavior test, mice are put in a new cage and given nest-building material (Nestlet) a few hours before the dark period of the day-night cycle of mice. This procedure has two major problems to overcome. The first is that mice do not complete nest-building in a short period of time (one to two hours). The second is that even by the next morning only a fraction of the animals built their nests, whereas we wanted all mice to build their nests within a short time after the Nestlet is given. In the present study we optimized two parameters so that mice reliably complete nest-building in a short period of time. The first parameter is the period of time during which mice are allowed to habituate to their new cage. The second is the time of the day at which Nestlet is presented to the subject. When we gave mice a 5-7 day habituation period and put the Nestlet into the cage 6 hours after the light cycle had begun, all test subjects completed nest building within an hour of receiving their Nestlet. Our future plans include identifying brain regions involved in the nest building behavior using our established platform. To achieve this, we plan to use c-Fos mapping of brain regions activated during nest building, as well as injections of the GABA agonist muscimol to inhibit candidate brain regions during the behavioral test.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-280 背側縫線核のグルタミン酸シグナルによるマウスの母性行動の制御 Yoshikage Muroi（室井喜景），Toshiaki Ishii（石井利明）帯畜大獣医薬理 Lactating female mice nurture their pups and attack intruders in their territory. These behaviors are the representatives of maternal behavior. When an intruder invades a dam's territory, she needs to switch her behavior from care to aggression to protect her pups and territory. Each behavior is displayed alternatively. Although the neuronal mechanisms underlying each distinct behavior have been studied, it is unclear how these behaviors are displayed alternatively. The dorsal raphe nucleus (DRN) regulates both nurturing and aggressive behaviors. In the present study, we examined whether the DRN is involved in regulating alternative display of maternal care and aggression. We first examined neuronal activity in the medial prefrontal cortex (mPFC) and lateral habenula (LHb), which send glutamatergic input to the DRN, in dams by injecting Fluorogold, a retrograde tracer, into the DRN. The number of c-Fos- and Fluorogold-positive neurons in the mPFC and LHb increased in the dams that displayed biting behavior in response to an intruder, but remained unchanged in the dams that displayed nurturing behavior. Injections of N-methyl-D-aspartic acid (NMDA) receptor antagonists or α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate receptor antagonists into the DRN inhibited biting behavior but not nurturing behavior. In contrast, injections of NMDA or AMPA into the DRN inhibited nurturing behavior. These results suggest that glutamatergic signals in the DRN, which may originate from the mPFC and/or LHb, regulate the preferential display of biting behavior over nurturing behavior in dams.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-281 雄マウスの攻撃行動の表出調節に果たすエストロゲン受容体β陽性細胞の役割 Kansuke Hasunuma（蓮沼寛介），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 Testosterone (T), a gonadal steroid secreted from testes, is known to regulate the expression of aggressive behavior. Besides acting through androgen receptors (AR), T can also act through estrogen receptor (ER), ERα and ERβ, after converted to estradiol by aromatase in the brain. Studies using knockout (KO) mice have suggested that activation of ERα is essential for the facilitation of aggressive behavior, while ERβ seems to play a modulatory role. However, how ERβ is involved in the modulation of aggressive behavior is not well understood. In this study, we investigated neuronal activity of ERβ positive neurons in various brain regions during aggressive behaviors. We used newly developed transgenic mice whose ERβ positive cells were labeled with red fluorescent protein (RFP). ERβ-RFP male mice were tested for aggression in a resident-intruder paradigm for 3 consecutive days and assigned to either Aggressor or Non-Aggressor groups based on their levels of aggression. 75 minutes after the last aggression test, mice were sacrificed and their brain samples were collected for double fluorescent immunohistochemistry for RFP and c-Fos, as a marker of neuronal activity. Distribution of double labeled cells was compared between the Aggressor and Non-Aggressor groups in 6 brain regions: 1) lateral septum (LS); 2) bed nucleus of stria terminalis (BNST); 3) medial amygdala posterodorsal area (MeApd); 4) medial preoptic area (MPOA); 5) paraventricular nucleus (PVN); and 6) dorsal raphe nuclei (DRN). We found that the number of c-Fos and RFP co-expressing cells was significantly higher in the Aggressor in comparison to the Non Aggressor groups in the PVN, even though no group difference was observed in the number of singly labeled cells for either c-Fos or RFP. A similar tendency was also found in the BNST but the actual number of double labeled cells was very low, compared to that found in the PVN. These results suggest that ERβ expressing cells in PVN may be involved in the regulation of aggressive behavior in male mice. (Supported by KAKEN #15H05724 to SO)

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-282 空腹状態におけるゼブラフィッシュ手綱核-脚間核神経回路の調節は闘争行動における勝利をもたらす Haruna Nakajo（中條暖奈）¹，Ming-Yi Chou（Chou Ming-Yi）³，Masae Kinoshita（木下雅恵）¹，Lior Appelbaum（Appelbaum Lior）⁴，Hitoshi Okamoto（岡本仁）^1,2 ¹理研CBS 意思決定回路動態 ²東京大院総合文化研生命環境科学 ³Dept Life Science, National Taiwan Univ, Taipei, Taiwan ⁴The Mina & Everard Goodman Faculty of Life Science, Bar-Ilan Univ, Ramat-Gan, Israel Animals fight with each other to monopolize food, keep territory, and increase reproductive probability. We previously suggested that two different pathways from habenula (Hb) to its downstream structure interpeduncular nucleus (IPN) can switch winner and loser states in zebrafish fighting (Chou et al., 2016, Science). However, it is unclear what kinds of factor can regulate a valance of these two Hb-IPN pathways. Considering that starved state modulates animal's behavior (Fokidis et al., 2013 etc.), animals in starved state may change their fighting behavior. To test this hypothesis, we observed zebrafish fighting behavior, which is an useful model of social conflict, in starved state. When we put two male zebrafish into a tank, they show fighting behavior in a stereotypic manner, and winner and loser clearly show different behaviors. We food-deprived from fish for 6 days and paired with the food-deprived fish with normally fed fish to fight with each other. As a result, starved fish tended to win the fight (75% of winning rate). We then observed Hb-IPN pathway activities by using acute slice Ca²⁺ imaging. Starved fish showed winner-like activity pattern in the Hb-IPN pathway before their fighting. Furthermore, by electrophysiological recording from the IPN region, we observed an elongation of AMPA-type glutamate receptor activity in starved fish. Orexin neuron is well known to be activated upon starvation and plays crucial role in feeding behavior. We therefore ablated orexin neuron in transgenic zebrafish and examined their fighting behavior. Orexin neuron-ablated fish did not show a tendency to win the fight and an elongation of AMPA receptor activity in the starved state. Collectively, our results suggest that starvation-activated orexin neuron triggers winner-like activity in the Hb-IPN pathway through elongation of AMPAR activity, resulting in the winning in social conflict. Because winner in the fight could have an advantage in obtaining foods, the behavioral change that we observed may be useful for hunger animals to increase probability to survive under limited resource situation.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-283 野生型とCD38ノックアウトマウスの子供を防御するための攻撃行動におけるオキシトシンの役割 Anna Shabalova（シャバロバアンナ）^1,2，Jin Zhong（鍾静）¹，Mingkum Liang（梁明坤）¹，Chiharu Tsuji（辻知陽）^1,2，Shigeru Yokoyama（横山茂）^1,2，Haruhiro Higashida（東田陽博）^1,2 ¹金沢大学子どものこころの発達研究センター 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 Oxytocin (OXT) is a nonapeptide, which is synthesized in hypothalamic paraventricular (PVN) and supraoptic (SON) nucleus. CD38 is a transmembrane glycoprotein, involved in OXT secretion. CD38 knockout mice display abnormal parental behavior. A lot of findings suggest a role of OXT in inducing maternal aggression as a part of maternal behavior, in sharp contrast of OXT's function for social bonding. Paternal aggression is less studied and a role of OXT in paternal aggression remains unclear. In my research, I analyzed the influence of family cues on aggressive behavior and activation of the oxytocinergic neurons within the PVN and SON of the hypothalamus in ICR and CD38 knockout sires. After delivery pups, by dams paired sires were divided into 4 groups and separated from their families. In the first group, the sires were housed alone. In the second and the third groups, the sires were exposed to their mates or the lactating paired dams, respectively. In the fourth group, biological pups were presented to their sires. Following one week of 6 hours daily family-cue-stimulation, aggression was assessed using the Resident-Intruder test. After the test brains were collected for analyzing OXT/c-fos positive cells within the PVN and SON. Sires, exposed to the pups demonstrated the highest level of aggression, whereas, in contrast, the sires spent alone showed the lowest level of aggression. Activation of the OXT-ergic neurons in the PVN and SON increased in the sires stimulated by their pups. In contrast, paternal experience did not affect aggressive behavior and OXT-ergic activation in CD38 knockout sires at all. These results suggest that direct physical contact with pups is likely to be the best for the induction of paternal aggression, and interesting, that attack behavior is associated with activation of OXT cell bodies within the PVN and SON.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-284 Perinatal stress: affiliative behaviors and neurohormones in rat pups Aldo B Lucion（Lucion Aldo B）¹，Celso R Franci（Franci Celso）²，Maiara L Lutz（Lutz Maiara L）¹ ¹Institute of Basic Health Sciences (ICBS), Federal University of Rio Grande do Sul (UFRGS) ²Faculty of Medicine (FMRP), University of São Paulo (USP) Several studies have shown the long-lasting effects of stress during the perinatal period. Differently than previous data that analyzed adult animals, present study focused on behavioral (maternal odor preference) and neuroendocrine (oxytocin - OT and prolactin - PRL) effects of pre (restriction of nest material in the home-cage from gestational day (GD) 14 to 21) and post-natal (presence of a live cat over the home-cage on post-natal day (PND 1) stress in the rat pups. In the maternal odor preference test, male and female pups (PND 9) had to choose between nest and clean sawdust. In female and male pups, post-natal stress abolished the preference for the maternal odor, while pre-natal stress induced no behavioral change in the pups' preference. The combination of both pre and post-natal stress affected the females but not the males. On PND 1, plasma hormones (corticosterone (CT) and PRL) and also cerebrospinal fluid (CSF) OT were measured after the last of the 3 predator exposures. In female pups, pre and post-natal stress decreased plasma CT, but the combination of both induced no significant change. In male pups, the 3 different environmental interventions (pre; post and the combination of both stressors) decreased plasma CT. Plasma PRL on PND 1 was not significantly affected by the stressors in both male and females. CSF OT was also not altered by the stressors. Previous study showed no significant changes in the maternal behavior in the stressed groups using the same stressors. The nature of the stressors is quite different, besides being applied at different periods. In the case of the predator stressor, the pups were actually present, while the reduction in nest material, its effect was through the mother. We may conclude that the presence of a natural predator as early as PND 1 may affect (decrease) the secretion of CT, in a period usually characterized by its hypo-responsiveness to stress. The pre-natal stress protocol (reduced nest material) was also effective to alter plasma CT, but not the behavioral preference. Curiously, stressors decreased plasma CT, instead of the increment observed in adults. Pups' affiliative behavior, measured 8 days after the last intervention, was compromised by the post-natal stress, but not by the pre-natal stress. Results indicate a dissociation between CT alteration on PND1 and the behavior on PND9, since CT responded to both stressors but only the post-natal stressors was effective to reduce infant attachment.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-285 マウスの社会挫折ストレスに対する抵抗性を担う内側前頭前皮質の出力経路の探索 Chisato Numa（沼知里），Hirotaka Nagai（永井裕崇），Tomoyuki Furuyashiki（古屋敷智之）神戸大院医薬理 Stress caused by aversive stimuli, if not excessive, causes both adaptive and maladaptive biological responses in brain. Dysregulation of the adaptive responses increases risks of psychiatric disorders. Previous studies have associated the mesocortical dopaminergic pathway with adaptation to stress. We have previously shown that single social defeat stress in mice activates dopamine D1 receptor in excitatory neurons of the medial prefrontal cortex (mPFC), leading to dendritic hypertrophy of these neurons and strengthening stress resilience. However, it remains elusive which brain regions mediate the action of the mPFC for stress resilience. In the present study, using c-Fos immunohistochemistry, we examined neuronal responses to single social defeat stress in multiple brain regions of male C57BL/6 mice and the involvement of dopamine D1 receptor in the mPFC. We found that the stress activated neurons in several brain regions related to emotional behaviors, including nucleus accumbens, bed nucleus of the stria terminalis, interstitial nucleus of the posterior limb of the anterior commissure (IPAC) and central amygdala. Next, we knocked down dopamine D1 receptor in the mPFC by adeno-associated virus expressing an artificial miRNA targeting this receptor prior to the stress, and found that the stress-induced c-Fos expression in the IPAC, a brain region which receives direct projections from the mPFC, was significantly decreased with the knockdown of D1 receptors in the mPFC. These findings show that social defeat stress activated the IPAC in a manner dependent on dopamine D1 receptor in the mPFC. Thus, our study paves the way for elucidating neuronal pathways which mediate the action of dopamine D1 receptor in the mPFC for facilitating stress resilience.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-286 性に関連した脳ネットワーク特性の変動 Tomohiro Donishi（堂西倫弘）¹，Masaki Terada（寺田正樹）²，Yoshiki Kaneoke（金桶吉起）¹ ¹和歌山県立医大医生理一 ²和歌山南放射線科クリニック Although mean value difference between the sex difference of the brain structures and functions has been intensively investigated, the difference has been shown to be too subtle to divide male and female groups. We examined if there exists sex-related variance difference in brain network parameters, using 200 right-handed young (100 males, 18-24 years old) healthy subjects' resting-state functional MRI data. Preprocessing was performed with SPM8 (http://www.fil.ion.ucl.ac.uk/spm) and in-house software developed on MATLAB (MathWorks, Natick, MA, USA). Functional connectivity was calculated by correlation coefficients between each gray matter voxel (6×6×6 mm) pair, converted into Z values with effective sample size correction, and then an adjacency matrix was created for network analysis. Using normalized α-centrality, we obtained two network property (nAC₀ and nAC₁) for each voxel, which corresponds to degree centrality and eigenvector centrality, respectively. For both male and female groups, we calculated the coefficient of variance (CV) of the two parameters to select voxels with lower CV values. Permutation test revealed that low CV regions specifically for males and females (p < 0.005). Most of the regions such as the anterior insular cortex, temporal lobe, and the precuneus were related to so-called social brain networks. Analogous to the vulnerability of the low genetic variability, low CV of the brain network parameters may explain sex difference of the prevalence of neuro-psychiatric diseases.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-287 X11L欠損マウスにおける社会的相互作用と動機づけ行動の低下 Kotomi Yamanaka（山中琴未），Shota Mizuno（水野翔太），Teiichi Furuichi（古市貞一），Yoshitake Sano（佐野良威）東京理科大院理工応用生物科学 X11-like (X11L)/Mint2 is a neuronal adaptor protein which interacts with several proteins such as amyloid precursor protein and Munc-18 involved in modulating neuronal activity. Previously, we reported that X11L-knockout (KO) mice show a deficit in some kind of motivated approach behavior: e.g., ones evaluated by decreased intruder exploration behavior in resident-intruder test and decreased digging and burrowing behaviors. Interestingly, X11L-KO mice also show significant weight loss under highly competitive feeding conditions, but not in non-competitive feeding conditions. However, detail features and neural basis of these behavioral phenotypes seen in X11L-KO mice is still unknown. To understand the neural basis underlying motivated approach behavior of X11L-KO mice, we comprehensively analyzed c-fos expression associated with digging behavior by immunohistochemistry and tested social behavior in more detail by using two different type of social interaction test. As a result, activity-dependent expression pattern of c-fos protein in several brain regions of X11L-KO mice during digging test was significantly different compared with that of wildtype (WT) mice. In social encounter test, X11L-KO mice showed reduced time of interaction with counterpart but increased number of interaction from counterpart compared to WT mice. Interestingly, social interaction in X11L-KO was indistinguishable from that in WT mice when counterpart was housed in a cup. These results suggest that identified brain regions decreased c-fos expression during digging behavior may be neural circuits responsible for regulating motivated approach behaviors of X11L-KO mice. Altogether, we also suggest that X11L-KO mice is a useful model for analyzing social-related motivated approach behavior under the conflicting situation.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-288 雄マウスにおける雄特異的なフェロモンを介した行動の受容体と神経基盤 Takumi Itakura（板倉拓海），Ken Murata（村田健），Kazushige Touhara（東原和成）東京大院農学生命科学応用生命化学専攻 In many animals including mice, chemical signals play pivotal roles in regulating sex dependent adaptive behaviors. However, little is known how such chemical signals are received and processed in the central nervous system. We have identified one murine vomeronasal receptor, here named VRX, which receives a male specific compound in male urine. Considering that VRX receives a male specific compound, a ligand for VRX may act as male specific pheromone and the sensory input from VRX can trigger a male-oriented behavior. In this study, we tested a hypothesis that the sensory input from VRX triggered aggression in male mice. We performed several behavioral experiments using a VRX-active purified fraction of male urine and VRX-knockout mice. We found that VRX-knockout male mice showed reduced aggression towards male mice and the VRX-active fraction enhanced male aggression. Furthermore, no enhanced aggression was observed in VRX-knockout male mice upon stimulation with this active fraction. These results indicate that the sensory input from VRX is necessary and sufficient for male aggression enhancement. Next, to elucidate the central processing of VRX-mediated neural signaling in male mice, we performed a fiber photometry imaging and investigated which brain regions are activated by the sensory input from VRX. We found that the VRX-active fraction activated genetically identified neuron subtypes in the ventrolateral part of the ventromedial hypothalamus (VMHvl). Furthermore, specific neuron subtypes in extended amygdalar and hypothalamic regions, which have a potential to regulate aggression, are activated by the VRX-active fraction. Taken together, our results suggest that VRX-mediated enhanced aggression is regulated by extended amygdala and hypothalamus including VMHvl.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-289 モテるオスはハゲるとモテなくなるのか？ Yoshinori N Ohnishi（大西克典），Yukie Kawahara（河原幸江），Yoko H Ohnishi（大西陽子），Akinori Nishi（西昭徳）久留米大医薬理 Baldness is one of fearful aging phenomenon for men. Complete baldness sometimes enhances masculinity, but partial baldness is believed to decrease sexual attractiveness in most cases. We have previously presented that attractiveness of male mice to female mice is dependent on their appearance, not voice nor smell, using behavior-based measurement with video camera tracking system (SfN 2017, SfN 2018). In those experiments, four littermate male mice were placed in four jail transparent boxes set on corner of a larger white box one by one. One female mouse was placed in the large box and allowed to move freely for 20 minutes. We calculated the sum of time around each male box for the last ten minutes. Surprisingly, attractive and/or unattractive male mice were existed despite the same genetic background. Analyses using small number of female mice reached significant levels, suggesting that female mice have a common preference for male attractiveness. This trend of preference disappeared by hiding male mice with four-layered air-permeable filter. Furthermore, genetically blind female mice showed completely different trend of preference against the same male mice set, indicating that appearance may be the main factor of male attractiveness. However, it is unknown how appearance affects the male attractiveness. So, in this report, we focus on the importance of hair appearance. We will examine whether hair cut or whisker trimming of attractive male mouse can affect his attractiveness to female mice. Additionally, we will perform in vivo microdialysis analyses under the experimental conditions, because dopamine levels in the nucleus accumbens of female mice responded to attractive male mice, but not to unattractive male mice. These experiments will reveal preference and mesolimbic dopamine reaction of female mice against male attractiveness defined as hair appearance.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-290 擬似的な社会ネットワークでの協調行動と信頼のメカニズム Rei Akaishi（赤石れい），Masahiko Haruno（春野雅彦）脳情報通信融合研究センター Social network, who we know and whom we communicate regularly, influences our behavior and well-being. Information transmitted through links in social network shapes our shared values and norms, which in turn determines our decisions in life. However, we know surprisingly little about the mechanisms of how we create and maintain our social networks. We address this important issue by examining sizes of real-world social networks of individuals, conducting controlled experiments of virtual social networks. Prisoner's dilemma (PD) task was performed within the social network of the subject; a social partner in each trial is chosen from the existing network of players connected to the subject. Subjects decided whether to cooperate or defect with a social partner in each trial. In some trials, subjects also decided whether to accept or reject a new comer who wished to join the network of the subject. If the subject accepted the new comer, the subject played the PD task with the new comer. After performing the several trials in the PD task, subjects decided whether to continue to play the game with the current partner. If the subject decided to stay with the partner, s/he remained in the network. If the subject decided to leave (switch) from the relationship, the partner was removed from the network. Comparing the behavior in the experimental room with the networks size in real life, we found that the probability of cooperation just before switch decision correlated with the social network sizes in the real world. In addition, the network size maintained in the task was also correlated with the social network size in the real world. We also analyzed behavioral data with computational models. The analysis of learning and decision making patterns revealed that the subjects are using some form of hypothesis testing mechanism whereby predictions of social partners' behaviors are actively used to form the impressions of the others. These results suggest that the behaviors in the experimental rooms are predictive of behaviors in real social world and that the mechanisms used in the social interactions may be rooted in the general computational principles of the brain.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-291 コモンマーモセット大脳皮質領域における上側頭溝の他者行動表現 Wataru Suzuki（鈴木航）^1,2，Misako Komatsu（小松三佐子）²，Takaaki Kaneko（兼子峰明）³，Yuri Shinomoto（篠本有里）³，Hideyuki Okano（岡野栄之）³，Noritaka Ichinohe（一戸紀孝）^1,2 ¹国立精神・神経セ神経研微細構造 ²理研CBS 高次脳機能分子解析 ³理研CBS マーモセット神経構造 Since information processing for social communication needs integration of various types of different modalities, different cortical areas in the brain would cooperatively participate in social behavior. In our previous studies, cells in the superior temporal sulcal (STS) region and the ventral premotor cortex (vPM) where mirror neurons existed in the common marmosets (Callithrix jacchus) strongly responded to the other's action as those of human and macaque monkeys, indicating that those regions participate in the social information processing (Suzuki et al., 2015). In this study, using the advantage of a lissencephalic and small brain of the marmoset with cortical areas comparable to those of human and macaque monkey, we investigated the neural representation of the other's action in whole-cortex and whether STS and vPM was the major areas for this representation by cortical-wide high-density electrocorticogram (ECoG). The 96 channel ECoG arrays covering wide range of the lateral surface of telencephalon were epidurally implanted on the either hemisphere in 3 marmosets. During recording, a human experimenter acted grasping behavior in front of the marmoset, consisting of reaching, grasping and retracting a target. There were 4 conditions of the grasping behavior with mixture of 2 factors: reaching direction (reaching from left or right side of the marmoset) and target condition (grasping a food or a vacant target). After and around the timing when the human actor touched either target, high gamma band activity in ECoG contacts, considered to be correlated with spike activity, was induced in visual cortices of the occipital and temporal lobe, including STS, and in the lateral frontal cortex, including vPM. Around timing of touching, the contacts on STS showed a clearer peak of the high gamma activity in comparison to the other contacts. After touching, a differential activity between conditions of grasping a food and a vacant target was observed at the contacts on STS and the lateral frontal cortex. Especially, the differential activity in STS was more prominent than in the lateral frontal cortex. These results are consistent with our previous unit-recording studies in that, while cells in STS and vPM in marmosets respond to the other's action, the goals of the actions are represented in STS rather than in vPM. This study also suggest that STS play a major role for representation of the other's action and its goal within whole lateral cortical region.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-292 性行動を起こした雌雄ラットの内側視索前野におけるカルビンディンD28K発現ニューロン活性の性差 Masahiro Morishita（森下雅大），Arisa Kamada（鎌田亜里紗），Shinji Tsukahara（塚原伸治）埼玉大院理工生命科学 The medial preoptic area is involved in the regulation of sexual behavior in both sexes. The medial preoptic area in rats contains a sexually dimorphic nucleus, which is mainly composed of neurons expressing calbindin-D28K (calb), a calcium-binding protein (hereafter this nucleus is termed the Calb-SDN). The Calb-SDN of male rats is larger in size and contains more calb neurons compared with the Calb-SDN of female rats, supposing a sex difference in the function of the nucleus. However, the physiological roles of the Calb-SDN are poorly understood. In the current study, to determine whether the Calb-SDN is involved in the regulation of sexual behavior in male and/or female rats, we examined the neuronal activity of the Calb-SDN in rats during sexual behavior. Female rats were ovariectomized (OVX) and treated with estradiol benzoate (EB) and progesterone (P) to induce estrous condition. They then copulated with sexually vigorous male rats. Sixty minutes after male rats ejaculated, the brains were sampled from the EB- and P-treated OVX females and the males ejaculated. EB- and P-treated OVX female rats that did not experience copulation and sexually vigorous male rats that did not ejaculate before brain sampling were used as the controls for each sex. After the brains were coronally cut, the brain sections were immunostained for calb and c-fos, a neural activity marker. Immunostained brain sections were used for morphological analyses to count the number of c-fos-immunoreactive (ir) cells in the Calb-SDN. As a result, in both EB- and P-treated OVX female rats and sexually vigorous male rats, the number of c-fos-ir cells in the Calb-SDN increased significantly with copulation. Although copulation induced c-fos expression of the Calb-SDN in both sexes, the percentage of c-fos expression in the total calb neuron in the female Calb-SDN was significantly higher than that in the male Calb-SDN. These results suggested that neurons in the Calb-SDN are stimulated during copulation in both sexes and that neurons of the female Calb-SDN are more activated during copulation than do neurons of the male Calb-SDN.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-293 ラットの性行動を制御する内側視索前野におけるカルビンディンニューロンの機能的性差 Kanna Ueno（植野寛菜），Sho Maejima（前嶋翔），Masahiro Morishita（森下雅大），Shinji Tsukahara（塚原伸治）埼玉大学大学院理工学研究科生命科学系専攻調節生理学研究室 Sex differences of brain structure underlie the control of sex-specific behaviors. The medial preoptic area (mPOA) of rats is involved in the regulation of sexual behavior in both sexes and contains a sexually dimorphic nucleus showing male-biased sex difference in the size and neuron number. The sexually dimorphic nucleus of the mPOA in rats is mainly composed of neurons expressing calbindin D-28K (calb), a calcium-binding protein, although the physiological roles of calb neurons in the sexually dimorphic nucleus of the mPOA are poorly understood. We recently found that calb neurons in the sexually dimorphic nucleus of the mPOA activate during sexual behavior in both male and female rats. In this study, we examined the effects of knockdown of calb in the mPOA on sexual behavior in male and female rats to determine whether calb neurons in the mPOA are required for the control of sexual behavior or not. An adeno-associated virus (AAV) vector to express a small hairpin RNA (shRNA) targeting calb mRNA was injected into the mPOA including the sexually dimorphic nucleus of male and female rats. Animals injected with an AAV vector to express shRNA targeting luciferase mRNA were used as the controls. After that, we analyzed sexual behavior of AAV vector-injected animals to evaluate the effects of calb knockdown on sexual behavior. In male rats, knockdown of calb in the mPOA had no significant effect on the expression of sexual behavior. On the other hand, knockdown of calb in the mPOA affected sexual behavior in female rats. The expression of lordosis, a female sexual behavior, in calb-knockdown females was significantly fewer than in the control females. These results suggest that calb neurons in the mPOA are involved in facilitation of sexual behavior in female rats. In male rats, calb expressed in mPOA may contribute less to the control of sexual behavior, although the roles of calb neurons require further investigation.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-294 社会ストレス反応における3種モノアミン神経の相互作用 Maki Kikawa（鬼川真季），Hidekazu Sotoyama（外山英和），Hisaaki Namba（難波寿明），Hiroyuki Nawa（那波宏之）新潟大学脳研究所分子神経生物学分野 Long-term social stress is well known as a risk factor for mental illness. As chronically stressed animals show depression-related behaviors such as reduced social behaviors, these animals are often employed as a model for post-traumatic stress disorder (PTSD) or depression. In order to investigate how chronic social stress regulates the neural activities of three monoaminergic neurons (dopamine, noradrenaline, serotonin) and their mutual interactions, neuronal firing of these cells was measured under an anesthetic condition from the rats receiving chronic social defeat stress. Firing frequency of dopaminergic neurons in the ventral tegmental area (VTA) was increased in the rats receiving chronic social defeat stress. Conversely, the firing frequency of the noradrenergic neurons in the locus coeruleus (LC) was reduced, compared with that in healthy control rats. There was no significant change in firing frequency of serotonergic neurons in raphe nucleus, however. To test the sensitivity of these monoaminergic alterations to an anti-depressant, a selective serotonin reuptake inhibitor (fluoxetine) was chronically administered during their exposure to social defeat stresses. The treatment with fluoxetine normalized the abnormal activities of dopaminergic and noradrenergic neurons. In contrast to the stressed rats, however, noradrenergic neurons in control rats failed to respond to the anti-depressant, although their dopamine neurons exhibited a similar reaction to the drug. These results suggest that the stress-driven behavioral changes are associated with increased firing of VTA dopaminergic neurons and decreased firing of LC noradrenergic neurons. Currently, their mutual regulation of these monoaminergic cells is under investigation by manipulating the dopaminergic firing with the DREADD protocol.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-295 社会的闘争行動の拮抗的な制御に関わる手綱核-脚間核-縫線核を並走する２つの神経回路 Takehsia Handa（半田剛久）^1,2，Taku Sugiyama（杉山拓）¹，Yoshie Itou（伊藤世志江）¹，Megumi Kobayashi（小林恵実）¹，Joshua P Johansen（Johansen P Joshua）³，Hitoshi Okamoto（岡本仁）¹ ¹理研CBS 意思決定回路動態 ²東京医科歯科大学　大学院医歯学総合研究科　分子神経科学分野 ³理研CBS 学習・記憶神経回路 The Medial Habenula (MHb) has an evolutionarily conserved topographic projection to the Interpeduncular Nucleus (IPN). For example in rodents, the dorsal medial habenula (dMHb) projects to the IPN lateral part (IPL) and the ventral medial habenula (vMHb) that projects to the central area of the IPN (cIPN). Our recent studies suggest that dMHb-IPL pathway and vMHb-cIPN pathway exert opposing effects on animal's behaviors under social conflict. To understand the neural mechanism by which the two pathways control opposing behaviors, it is necessary to elucidate specific anatomical connections of each pathway. In current study, using genetical and viral tracing techniques, we investigated the downstream structures of dMHb-IPL and vMHb-cIPN pathways. To specifically label IPL neurons we employed DRD3:cre mouse that has been reported to express cre recombinase in the IPL. We first confirmed that DRD3:cre positive IPN neurons have synaptic connection with afferents from the dMHb using cre-dpendent monosynaptic rabies virus. Neurons in the dMHb were labeled by the retrograde transsynaptic tracing starting from DRD3:cre(+) IPL neurons. Thus DRD3:cre mouse is an appropriate tool to study the downstream of the dMHb. Next, to visualize presynaptic terminals of IPL efferents, we injected adeno-associated virus (AAV) cre-dependently expressing synaptophysin fused mCherry (syp-mCherry) into the IPN of DRD3:cre mice. For comparison, to visualize presynaptic terminals of cIPN efferents, AAV constitutively expressing syp-mCherry was carefully injected so that infection was confined in the cIPN. IPL efferents labled with syp-mCherry were dominant in the paramedian raphe (PMnR) .In contrast, cIPN efferents labeld with syp-mCherry were dominant in the median raphe (MnR) that contains serotonin neurons. PMnR is rich in GABA neurons and closely adjacent to the MnR. As they are very close, the PMnR may send GABAergic projection to MnR and exert the antagonistic function. Thus our data suggests that the PMnR and the MnR are the output of dMHb-IPL and vMHb-cIPN pathway respectively, and this may explain the antagonistic functions of these pathways. Further results will be discussed in the poster.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-296 マウスにおける同種個体に対する救助様行動 Hiroshi Ueno（上野浩司）¹，Shunsuke Suemitsu（末光俊介）²，Motoi Okamoto（岡本基）³，Takeshi Ishihara（石原武士）² ¹川崎医療福祉大医療技術臨床検査 ²川崎医科大精神 ³岡山大院保健 Prosocial behavior signifies behavior that benefits others without the expectation of external compensation. Prosocial behavior was considered an idiom of only humans and primates. In recent years, prosocial behavior has been reported in rats. However, the findings of prosocial behavior in rats are somewhat controversial. To investigate whether mice, another rodent species, also show prosocial behavior, we examined if they would help conspecifics trapped in tubes. Subject mice were allowed to move freely in cages with tubes containing constrained conspecifics. We observed the helping behavior of subject mice towards constrained mice. The subject mice released both cage-mate and stranger mice but showed no interest in opening empty tubes. Furthermore, the same behavior was observed under aversive conditions and when the conspecifics were anesthetised. Neither vocalising nor distress is necessary for helping behavior, as mice also showed rescue behaviou towards anesthetised cage mates. Interestingly, hungry mice first opened the tubes containing food before engaging in helping behavior. Mice showed a similar degree of interest towards constrained and freely moving conspecifics, which showed that they neither avoided nor had increased interest towards the constrained mice. We showed that mice show helping behavior. Our results do not prove that mice show prosocial behavior; they indicate that a new experimental method is necessary to prove or disprove that mice show prosocial behavior. The findings of this study suggested that helping behavior in mice is more readily explained as pursuit of social contact rather than as empathic action.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-297 闘争行動における敗北の経験は腹側脚間核でのグルタミン酸作動性神経伝達の増強を引き起こす Masae Kinoshita（木下雅恵），Ming-Yi Chou（周銘翊），Hitoshi Okamoto（岡本仁）理研CBS 意思決定回路動態 In the social conflicts of diverse animals, experience of defeat reduces winning rate in the following fight. This phenomenon is called "loser's effect". Previously, we demonstrated that inhibition of transmission from the medial part of dorsal habenula (dHbM) to the ventral interpeduncular nucleus (vIPN) increases winning rate in the social conflict of zebrafish, and hypothesized that potentiation of the dHbM-vIPN transmission is neural basis for loser's effect. In the present study, we recorded postsynaptic currents from vIPN neurons with slice patch-clamp recording techniques, and compared properties of recorded currents between winner and loser. The dHbM-vIPN transmission is mainly glutamatergic, but we found that only a part of cells in the vIPN was responsive to glutamatergic inputs, and that responsive cells were distributed in the dorsal part of vIPN. All of recorded cells in dorsal vIPN of the loser were responsive to glutamatergic inputs, but significantly smaller proportion of cells was responsive in the winner. These results indicate that increase in proportion of responsive cells causes potentiation of dHbM-vIPN transmission in the loser. Our further study revealed that acetylcholine promotes recruitment of GluA1 AMPA receptor subunit onto the postsynaptic membrane via α7 nicotinic acetylcholine receptor. Our study suggests that experience of defeat induces acetylcholine-mediated potentiation in the vIPN, and that this plasticity is neural basis for loser's effect.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-298 海馬台から投射先領域への経路選択的な情報伝達 Takuma Kitanishi（北西卓磨）^1,2,3，Ryoko Umaba（馬場良子）^2,4，Kenji Mizuseki（水関健司）^1,2 ¹大阪市大院医神経生理 ²大阪市大院医脳科学研究センター ³JSTさきがけ ⁴大阪市大院医脳外科 Higher brain areas often process multimodal information. The hippocampus is one such area, in which principal cells convey a variety of information including space, time, emotion, and memory. However, how such information is distributed to multiple downstream areas remains poorly understood. The subiculum is the major hippocampal output structure that receives the hippocampal CA1 output and projects to multiple cortical/subcortical areas. Despite its anatomical importance, the nature of information distribution from the subiculum to downstream areas is largely unknown. To investigate this issue, we devised a method to optogenetically identify the projection targets of individual subicular neurons during large-scale extracellular recordings in freely behaving rats. We first stereotaxically introduced channelrhodopsin-2-expressing viral vector and a 256-channel silicon probe into the dorsal subiculum, and also implanted up to four optical fibers targeting each of the subicular projection targets (i.e. nucleus accumbens, anteroventral thalamic nucleus, retrosplenial cortex, and medial mamillary body) in single rats. Then, two to three weeks after the surgery, while monitoring the firing activity of dozens of subicular neurons, we identified projection targets of the activity-monitored neurons by giving blue light pulses sequentially to each of the target areas. A proportion of subicular neurons reliably generated short-latency, low-jitter, antidromic spikes in response to the light irradiation, and were identified as projection neurons targeting the irradiated area. We then measured the information encoded by the projection neurons (e.g., spatial, head-direction, and working memory information), by monitoring neuronal activity during multiple behavioral tasks (open field, linear track, T-maze, and zigzag maze) and sleep. Preliminary analysis suggests that several types of information conveyed by subicular neurons are distributed in a target-specific manner during the tasks and sleep. Thus, our large-scale opto-electrophysiological method allows us to uncover how multimodal information in the hippocampus is distributed to multiple downstream areas through the subiculum.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-299 恐怖記憶を支える局所回路ならびに大域回路のダイナミクス Hiroyuki Miyawaki（宮脇寛行），Kenji Mizuseki（水関健司）大阪市大院医神経生理学 Animals acquire a memory through experiences and retrieve the memory upon demands. Memories are enabled by global neuronal circuits across brain regions as well as local circuits within each brain region. In this study, we investigated how dynamics of neuronal circuits in different anatomical scales are involved in the memory process. Hypothesized neuronal dynamics in local and global circuits depict quite different pictures. In the local circuit level, it is proposed that a subset of neurons is responsible for a memory, and the same subset is activated during memory acquisition and retrieval. On the other hand, in the global circuit level, it is suggested that an acquired memory is consolidated through sleep, which involves shifting responsible neuronal circuits. As the consequence, the activated neuronal circuits should be different during memory acquisition and retrieval. Thus, constancy and changeability in neuronal dynamics underlying memory are suggested in local and global circuit levels, respectively. To seek how these discrepant views are incorporated, we recorded activities of hundreds of neurons in multiple brain regions simultaneously though the entire memory process. We used fear conditioning as a model system, because the timing of memory acquisition and retrieval are clearly dissociated. Accumulating evidence suggests that fear memory involves amygdala, prefrontal cortex, and ventral hippocampus. Thus, we performed simultaneous large-scale electrophysiological recordings in these brain regions of freely moving rats. Recordings started prior to memory acquisition and continued until post-retrieval period including sleep interleaved behavioral sessions. For behavioral evaluation and sleep scoring, electrocardiogram, electrocorticogram of olfactory bulbs, electromyogram of nuchal muscles, and head acceleration were monitored during the recordings. We observed different modulations of neuronal firing by freezing behavior during memory acquisition and retrieval. Interestingly, changes of the modulation were variable across the brain regions. These results suggest that neuronal dynamics changes through memory process both in local and global levels. We will present how neuronal coding changes in single cells, local circuits, and global circuits, and how the coding evolves across sleep interleaved memory acquisition and retrieval.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-300 大脳皮質の機能的連関とストレス誘発性心拍変動の関連 Ryota Nakayama（中山亮太）¹，Yuji Ikegaya（池谷裕二）¹，Takuya Sasaki（佐々木拓哉）^1,2 ¹東京大院薬薬品作用 ²JSTさきがけ Mental stress induces a variety of biological responses across the body. Although many early studies revealed stress-induced biological factors at the molecular and cellular levels, relatively few systemic physiological studies provided insights into stress responses, especially for functional interactions across the central and peripheral organs. It has been shown that stress-induced physiological responses in animals considerably differ across individual rats. We hypothesized that variability in stress-induces response may be explained by intrinsic brain activity patterns. To address this hypothesis, we recorded local field potential signals from six cortical areas, electrocardiograms, and electromyograms from freely moving rats exposed to acute social defeat stress. Based on their stress-induced changes in cardiac signals, individual defeated rats were classified into stress susceptible and resilient groups. Rats with higher correlations in delta power and lower correlations in theta power across wide ranges of cortical regions before the stress challenge had higher probability to be stress-susceptible rats as defined based on the irregularity of heartbeat signals. A combination of principal component analysis and the support vector machine algorithm revealed that functional connections across cortical regions could be predictive factors accounting for individual differences in future stress susceptibility. These results suggest that individual differences in cortical activity may be a mechanism that causes abnormal activity of peripheral organs in response to mental stress episodes. The evidence will advance the understanding of the neurophysiological correlates of mind-body associations during mental stress exposure.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-301 海馬における優先順位付けされた経験のリプレイ Hideyoshi Igata（井形秀吉）¹，Yuji Ikegaya（池谷裕二）¹，Takuya Sasaki（佐々木拓哉）^1,2 ¹東京大院薬薬品作用 ²JSTさきがけ Animals learn appropriate spatial navigation strategies to a goal through multiple experiences. During this learning process, activity patterns of brain circuits are gradually optimized. Accumulated evidence has shown that the hippocampus plays an important role in spatial learning by representing current and future episodes. Especially, memory consolidation and future planning are considered to be supported by awake hippocampal replay in which temporally compressed sequential patterns of place cells are reactivated corresponding with running trajectories. To further reveal detailed physiological characteristics of hippocampal awake replay during learning, we designed a spatial task in which rats learn to take a specific route point to a fixed goal in a two-dimensional task field. In this task, a trial began when the animals performed an active nose poke in a start box where sucrose water was presented for 10 seconds. A start door was then opened so that the rats entered into the field. The rats could obtain reward at a goal box if they correctly stopped at a specific point where a small amount of chocolate milk was placed on the way to the goal box. After the rats continuously performed the same task for 3-4 days, a reward point was replaced to a different location. The rats first showed exploratory behavior throughout the field after the rewarding rule was changed but could learn a new optimal route through the trial-and-error exploration. During this learning process, a multiunit recording was performed from hippocampal CA1 neurons. We found that the number of synchronous events more frequently emerged and correlation of synchronous events became higher after the rewarding rule was changed, demonstrating that active neuronal ensembles within synchronous events were pronouncedly reorganized associating with a change in behavioral strategy. Furthermore, Bayesian decoding analysis revealed that synchronous events over-represented the trajectory to new reward location. These results suggest that the contents of hippocampal neuronal reactivation are prioritized for learning, which supports the reinforcement of a specific behavior pattern.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-302 接近-回避の葛藤における背側及び腹側海馬の活動 Sakura Okada（岡田桜）¹，Yuji Ikegaya（池谷裕二）¹，Takuya Sasaki（佐々木拓哉）^1,2 ¹東京大院薬薬品作用 ²JSTさきがけ Animals evaluate the influences of external events and decides whether to take avoidance behavior. In some circumstances, animals need to actively accept an aversive situation depending on the balance between the amount of expected benefit and the strength of negative emotion. In our previous research, we designed a behavioral task in which rats performed an acceptance/aversion conflict behavior against an airpuff stimulation during a continuous version of the T-maze alternation task. By recording neuronal spike patterns from the dorsal hippocampus, we found that the application of airpuff altered spatial firing activity of a certain population of neurons. This result suggests that hippocampal spatial maps are flexibly reorganized to represent particular aversive events. On the other hand, the ventral side of the hippocampus is considered to be crucial for affective behavior. In addition, a previous study has shown that ventral hippocampal neurons projecting to different extrahippocampal areas are activated in different emotional states such as reward anticipation and fear. In this study, we hypothesized that ventral hippocampal activity is related to an acceptance/aversion conflict behavior. Our previous task was improved so that it allows stable recordings for consecutive several days. In addition, we developed new devices using a 3D printer for simultaneously implanting tetrode assembly and multisite silicon neural probes to the dorsal and ventral parts of the hippocampus, respectively. We present some typical spike patterns of hippocampal neurons obtained from rats performing the improved task using these recording configurations. The datasets will provide a new insight about neuronal mechanisms underlying information processing of spatial and aversive events along the dorso-ventral axis of the hippocampus.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-303 覚醒ラットにおける海馬場所細胞群の発火活動と前頭皮質ニューロンの膜電位変動の解析 Yuya Nishimura（西村侑也）¹，Yuji Ikegaya（池谷裕二）¹，Takuya Sasaki（佐々木拓哉）^1,2 ¹東京大院薬薬品作用 ²JSTさきがけ The hippocampus plays an important role for processing of episodic memory. During sleep states, the hippocampus generates a characteristic high-frequency oscillation, termed sharp wave-ripple (SWR), during which hippocampal cells are sequentially activated. Selective suppression of SWRs during sleep after memory acquisition has been shown to interfere with memory consolidation. A number of electrophysiological studies have revealed that spike timing of neurons in the neocortex is modulated by hippocampal SWRs, implying that SWRs serve as a neural substrate for information transfer from the hippocampus to the neocortex for systems memory consolidation. It remains unclear how hippocampal SWR-induced synchronous spike patterns modulate subthreshold membrane potential dynamics of downstream neocortical neurons. To address this question, we simultaneously performed a multiunit recording of hippocampal neurons and an in vivo whole-cell patch-clamp recording of a single neuron in the prefrontal cortex (PFC) from an awake head-fixed animal. First, spike patterns of hippocampal cell populations were recorded as rats performed a U-maze spatial task. After the multiunit recording, the animals were head fixed and a whole-cell patch-clamp recording was obtained from a PFC neuron. We found that subthreshold depolarization was evoked in a subset of patched PFC neurons during SWR-induced reactivation of hippocampal place cells. Especially, place cells encoding the same space tended to evoke stronger depolarization in the PFC neurons. These results suggest that specific reactivation patterns of hippocampal place cell ensembles are effectively transferred in the hippocampal-neocortical circuit.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-304 海馬場所細胞における空間の情報と目的地に関する情報の統合 Yuki Aoki（青木勇樹）¹，Hideyoshi Igata（井形秀吉）¹，Yuji Ikegaya（池谷裕二）¹，Takuya Sasaki（佐々木拓哉）^1,2 ¹東京大院薬薬品作用 ²JSTさきがけ The hippocampus is composed of place cells that fire preferentially when an animal visits a restricted area of an environment. Spatial firing of hippocampal place cells varies depending on the animal's moving directions relative to its goals, as demonstrated by spatial tasks with linearly constrained paths. Here, we systematically examined how single hippocampal neurons represent a combination of goal-directed and place-selective signals in a two-dimensional open field. Rats were well trained to approach visually guided reward ports attached to individual walls of the field. In this two-dimensional condition, hippocampal place cells encode two independent pieces of information, spatial representation and goal-directed representation by amplifying firing rates within their place fields specifically while the animal is moving toward a specific goal location defined for individual cells. The in-field firing rates of place cells were higher than out-of-field firing rates irrespective of running speed and direction, indicating the presence of substantial place-selective excitatory drives that sustain a basal spatial map independent of goal-directed signals. When animals were allowed to freely forage in the same field, in-field firing rates similarly increased when the animals transiently ran toward remembered goal locations, suggesting that internally driven behavior without visual cues is sufficient to boost place-selective firing. Disruption of medial septal activity significantly decreased goal-directed firing while maintaining spatial representation patterns. Overall, such integrated encoding of spatial and goal-directed signals by hippocampal circuits is considered to underlie flexible spatial navigation to a goal location.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-305 海馬CA1野シータ波を生み出す錐体細胞の閾値下膜電位ダイナミクス Asako Noguchi（野口朝子），Nobuyoshi Matsumoto（松本信圭），Yuji Ikegaya（池谷裕二）東京大院薬薬品作用 Local field potentials (LFPs) in the hippocampal CA1 subregion are associated with behaviors of animals, including memory, learning, and place representation. For example, theta (θ) oscillations (3-10 Hz) are believed to play an important role in active exploration and memory encoding; however, little is known about how θ oscillations of LFPs (θ_LFP) emerge collectively from activity of individual neurons. LFPs is thought to mainly reflect synaptic transmembrane currents (Buzsàki et al., 2012), which also is the main source of subthreshold fluctuations in membrane potentials (Vm) of individual neurons. In the present study, we investigated how subthreshold Vm dynamics of neuron ensembles are associated with extracellular θLFP. In the present study, we recorded CA1 LFPs simultaneously with Vm of up to four CA1 pyramidal neurons in anesthetized mice. We found that individual neurons intermittently exhibited θ oscillations in subthreshold Vm (θ_Vm), often together with θ_LFP. The simultaneously occurring θ_LFP and θ_Vm and the simultaneously occurring θ_Vm of CA1 cell pairs tended to have the identical frequencies, although the frequencies of θ_LFP and θ_Vm spontaneously fluctuated over time. We also showed that when multiple CA1 pyramidal neurons emitted more similar θ_Vm frequencies among them, θ_LFP became greater in power, and its frequency became closer to the mean of the θ_Vm frequencies of those neurons. Therefore, CA1 θ_LFP is predictable from θ_Vm of only a few surrounding CA1 pyramidal neurons.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-306 海馬-線条体ネットワークにおける分単位の経過時間をエンコードする神経発火 Yu Shikano（鹿野悠）¹，Yuji Ikegaya（池谷裕二）¹，Takuya Sasaki（佐々木拓哉）^1,2 ¹東京大院薬薬品作用 ²JSTさきがけ To plan future actions, animals need to recognize the temporal context of the events according to previously encoded memories with a variable time range. Previous studies have reported that neuronal spike activity in the hippocampus, neocortex, and striatum, underlies temporal processing on the timescale of tens of seconds. However, neuronal mechanisms for processing longer timescales remain unknown. In this study, we developed a novel behavioral task in which rats were placed in a small behavioral box with a feeding port at a specific location. The animals were rewarded if they peeked in the reward port to collect single 45-mg food pellets that were presented for 2-3 s every five minutes. This paradigm provided the opportunity to examine how animal's explicit timing behavior for predictable events are associated with temporal patterns of neuronal spiking activity over minutes-range timescale. To minimize the effects of variable locomotion and locations on neural signals, the flexibility of animal's locomotion was held relatively constant in a small box. Temporal changes in the probability of peeking behavior was quantified as a behavioral correlate of the temporal estimate from the animals. Bilateral injection of muscimol into the hippocampus decreased temporal information. Next, we recorded multiunit spike activity from the dorsal hippocampal CA1 area and dorsal striatum as rats performed the task. A subset of the recorded neurons showed temporal changes in firing patterns in each 5-min trial. Such characteristic spike patterns were not simply explained by animals' peeking behavior, positions, or running speed. Our results provide evidence that hippocampal and striatal neurons periodically and repeatedly represent the elapsed time to cover a duration on a time scale of minutes.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-307 単一動物からの脳波の多領域計測と脳組織サンプル回収法の開発 Daichi Konno（紺野大地）^1,2，Ryota Nakayama（中山亮太）¹，Yuji Ikegaya（池谷裕二）^1,4，Takuya Sasaki（佐々木拓哉）^1,3 ¹東京大院薬薬品作用 ²東京大院医 ³JSTさきがけ ⁴JSTERATO For understanding brain functions in health and disease, a large number of studies have been performed at molecular, genetic, and network system levels. To further comprehensively connect the pieces of the insights obtained from these independent studies, experimental techniques are required to collect as much information as possible from a single living animal, including whole-brain functional connection patterns, the concentration of bioactive substances, and gene expression patterns. This technical requirement has not been easily attainable for small experimental animals due to the space and weight limitations of experimental devices. To address this technical issue, we developed a novel recording technique to monitor multiple extracellular local field potential and electrocorticography signals from wide brain areas in one hemisphere while biochemical samples including genes and extracellular fluid are collected from the other hemisphere from a freely moving rodent. We describe technical procedures of our novel method and representative recording examples from rats and mice. Additionally, the recordings were chronically stable for at least 2 months after implantation of the device. The methodological concept is applicable to a wide range of biological reactions at various spatiotemporal scales, allowing us to integrate an idea of physiolomics into existing omics analyses and help connect insights at a variety of spatiotemporal scales from microscopic to macroscopic studies.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-308 ラット海馬の機能的左右半球非対称性および半球間相互作用 Yukitoshi Sakaguchi（阪口幸駿），Yoshio Sakurai（櫻井芳雄）同志社大院脳科学 Functional asymmetry in the left/right hemisphere is a well-known feature in human, e.g., the left-sided function of the language area and the right-sided spacial cognitive function. However, such asymmetrical function is not specific in human but also in the various animals ranging from fish, birds, and reptiles. Recently, an anatomical asymmetry has also been reported in some areas of the brain of rodents such as rats and mice. Among them, the brain region in which a large number of reports have been reported is the hippocampus, and the left and right difference such as the volume, the expressed gene, and protein, the type, and density of receptors contained in the synapse, etc. are repeatedly suggested (Kawakami et al. 2003; Shinohara et al., 2008). However, it is not known in detail whether such anatomical difference will affect the functional differences. Therefore, we investigated the functional asymmetry of the rat hippocampus and the actual condition of the hemispheric interaction using the hippocampus-dependent memory tasks. We performed two types of experiments; a group for electrically damaging the left or right hippocampus and a group which injected virus into the hippocampus for introducing the photoactivated protein (hSyn-hChR2(H134R)-EYFP, hsyn-Jaws-KGC-GFP-ER2) and the projection pathway through the ventral hippocampal commissure, from the left CA3 to the right CA3 or the right CA3 to the left CA3 was activated/inactivated by the optogenetic method. For behavioral tests, spontaneous alternative test (SAT), place preference test (SPT) and plus maze test (PMT) was used. In this presentation, from the results obtained from these experiments, we will introduce the possible contribution of the functional left and right hemispherical asymmetry and the actual condition of hemisphere interaction to the memory formation.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-309 海馬-嗅内皮質における細胞種、領域特異的な速度表現 Motosada Iwase（岩瀬元貞），Takuma Kitanishi（北西卓磨），Kenji Mizuseki（水関健司）大阪市大院医神経生理学 Place cells in the hippocampus and grid cells in the medial entorhinal cortex (MEC) represent the current location of an animal. To reflect on-going self-motion, place and grid cells update spatial representation by accessing information regarding current direction and speed of an animal's movement. Speed is encoded by context-invariant, speed-responsive cells in the MEC and hippocampus (Kropff et al., 2015). However, complete quantitative characterization of speed representation in the hippocampal-entorhinal circuit is yet to be achieved. Using electrophysiological data recorded simultaneously from the hippocampus and MEC during foraging in the open-field (Mizuseki et al., 2014), we compared speed representation in distinct cell types across sub-regions and layers in the hippocampal-entorhinal circuit. We demonstrated that, in comparison with principal cells, a larger proportion of putative interneurons were significantly speed-modulated in CA1, CA3, MEC layer 3 (EC3), and EC5; these results align with those of previous reports (Kropff et al., 2015; Ye et al., 2018). In contrast, the proportion of principal cells in EC2 that were significantly speed-modulated was as large as that of EC2 interneurons. The slopes of regression lines calculated for speed and instantaneous firing rate of CA1, CA3, and EC5 principal neurons were larger than those for principal neurons of EC2 and EC3 and interneurons of CA1, CA3, and EC5. In congruence with Kropff et al., 2015, we found that most speed-modulated cells in EC2 encode speed prospectively, whereas those in the CA1 and EC3 encode speed retrospectively. As a population, CA3 and EC5 neurons did not show any significant bias toward prospective or retrospective coding. EC2 and EC3 neurons, but not hippocampal or EC5 neurons, exhibited significant correlations between the magnitude of theta modulation and prospective vs. retrospective coding. Additionally, we identified hippocampal interneurons that were putative parvalbumin- (PV) and somatostatin- (SOM) positive by using physiological criteria based on burst tendency and refractory period (Fernandez-Ruiz et al., 2017). Majority of hippocampal interneurons exhibiting positive speed-modulation were putative PV-positive interneurons, whereas those exhibiting negative speed-modulation were putative SOM-positive interneurons. Our results suggest that distinct cell types in each anatomical station in the hippocampal-entorhinal circuit distinctly represent speed information.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-310 特定神経回路の分子遺伝学的機能操作による行動制御 Shigeki Kato（加藤成樹），Masateru Sugawara（菅原正晃），Kazuto Kobayashi（小林和人）福島県立医大医生体情報伝達研生体機能 Various behaviors are based on the complex neural circuitry functions. Although the reports of the correlation between a specific behavior and a selective neural pathway become increasing, a lot of unexplained mechanisms are still remained. In order to understand the systems for the information processing and those regulations in the view of neural circuitry function, gene transfer and expression via retrograde axonal transport of the viral vectors enables us to introduce the transgene into neuronal populations that innervate the brain regions where the vectors are injected. In recently, our group established new vector for neuron-specific retrograde gene transfer (NeuRet) with pseudotyping of fusion glycoprotein type E with mutated P440E (FuG-E (P440E)), which consists of the N-terminal region (440 amino acids) of the extracellular domain of rabies virus glycoprotein (RVG, CVS strain) and the membrane-proximal region (15 amino acids) a short C-terminal portion of the extracellular domain and the transmembrane/cytoplasmic domains of vesicular stomatitis virus glycoprotein (VSVG) (Kato et al., 2019). By combining with some molecular genetic approaches such as cell targeting, opto/chemogenetics methods, we addressed the roles of the pathways originating from the parafascicular nucleus (PF) and central lateral nucleus (CL) in the intralaminar thalamic nuclei in mice. Our data suggest that the PF and CL thalamostriatal systems are involved in cognitive function of basal ganglia circuitry, and that these two circuits possess distinct roles in the control of behavioral selection and flexibility. Furthermore, the subcircuits of basal ganglia including the thalamic nuclear groups that mediate discriminative learning appear to shift during the processes of learning (Kato et al., 2018). We would present a preliminary new data focusing on another neural pathway regulated animal model.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-311 記憶の固定化と想起における海馬と聴覚野のセルアセンブリのダイナミクス Shogo Takamiya（高宮渉吾），Shoko Yuki（結城笙子），Junya Hirokawa（廣川純也），Yoshio Sakurai（櫻井芳雄）同志社大学大学院脳科学研究科 Cell assemblies are populations of functionally connected neurons that encode memory and can be memory engrams. Recently, experiments utilizing optogenetics revealed reactivation of engram cells in hippocampus is necessary for retrieval of recent memory while in neocortex is necessary for retrieval of remote memory. However, these experiments used behavioral tasks that can be learned in only one experience, such as contextual fear conditioning. Therefore, the activation and reactivation of engram cells which gradually encode memory over a longer time span is unclear. The objective of this study is to reveal the dynamic change of cell assemblies in gradual processes of memory consolidation and rapid processes of memory recall using a complex behavior task and multineuronal recording. We used a conditional discrimination task with tones where rats were required to discriminate between high and low pitches. Multineuronal activities were recorded from the hippocampus during the learning processes of the task. After an interval following completion of learning, we retrained the rats with the same behavioral task to make them recall the memory of the task. In the processes of learning and recalling, we detected changes in the synchronous firing of neurons which is characteristic of cell assemblies, and we analyzed the interaction between hippocampus and auditory cortex. We discuss the relation between dynamic activities of cell assemblies in the hippocampus and auditory cortex, and degrees of memory consolidation and recall.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-312 睡眠時の皮質樹状突起活動による知覚記憶の固定化 Daichi Hirai（平井大地）^1,2，Daisuke Miyamoto（宮本大祐）¹，Yasuhiro Oisi（大石康博）¹，Maya Odagawa（小田川摩耶）¹，Chie Matsubara（松原智恵）¹，Kanako Ueno（上野加奈子）¹，Kenta Kobayashi（小林憲太）³，Akiko Hayashi-Takagi（林（高木）朗子）⁴，Masanori Murayama（村山正宜）¹ ¹理研CBS 触知覚生理学 ²日本学術振興会特別研究員 ³生理学研究所ウイルスベクター開発室 ⁴群馬大学生体調節研究所脳病態制御分野 Non-rapid eye movement (NREM) sleep after learning promotes the formation of postsynaptic dendritic spines and plays a vital role in memory consolidation. Recently, we found that top-down projection from the secondary motor cortex (M2) neurons to the primary somatosensory cortex (S1) initiated dendritic activity and persistent firing of S1 layer 5 (L5) neurons (Manita et al., 2015, Neuron), and that the top-down cortical information flow during NREM sleep is required for perceptual memory consolidation (Miyamoto et al., 2016, Science). The dendritic activity may cause an increase in intracellular concentrations of Ca²⁺, which acts as a secondary messenger in neurons and induces an activity-dependent increase or decrease in synaptic strength. Therefore, we hypothesized that the activation of such top-down circuits during NREM sleep induces dendritic activities and subsequent growth of dendritic spines in individual pyramidal neurons. To test this hypothesis, transgenic mice with cortical layer 5-specific expression of GCaMP6s (Rbp4-Cre; syn-flex-GCaMP6s) were used to perform a perceptual memory task. We measured Ca²⁺ activity during NREM sleep in single dendrites of L5 pyramidal neurons in these animals. In order to examine the effects of top-down control on dendritic activation, subsequent spine growth, and perceptual memory-based behavior, we chemogenetically inactivated the top-down inputs and observed the resulting perception inaccuracies, dendritic activity, and spine dynamics. Furthermore, the acquired perceptual memory was disrupted by the optical shrinkage of the newly potentiated spines of L5 pyramidal neurons. These data demonstrated that top-down inputs to the S1 are necessary for the dendritic dynamics underlying perceptual memory consolidation during NREM sleep.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-313 海馬神経新生エンハンサーの記憶忘却効果による社会的敗北ストレス後のPTSD様行動異常の改善 Yuka Takyu（武生優花），Chiaki Uchida（内田千晶），Rie Ishikawa（石川理絵），Satoshi Kida（喜田聡）東京農大院農バイオ Post-traumatic stress disorder (PTSD) is a mental disorder associated with traumatic memory such as fear memory. Our previous study showed that increasing adult hippocampal neurogenesis by treatment with memantine (MEM), an uncompetitive antagonist of the N-methyl-D-aspartate glutamate receptor, promotes hippocampus-dependent fear memory forgetting in mice (Ishikawa et al., 2016), raising the possibility that fear memory forgetting is a novel therapeutic target of PTSD (Kida, 2018). In rodents, repeated exposures to social defeat (SD) stress by aggressor mice induce social avoidance behaviors to the aggressor and chronic anxiety-like behaviors. In this study, we used SD stress paradigm as a new PTSD model and examined effects of MEM treatment on SD stress-induced PTSD-like behaviors. Male C57BL/6 mice were exposed to an aggressor ICR mouse for 10 min per day for 10 consecutive days (SD treatment) and then were assessed time spent in interaction and avoidance zones, respectively, in the open field in which encaged the aggressor stayed in the center of interaction zone. SD mice exhibited significantly more or less time spent in the avoidance and interaction zones, respectively, compared with control mice. Furthermore, SD group exhibited significantly less entries into open sections and time spent in open sections in the elevated zero maze task compared with control group. These observations confirmed previous findings that SD mice showed social avoidance memory and anxiety-like behaviors. Therefore, we next asked whether forgetting of social avoidance memory by neurogenesis enhancer improves anxiety-like behaviors. SD mice received systemic injections of MEM (50 mg/kg) or vehicle once a week for four weeks (SD-MEM or -VEH mice, respectively). SD-MEM mice showed less social avoidance behaviors than SD-VEH mice. Moreover, SD-MEM mice did not show reinstatement of social avoidance memory following single re-exposure to the aggressor mouse. Interestingly, SD-MEM mice showed more entries into open sections and time spent in open sections than SD-VEH mice. These observations suggest that MEM treatment showed forgetting of social avoidance memory and improved anxiety-like behaviors. Therefore, we suggest that neurogenesis enhancer improves PTSD (anxiety)-like behaviors through forgetting of traumatic (social avoidance) memory.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-314 ドネペジルとシロスタゾールの低用量併用が老齢マウスの記憶機能改善に及ぼす効果 Syuichi YANAI（柳井修一），Tomoko Arasaki（新崎智子），Shogo Endo（遠藤昌吾）東京都健康長寿医療センター研究所 The combination of two or more drugs is often more effective than using a single drug for treating diseases. Donepezil, which inhibit the hydrolysis of acetylcholine, is now considered to be the first-line treatment in patients with mild-to-moderate Alzheimer's disease, though, the combination strategy has not been established to cognitive impairment. As cAMP pathway play essential roles on learning and memory in many species, recent studies have focused on the cAMP pathway as potential therapeutics for cognitive impairment. Among them, inhibitors of phosphodiesterases (PDEs), which elevate and/or maintain intracellular cAMP concentration, are promising targets for cognitive enhancement drugs. In the present study, we examined the combination effect of sub-effective doses of donepezil and cilostazol, a PDE3 inhibitor, on age-related memory impairment in aged C57BL/6J mice. Mice (C57BL/6J) were divided into 4 groups at 22 months of age: (1) control group, (2) cilostazol group received feed containing 0.3% cilostazol, (3) donepezil group administered 0.3mg/kg/day donepezil via drinking water, (4) concurrent administration group received both cilostazol and donepezil as mentioned above. Mice were allowed feed and water ad libitum for 1 month, and then were subjected to the behavioral test battery at the age of 23 months. In the Morris water maze probe test, the control, the cilostazol, and the donepezil group performed poorly compared to young mice. This demonstrated the impaired spatial memory in 23-month-old mice, in addition, drug doses used had no beneficial effect on age-related memory impairment. However, the concurrent administration group performed significantly better than age-matched three groups, allowing them to achieve a level of memory performance similar to young mice. In the open field test, the concurrent administration group showed no significant side effects on locomotor activity and anxiety. Furthermore, the safety evaluations of cilostazol and donepezil are well established, concurrent administration of cilostazol and donepezil at low doses may lead to a new treatment for age-related cognitive impairment.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-315 ニューロプシン欠損マウスにおける空間作業記憶障害 Shoya Ide（井出翔也），Shotaro Noguchi（野口翔太郎），Hina Tajika（田近妃奈），Yoshiki Kanai（金井良樹），Yasuyuki Ishikawa（石川保幸）前橋工科大学システム生体工学科 Neuropsin (kallikrein-related peptidase 8) is concentrated in the hippocampus, amygdala, olfactory bulb, and prefrontal cortex, where it regulates memory formation. Enzymatically active neuropsin is necessary to establish the early phase of long-term potentiation (LTP) and synaptic tagging/capture in the Schaffer-collateral pathway of hippocampus, is required for spatial working memory (Y-maze) and spatial learning and memory (Morris water maze). Although evidence from genetic manipulation mice suggests an important role of neuropsin in spatial working memory, it remains unclear whether or not the requirement of neuropsin for spatial working memory depends on the time of spatial information maintained. The purpose of this study was to assess the effects of neuropsin deficiency on working memory. We found that neuropsin-deficient mice are impaired in learning and working memory as tested in spontaneous alternation and delayed non-match to sample test ( delayed time : 0, 10, 60, 180, 300 (sec)) in the modified T-maze. The major advantage of the modified T-maze protocol is its relatively simple design that is powerful enough to assess functional learning/memory. Results from the spontaneous alternation and delayed non-match to sample test tasks showed neuropsin-deficient mice exhibit slower learning in acquisition phase and slightly reduced memory performance in short-delay task ( 0, 10 (sec)). These findings indicate that the neuropsin system plays an important role in spatial working memory processes.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-316 T迷図の作業記憶減衰に関する前頭前野と海馬の局所電場電位の関係 Masatoshi Takita（瀧田正寿）^1,2，Seietsu Fujiwara（藤原清悦）³，Yukio Ichitani（一谷幸男）⁴ ¹産総研　人間情報RI ²電通大脳科学ライフサポートRC ³聖マリアンナ医大医生理 ⁴筑波大人間系 We investigated the neural mechanism of working memory decay, which is thought to be related to temporal decay and/or interference. We conducted a T-maze experiment that we designed in a previous study (Suenaga, Kaku and Ichitani, 2008). Rats were trained to choose one of two arms of a T-maze following a 75- or 150-s stay (i.e., delay period) at the start box. Their performances reached 85-95% and 80-90% correct choices for a 75- and 150-s delay, respectively, before the over-training stage. After arriving at a hole with or without a reward pellet, the rats had to return to the start box by their own volition. In previous studies of cross-lesioning and delayed response alternation, the ventral hippocampo-prefrontal pathway was found to be essential for working memory (e.g., Floresco, Seamans and Phillips, 1997; Wang and Cai, 2006), whereas our previous study suggested that the intermediate hippocampo-prefrontal pathway was also essential (Izaki, Takita and Akema, 2008). Therefore, bipolar electrodes were implanted into the medial prefrontal cortex and the intermediate and ventral hippocampus to measure differential voltages between these regions using a TBSI wireless system. Our subsequent analysis will examine the relationships between measured local field potentials in these regions of the brain (c.f. Fujiwara, Akema and Izaki, 2010) and the selection of correct or incorrect points visited by rats while they pass through a T-maze.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-317 味覚嫌悪学習によって形成されるヨーロッパモノアラガイの長期記憶と神経活動に対するエピカテキンの効果（II） Ayaka Itoh（伊藤綾香）¹，Shogo Nakada（仲田正吾）²，Yoshimasa Komatsuzaki（小松崎良将）³，Minoru Saito（斎藤稔）^1,2 ¹日本大学大学院総合基礎科学研究科 ²日本大学文理学部自然科学研究所 ³日本大学理工学部 Associative learning in the pond snail Lymnaea stagnalis has been used an important system for investigating the mechanism of learning and memory. It includes classical and operant conditioning using feeding behavior. On the other hand, modulation of memory and neurocognitive performance by flavonoids (plant phytochemicals) has been attracting much attention. In the preset study, we first investigated the effects of epicatechin, which is abundant in green tea, on long-term memory (LTM) formed by taste-aversive conditioning in Lymnaea. In this conditioning, the snail was immersed in an appetitive (10 mM sucrose) solution, which increases the feeding behavior, for 15 s paired with an aversive (10 mM KCl) solution, which inhibits it, for 15 s, and then immersed in the pure water or epicatechin solution (15 mg/L) for 9.5 min. This procedure was repeated 5 times. Before and after the conditioning, the feeding behavior was counted in the appetitive solution for 5 min in the test session. 24 hours after the conditioning, the feeding behavior decreased to over 40% and the LTM was formed in about 75% snails (good learners), but not in the other snails (poor learners). In the good learners, the LTM gradually disappeared by 72 hours in those conditioned in the pure water, while it lasted 72 hours in those conditioned in the epicatechin solution. This result shows that the LTM formation can be enhanced by epicatechin. We next measured the activities of the central giant cell (CGC), which is involved in the feeding behavior, by intracellular recording. As a result, the spontaneous activity (firing rate) in the epicatechin-included saline decreased than that in the saline. In the good learners, the spontaneous activity in the saline decreased than that in the naïve snails or the poor learners. We will also examine the activities of the neural network involved in the feeding behavior using fluorescent voltage imaging.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-318 記憶強化と想起は異なるドーパミン神経が担う Shintaro Nagano（長野慎太郎），Minoru Saitoe（齊藤実）（公財）東京都医学総合研究所　学習記憶プロジェクト Dopamine is a monoamine neurotransmitter playing critical roles in various high-order brain functions, including learning and memory. Activities of dopaminergic neuron during associative learning are relatively well identified in mammalians. However, recent studies demonstrate that dopamine release is not necessarily consistent with neural activities, suggesting that direct detection of dopamine release during associative learning is essential to understand the role of dopamine signaling in learning and memory. In Drosophila, twelve dopaminergic neurons in the fly brain, which forms the PPL1 cluster, project their axons to the mushroom bodies, an olfactory memory center of Drosophila, and concern with olfactory aversive memory. We employed genetically encoded fluorescent dopamine sensor, GRAB_DA, and fluorescent calcium sensor, G-CaMP, into in vivo imaging system to detect signaling of PPL1 dopaminergic neurons during olfactory aversive conditioning at single-cellular level using the split-GAL4 system. We found that dopaminergic neurons in PPL1 cluster released dopamine to the vertical lobes of mushroom bodies in response to either CS or US presentation. We also conducted olfactory associations and found that distinct PPL1 dopaminergic neurons regulate either memory reinforcement or memory retrieval via dopamine release to the vertical lobes of the mushroom bodies. Interestingly, dopamine release is not necessarily consistent with calcium responses in PPL1 neurons, suggesting that calcium independent cellular mechanisms regulate dopamine release. Our results demonstrate the first details of dopamine release to a memory center at single-cellular level and provide the unique properties of dopaminergic neurons in associative learning.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-319 シナプス開口放出調節因子CAPS1の海馬三シナプス経路と海馬学習における役割 Tomoki Arima（有馬知輝）¹，Chiaki Ishii（石井千晶）¹，Yuki Ishii（石井佑季）¹，Natsumi Shibano（柴野奈津美）¹，Mio Yamazaki（山﨑実緒）¹，Yuna Kato（加藤優奈）¹，Ami Yamato（大和綾巳）¹，Yo Shinoda（篠田陽）²，Tetsushi Sadakata（定方哲史）³，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東京薬科大学公衆衛生学教室 ³群馬大先端科学研究指導者育成ユニット Presynaptic exocytosis of synaptic vesicles (SVs) is a key event in synaptic transmission and plasticity and comprises three steps: docking, priming and fusion which are finely regulated by many kinds of presynaptic molecules in an orchestrated fashion. Recent studies have demonstrated that Ca2+-dependent activator protein for secretion 1 (CAPS1) that is thought to be as a regulator for the priming of exocytosis of dense-core vesicles (DCVs), is now also recognized as one of the regulatory molecules that are involved in exocytosis of SVs in hippocampus. However, the physiological role of CAPS1 in exocytosis of SVs still remains unknown. Previously, we generated forebrain-specific CAPS1 conditional knockout mice (cKO). And we showed that the basal excitatory synaptic transmission and presynaptic release were severely impaired in hippocampal CA3-CA1 synapses of CAPS1 cKO mice. Electron-microscopic analysis revealed decreased number of SVs docked to the presynaptic active zone membrane of cKO mice compared to control mice. From these findings, we proposed that CAPS1 stabilizes the state of readily releasable pool, thereby enhancing synaptic transmission efficacy. To further address the physiological role of CAPS1, we analyzed function in trisynaptic circuit and learning and memory abilities of CAPS1 cKO mice. We found that presynaptic short-term plasticity and theta burst-induced long-term potentiation (LTP) were greatly impaired in DG-CA3 synapses of cKO mice compared to control mice. Interestingly, LTP in CA3-CA1 synapse was not inhibited in CAPS1 cKO compared to control mice. Thus we suggest that the contribution of CAPS1 to synaptic plasticity in the hippocampal trisynaptic circuit is pathway-specific. In the behavioral analysis, CAPS1 cKO mice showed deficit in contextual fear conditioning (CFC) but not in auditory cued fear conditioning. In CFC, both of short-term and long-term memory formation were impaired in CAPS1 cKO mice. We also found that memory formation in CFC was impaired when caps1 was deleted in the adult hippocampus. Consequently, we again suggest that CAPS1 plays an essential role not only in exocytosis of SVs but also in short-term and long-term plasticity and hippocampal memory formation, although the significance of the role of CAPS1 in the hippocampal trisynaptic circuit is synapse-type-specific.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-320 Alcohol – nicotine interactive effect on hippocampal-dependent cognitive functions in mice Dong Kwan Kim（Kim Dong Kwan），Jaehun Jung（Jung Jaehun），Daeryong Kim（Kim Daeryong），Suaa Shin（Shin Suaa），Jinha Kim（Kim Jinha），So Young Choi（Choi So Young） Department of Physiology, Konyang University College of Medicine, Daejeon, Korea Alcohol – nicotine interactive effect on hippocampal-dependent cognitive functions in mice 抄録：Alcohol and nicotine are commonly co-abused. It is still controversial to improve the alcohol induced cognitive deficits by nicotine, one possible explanation for co-abuse. Thus, this study simultaneously examined the interactive effects of 1 week’s ethanol and nicotine co-administration on Hippocampal long-term potentiation (LTP) and memory behaviors. C57BL/6 mice were administered nicotine (0.5 mg / kg) or saline twice daily by subcutaneous injection, and once daily with ethanol (3 g / kg, 25% v / v) or tab water via a feeding needle for 7 days. In this experiment, we divided into four groups: sham (saline + tab water), nicotine (nicotine + tab water), ethanol (saline + ethanol) and nicotine + ethanol. At 24 hours after 1 week’s administrations, the hippocampal memory behaviors were checked with the Y maze test and passive-avoidance test. To induce LTP, we used theta burst in CA1 field recording with hippocampal slice. Alcohol alone administration caused memory deficit and LTP impairment, but nicotine alone administration did not alter memory behavior and hippocampal LTP. There were no the negative effects on hippocampal LTP and memory behaviors in combination group (nicotine + ethanol). This study clearly demonstrated the relationship between alcohol and nicotine in hippocampal-dependent cognitive functions and the ameliorative effect of nicotine on ethanol-induced memory deficits have been simultaneously reproduced at the hippocampal LTP. These results could be helpful to understand the interactive effects of both drugs and how they are altering behavior and synaptic plasticity, and to develop treatments for drug addiction and memory impairment.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-321 線虫早期順応での感覚および介在神経細胞間関係のイメージングによる解明 Keita Ashida（芦田慶太）¹，Hisashi Shidara（設樂久志）^1,2，Kohji Hotta（堀田耕司）¹，Kotaro Oka（岡浩太郎）¹ ¹慶應大理工生命情報 ²北海道大院理 The input-output relationship and its modulation are fundamental factors for understanding neuronal information processing. To understand the relationship and the modulation, detailed dissection of synaptic transmission between pre- and postsynaptic neuron is required. However, the relationship between pre- and postsynaptic neurons in vivo is still largely unknown due to the complexity of the involved networks. The nematode, Caenorhabditis elegans is suitable for such investigations due to its simple neuronal network, detailed information about neuronal connectivity, and transparent body for imaging. Although previous imaging studies have visualized synaptic-vesicle release and input, no research has been reported on modulations of both pre- and postsynaptic relationship directly. In C. elegans, pre-exposure to odor reduces chemotaxis to the odorant. A specific adaptation, induced by five minutes of pre-exposure, is known as early adaptation. AWC sensory neurons and AIY interneurons play crucial roles in the adaptation. AWC neurons sense volatile odors, and AIY neurons receive glutamatergic inputs from AWC. In addition, these neurons are necessary for odor attraction. While the molecular basis of early adaptation is well investigated, modulations of neuronal activity in AWC and AIY neurons are still unknown. To understand such modulations, both the neural activities of AWC and AIY, and the synaptic transmission between these neurons must be investigated in detail. Here, to use various fluorescent imaging techniques, we show that the reduction of synaptic-vesicle release without Ca²⁺ modulation in AWC suppresses Ca²⁺ response in AIY by early adaptation. Our results demonstrate modulation of the pre-post synaptic relationship in vivo by imaging techniques and provide new type of modulations of neurotransmitter release without Ca²⁺ modulation in vivo.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-322 加齢による記憶・学習能力の低下と睡眠の関係 Masahiro Fujimura（藤村将大）¹，Takaaki Yamada（山田貴亮）¹，Yuichiro Ogata（大形悠一郎）¹，Hisashi Yoshioka（吉岡寿）¹，Seiji Hasegawa（長谷川靖司）^1,2，Satoru Nakata（中田悟）¹ ¹日本メナード化粧品株式会社　総合研究所 ²名古屋大学　メナード協同研究講座 Neurogenesis from neural stem cells in the hippocampal dentate gyrus in the brain has a critical role for memory and learning ability. However, both the number of neural stem cells and neurogenesis in the dentate gyrus have been reported to decline with aging. Previously, we reported that sleep deprivation impaired memory and learning ability in mice, and it was suggested to be caused by elevated expression levels of inflammatory cytokines and a decrease in neurogenesis in the brain. In this study, we tested a hypothesis that poor sleep quality in older adults might affect memory and learning ability, and also examined the changes in the number of neural stem cells and neurogenesis with aging. Male HR-1 mice (Japan SLC) were categorized as younger (12 week old) and older (55 week old) adult groups. Activity-sleep cycle was monitored using SUPERMEX. To evaluate memory and learning ability of mice in each group, Morris water maze test was conducted, and inflammatory cytokines in the brain were quantified using a Cytokine Array. We also performed immunostaining in the frozen sections of the brains from younger and older adult mice to evaluate neurogenesis. Moreover, in order to examine the effects of inflammatory cytokines on neurogenesis, a variety of cytokines were added to in vitro cultures of MEB5 cells, a mouse neural stem cell line, during neural induction, and the changes in the gene expression were analyzed. Total sleep time (TST) during the light/sleeping period was significantly reduced in the older group compared to the younger group. In Morris water maze, the older group had a lower score than the younger group. Immunostaining against doublecortin (DCX), a neurogenesis marker, revealed a reduction in the number of DCX-positive cells in the older group. Furthermore, we found that the expression levels of some cytokines including fractalkine were higher in the brains of the older group. When we added these cytokines to MEB5 cells during neural induction, expressions of differentiation-related genes were observed to be downregulated. Given that in older mice TST was reduced and the expressions of some inflammatory cytokines such as fractalkine were increased in the brain, it was suggested that poor sleep quality might be one of the causes of age-related decreases in memory and learning ability. We will further analyze how sleep affects memory and learning ability in detail.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-323 MPTPパーキンソン・モデルマウスの3レバー・オペラント課題成績の低下 Mitsugu Yoneda（米田貢），Akari Usuta（臼田明莉），Masataka Nakagawa（中川雅嵩），Hayate Tanigami（谷上颯），Ryo Sakata（坂田嶺），Takako Ohno-Shosaku（少作隆子）金沢大学医薬保健研究域保健学系リハビリテーション科学 We have developed a three–lever operant task to evaluate reward–based learning of a motor sequence in rats and mice. To examine the dependence of this task on basal ganglia and dopaminergic signal, we applied this task to an MPTP mouse model of Parkinson´s disease. We used eight MPTP–treated mice (MPTP mice) and seven control mice of the C57BL/6 strain. Body weights were not different between the two groups. In the operant box, three levers (A–C) were positioned 2 cm (right and left levers) and 4 cm (center lever) above the floor. One training session lasting 60 min was given once a day and five times a week. The mice were trained to press any one of the active levers for a food reward (reinforcement) as shaping (1–lever task). When the mice pressed the same active lever more than 100 times per session in two consecutive sessions, the lever was inactivated in the subsequent sessions. When all three levers were inactivated, 1–lever task was completed. The mice were then trained to press three levers in a given sequence (ABC) (3–lever task). After the mice showed good performance of the three-lever task, the order was reversed to CBA (reverse 3–lever task). In 1–lever task, the total number of sessions required for completion of 1–lever task was not different between the two groups. The number of lever presses and reinforcements during the first several sessions and the rate of win–stay (pressing the same lever following reinforcement) under the condition of one active lever were higher in MPTP mice than in control mice, whereas the inactive lever press ratio ("the number of inactive lever presses" / "the number of lever presses") under the condition of one active lever was lower in MPTP mice. In 3–lever and reverse 3–lever tasks, the total number of sessions required for completion of the task was not different between the two groups. The success rate was lower in MPTP mice, whereas the number of AA/BB/CC patters was higher in MPTP mice. These results suggest that MPTP mice tend to press the same lever repeatedly, which interferes with the learning of a motor sequence. The impairment of reward–based learning of a motor sequence in the three–lever operant task in MPTP mice clearly shows the dependency of this task on basal ganglia and dopaminergic signal.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-324 歩行する海鳥および遊泳するサケ科魚類の脳における空間ナビゲーションの神経相関 Kaoru Ide（井出薫）¹，Takumi Hombe（本部拓未）²，Shinichiro Okamoto（岡本慎一郎）⁴，Sakiko Matsumoto（松本祥子）²，Ken Yoda（依田憲）²，Yuya Makiguchi（牧口祐也）³，Susumu Takahashi（高橋晋）¹ ¹同志社大院脳科学 ²名古屋大院環境学 ³日本大学生物資源科学部 ⁴順天大院　医神経生物・形態 The discovery of place cells in the hippocampus provided an insight into the neuronal mechanism on spatial navigation. However, whether place cell relatives exist in the brain of other species such as fish, amphibians, reptiles and birds remains unclear. The divergence times between mammals and fish and between mammals and birds are incredibly long. Besides, they have much different brain structure. Especially, seabird and salmonid fish can navigate through the ways over 100 kilometers long distance. Can the accumulating lines of evidence on the place cell and its relatives in the mammalian brain account for such amazing navigational ability? On the other hand, leading hypotheses in the environmental science field propose that geomagnetic and/or odor cues guide the long-distance navigation. However, such hypotheses never account for the neuronal mechanisms for spatial navigation in the brain. Here, to address the question, we are recording neuronal activity from the brain of seabirds and salmonid fish. To examine the neuronal underpinning for long-distance navigation, we constructed an experimental setup for simultaneously recording single-unit activity and local field potential from the brain region homologous to the hippocampus of mammals, of walking seabirds and swimming salmonid fish using a tiny, lightweight logger for neuronal activity. We implanted four tetrodes into the most caudal, dorsomedial regions of the telencephalon of young seabirds called Streaked Shearwater freely walking in an open field, and the telencephalon of adult salmonid fish called rainbow trout freely swimming in a square-shaped water tank. For seabirds, complex spike bursts could be observed, suggesting that the recorded region is homologous to the hippocampus of mammals. However, inhibitory interactions between two putative interneurons simultaneously recorded from a tetrode could be observed. A few neurons transiently fired at a specific time or location during walking, might suggesting the existence of place cell relatives in the brain of seabirds. For salmonid fish, we found a head direction-like neuron which fires specifically when the heading directs to a certain direction. We report preliminary results on such neural correlate between neuronal activity and spatial navigational behavior.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-325 マウスの3レバーオペラント課題を用いた報酬に基づく運動学習に対するチョコレート摂取の効果 Hayate Tanigami（谷上颯），Masataka Nakagawa（中川雅崇），Akari Usuda（臼田明莉），Mitsugu Yoneda（米田貢），Takako Ohno-Shosaku（少作隆子）金沢大学・医薬保健学・リハビリテーション科学 Theobromine is the primary methylxanthine in the products of Theobroma cacao, and found in chocolate. It works as a phosphodiesterase inhibitor to increase cAMP, and activates cAMP-response element-binding protein (CREB). Activation of CREB is involved in a variety of processes in the brain, including induction of brain-derived neurotrophic factor (BDNF) and higher brain functions such as learning and memory. We previously reported that orally-administered theobromine up-regulates CREB/BDNF pathways, and promotes reward-based learning of a motor sequence in mice. In the present study, we examined effects of chocolate on motor learning, by using the same learning task, a three-lever operant task, where animals are trained to press three levers in a given sequence. Before training, mice had normal diet (control mice) or a diet supplemented with chocolate (chocolate mice) for 30 days. Body weights were not different between the two groups. In the operant box, three levers (A-C) were positioned 2 cm (right and left levers) and 4 cm (center lever) above the floor. One training session lasting 60 min was given once a day and five times a week. The mice were trained to press any one of the active levers for a food reward (reinforcement) as shaping (1-lever task). When the mice pressed the same active lever more than 100 times per session in two consecutive sessions, the lever was inactivated in the subsequent sessions. When all three levers were inactivated, 1-lever task was completed. The mice were then trained to press three levers in a given sequence (ABC) (3-lever task). After the mice showed good performance of the three-lever task, the order was reversed to CBA (reverse 3-lever task). In 1-lever task, the total number of sessions required for completion of 1-lever task and the total number of lever press at each session were not different between control and chocolate mice. In 3-lever task, the total number of sessions required for completion of 3-lever task was not different. The number of reinforcements and ABCABC patterns were significantly higher in chocolate mice than in control mice. The success rate tended to be higher in chocolate mice. In reverse 3-lever task, neither the total number of sessions required for completion of reverse 3-lever task nor the success rate were different between the two groups. These results suggest that chocolate ingestion might have positive effects on motor learning in mice.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-326 ニューロステロイドによる時刻依存的な空間記憶の長期維持 Kimiko Shimizu（清水貴美子）¹，Kanako Maehata（前畑佳納子）¹，Tomoko Ikeno（池野知子）¹，Qiuyi Wang（WANG Qiuyi）²，Toshifumi Takao（高尾敏文）²，Yoshitaka Fukada（深田吉孝）¹ ¹東京大院理生物科学 ²大阪大蛋白研 Neurosteroids are neuroactive steroids that are synthesized in the brain and they regulate a variety of biological functions including cognition and behavior. These neurosteroids are synthesized from cholesterol by a series of cytochrome P450 enzymes, among which a new member of P450 hydroxylase, cytochrome P450-7b1 (CYP7B1), has been shown to catalyze formation of 7-hydroxylated neurosteroids, 7α-hydroxypregnenolone (7α-OH-Preg) and 7α-hydroxydehydroepiandrosterone (7α-OH-DHEA). Here we found that the mRNAs encoding the enzymes required for the synthesis of 7α-OH-Preg and 7α-OH-DHEA from cholesterol were detected in all the brain regions we examined. Higher levels and day-night change of Cyp7b1 mRNA expression were determined in the mouse hippocampus among various brain regions. We identified 7α-OH-Preg and 7α-OH-DHEA in the mouse hippocampal extract by using liquid chromatography coupled tandem mass spectrometry. In the Morris's water maze test, we found a dawn-dusk change in remote spatial memory in WT mice, the memory performance at ZT1 (dawn) was significantly higher than that at ZT11.5 (dusk). The remote memory performance at ZT1 was impaired by Cyp7b1 deficiency. In Cyp7b1-deficient mice, chronic intracerebroventricular administration of 7α-OH-Preg and 7α-OH-DHEA improved remote spatial memory performance. Furthermore, the administration of mixture of these neurosteroids was more effective than the single administrations. We conclude that 7α-OH-Preg and 7α-OH-DHEA play a cooperative role for the long-term maintenance of remote spatial memory in mice.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-327 外側嗅内皮質と内側嗅内皮質Vb層が形成する局所回路の構造的差異 Shinya Ohara（大原慎也）^1,2，Rajeevkumar R Nair（Nair R Rajeevkumar）²，Stefan Blankvoort（Blankvoort Stefan）²，Clifford Kentros（Kentros Clifford）²，Menno P Witter（Witter P Menno）² ¹東北大院生命科学脳神経システム ²Kavli Institute for Systems Neuroscience, NTNU, Trondheim, Norway The entorhinal cortex (EC), which is the major gateway between the hippocampus and the neocortex, is composed of functionally distinct subdivision, lateral and medial EC (LEC and MEC). A large proportion of neurons in MEC code spatial information, while LEC processes information of odor and objects. One factor which may contribute to these functional differences is the unique local circuit of these two regions. Here, we focused on the local circuits constituted by layer V neurons in deep-sublayer Vb (LVb). LVb neurons are main recipients of hippocampal projections, and project intrinsically, mediating two circuits in the hippocampus-memory system: 1) a feedback projection, sending information back to the EC-hippocampal loop via neurons in layer II/III (LII/III), and a hippocampal output circuit to telencephalic areas by projecting to the superficial-sublayer a of layer V (LVa) (Ohara et al., 2018). To examine the differences of these local circuits between MEC and LEC, we used a transgenic mouse which expresses tetracycline-controlled transactivator (tTA) mainly in EC LVb excitatory neurons (MEC-13-53D, Blankvoort et al., 2018). We first examined the axonal distribution of LVb neurons by specifically labeling them with tTA dependent virus, AAV2/1-TRE-Tight-GFP. We next examined the differential innervation of LVb neurons to LII/III/Va neurons by expressing channelrhodopsin variant (oChIEF) in LVb neurons with AAV2/1-TRE-Tight-oChIEF-citrine, and performing whole-cell patch-clamp recordings from the targeted cells while stimulating LVb fibers in acute slices. The projections from LVb-to-LII/III were similar in LEC and MEC, and GFP-labeled axons were more numerous in LIII than in LII. In line with this anatomical observation, LIII neuronal excitatory responses were larger than those in LII in both regions. This indicates that the EC-hippocampal loop favors the LIII-to-CA1/subicular projection rather than the LII-to-DG/CA3 one in both regions. In contrast, the LVb-to-LVa projection differed between LEC and MEC: the LVb axons of LEC were densely distributed in LVa, whereas in MEC they avoided LVa. The excitatory response after LVb stimulation was significantly lower in LVa neurons than in LIII neurons in MEC but not in LEC. This result indicates that hippocampal output circuit is more prominent in LEC than in MEC. These findings may provide new insights to unravel the neuronal basis underlying the phenotypical differences between MEC and LEC.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-328 ラット前海馬台3層ニューロンの軸索形態 Yoshiko Honda（本多祥子）¹，Takahiro Furuta（古田貴寛）² ¹東京女子医大院医解剖 ²大阪大院歯口腔解剖 The presubiculum plays a key role in processing and integrating spatial and head-directional information. Layer III neurons of the presubiculum provide strong projections to the superficial layers of the medial entorhinal cortex in the rat. To further elucidate the functional roles of the presubiculum, it is important to identify the axonal branching patterns of layer III neurons of the presubiculum. In the present study, we visualized the entire axonal processes and terminal distributions of single presubicular layer III neurons in the rat, by using in vitro injection of a viral vector expressing membrane-targeted palmitoylation site-attached green fluorescent protein. We found that layer III of the rat presubiculum (n=12) comprised multiple types of neurons with characteristic patterns of axonal collateralization, including cortical projection neurons and several types of intrinsic connectional neurons. Two cortical projection neurons (including both pyramidal and non-pyramidal neurons) provided two or three major axonal branches to the medial entorhinal cortex. These neurons formed elaborate terminal arbors that seemed to face each other and made plexus within the superficial layers of the medial entorhinal cortex. The other two cortical projection neurons provided axonal branches to the medial entorhinal cortex without any plexus, and sent many collaterals to the subiculum. We also found the neurons with axonal branches terminating only in the subiculum or parasubiculum. They also provide recurrent axons to the presubiculum. The intrinsic connectional neurons, including both pyramidal and basket type cells, extended axonal branches to various parts and layers of the presubiculum. Our data demonstrate that layer III of the rat presubiculum consists of multiple types of cortical projection neurons, suggesting that single neurons simultaneously distribute their output signal to various areas and recurrently to the presubiculum.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-329 マーモセット前頭連合野の機能マッピングのための記憶課題 Daisuke Koketsu（纐纈大輔）¹，Atsushi Nambu（南部篤）^1,2 ¹生理研生体システム ²総研大生理科学 Common marmosets (Callithrix jacchus) are small primates (300-400 g body weight) and easy to handle. They have been expected as experimental model animals in the field of cognitive neuroscience and medical research for psychiatric and brain disorders. The prefrontal cortex (PFC) is one of the main target brain areas to investigate cognitive functions and psychiatric disorders. While anatomical connections of marmoset PFC have been reported, neurophysiological studies are scanty. Therefore, it is necessary to accumulate basic neurophysiological knowledge on marmoset PFC. For functional mapping of the PFC, the marmoset should be placed on a marmoset chair with its head restrained and perform tasks, such as the working memory task, which activates PFC. In the present study, we trained marmosets to perform delayed matching-to-sample tasks (DMTS). In DMTS, the first visual cue stimulus indicating location (left/right), color (yellow/blue), or shape (rectangle/cross) was presented on the screen. After a delay period of 0.5-1.5 s, two target stimuli were presented, and marmosets were required to touch one of the target stimuli, which matched the first cue stimulus. In DMTS with the location-cue, marmosets showed an about 80% success rate, while success rates decreased to about 60% in DMTS with color- or shape-cues. We increased the delay periods, and marmosets showed significant correct rates in DMTS with 15-30 s delay, suggesting that marmosets can hold cue information for 15-30 s as macaque monkeys do. We also increased the number of target stimuli to three in DMTS, and marmosets showed an about 80% success rate in the difficult memory task. These results have shown that we can train marmosets to perform the working memory task enough to examine PFC neuronal activities during the memory information processing.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-330 恐怖記憶再固定化と消去エングラムニューロンの同定と機能解析 Rie Ishikawa（石川理絵），Satoshi Kida（喜田聡）東京農大生命科学バイオ Brief fear memory retrieval triggers fear responses followed by memory reconsolidation, whereas long-time or repeated retrieval extinguishes fear memory. Memory circuits contributing to fear and extinction have been identified, respectively. However, discrimination and interaction of fear and extinction neurons have not well examined. We have identified and characterized "fear" and "extinction" ; neuron using contextual fear conditioning and inhibitory avoidance tasks; we found that c-fos positive neurons are increased in the mPFC, hippocampus and amygdala when fear memory is reconsolidated, while these neurons are increased in the mPFC and amygdala when the memory is extinguished (Suzuki et al., 2004; Mamiya et al., 2009; Fukushima et al., 2014). In this study, we identified fear and extinction neurons using catFISH by detecting Arc and Homer1a (H1a) mRNAs following retrieval of inhibitory avoidance memory. Amygdala showed distinct populations that were activated at the reconsolidation and extinction phases; reconsolidation neurons express H1a mRNA in the nucleus, while extinction neurons express Arc mRNA in the nucleus, respectively, suggesting that reconsolidation and extinction are regulated by distinct neuronal populations in the amygdala. More interestingly, mPFC showed single neuronal population that was activated at the both reconsolidation and extinction phases; these neurons express both Arc and H1a mRNAs in the nucleus. These results raised the possibility that reconsolidation and extinction are regulated by single neuronal population in mPFC. To further examine functional roles of neuronal populations in the mPFC and amygdala, we are now trying to characterize reconsolidation and extinction engram neurons. We labeled these engram neurons activated following the retrieval of inhibitory avoidance memory by channelrhodopsin-2 (ChR2) using c-fos-tag system. Optogenetic activation of reconsolidation neurons increased freezing level in an unconditioned context, whereas optogenetic activation of extinction neurons enhanced memory extinction. Collectively, we identified functional populations of reconsolidation and extinction engram neurons.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-331 A brainstem-to-amygdala circuit provides aversive instructive signals for fear learning Li-Feng Yeh（Yeh Li-Feng）^1,2，Akira Uematsu（Uematsu Akira）¹，Tomoya Dunki（Dunki Tomoya）¹，Yusuke Kasuga（Kasuga Yusuke）^1,2，Joshua Johansen（Johansen Joshua）^1,2 ¹RIKEN Center for Brain Science ²Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo Learning to avoid threats depends on fear memory formation, an adaptive associative process in which animals learn that sensory cues predict subsequent aversive events. Synaptic plasticity in the lateral and basal nuclei of the amygdala (LA/B), which receives converging sensory and aversive inputs, is believed to underlie such memory formation (Johansen et. al., 2011; Herry and Johansen, 2014; Janak and Tye, 2015). While the sensory pathways to LA/B are well characterized, the aversive input circuits are largely unknown. Identifying these pathways is critically important- they are thought to provide emotional valence to the LA/B and instruct fear learning there. Here we show that LA/B projecting neurons in the brainstem cuneiform nucleus (CnF) conveys aversive information during auditory fear conditioning in rats. Optogenetic inhibition of LA/B projecting CnF neurons suppresses aversive stimulus evoked spiking activity in LA/B neurons and prevents fear memory formation. Activating this circuit elicits defensive behaviors, and is sufficient in the absence of an external aversive stimulus to produce fear conditioning. In sum, we identify an aversive signaling circuit which triggers emotional fear learning.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-332 Altered hippocampal replay underlies memory impairments in SCN2A heterozygous mice Steven James Middleton（Middleton Steven James）¹，Emily M Kneller（Kneller Emily）¹，Shuo Chen（Chen Shuo）¹，Ikuo Ogiwara（Ogiwara Ikuo）^1,2，Kazuhiro Yamakawa（Yamakawa Kazuhiro）¹，Thomas J McHugh（McHugh Thomas J）¹ ¹RIKEN Center for Brain Science ²Nippon Medical School, Tokyo The SCN2A gene encodes the Na_v1.2 α-subunit protein and is a site frequently mutated in human neurological disorders including, but not limited to epilepsy, severe intellectual disability, autism spectrum disorder and schizophrenia. The hippocampal formation has been identified as a crucial circuit for spatial memory (O'Keefe & Dostrovsky, 1971) and in concert with other cortical circuits has been implicated in working memory tasks (Jones & Wilson, 2005). It achieves this in multiple steps by initially encoding spatial information sequentially during exploratory behavior through place specific modulation of pyramidal cell firing rates. Place cell sequences are subsequently time compressed to a scale presumably invoking synaptic plasticity during their repetitive replay during both behavior, as theta sequences (Middleton & McHugh, 2016) and during sharp wave ripples (SPW-Rs) in goal-directed behavior (Pfeiffer & Foster, 2013) and sleep (Diba & Buzsaki, 2007). Notably, the formation of cell assemblies (groups of anatomically distributed neurons exhibiting transient periods of enhanced synchrony) during behavior and their subsequent reactivation in SPW-Rs (van de Ven et al., 2016) has often been highlighted as a key mechanism underlying memory. Although artificial SPW-R disruption has been demonstrated to degrade performance in memory dependent tasks (Girardeau et al., 2009) and altered SPW-Rs have been observed in several disease models, how this impacts the information content during SPW-R episodes is unclear. Here we took advantage of the SCN2A^+/- mouse to address how changes in SPW-R activity might relate to neurological dysfunction.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-333 結節乳頭核ヒスタミン神経細胞の活性化による物体認識記憶の想起増強 Ayame Kubo（久保絢女），Shohei Ohmori（大森翔平），Kyoka Nishimura（西村京華），Masabumi Minami（南雅文），Hiroshi Nomura（野村洋）北海道大学大学院薬学研究院薬理学研究室 Histamine in the central nervous system is produced mainly in the tuberomammillary nucleus (TMN) and is implicated in learning and memory as well as sleep and wakefulness, feeding and drinking, and neuroendocrine regulation. Previously, we demonstrated histamine H3 receptor inverse agonists upregulate histamine release in the perirhinal cortex and promote the recall of forgotten object memories. However, since histamine H3 receptors are expressed non-histaminergic neurons as well as histaminergic neurons, other neurotransmitter systems could be also involved in the memory recovery which is induced by H3 receptor inverse agonists. In this study, we examined whether chemogenetic activation of histamine neurons in the TMN enhances retrieval of forgotten long-term object memory in mice. We virally targeted hM3Dq, the Gq-coupled excitatory designer receptor exclusively activated by designer drugs (DREADD), to histaminergic neurons in the TMN of HDC-Cre mice. The HDC-Cre mice express a codon optimized Cre recombinase from the histidine decarboxylase (HDC) promoter/enhancer elements. In a training session of the novel object recognition task, mice were placed in the field, in which two identical objects were positioned. One week later, they underwent a test session where one familiar and one novel object were presented. In the first experiment, mice received intraperitoneal clozapine-N-oxide (CNO) or saline injection before the test session. In the second experiment, mice received CNO or saline injection before the training session. The pre-test injection of CNO to the mice receiving AAV-DIO-hM3Dq led to a significant increase in the discrimination between novel and familiar objects as compared to controls. In addition, we investigated the effect of activation of histamine neurons on anxiety-like behavior and fear memory. The activation of histamine neurons had no effect on anxiety-like behavior in the elevated plus maze test. These findings indicate that activation of histamine neurons enhances retrieval of a forgotten long-term object memory.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-334 想起後の恐怖記憶制御に対する海馬記憶エングラムの役割の解析 Taikai Nagayoshi（長葭大海），Hotaka Fukushima（福島穂高），Satoshi Kida（喜田聡）東農大院・農・バイオ Memory engram cells (ECs) are neural networks storing information acquired during learning. Reactivation of the ECs leads to retrieval of stored memory. Retrieval of contextual fear memory by short (3-6 min) or long re-exposure (30 min) to conditioned stimulus (CS; context) initiates reconsolidation and extinction, respectively. Our previous study showed that reconsolidation of contextual fear memory following short re-exposure to the CS requires the activation of gene expression in the hippocampus (HP) but that the acquisition of contextual fear extinction by prolonged re-exposure cancels this gene expression activation (Mamiya et al, 2009). These findings suggest critical roles of HP in positive and negative regulation of retrieved contextual fear memory, raising the possibility that the activation and inactivation of hippocampal ECs enhance or weakens, respectively, contextual fear memory. To examine this hypothesis, we examined the effects of manipulation of hippocampal ECs during retrieval on contextual fear memory using c-fos-tag system. c-fos-tTA transgenic mice received hippocampal micro-infusion of adeno-associated virus expressing channelrhodopsin-2 (ChR2) or archaerhodopsin-T (ArchT) under the control of tetracycline-dependent promoter. Mice were contextual fear conditioned with a single footshock to label engram cells by ChR2 or ArchT. Twenty-four hrs later, these mice were re-exposed to the context for 6 or 30 min (re-exposure) and tested at 24 hrs or 30 days after the re-exposure. Optogenetic activation of hippocampal ECs for last 25 min of 30 min re-exposure blocked the acquisition of memory extinction during re-exposure (inhibited decreases in freezing) and also blocked long-term extinction of contextual fear memory. These results suggested that memory extinction requires inactivation of hippocampal ECs during the re-exposure. On the other hand, optogenetic inactivation of hippocampal ECs for last 3 min of 6 min re-exposure blocked retrieval of contextual fear memory (decreased freezing). More interestingly, impaired contextual fear memory was observed when tested at 24 hrs and 30 days after the re-exposure. These observations suggested that hippocampal ECs inactivation during retrieval erases contextual fear memory, perhaps, by blocking memory reconsolidation. Taken together, our findings suggest that hippocampal ECs plays critical roles in positive and negative regulation of retrieved contextual fear memory.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-335 Htr3aを介したセロトニンによる腹側海馬の抑制は目的指向型行動の維持を担う Keitaro Yoshida（吉田慶多朗），Masaru Mimura（三村將），Kenji F Tanaka（田中謙二）慶應大医精神・神経科学 The inability to sustain and complete goal-oriented behavior is a shared symptom of numerous psychiatric conditions. The neural substrates underlying goal-oriented action preparation and action initiation have been well characterized, but those underlying action sustainment are poorly understood. One requirement for sustained goal directed action is suppression of competing negative emotional states. Food seeking, for instance, is potently suppressed by threatening stimuli that evoke fear or anxiety. Successful goal-directed behavior is likely to require gating of the behavioral inhibition associated with negative emotional states. The hippocampus is known to monitor the environment, with dorsal hippocampus contributing to spatial and contextual processing and ventral hippocampus (vHP) regulating emotional responses to internal and external contextual cues. In particular, vHP modulates threat-related responses, such as anxiety-like behavior and fear. In addition to projecting to subcortical structures classically associated with these behaviors, vHP densely projects the ventral striatum, a key orchestrator of goal-oriented behavior. vHP is therefore a potential interface between emotion and goal-oriented action. To investigate potential contributions of vHP to goal-oriented behavior, we monitored and manipulated vHP activity during appetitive and aversive operant tasks. Our results demonstrate that a vHP-dependent mechanism gates sustainment versus termination of goal-directed behavior.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-336 ケタミン投与による採餌行動の変化 Ryo Sawagashira（澤頭亮），Masaki Tanaka（田中真樹）北海道大学　医学研究院・医学院・医学部医学科　神経生理学教室 Working memory deficits are commonly observed in a variety of psychiatric and neurodegenerative disorders, whereas quantitative evaluations of these deficits are usually difficult in clinical cases. Recent studies in experimental animals use low-dose ketamine (NMDA receptor antagonist) to produce working memory deficits that resemble to those observed in schizophrenia. Here, we developed a new behavioral paradigm to assess memory capacity and utility, and adapted it to monkeys with and without ketamine administration. In the oculomotor foraging task, the animals were presented with 15 identical objects (0.6° white squares, > 4° apart each other) on the screen. One of the objects was associated with a liquid reward, and monkeys were trained to search for the target by making sequential saccades for up to 6 seconds. We assume that the rate of perseverative behavior (redirecting eyes to the same objects) may depend on the amount of memory capacity and utility. We constructed the ""foraging model"" incorporating the following three parameters: 1) memory capacity, 2) exploratory rate and 3) memory decay, which could explain several features of behavioral data. When we fit the data obtained from two monkeys with those expected from the model, the goodness-of-fit evaluated by coefficient of determination was always greater than 0.9. The resultant estimates of memory capacity were 11 and 7 items, exploratory rate were 24% and 28% and the memory decay were 6 and 2 initial items in memory for monkeys O and E, respectively. Following the systemic administration of a subanesthetic dose of ketamine (<= 1.5 mg/kg), all these parameters dramatically changed in a dose-dependent and time-dependent manner. In contrast, systemic administration of medetomidine (α2 adrenoceptor agonist, 10 μg/kg) reduced saccade velocity, but failed to alter the model parameters. Thus, our oculomotor paradigm and the foraging model appear to be useful to quantitatively evaluate working memory capacity and utility, which might be applicable to clinical testing in psychiatric and neurodegenerative disorders in future studies.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-337 ワーキングメモリにおける実行機能の個人差と前頭・頭頂間シータ帯域コヒーレンスの関係性 Takuya Ideriha（出利葉拓也）¹，Junichi Ushiyama（牛山潤一）^1,2 ¹慶應大環境情報 ²慶應大医リハビリ Introduction: Working memory is the system of short-term memory (STM), which is just to memorize something such as numerical sequences for a few seconds, and executive functions (EF), which are other cognitive functions like processing the STM. Executive functions are related to academic performances, reading disorder and other mental disorders. This study aimed to elucidate the neural basis of individual differences in executive functions of working memory by conducting neurophysiological experiments. Methods: Electroencephalogram (EEG) signals were recorded from 19 electrodes while 13 healthy young individuals performed four cognitive tasks. The tasks included two types (verbal and spatial) and two cognitive functions (STM and EF) resulting in four tasks. In short, they were two digit-span tasks and two delayed match to sample tasks. We assessed EEG power from every electrode and coherence increase within all electrode pairs and their correlation with individual differences in executive functions of working memory. Results: It was revealed that high-alpha and low-beta power(11-20Hz) increase above right temporal area and the rate of increase in fronto-parietal coherence in theta-band (4-7Hz) is well correlated with the ability of executive functions(p < 0.01). Discussion: This is the first study to demonstrate the significant relationship between fronto-parietal theta coherence and executive functions of working memory across individuals. In previous researches, it was revealed that fronto-parietal theta coherence increases during tasks requiring executive functions. Thus, the present results would be reasonable. Moreover, the current research showed that the more the participants can increase fronto-parietal theta coherence, the better they perform tasks requiring executive functions. As theta phase-synchronization has been suggested to promote the information transmission between the regions, it is suggested that those who are good at working memory can transmit information among fronto-parietal regions and make it easier to realize working memory.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-338 後部頭頂皮質は反応抑制に因果的に寄与する：fMRI・TMSによる検証 Takahiro Osada（長田貴宏）¹，Shinri Ohta（太田真理）¹，Akitoshi Ogawa（小川昭利）¹，Masaki Tanaka（田中政輝）¹，Akimitsu Suda（須田晃充）^1,2，Koji Kamagata（鎌形康司）³，Masaaki Hori（堀正明）³，Shigeki Aoki（青木茂樹）³，Yasushi Shimo（下泰司）²，Nobutaka Hattori（服部信孝）²，Takahiro Shimizu（清水崇宏）⁴，Hiroyuki Enomoto（榎本博之）⁵，Ritsuko Hanajima（花島律子）⁴，Yoshikazu Ugawa（宇川義一）⁵，Seiki Konishi（小西清貴）¹ ¹順天大院　医神経生理 ²順天大院　医脳神経内科 ³順天大院　医放射線医学 ⁴鳥取大医神経内科 ⁵福島県立医大神経再生医療学 The posterior parietal cortex (PPC) features close anatomical and functional relationships with the prefrontal cortex. However, the necessity of the PPC in executive functions has been challenged by previous negative neuropsychological results. In the present study, activation and functional connectivity were measured to analyze a parcellation-based network that aimed at identifying a candidate PPC region essential for response inhibition, an executive function that inhibits prepotent response tendency. FMRI scans were conducted during performance of a stop-signal task and during a resting state using a 3-T MRI scanner (Siemens Prisma, TR 1 s, TE 30 ms, voxel 2 mm). The cortical surface was parcellated based on boundary mapping using the resting-state data, and then parcel-wise task activation and functional connectivity were calculated to reconstruct a network for response inhibition. The target PPC region was identified using the following criteria: (1) activated during response inhibition (Stop success minus Go success), and (2) connected with the right inferior frontal cortex (IFC) and right presupplementary motor area (preSMA), the two frontal regions necessary for response inhibition. The candidate PPC region was successfully identified as the right intraparietal sulcus (IPS) region in 12 out of 14 subjects. As the control that lacks the connectivity, the TPJ region was additionally identified to be activated but not connected with the IFC or preSMA. Next, the necessity of the identified IPS region was tested by using transcranial magnetic stimulation (TMS). Single-pulse stimulation was delivered to the PPC regions in one half of Go and Stop trials (Stim trials), with No-stim trials intermixed within run. Prolonged stop-signal reaction time (SSRT) was observed in the IPS region at the time window of 30 to 0 ms prior to stopping [mean difference: 26.3 ms, t(11) = 6.7, p = 3.4 × 10^-5, paired t-test, Stim vs No-stim], but no significant changes were observed in the TPJ region [t(11) = 0.4, p = 0.73]. In the ten further recruited subjects, prolonged SSRT was reproducibly observed in the IPS region [mean difference: 23.8 ms, t(9) = 6.3, p = 1.4 × 10^-4], but not in the TPJ region [t(9) = 0.2, p = 0.88]. These results indicated that the IPS plays an essential role in executive functions and suggest that the parcellation-based network can predict essential regions that are yet identified.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-339 認知課題中のヒト前頭前野における局所グルコース代謝変化に対する抗ヒスタミン薬の影響 Nobuya Suzuki（鈴木頌也）¹，Asuka Kikuchi（菊地飛鳥）¹，Akie Inami（稲見暁惠）¹，Shoichi Watanuki（四月朔日聖一）¹，Masayasu Miyake（三宅正泰）²，Kazuko Takeda（武田和子）¹，Kotaro Hiraoka（平岡宏太良）¹，Kazuhiko Yanai（谷内一彦）^1,3，Hiroshi Watabe（渡部浩司）²，Manabu Tashiro（田代学）¹，Fairuz B.M. Nasir（ナシールファイルーズ）²，Attayeb Mohsen（モーセンアタイエフ）^1,3,4 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東北大学サイクロトロン・RIセンター　放射線管理研究部 ³東北大学大学院医学系研究科機能薬理学分野 ⁴医薬基盤・健康・栄養研究所 Abstract Objective: Antihistamine drugs may often induce sedative side effects. We previously demonstrated, using positron emission tomography (PET), that antihistamine treatment was associated with increased brain glucose (energy) metabolism during cognitive tasks in healthy young adults. This study aimed at elucidation of the brain mechanisms of the finding of increased glucose consumption, clarifying whether the metabolic change was more strongly associated with higher energy demands during tasks than the changes in the baseline glucose consumptionn level in the brain. Methods: Healthy young men were recruited for cognitive study (n=18) and for resting study (n=12), in a double-blind, placebo-controlled, three-way crossover study. Subjects were scanned with PET with [18F]fluorodeoxyglucose (FDG PET) for measurement of baseline level. Then, these subjects received single doses of a non-sedative antihistamine (levocetirizine 5 mg) or a sedative antihistamine (diphenhydramine 50 mg), and were scanned again about 2 hours later. Regional cerebral glucose consumption changes were calculated based on the 2 PET scans, in terms of standardized uptake values (SUVs) normalized for body weight and injected radioactivity doses, using PMOD software package. Results and Discussion: In comparison to the baseline level, brain glucose consumption in the frontal regions of the subjects increased during cognitive study by approximately 8% after placebo treatment and 10% to 13% after antihistamines treatment. In contrast, glucose consumption in the frontal regions of the subjects showed no changes during resting condition after treatment with both placebo and antihistamines. Conclusions: The present findings demonstrated that the brain glucose consumption is stable in resting condition even after treatment with placebo and antihistamines while the brain glucose consumption in task conditions increased more intensely after antihistamines treatment than after placebo treatment. The present findings suggested that the increased glucose consumption was more associated with higher energy demands during cognitive tasks than the baseline level in brain glucose consumption.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-340 実行機能における視床下核の因果的役割 Tadaaki Nishioka（西岡忠昭）^1,2，Kosuke Hamaguchi（濱口航介）¹，Satoshi Yawata（矢和多智）¹，Takatoshi Hikida（疋田貴俊）²，Dai Watanabe（渡邉大）¹ ¹京都大院医 ²大阪大 The subthalamic nucleus (STN), a key component of the basal ganglia, is a potential contributor to executive functions such as impulse control. However the causal relationship between the STN and executive functions remains controversial. Here we chemogenetically suppressed the STN neuronal activities and tested the effects on executive functions including attentional control and impulse control by using mice performing the five-choice serial reaction time task (5-CSRTT). We found that chemogenetic inactivation of the STN impaired attentional performance by decreasing accuracy. Moreover, the attentional performance was severely impaired when the attentional demands were increased, indicating that the STN plays an important role in response control under the conditions of high attentional demand. Also, we found that inactivation of the STN increased impulsive responding in delay period but not compulsive responding. These selective effects on impulsive response demonstrate that the STN suppresses an inappropriate response in a specific context but not a general motor response. Furthermore, correct response latency and reward collection latency were comparable to those treated with vehicle, suggesting that the decrease in accuracy were not attributable to motor deficits or motivational deficits. These findings elucidate the causal role of the STN in executive functions and offer novel insights to understand neural mechanisms by which the basal ganglia is contributed to executive functions.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-341 マラウィ湖産ヒレ食性シクリッド科魚類の捕食行動の特殊性と左右性 Yuichi Takeuchi（竹内勇一）¹，Hiroki Hata（畑啓生）²，Atsushi Maruyama（丸山敦）³，Takuto Yamada（山田拓人）¹，Takuma Nishikawa（西川巧馬）³，Makiko Fukui（福井眞生子）²，Zatha Richard（Richard Zatha）⁴，Rusuwa Bosco（Bosco Rusuwa）⁴，Yoichi Oda（小田洋一）⁵ ¹富山大院医解剖・神経科学 ²愛媛大院理工 ³龍谷大院理工 ⁴マラウィ大生物 ⁵名古屋大院理脳機能構築 Several vertebrates, from fish to mammals, exhibit behavioural laterality and associated morphological asymmetry. Laterality may increase individual fitness, and foraging strength, accuracy, and speed. However, little is known about which behaviours are affected by laterality or what fish species exhibit obvious laterality. Previous research on the predatory behaviour of the scale-eating Lake Tanganyika cichlid Perissodus microlepis indicates behavioural laterality that reflects asymmetric jaw morphology. The Lake Malawi cichlid Genyochromis mento feeds on the fins of other fish, a behaviour that G. mento developed independently from the Tanganyikan Perissodini scale-eaters. We investigated stomach contents and behavioural laterality of predation in aquarium to clarify the functional roles and evolution of laterality in cichlids. We also compared the behavioural laterality and mouth asymmetry of G. mento and P. microlepis. The diet of G. mento mostly includes fin fragments, but also scales of several fish species. Most individual G. mento specimens showed significant attack bias favouring the skew mouth direction. However, there was no difference in success rate between attacks from the preferred side and those from the non-preferred side, and no lateralized kinetic elements in predation behaviour. G. mento showed weaker laterality than P. microlepis, partly because of their different feeding habits, the phylogenetic constraints from their shorter evolutionary history, and their origin from ancestor Haplochromini omnivorous/herbivorous species. Taken together, this study provides new insights into the functional roles of behavioural laterality: Predatory fish aiming for prey that show escape behaviours frequently exhibit lateralized behaviour in predation.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-342 自発電気活動に基づくドーパミンと非ドーパミンニューロンの判別 Keiji Miura（三浦佳二）¹，Tsuyoshi Ishikawa（石川剛史）¹，Hideyuki Matsumoto（松本英之）² ¹関西学院大 ²大阪市大院医神経生理 In the past studies in neuroscience, the electrophysiological recordings played a powerful role and revealed the nature of midbrain dopamine neurons and related reward systems. Although dopamine neurons are considered essential in decision making and motor controls, their specific roles are under debate, partially due to the difficulty to identify dopamine neurons among surrounding neurons deep in the brain. Thus, a handy method to discriminate dopamine neurons based on the spontaneous activity patterns is desired. Here we tried to discriminate optogenetically identified dopamine/non-dopamine neurons. We optogenetically identified dopamine neurons, which enabled us to assess the performance as a hit rate. We want to discriminate only based on the spontaneous electrical activities without using the responses to rewards or sensory stimuli. This is because in the neuroscientific researches we want to characterize the responses of (correctly identified) dopamine neurons and, thus, we have to discriminate them based on the other clues. Here we used baseline firing rates, spike widths and logCV as features in the support vector machines, which was performed by MATLAB fitcsvm() function. We successfully discriminated 86.0% of dopamine and non-dopamine neurons based on the three spontaneous electrical activities: firing rates, spike widths and logCV. When we only used a single feature from the three, the classification was rather low (~68.6%). In future works, we will explore further features and pattern classifiers for improvement.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-343 「することをしない」か「しないことをする」か：行動試験と強化学習による解析 Sai Tanimoto（谷本彩）^1,2，Masashi Kondo（近藤将史）^2,3，Kenji Morita（森田賢治）⁴，Masanori Matsuzaki（松崎政紀）^1,2 ¹東京大医 ²東京大院医細胞分子生理 ³日本学術振興会 ⁴東京大院教育身体教育 Animals choose something and act appropriately in their daily life. They get something good and bad at the same time through their choice. The former is the reward such as food and water and the latter is the cost and/or energy that inevitably accompany body movements. Animals have to compute both of them before they decide whether they act or not. However, it is poorly understood how the rewards obtained by animals' action and the costs accompanying their action affect their decision-making and whether doing nothing is actively chosen or not. To address these issues, we developed a simple behavioral task focusing on water reward and physical effort (forelimb movement) and performed a model-based analysis with reinforcement learning, a framework using animal behavior and the outcomes. In our task, mice were let to choose whether to pull the lever or not in response to two different tones associated with two different probabilities of water delivery (reward probabilities) following a successful lever pull. No reward was delivered after either tone when the lever was not successfully pulled. After two-week training sessions, most of the mice frequently pulled the lever after tones with higher reward probability but infrequently did after tones with lower one. This tendency was similarly observed when different sets of reward probabilities were used. We then fit the changes in their behavior during the training sessions by several models with different free-parameter sets. In terms of the physical effort, we assumed two cases: `cost model' and `saving-energy model'. `Cost model' is that the mice get the cost (negative reward) when they choose `to pull' because they should get tired. `Saving-energy model' is that they get `rewarded' when they choose `not to pull' because the mice could save the physical effort. We compared these models with the use of BIC (Bayesian information criterion) and found that `saving-energy model' described their learning process better than `cost model'. These results suggested that the mice found something good in the choice of `not to pull' even though they got no water reward. The mice might regard avoiding the costs as `reward', and they might actively choose `not to pull' rather than they just didn't choose `to pull'. This paradigm will help us to understand the active choice of `doing nothing' and how animals evaluate the cost or energy.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-344 他者観察による数の認知への社会的影響の神経相関 Akitoshi Ogawa（小川昭利）¹，Tatsuya Kameda（亀田達也）²，Hironori Nakatani（中谷裕教）³ ¹順天大医 ²東京大院人文社会 ³東京大進化認知科学研究センター Observing others' decisions influences our own decision-making not only in behavioral domains but also in perceptual domains. To reveal the neural basis for social influence on numerical cognition that is typical processing in perceptual domains, this functional magnetic resonance imaging study investigated that when estimating the number of presented dots, how the numerical cognition and the activation of number-associated regions were modulated after observing over- and under-estimation in numeric discrimination by others. The participants (N=21) performed a numeric discrimination task that asked them to answer whether the number of presented dots was larger than a criterial number. The participants performed this task in the first, third, and fifth runs by themselves (self-choice runs). In the second and fourth runs, the participants observed that others, i.e., over-estimator (OE) and under-estimator (UE), performed the same task. As expected, the participants answered ""larger "" more often after OE run [SO (Self-choice after Over-estimator) hereafter] than after UE run [SU (Self-choice after Under-estimator)]. The generalized-linear-mixed-model (GLMM) analysis showed that the number estimation in SO run was significantly more over-estimated than in SU run. The whole brain analysis showed that the medial prefrontal cortex (MPFC) was activated for the incongruity between the others' choices and the choices estimated individually from the psychometric function in the first self-choice run. The number-associated region in right intraparietal sulcus (IPS) that previous studies often reported was sensitive to the lower range of dot number in SO run but not in SU run. The result of representational similarity analysis showed that the activation patterns in the right IPS in SO and SU runs were significantly modulated for the presented dot numbers. These results suggest that the MPFC detects the incongruity between the participant's own discrimination and other's discrimination, and the right IPS is associated with the social conformity of numerical cognition after observing the others' numeric discrimination.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-345 意思決定中のゼブラフィッシュの大脳基底核―皮質経路の神経活動ダイナミクス Yuki Tanimoto（谷本悠生）¹，Makio Torigoe（鳥越万紀夫）¹，Tanvir Islam（イスラムタンビル）¹，Ryo Aoki（青木亮）²，Hisaya Kakinuma（柿沼久哉）¹，Hitoshi Okamoto（岡本仁）¹ ¹理研CBS 意思決定回路動態 ²理研CBS 視覚意思決定 Animals make behavioral decisions based on their prediction of future reward or punishment. To elucidate computational mechanisms as to how such predictions affect decision-making, large-scale calcium imaging of neuronal activities during decision-making is ideal. For this, we recently have established a closed-loop virtual-reality system for 2-photon calcium imaging of adult zebrafish (Torigoe et al., in prep). Neuronal activities from dorsal pallium, a zebrafish cerebral cortex, are recorded in the course of learning of Go/No-go task in virtual reality. In this task, a fish needs to move from blue- to red-region in Go trials and needs to stay in red-region in No-go trials to avoid electrical shock. Using this system, we observed emergence of neural ensembles encoding ""state prediction errors"", deviance of a current perceptual state from an expected ideal perceptual state to avoid the shock (Torigoe et al., in prep). These errors are thought to guide the fish's behavior to achieve ideal perceptual states by facilitating actions that reduce the errors. Theoretically, the state prediction errors can be calculated by subtracting current perceptual neural activity from corresponding predictive neural activity. I hypothesized that the predictive neural activity is generated by basal ganglia because of its crucial role in reward and punishment prediction. Zebrafish has partially equivalent structures to the mammalian basal ganglia, such as dorsal entopeduncular nucleus (dEN), a putative zebrafish globus pallidus interna. In the presentation, I report activity patterns of axonal projections from dEN to dorsal pallium during the Go/No-go task in the virtual reality. In the beginning of learning, dEN axons started to respond to a blue-region where they receive shocks in failure-trials. After learning, dEN axons started to be suppressed by red-region where they experience shock omission in success-trials. These results indicate that dEN encodes prediction of shock or shock omission in a current perceptual state and output it to the cortex. In the end of my presentation, I will discuss a neuronal circuit model for generation of the dEN output in the basal ganglia, calculation of the state prediction errors in the cortex, and choice of appropriate behaviors based on the prediction errors.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-346 Taking advantage of in vivo electrophysiological recording and model fitting approaches to investigate the role of Akt1, a schizophrenia susceptibility gene, in reward-based decision making in mice Chia-Yuan Chang（Chang Chia-Yuan）¹，Ching Chen（Chen Ching）¹，Wen-Sung Lai（Lai Wen-Sung）^1,2,3 ¹National Taiwan University ²Graduate Institute of Brain and Mind Sciences, National Taiwan University ³Neurobiology and Cognitive Science Center, National Taiwan University Schizophrenia is a severe neuropsychiatric disorder in which cognitive impairment features prominently. Accumulating evidence from human genetic studies suggests that multiple susceptibility genes might contribute to the pathogenesis of schizophrenia, including AKT1 (protein kinase B α), a key kinase intermediate downstream of dopamine D2 receptor. Alterations of dopaminergic transmission have been implicated in the pathogenesis of schizophrenia and patients with schizophrenia also show worse performance than healthy controls in many decision-making tasks. Recent finding further revealed that Akt1 might play a crucial role in the modulation of reward-based decision making, especially in the striatum. However, so far, the importance of Akt1 in reward-based decision making and its specific role in the dorsomedial striatum (DMS) during this process remain elusive. To this end, we examined the role of Akt1 in a probabilistic two-choice foraging task and the relationship between electrophysiological properties in the DMS and behavioral performance in Akt1 heterozygous mutant mice and their wild-type littermates. In this task, each food-deprived mouse had to discover which choice has a higher reward rate by trial and error and to maximize the total amount of reward. Once the pre-set criterion was achieved, each mouse moved on to reversal phase and the reward rates of the two choice arms were switched. Simultaneously, in vivo electrophysiological recording was conducted in the DMS of the mouse brain during the whole acquisition and reversal phases. Our results indicated that 1. Akt1 mutant mice required significantly fewer trials to achieve the criteria compared to their controls in both acquisition and reversal phases. 2. The evoked power in the DMS is time-locked while receiving the outcome of each choice. In the trials of no-reward outcomes, the evoked theta and gamma powers had a high correlation with their behavioral outcomes whereas no correlation was found in the trials of reward outcomes. 3. Taking advantage of a Bayesian approach to estimate the parameters in a modified reinforcement learning model, we found that Akt1 mutant mice have a higher learning rate in the no-reward outcomes compared to controls. Collectively, our results suggest that the neural oscillation in the DMS contributed to the learning rate which resulted in the differential behavioral performances in Akt1 mutant mice during decision making. Further analysis is still in progress.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-347 The significance of hippocampal-entorhinal pathway in the Differential-Reinforcement-of-low-Rate (DRL) Tasks Wan-Ting Liao（Liao Wan-Ting），Yi-Tse Hsiao（Hsiao Yi-Tse） National Taiwan University, School of Veterinary Medicine Impulsivity as a hallmark covers a range of neuropsychiatric disorders. How impulsivity becomes the behavioral expression in these disorders is a key question. One possible mechanism is the failure of the "top-down" and inter-connected control networks. Neuroanatomical studies demonstrate that abundant projections from the hippocampus and medial entorhinal cortex (MEC) circuitries could participate in the control of behavior. However, the functional interaction between these structures has not been experimentally addressed with behavioral testing. Thus, in our study, the DRL tasks were conducted to investigate the neural strategies between the impulsive and normal responses by observing neurophysiological substrates, the oscillations among the hippocampal-entorhinal circuitries. Interestingly, evaluating the coherence and causality of the LFP signals, it showed the synchrony of the directional brain signals was mostly from the CA1 to the MEC, and this causal interaction striking declined in the impulsive states. Then theta phases modulated gamma amplitudes from the CA1 to MEC direction, significantly vary from the impulsive epochs both in fast and slow gamma bands. These significant alternations during the impulsive epochs suggested that the consequence may result from the strong but unorganized signals, which contributes to the inefficient communication between the CA1 and MEC. Overall, this study helps us conceptualize more how the hippocampal-entorhinal pathway contributes to the impulsivities.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-348 異種感覚刺激の時間弁別におけるサル内側運動前野での時間情報表現 Atsushi Chiba（千葉惇），Kazunori Morita（守田和紀），Ken-ichi Oshio（生塩研一），Masahiko Inase（稲瀬正彦）近畿大医生理 Medial premotor areas (MPAs) are considered to be included in a timing neural network. In the present study neuronal representation of temporal information of visual and auditory stimuli was examined in MPAs. Neuronal activity was recorded from MPAs while a monkey was performing a duration discrimination task using visual and auditory cues. In the task, two cues (first cue, C1 and second cue, C2) were presented consecutively for different duration ranging from 0.2 to 1.8 sec. Each cue was followed by a delay period (first delay, D1 and second delay, D2) of 1 sec. The subject was required to choose the longer presented cue after the D2 period. Cues were either visual (a green square) or auditory (a 2000Hz tone). Four types of modality combinations of the C1 and C2 were adopted; visual-visual, visual-auditory, auditory-visual, and auditory-auditory. Cue modality combinations and orders of relative cue duration (long-short, LS or short-long, SL) were pseudo-randomly determined in each trial. Correct response rates were significantly different among the four modality combinations. Those for the auditory-auditory trials were lower than other three types. Out of 168 neurons examined in MPAs, 3, 9, 13 and 41 neurons were C1, D1, C2 and D2 responsive, respectively. C1 response neurons responded to either visual or auditory cues, and C2 response neurons also responded to single mode of sensory cues. However, D1 response neurons increased their activity after longer C1 regardless of the sensory modality, visual or auditory. D2 response neurons showed differential activity depending on the order of relative cue duration. Nine neurons exhibited greater D2 activity in the LS trials while other 9 neurons did so in the SL trials. These differential D2 activity was found regardless of the cue modality combinations. Thus, the timing-related activities during the D1 and D2 periods, which can be related to estimation, memory or discrimination of stimulus durations, commonly represented duration information about both visual and auditory stimuli. These results suggest that temporal information of different modes of sensory stimuli was processed in an overlapping timing network, which includes MPAs.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-349 大脳皮質における価値情報の巨視的・微視的表象 Masashi Kondo（近藤将史）^1,2，Masanori Matsuzaki（松崎政紀）² ¹日本学術振興会 ²東京大院医 Transformation from sensory signal to value-related information is crucial for decision-making in animals. Subcortical regions, such as the basal ganglia, are known to be the primitive regions to handle value-related information. However, it is partially understood how the sensory and value-related information is represented in each cortical area to generate accurate movements. Here, we trained head-fixed mice for reward probability (value)-biased Pavlovian conditioning task. In this task, one of two sound cue was presented and followed by delivery of a water reward at high or low probability. During mice performed the task, we conducted wide-field calcium imaging of the entire dorsal cortex as a ""macroscopic view"" and two-photon calcium imaging of four frontal areas (frontal association area, anterior-lateral motor area, primary motor cortex, and medial prefrontal cortex) as a ""microscopic view"", with red genetically-encoded calcium indicators. In the wide-field calcium imaging, we found that in all dorsal cortical areas, even in auditory and visual cortices, licking-related neuronal activity dominated the other modalities including sound cue-related activity throughout all training stages. As the learning stage progressed, however, cortex-wide neuronal ensemble during the delay period at the trials of high reward-probability became more stable. On the other hand, the two-photon imaging with a single-cell resolution revealed that many neurons in the frontal cortices clearly showed not only licking-related activity but also task-related activity. The neuronal activity in each frontal area was also modulated by a gain of the reward itself. Especially, in the frontal association area, the fraction of neurons whose activity in each trial was modulated by the cue at that trial was higher than those in the other frontal areas. These results suggest that the frontal association area may bridge between the cue and its value-related information and engage in decision-making. Our macroscopic and microscopoic imaging also revealed that even slight movement such as licking strongly affects the neuronal activities across the whole cerebral cortex and the cortical representation of decision-making in awake animals is frequently masked by the movement-related representation.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-350 意思決定時のサル中脳ドーパミンニューロンと前頭眼窩ニューロンの信号ダイナミクスは価値から選択への変換過程と一致する Mengxi Yun（惲夢㬢）¹，Takashi Kawai（川合隆嗣）²，Masafumi Nejime（禰占雅史）²，Hiroshi Yamada（山田洋）²，Masayuki Matsumoto（松本正幸）² ¹筑波大院　人間総合科学生命システム医 ²筑波大　医学医療系 In economic decision-making, individuals decide to choose or not to choose an option based on its value. However, how the brain transforms the value information into a choice command remains elusive. Although some prefrontal regions (e.g., the orbitofrontal cortex, OFC) have been proposed to be involved in this transformation process, little is known about whether subcortical regions also contribute to this process. Here we demonstrate that midbrain dopamine neurons, a subcortical center for reward processing, exhibit a signal dynamics corresponding to the value-to-choice transformation. To investigate the role of dopamine neurons in the value-to-choice transformation, we recorded single-unit activities from dopamine neurons in monkeys while performing an economic choice task, in which the monkey needed to choose or not to choose an option based on the option's value. Additionally, owing to its reported role in value- and choice-related processes, we also recorded from neurons in the OFC for comparison. As well as OFC neurons, we found that dopamine neurons represented diverse signals related not only to the option's value, but also to the animal's choice; some neurons represented the value of the offered option, some represented whether the animal would choose or not choose the option, and some represented the combination of the value and choice (i.e., these neurons represented the value of the option only when the option was chosen by the monkey). We next analyzed the time course of these dopamine signals, and found that these signals were represented at different times. Shortly after the onset of the option, the value signal rapidly appeared, followed by the combination signal of value and choice. The choice signal arose at last. This time course of the three signals, which was also observed in the OFC, corresponds to that of value-to-choice transformation. Notably, the last-arising choice signal appeared before the monkey executed an action to choose the option in both dopamine neurons and the OFC, suggesting the value-to-choice transformation processes in these structures have completed before the animal's final motor action. Our findings show that dopamine neurons and OFC neurons share the same signal dynamics corresponding to the value-to-choice transformation, and provide evidence suggesting that not only prefrontal regions but also the subcortical dopamine system regulates the value-based choice formation.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-351 ラット超音波発声の自他の識別メカニズムの神経生理学的研究 Jumpei Matsumoto（松本惇平），Hiroshi Nishimaru（西丸広史），Yusaku Takamura（高村雄策），Taketoshi Ono（小野武年），Hisao Nishijo（西条寿夫）富山大院(医) システム情動科学 Discrimination between vocalizations of self and other is a fundamental function for normal communication. The auditory hallucinations in schizophrenia is suggested to be ascribed to misidentification of inner speech as external voices. We investigated neural responses to ultrasonic vocalizations (USVs) as affective communication in rodents. In the first experiment, we recorded neuronal response to self and other USVs in the amygdala, which is an important brain region for recognizing biological significance of stimuli, in male rats freely interacting with a conspecific male. EMG activities of the thyroarytenoid muscles around observation of a USV call was used to estimate which rat was emitted the USV. The results indicated that one quarter of the recorded amygdalar neurons (15/60) responded to 50 kHz USV calls emitted by subjects and/or conspecifics. Among the responsive neurons, most neurons (73%, 11/15) responded selectively to other's vocalizations, suggesting the amygdala involvement in self/other discrimination. Similar response preference to the other's vocalizations has been reported in the other brain areas in the auditory system and hypothesized to be due to the corollary discharges from the motor execution system that might inhibit the responses in the auditory system to the own vocalizations. However, it remains unknown how those brain regions interact during vocalizations. In the second experiment, we are presently performing simultaneous large-scale neural recording from the multiple brain regions during vocalizations, with a new experimental system incorporating 128-ch neural recording and a state-of-art non-invasive USV assignment using an ultrasonic microphone array. The present study on the neural mechanism of self-other discrimination in USVs should contribute to understanding of the neural mechanisms of auditory hallucination.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-352 サル上丘の中心視野ニューロンは、周辺視野ニューロンより顔様刺激に高い選択性を有する Le Van Quang（Quang Van Le），Quang Van Le（Le Van Quang），Quan Van Le（Le Van Quan），Hiroshi Nishimaru（西丸広史），Junpei Matsumoto（松本惇平），Yusaku Takamura（高村雄策），Etsuro Hori（堀悦郎），Taketoshi Ono（小野武年），Hisao Nishijo（西条寿夫） Dept System Emotional Science, Grad Sch Med Pharmaceu Sci, Univ Toyama, Japan Previous developmental studies suggest that the superior colliculus (SC) is implicated in innate face detection as a prototypical face template. However, there is no neurophysiological evidence that activity of primate SC neurons is selective to faces. In the present study, we recorded SC neuronal activities from monkeys during discrimination of various monochrome face-like and non-face patterns in a delayed non-matching to sample (DNMS) task. Visual stimuli included; 1) upright and inverted face-like patterns consisting of one of face contours (rice scoop, star, circle and square) and 5 facial features (2 eyes, 2 eyebrows and 1 mouth), 2) non-face patterns consisting of one of the same facial contours and 5 facial features in which the facial features were randomly positioned within the face contours, and 3) 4 face contours only (another non-face patterns). Furthermore, the same visual stimuli with opposite contrast polarity were tested. SC neurons were classified into 3 types based on their receptive fields (RFs); SC neurons with upper, lower and central RFs. The results showed that SC neurons responded stronger and faster to upright and inverted face-like patterns than to non-face patterns regardless of contrast polarity and face contours, and responses to original images were significantly correlated to those to negated images in more than half of the SC neurons. Furthermore, SC neurons with the central RF showed responses more selective to the face-like patterns; 1) relative response magnitudes to the face-like patterns were larger in the SC neurons with the central RFs than those with the peripheral RFs, and 2) more SC neurons with the central RF showed significant correlation between responses to the white and black stimulus sets than those with the peripheral RFs. Population activity of SC neurons with central RF discriminated face-like patterns from the other stimuli in the first 50-ms period after stimulus onset, and upright and inverted face-like patterns in the second 50-ms period. These results suggest existence of a broadly tuned face template in the midbrain, and provide neurophysiological evidence for SC involvement in face detection.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-353 主観的見えが空間的注意に与える影響 Momoko Hishitani（菱谷桃子）¹，Yuma Osako（大迫優真）²，Shota Murai（村井翔太）³，Kohta I Kobayasi（小林耕太）³ ¹同志社大学　生命医科学部 ²同志社大学大学院脳科学研究科神経回路情報伝達機構部門 ³同志社大学大学院生命医科学研究科神経行動工学研究室 Generally our environment is littered with large amount of visual information, and information in the same of neighboring visual field compete to be perceived. This competition predicts that only limited visual information reaches our visual recognition. In other words, our brain could selectively process information depending on their relevancy. This selective processing could be conducted by visual attentional mechanism, and the mechanism could be endogenously or exogenously oriented to a specific spaces or features. According to previous studies, subjective visibility and visual attention were independent but seem to be intimately related each other. However, how subjective visibility modulates visual attentional effect remains enigmatic. Clarifying effects of subjective visibility on visual attentional modulation will give a critical insight of generating executive function under our fluctuating subjective states. In this study, we modified classical Posner cueing task by introducing visual masking methods to quantify the attentional effect. Participants have to discriminate the location, where tilted target (in left or right) was presented, while cues presented before the target presentation instructed them to orient their attention to peripheral visual space. To clarify the effect of subjective visibility, inter-stimulus interval (ISI) between cue and visual masking were varied as they reported three graded subjective visibilities. The cue was presented either at peripheral location, where target stimulus was presented (peripheral condition), or at central location, where target stimulus was not presented (central condition). We revealed that subjective visibility shortened participant's reaction time when the cue was informative (valid cue) in both peripheral and central conditions. Interestingly, reaction time was not affected by subjective visibility when the cue was not informative (invalid cue) in both conditions. Moreover, we analyzed attentional effect as difference of reaction time between invalid and valid cue (Delta). The delta was increased with increasing subjective visibility in both conditions, indicating that subjective visibility enhanced visual attentional modulation. We will discuss these findings and preliminary fMRI data as to how subjective visibility enhance the visual attentional modulation.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-354 アセチルコリンによる睡眠覚醒制御の分子細胞学的理解 Yasutaka Niwa（丹羽康貴）¹，Hiroki R Ueda（上田泰己）^2,3，Takeshi Sakurai（櫻井武）¹ ¹筑波大　国際統合睡眠医科学研究機構 ²理研BDR合成生物学研究グループ ³東京大院医システムズ薬理学 Acetylcholine is the first molecule that was identified as a neurotransmitter. Although huge amount of data suggested that acetylcholine is important for wakefulness and REM sleep, the exact mechanisms of cholinergic regulation of sleep-wake states are still to be determined due to the complexity of its circuitry and receptors. Here we performed sleep analysis by using genetically modified mice where basal forebrain cholinergic neurons were inhibited or receptor genes were comprehensively deleted by CRISPR/Cas9, and found that muscarinic receptors Chrm1 and Chrm3 are essential for REM sleep and the amount of sleep during dark phase. Briefly we first made genetically modified mice where TrkA positive neurons were specifically inhibited by tetanus toxin. Interestingly those mice were severely affected on the amount of sleep only during dark phase. Next we developed tTR system which allows us to perform rescue experiments with Tet system in vivo. Taking advantage of this system, we found that TrkA positive cholinergic neurons are responsible for regulating the amount of sleep during dark phase. Finally we generated almost all acetylcholine receptor KO mice by using CRISPR/Cas9. We found that Chrm1 and Chrm3 are the receptors responsible for this regulation, but, surprisingly, DKO of Chrm1 and Chrm3 deprived mice of REM sleep. Our finding of unexpected viability despite the almost completely abolished REM sleep will allow us to rigorously verify whether REM sleep plays crucial roles in fundamental functions of organism, such as learning and memory and open a way to study its underlying molecular and cellular mechanisms.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-355 How attention shapes song perception in juvenile zebra finches during song learning Jelena Katic（Katic Jelena），Yoko Yazaki-Sugiyama（Yazaki-Sugiyama Yoko） Okinawa Institute of Science and Technology Graduate University Juvenile zebra finches learn to sing via vocal communications with their fathers (tutors) during development. Song learning improves more effectively through social interactions with tutors, compared to passive listening to recorded tutor songs. This suggests that high attention level, induced by social interactions with tutors, enhances song learning. Here we investigated how attention regulates zebra finch song learning by recording activities of neurons in the attention control brain area, the nucleus locus coeruleus (LC), and the higher auditory area, the caudomedial nidopallium (NCM), where tutor song memories appear to be stored (Yanagihara & Yazaki-Sugiyama, 2016). We chronically recorded extracellular, single-unit activity from LC and NCM neurons of freely behaving juvenile zebra finches for three days before and then three days after tutoring. LC and NCM neurons were tested for auditory responsiveness with playbacks of several song stimuli: tutor songs, songs of conspecific and heterospecific birds, and live tutors singing songs. LC neurons showed unbiased responses to various song playbacks during song learning. However, LC neurons responded more intensely to live tutor singing than to tutor song playbacks. LC neurons that showed high spontaneous tonic spiking activities, exhibited offset responses to live tutor singing, evidenced by decreased firing rates, but not to playbacks of tutor songs. The duration of decreased neural firing to song offset correlates with the number of song motifs (lasting up to 0.5 sec for 5 or more song motif repetitions). Anatomical analysis showed that LC neurons expressing GFP under the c-Fos promoter that were activated by tutor song exposure, project to the NCM. Proportions of NCM neurons that responded to more than two types of song playbacks decreased with tutoring (73.5% on the first day and 12.5% on the third day of tutoring), whereas the proportions of NCM neurons that showed selective responses to a specific song type increased (17.6% first day and 22.5% on the third day). A significant fraction of selective neurons (18%) showed biased responses to tutor songs. As in LC neurons, tutor-selective NCM neurons showed an offset response to live tutor singing by decreasing their firing rates. Taken together, we suggest that social interactions with tutors modulate neuronal activity of the LC, which affects auditory responses of the NCM, resulting in tutor song memory formation.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-356 To investigate the protective effect of fisetin on PM2.5-induced the inflammatory responses and its mechanism in murine microglia cells Hui-Wen Lin（Lin Hui-Wen）¹，Yu-Hsiang Kuan（Kuan Yu-Hsiang）²，Yuan-Yen Chang（Chang Yuan-Yen）³ ¹Department of Optometry, Asia University ²Department of Pharmacology, School of Medicine, Chung Shan Medical University ³Department of Microbiology and Immunology, School of Medicine, Chung-Shan Medical University Aim: Fisetin, 3,7,3,4-tetrahydroxyflavone, is a polyphenol and naturally occurring flavonoid that is abundantly found in several fruits and vegetables. Fisetin has anticancer, anti-proliferative, neuroprotective, and antioxidant activities. In this study, the anti-PM2.5 inducted inflammatory capacity of fisetin and its molecular mechanisms of action were analyzed. Methods and Results: The cytotoxic effects of fisetin were assessed in the presence or absence of PM2.5 via LDH and MTT assays. The results showed that fisetin (< 50 μM) had no toxic effects. In PM2.5 treated murine microglia (BV-2) cells, fisetin potently inhibited the production of NO, iNOS, COX-2 and inflammatory cytokine production via ELISA and Western blotting analysis. We found that PM2.5-induced NF-κB activation is regulated through inhibition of JAK, STAT1and STAT3 phosphorylation in response to fisetin. Additionally, fisetin caused the induction of HO-1. Conclusions: Taken together, our data indicate that fisetin diminishes the proinflammatory mediators NO, inflammatory cytokines and the expression of their regulatory genes, iNOS and COX-2, in PM2.5-infected BV-2 cells by inhibiting JAK-STAT pathway and activation of HO-1. Keywords: Fisetin, anti-inflammatory, proinflammatory mediators, PM2.5		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-357 Early-life regulator of astrocyte reactivity attenuates scarring and improve neuronal sparing after focal ischemia in adult marmoset monkeys Leon Teo（Teo Leon），Anthony G Boghdadi（Boghdadi Anthony），Jihane Homman-Ludiye（Homman-Ludiye Jihane），Inaki Carril-Mundinano（Carril-Mundinano Inaki），Mitchell deSouza（deSouza Mitchell），William C Kwan（Kwan William C），James A Bourne（Bourne James A） Australian Regenerative Medicine Institute Over the past two decades, thousands of preclinical drugs developed for the treatment of CNS injuries have ultimately failed in clinical trials. However, persisting evidence demonstrates that neurodevelopmental processes are inexorably linked to neuroregeneration. Therefore, the key to unlocking regenerative capacities after CNS injuries may lie in recapitulating developmental processes through novel pharmaco-therapeutic strategies. Glial scarring and reactive astrogliosis after stroke is a major impediment to neuroregeneration. However, chronic scarring in the infant primate neocortex is markedly less severe compared to adults, allowing greater permissiveness towards functional recovery after injuries. EphA4 is a major modulator of astrogliosis after CNS injuries; however, the ephrin ligands involved in the injured brain remains unknown. A clinically translatable non-human primate (NHP) model of stroke in the infant and adult neocortex was utilised in this project. We discovered that astrogliosis in the infant and adult brains are regulated differentially and are reflected in the ephrin ligands expressed on reactive astrocytes after injury. Specifically, ephrin-A1 in infants and ephrin-A5 in adults. In experiments using human astrocytes, ephrin-A1 induced astrocyte repulsion, suppressed proliferation and prevented wound closure, accounting for discrete scarring in infants. Conversely, ephrin-A5 elicited cell attraction, increased proliferation and induced rapid wound closure, contributing to severe scarring in adults. Surprisingly, ephrin-A1 treatment did not suppress astrocyte reactivity, instead induced a more neuroprotective phenotype through GAP43 dependent upregulation of glutamate transporter (EAAT2) that likely contributes to improved neuronal sparing. Most importantly, reintroduction of ephrin-A1 forward signalling in vivo through local infusion into the injured adult brain resulted in a significant reduction in the volume, density and severity of glial scarring as well as concomitant improvements in neuronal survival and sparing of local circuitry and function. Here we show that ephrin-A1 signalling contributes to discrete scarring in infants and can be recapitulated in the injured adult brain to attenuate adult scarring and improve neuronal survival and functional sparing. Thus, we provide evidence that beneficial infant repair processes can be recapitulated in the adult brain to improve outcomes after CNS injuries.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-358 in vitro脳内出血モデルにおける血腫への好中球浸潤に対するロイコトリエンB₄およびリポキシンA₄の機能解析 Masanori Hijioka（肱岡雅宣），Risa Futokoro（懐理紗），Yoshihisa Kitamura（北村佳久）立命館大薬薬効解析科学 Intracerebral hemorrhage (ICH) is a devastating neurodegenerative disease result from the leakage of blood constituents into brain parenchyma. In ICH brain, numerous leukocytes invade into the hematoma. Among the leukocytes, neutrophil is assumed to be detrimental for the ICH pathogenesis. Our previous research showed that leukotriene B₄ (LTB₄), known as inflammatory lipid mediator, promotes neutrophil invasion into hematoma and exacerbates motor dysfunctions of ICH model mice (Hijioka M. et al., 2017). Here we focused on microglia as the source of LTB₄ production. First, we checked the expression profiles of LTB₄ and other arachidonic acid metabolites in ICH brain. Male C57BL/6J mice received the injection of typeVII collagenase (0.025 U) into the striatum. Lipidomics analysis based on LC-MS/MS revealed the increase of LTB₄ content at 12 h after ICH induction. Then, we examined the effect of LTB₄ secreted from microglia. Human promyelocytic leukemia cell line HL-60 cells were differentiated into neutrophil-like cells by 1 μM all-trans retinoic acid. Differentiated HL-60 (dHL-60) cells showed polymorphonuclear phenotypes revealed by Giemsa staining and mRNA expression levels of LTB₄ receptor 1 (BLT1) were increased by the differentiation into dHL-60 cells. dHL-60 cells were seeded on the upper layer of cell culture inserts with 3.0 μm pores and conditioned medium of BV-2 cells treated with thrombin for 12 h were applied to the bottom layer. Conditioned medium of thrombin-treated BV-2 cells promotes infiltration of dHL-60 cells into the bottom layer, but U75302, a BLT1 antagonist, suppressed the infiltration. Finally, we evaluated the effect of lipoxin A₄ (LXA₄) known as anti-inflammatory lipid mediator synthesized by the same enzymes that producing LTB₄. LXA₄ treatment significantly suppressed the infiltration of dHL-60 cells into the bottom layer. These results suggest that LTB₄ secreted by microglia promote neutrophils invasion in ICH and LXA₄ can suppress the neutrophil invasion into hematoma.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-359 低負荷走行運動が脳卒中モデルラットの海馬におけるシナプス関連因子の遺伝子発現に与える影響について Misato Okamura（岡村未里）¹，Takahiro Inoue（井上貴博）²，Akari Kako（加古明里）¹，Yasuyuki Takamatsu（高松泰行）³，Hiroshi Maejima（前島洋）³ ¹北海道大学医学部保健学科 ²北海道大学大学院保健科学院 ³北海道大学大学院保健科学研究院機能回復学分野 Post-stroke cognitive impairments severely disturb the patients to rehabilitate sensory-motor functions. Synaptic plasticity in the hippocampus has a crucial role for memory and cognitive function. In particular, brain-derived neurotrophic factor (BDNF) plays an important role in neuroplasticity, neurogenesis and neuroprotection in the hippocampus. It has been well recognized that low-intensity exercise beneficially contributes to improve cognitive impairments after stroke accompanying the upregulation of BDNF protein expression in the hippocampus. However, the effects of low-intensity exercise on the gene expressions in the post-stroke hippocampus are poorly understood. Therefore, the objective of this study was to examine the effects of low-intensity exercise on the gene expression relating to synaptic plasticity in the post-stroke hippocampus using model rats for stoke. Male Sprague-Dawley rats were divided into 4 groups based on the two factors of stroke and exercise: a sham group (SHAM, n=8), a sham-exercise group (SHAM-Ex, n=8), a stroke group (Stroke, n=7), and a stroke-exercise group (Stroke-Ex, n=7). The Stroke and Stroke-Ex groups received the middle cerebral artery occlusion (MCAO) surgery to induce an ischemic stroke. The SHAM-Ex and the Stroke-Ex groups were forced to run on a treadmill (11 m/min for 30 min) every day for 2 weeks. Neurological deficits were evaluated by 5-point scale. The mRNA expressions of BDNF, TrkB, Synaptophysin, and PSD-95 in the ipsilesional hippocampus were assayed using quantitative RT-PCR method. Low-intensity exercise had a marginal effect on the neurological deficits after stroke, and there was no significant effect of low-intensity exercise on gene expression in the ipsilesional hippocampus after stroke. Meanwhile, the mRNA expression of TrkB was significantly greater in Stroke and Stroke-Ex groups compared with the expression in SHAM and SHAM-Ex groups, indicating that stroke increased the mRNA expression of TrkB in the ipsilesional hippocampus. Altogether, it was suggested that stroke could induce the plastic change to enhance the association between BDNF and TrkB in the ipsilesional hippocampus. The present study showed novel findings regarding the neuroplastic changes in the hippocampus after stroke.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-360 ラット一過性中大脳動脈閉塞モデルにおけるプロゲステロン受容体アゴニストnestoroneによる長期神経保護作用 Motoki Tanaka（田中基樹）¹，Takunori Ogaeri（魚返拓利）¹，Mikhail Samsonov（Samsonov Mikhail）²，Masahiro Sokabe（曽我部正博）¹ ¹名古屋大学大学院医学系研究科メカノバイオロジー・ラボ ²R-Pharm, Moscow, Russia Post-ischemic administration of progesterone (P4) has been shown to improve short- and long-term histological and functional outcomes in animal models of stroke, and P4 receptors play a crucial role in this neuroprotection. However, it currently remains unclear whether the activation of P4 receptors alone is sufficient to exert long-term neuroprotection against stroke because P4 exhibits androgenic, estrogenic, and GABAergic activities via several of its metabolites. Nestorone is a highly potent selective P4 receptor agonist without such steroidogenic and GABAergic activities. Therefore, we evaluated the effects of nestorone, its dose-response relationship, and long-term histological and functional outcomes in male rats subjected to transient middle cerebral artery occlusion (MCAO). The dose-response relationship of nestorone showed that the 6-h post-ischemic administration of 10 µg/kg nestorone resulted in greater reductions in infarct sizes 48 h after MCAO than the other two doses tested (5 and 80 µg/kg), and this dose of nestorone coincidently significantly decreased astrocyte activation in the peri-infarct cortical region. Moreover, 10 µg/kg nestorone significantly prevented impairments in spatial working memory, motor coordination, and somatosensory function on the 28th and 29th days and slightly reduced infarct size on the 30th day after MCAO. Our present results suggest that sole activation of P4 receptors by nestorone is sufficient to exert short- and long-term neuroprotection against transient focal cerebral ischemia in male rats. Stroke patients may have androgen or estrogen-related health issues, and, thus, natural P4 treatments may not be applicable due to the androgenic and estrogenic activities of P4 metabolites. Nestorone may be therefore applicable as a therapeutic agent against stroke in a wider range of stroke patients than natural P4 due to its potent progestational effects without other steroid-related activities.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-361 くも膜下出血後脳血管攣縮/神経機能不全の本態は白血球の脳血管浸潤である Hiroshi Ishii（石井宏史）¹，Yasuhiro Aida（会田泰裕）²，Yasuyuki Ohkuma（大熊康祐）¹，Seiichi Munesue（棟居聖一）³，Jureepon Roboon（ローブンジュリーポン）¹，Tsuyoshi Hattori（服部剛志）¹，Mika Takarada（寳田美佳）¹，Mitsutoshi Nakada（中田光俊）²，Yasuhiko Yamamoto（山本靖彦）³，Osamu Hori（堀修）¹ ¹金沢大学医学系　神経解剖学 ²金沢大学医学系　脳神経外科学 ³金沢大学医学系　血管分子生物学 Purpose: Subarachnoid hemorrhage (SAH) suddenly occurs predominantly by ruptured cerebral aneurysm. The severity of SAH is higher than other types of stroke such as cerebral ischemia and intracranial hemorrhage. It is critical to prevent disability following cerebral vasospasm (CVS) after SAH. Our research purpose is to elucidate the mechanism underlying CVS and find novel target of brain injury after SAH. Method: In BL6/J background mice, a micro-filament with a tip cut at a right angle was inserted from left external carotid artery stump into internal carotid artery, and ruptured at the bifurcation of anterior cerebral artery and middle cerebral artery. Results: In a damage-associated molecular patterns (DAMPs) receptor-deficient mice, neurological score and cerebral vasospasm was markedly improved compared with control mice. The DAMPs receptor was expressed in endothelial cells of internal carotid artery after SAH. However, vascular specific DAMPs receptor-deficient mice did not show improvement of CVS after SAH. Instead, bone marrow transplantation of wild type mouse into DAMPs receptor-deficient mice did not show improvement these phenotypes compared with control one into wild type mice. Therefore, DAMPs receptor in immune cells may contribute to CVS after SAH. Indeed, neutrophils accumulated around cerebral artery after SAH in DAMPs receptor-dependent manner. Conclusion: We newly identified a DAMPs receptor as an inducer molecule of CVS/brain injury after SAH. The DAMPs receptor in leukocytes is attributed to accumulation around cerebral vessels, which contributes to critical pathology of CVS after SAH.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-362 A novel cell-penetrating peptide protects against neuron apoptosis after cerebral ischemia by inhibiting the nuclear translocation of annexin A1 Xing Li（Li Xing）¹，Lu Zheng（Zheng Lu）¹，Qian Xia（Xia Qian）¹，Lu Liu（Liu Lu）¹，Meng Mao（Mao Meng）¹，Huijuan Zhou（Zhou Huijuan）¹，Yin Zhao（Zhao Yin）²，Jing Shi（Shi Jing）¹ ¹Huazhong University of Science and Technology ²Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology Nuclear translocation of annexin A1 (ANXA1) has recently been reported to participate in neuronal apoptosis after cerebral ischemia. Prevention of this process should therefore inhibit neuronal apoptosis and protect against cerebral stroke. Here, we identified the amino acid residues from Arg228-Phe237 function as a nuclear translocation signal (NTS) and are responsible for nuclear translocation of ANXA1. Intriguingly, we synthesized a cell-penetrating peptide derived by conjugating the trans-activator of transcription (Tat) domain to the NTS sequence. This Tat-NTS peptide specifically blocked the interaction of ANXA1 with importin β and, consequently, the nuclear translocation of ANXA1. The Tat-NTS peptide improved the survival of hippocampal neurons subjected to oxygen-glucose deprivation and reperfusion in vitro. Moreover, using a focal brain ischemia animal model, we showed that the Tat-NTS peptide could be efficiently infused into the ischemic hippocampus and cortex by unilateral intracerebroventricular injection. Importantly, further work revealed that administration of the Tat-NTS peptide alleviated neuronal apoptosis, resulted in a dramatic reduction in infarct volume and that this was correlated with a parallel improvement in neurological function after reperfusion. Based on its profound neuroprotective and cognitive-preserving effects, the Tat-NTS peptide represents a novel and potentially promising new therapeutic candidate for the treatment of ischemic stroke.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-363 視神経障害に対する分子状水素の保護効果 Mami Noda（野田百美），Tetsushi Niiyama（新山哲士），Ayaka Fukuo（福尾彩香）九州大院薬病態生理 The therapeutic potential of molecular hydrogen (H₂) is emerging in a number of human diseases and in their animal models. We have previously reported that H₂ has protective effect on optic nerve loss due to ischemia-related injury in type 1 diabetic model mice even though there was no inhibitory effect on the increase in blood glucose level. Here we tested whether H₂ has neuroprotection against lifestyle-related disease with or without diabetes. Lifestyle-related disease was mimicked by feeding high fat diet (HFD) and diabetic model was made by injection of streptozotocin (STZ). Using male C57BL/6J mice (7 weeks old~), HFD (Quick fat, CLEA Japan, Inc.)was given for 42 days. STZ (150 mg/kg BW) was injected (i.p.) 7 days after starting HFD. Saturated H₂ water (~0.8 ppm) or normal tap water (control) was given for the same period of HFD. Body weights and blood glucose levels were monitored once a week. After 42 days, the mice were sacrificed and their optic nerves were subjected for immunostaining of nerve fiber and glial cells. The body weights were significantly decreased in HFD/STZ group, which was significantly prevented by H₂. The blood glucose levels were significantly increased in HFD/STZ group, which was partially attenuated by H₂. The loss of optic nerves was significant in HFD group, which was significantly prevented by H₂. HFD/STZ group also showed significant loss of optic nerves, which was partially prevented by H₂. The loss of oligodendrocytes was also observed in HFD group but the protective effect of H₂ was not significant. These results show intriguing pathological findings that HFD does not increase body weight or blood glucose level but induces significant loss of optic nerve and oligodendrocytes, which can be prevented by continuous consumption of H₂. On the other hand, significant increase in blood glucose level and loss of optic nerves by HFD/STZ were only partially attenuated by H₂, though decreased body weights were significantly prevented by H₂.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-364 脳梗塞発症前予防運動は、ラットの脳虚血後、内在性14-3-3gammaの増強を介して脳損傷および神経細胞のアポトーシスを減少させる。 Shotaro Otuka（大塚章太郎）^1,2，Seiya Takada（高田聖也）¹，Kazuki Nakanishi（中西和毅）¹，Harutoshi Sakakima（榊間春利）¹ ¹鹿児島大医保理学 ²鹿児島大院医歯システム血栓 Ischemic stroke is one of the leading causes of mortality and severe long-term morbidity worldwide. Preconditioning exercise can exert neuroprotective effects after stroke. However, the mechanism underlying these neuroprotective effects by preconditioning exercise remains unclear. 14-3-3gamma is an important early ischemia-inducible protective factor against ischemic cell death in cerebral cortical neurons. We investigated the anti-apoptosis mechanism of enhanced 14-3-3gamma mediated by preconditioning exercise-induced brain ischemic tolerance after stroke. Rats were assigned to four groups: exercise and ischemia (Ex), ischemia and no exercise (No-Ex), exercise and no ischemia (Ex-only), and no exercise and ischemia (control). Rats were trained on a treadmill for 5 days a week for 3 weeks (running speed, 25 m/min; running duration, 30 min/day). After the exercise program, stroke was induced by a 60 min left middle cerebral artery occlusion using an intraluminal filament. The infarct volume, neurological deficits, and motor function, as well as expression levels of hypoxia-induced factor-1α (HIF-1α), 14-3-3gamma, P2X7 receptors, p-β-catenin Ser37, Bax, and caspase 3 were evaluated by immunohistochemistry and western blotting. The expression of HIF-1α and 14-3-3gamma significantly increased in neurons and astrocytes in the Ex-only group. HIF-1α was co-expressed with P2X7 receptor- and GFAP-positive astrocytes. After stroke, the Ex group had significantly reduced brain infarction. HIF-1α and 14-3-3gamma significantly increased in the Ex group compared to the No-Ex group. In addition, p-β-catenin Ser37 significantly increased following elevated 14-3-3gamma; in contrast, Bax and caspase 3 were significantly reduced in the Ex group. Our findings indicate that preconditioning exercise induces ischemic tolerance through upregulation of HIF-1α and 14-3-3gamma in the cortical neurons and astrocyte; preconditioning exercise reduces infarct volume and sensorimotor deficits after stroke. Additionally, our findings suggest that upregulated 14-3-3gamma by preconditioning exercise reduce ischemic neuronal cell death via the 14-3-3gamma/p-β-catenin Ser37/Bax/caspase 3 anti-apoptotic pathway, one of the underlying neuroprotective mechanisms after ischemic stroke.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-365 GABA_A受容体阻害と運動の併用は脳卒中モデルラットの機能回復と損傷側一次運動野におけるBDNF発現を増強する Takahiro Inoue（井上貴博）¹，Yasuyuki Takamatsu（高松泰行）²，Hiroshi Maejima（前島洋）² ¹北海道大学大学院保健科学院 ²北海道大学大学院保健科学研究院機能回復学分野 Stroke is one of the leading causes of physical disability in the world. It has been well recognized that brain-derived neurotrophic factor (BDNF), a member of neurotrophins, has a crucial role for the recovery of motor function after stroke. Our previous study using regular mice showed that exercise combined with low-level GABA_A receptor antagonist increased the protein abundance of BDNF in the motor related brain region. Thus, it was expected that exercise combined with GABA_A receptor antagonist could facilitate stroke recovery. The objective of this study was to examine the interactive effects of low-level GABA_A receptor antagonist and exercise on BDNF expression in the ipsilesional primary motor cortex (M1) of stroke model rats. 10-week-old male Sprague-Dawley rats were divided into 4 groups based on the two factors of exercise and drug administration: a control group (CON, n=7), an exercise group (EX, n=7), a bicuculline group (BIC, n=7), and a bicuculline and exercise group (BIC-EX, n=8). All rats received the middle cerebral artery occlusion (MCAO) surgery to induce an ischemic stroke. We administered the GABA_A receptor antagonist bicuculline (0.25 mg/kg, i.p.) to the BIC and the BIC-EX groups. The EX and the BIC-EX groups exercised on a treadmill (11m/min for 30 min). Each intervention started 3 day after MCAO surgery and was carried out every day for 2 weeks. For behavioral assessments, adhesive removal test and cylinder test were performed. The mRNA expression and protein abundance of BDNF in the ipsilesional M1 were assayed using RT-PCR and ELISA. No Significant motor recovery was found in the EX and the BIC groups, whereas exercise in the presence of bicuculline administration significantly enhanced the recovery of motor function specifically in the BIC-EX group. Furthermore, BDNF protein level in the ipsilesional M1 was significantly greater in the BIC-EX group compared with those in the CON, EX, and BIC groups. This study indicated that low-level inhibition of GABA_A receptors after stroke could beneficially facilitate functional recovery accompanying the upregulation of BDNF expression in the ipsilesional M1. Therefore, this study provides a novel insight of pharmacological neuromodulation into stroke rehabilitation.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-366 血管性認知症モデルとしてのマイクロスフェア脳塞栓ラットの特性 Naoyuki Himi（氷見直之）¹，Naohiko Okabe（岡部直彦）¹，Hisashi Takahashi（高橋尚）²，Emi M Nakamura（中村 M 恵美）¹，Norito Hayashi（林範人）¹，Issei Sakamoto（坂本一晴）¹，Tomoshige Koga（古我知成）²，Osamu Miyamoto（宮本修）¹ ¹川崎医科大基礎医生理２ ²川崎医療福祉大リハビリテーション [Introduction] There are many types of animal models representing the cerebrovascular disease, however, there is no multiple microinfarction model showing cognitive dysfunction by small vessels dysfunction as like the vascular dementia (VD). Because microspheres (MS) induce the brain embolism in small vessels and memory dysfunction, MS embolism is thought to be a suitable VD animal model. In this study, we investigated the optimal condition in MS embolism model and their characteristics of cerebral blood flow (CBF), motor and memory function. [Methods] Embolism were produced by MS (from 2,500 to 4,000 particles) injection into right internal carotid artery of rats with measurement of CBF in motor cortex and hippocampus. The pathological output including motor functions (physical deficit score and rotarod test) and memory functions (Morris water maze test, MWM) were observed. [Results] CBF in both hippocampus and cortex decreased by MS injection, and motor functions (physical deficit score and rotarod test) and memory function (MWM) were aggravated dose-dependently. In this experiment, the injection of 3,000 particles of MS decreased in CBF to 74.7 ± 11.8 % and 73.9 ± 4.0 % vs control at hippocampus and motor cortex, respectively. Moreover, 3,000 particles of MS caused significant memory dysfunction but not motor dysfunction. Therefore, 3,000 particles of MS injected rat would be more appropriate as a VD model. [Conclusion] 3,000 particles of MS injection decreased CBF in cortex and hippocampus, and induced memory dysfunction like VD without severe motor dysfunction.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-367 リコペンおよび松葉エキスによる脳虚血後のミクログリアの活性化抑制はTNF-αの放出を制限する Kimikazu Fujita（藤田公和）¹，Nobuko Yoshimoto（芳本信子）²，Chiyuki Kaneko（金子千之）³，Mahito Ohkuma（大熊真人）¹，Soh Hidaka（日高聡）¹，Hiroyuki Suganuma（菅沼大行）⁴，Eiichi Miyachi（宮地栄一）¹ ¹藤田医科大学医学部生理学Ⅱ ²橋本内科クリニック ³藤田医科大学医療科学部 ⁴カゴメ株式会社総合研究所 Selective neuronal loss (apoptosis) was observed in the hippocampal CA1 region several days after transient occlusion of the bilateral carotid arteries restricted blood supply to the forebrain of Mongolian gerbils. As cerebral ischemia increases microglial activity and release of cytokines, we hypothesized that this increased activity may play a key role in the progression of brain injury. Several studies have revealed that lycopene inhibits microglial activity and that pine needle extract has anti-inflammatory effects. Therefore, in this study, using western blotting, we investigated changes in hippocampal expression of a marker protein for microglial activity (ionized calcium-binding adapter molecule 1: Iba-1) in the brains of Mongolian gerbils exposed to lycopene or pine needle extract-induced neurodegenerative depression. Moreover, we performed immunohistochemical staining for Iba-1 in the hippocampus and analyzed inflammatory cytokines (TNF-α and IL-1β) expression using ELISA. Animals were fed either a normal diet (MF feed) or a lycopene or pine needle extract-containing MF feed diet (5% lycopene or pine needle extract). After attaining a body weight of 60-65g, the animals were subjected to transient cerebral ischemia for 10 min or to a sham operetion. Animals with ischemic or sham treatments were decapitated at 3 h, and 1,3, and 7 days post-ischemia, and the hippocampal regions were dissected from the brain. The number of Iba-1 immumoreactive cells was found to have decreased in the CA1 region of the animals in the lycopene or pine needle extract-treated group at 7 days post-ischemia. Iba-1 protein expression in the hippocampus gradually increased in the control group after ischemia/reperfusion. In contrast, Iba-1 protein levels in the lycopene or pine needle extract-treated group were significantly lower than those in the control group at 3 h post-ischemia. In addition, TNF-α and IL-1β levels increased gradually in the control group after ischemia/reperfusion. TNF-α levels were significantly lower in the lycopene or pine needle extract-treated group than in the control group at 3 h after ischemia. Our results indicate that lycopene and pine needle extract exert a neuroprotective effect on the hippocampus through their ability to suppress post-ischemic microglial activity, and to inhibit inflammation by reducing TNF-α levels.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-368 C9orf72 FTLD/ALS細胞モデルにおけるリピート翻訳の調整機構 Shiho Gotoh（後藤志帆），Kohji Mori（森康治），Yuya Kawabe（河邉有哉），Tsubasa Omi（近江翼），Tomoko Yamashita（山下智子），Manabu Ikeda（池田学）大阪大院医 Frontotemporal lobar degeneration (FTLD) is a neurodegenerative disease that causes dementia, aphasia and behavior disorders with or without psychiatric symptoms. Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease in which motor neurons degenerate and muscle strength decreases. Recent studies have revealed that these clinically distinct diseases share common molecular mechanism. For the onset of genetically inherited version of FTLD and ALS, abnormally extended GGGGCC repeat sequence in noncoding region of C9orf72 gene is a most frequent causative molecule. The GGGGCC repeat sequence is translated into accumulating dipeptide repeat proteins (DPR) via repeat-associated non-ATG (RAN) translation. We consider DPR is a primary driver for neurodegeneration in C9orf72-related FTLD and ALS although the other disease mechanisms including haploinsufficiency of C9orf72 protein and repeat RNA toxicity are not exclusive. Inhibition of repeat translation would lead to reduction of the toxic dipeptide repeat protein expression; therefore, selective inhibition of RAN translation would be potentially therapeutic. To date; however, it is still unclear how the unusual repeat translation is regulated and also how we could manipulate it. In this study by using a cellular model of C9orf72-FTLD/ALS, we analyzed how DPR translation is regulated. Our strategies were 1) How DPR translation is regulated through C9orf72 RNA binding proteins, 2) How DPR translation is regulated through translation regulating molecules. In this poster, we will show and discuss our latest data. Inhibition of repeat translation would be a promising therapeutic strategy for FTLD, ALS and also for the other neurodegenerative disorders with expanded repeats. Therefore, we have to understand the unconventional repeat translation in more detail.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-369 シナプス前のILEI/FAM3CはAβのエキソサイトーシスと同様に神経活動依存的に分泌される Masaki Nakano（中野将希）¹，Yachiyo Mitsuishi（三ツ石弥千代）¹，Naoki Watanabe（渡邊直希）¹，Emi Hibino（日比野絵美）¹，Takuma Sugi（杉拓磨）¹，Takashi Saito（斉藤貴志）²，Takaomi C Saido（西道隆臣）²，Toshiharu Suzuki（鈴木利治）³，Masaki Nishimura（西村正樹）¹ ¹滋賀医大神経難病研究センター ²理研CBS 神経老化制御 ³北海道大院薬神経 Alzheimer's disease is characterized by extracellular deposition of amyloid-β peptide (Aβ), intracellular neurofibrillary tangle formation, and neuronal cell death. Accumulation of Aβ in the brain triggers the pathological process of Alzheimer's disease. Aβ is produced by proteolytic cleavage of amyloid-β precursor protein (APP) and secreted mainly from neurons. Previously, we reported a novel role of a secretory protein ILEI (interleukin-like epithetial-mesenchymal transition inducer, also known as FAM3 superfamily member C) in modulation of Aβ metabolism. ILEI binds to the γ-secretase complex and destabilizes APP C-terminal fragments to suppress Aβ production without inhibiting γ-secretase activity. Neuronal ILEI resides in the trans-Golgi network and presynaptic terminal. Synaptosome fractionation analysis further indicated that ILEI is colocalized with APP and the γ-secretase complex at the active zone-docked synaptic vesicles. In this study, we investigated the relationship between ILEI release and synaptic activity. [Methods] We generated monoclonal antibodies against recombinant ILEI and setup sandwich ELISA for mouse ILEI. Using the established ELISA, we measured ILEI in brain soluble fractions and cerebrospinal fluid (CSF). We also applied microdialysis to monitor ILEI and Aβ concentrations in the interstitial fluid of cerebral cortex of freely moving humanized mutant APP-knockin mice. [Results] Soluble fractions of brain homogenates contain a secreted form of ILEI/FAM3C. Three mice in a same cage were sacrificed every 3 hours for a day. Brain ILEI level showed phasic alteration; low in daytime and high in nighttime. Fluctuation of CSF ILEI was not synchronized with that of brain ILEI. Microdialysis study using APP-knockin mice revealed that interstitial fluid concentration of ILEI was roughly parallel with a level of physical activity (amount of movement) but not always with that of Aβ. Treatment with anesthetics, tetrodotoxin or tetanus toxid obviously decreased both of ILEI and Aβ in the interstitial fluid. [Conclusion] Our results suggest that ILEI is excytosed into the extracellular space in a synaptic activity-dependent manner and negatively regulates Aβ production at the presynaptic terminal.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-370 無麻酔マウスfMI法による自閉症モデルマウスの脳内ネットワ－ク解析 Tomokazu Tsurugizawa（釣木澤朋和）¹，Tomokazu Tsurugizawa（釣木澤朋和）¹，Kota Tamada（玉田紘太）²，Akihiko Kitamura（北村明彦）³，Toru Takumi（内匠透）² ¹Neurospin, CEA-Saclay, Gif-sur-Yvette, France ²理研CBS 精神生物学 ³味の素(株) The autism model mouse (15q dup, a copy number variation model of autism spectrum disorder (ASD)) shows the abnormal behavior in social interaction. In the current study, we aimed to investigate the altered brain network using resting state functional connectivity and task-based fMRI using fMRI. Although that the mouse functional MRI (fMRI) usually uses the anesthetics for suppression of motion and physiological noise, the anesthesia causes the suppression of the consciousness of mice as well as the modification of neurovascular coupling. Here, we developed the awake fMRI protocol in the mouse to perform the fMRI study (resting state fMRI and odor cognition task in the MRI bore) in 15q dup mice. The alteration of functional connectivity in the whole brain and the suppression of the activation during odor cognition to the stranger odor was observed in 15q dup mice. In parallel to the fMRI study, diffusion tensor imaging was performed on fixed brain of 15q dupmice. Widespread reductions in fractional anisotropy were also evident in 15q dup mice relative to wild type mice. The profile of amino acid concentration in the forebrain was altered in 15q dup mice. The D-cycloserine (DCS)-treatment partially normalizes hypo-connectivity and odor-evoked fMRI responses in the frontal cortex of 15q dup mice as well as the social behavior. These results indicate that awake mouse fMRI is useful for translational research and the alteration of functional/structural brain network could link to the abnormal behavior in 15q dup mice.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-371 ヒトIQおよびアミロイド蓄積に及ぼすACE抑制の影響 Kun Zou（鄒鶤）¹，Shuyu Liu（劉姝余）²，Fujiko Ando（安藤富士子）³，Yu Fujita（藤田融）²，Junjun Liu（劉俊俊）²，Tomoji Maeda（前田智司）²，Xuefeng Shen（沈雪峰）²，Kota Kikuchi（菊池光太）²，Hiroshi Shimokata（下方浩史）⁴，Hiroto Komano（駒野宏人）²，Makoto Michikawa（道川誠）¹ ¹名古屋市立大学　医学研究科　病態生化学分野 ²岩手医科大学　薬学部　神経科学分野 ³愛知淑徳大学　健康医療科学部 ⁴名古屋学芸大学　大学院栄養科学研究科 Inhibition of angiotensin-converting enzyme (ACE) is a worldwide treatment strategy for hypertension. In addition to converting angiotensin I to angiotensin II, ACE also converts amyloid β-protein 42 (Aβ42) to Aβ40. Neurotoxic Aβ42 is believed to play a causative role in the development of Alzheimer's disease (AD), whereas Aβ40 has neuroprotective effects against Aβ42 aggregation and against metal-induced oxidative damage. Whether ACE inhibition enhances Aβ42 aggregation or impairs human cognitive ability are very important issues for preventing the onset of AD and for optimal treatment of hypertension. In an 8-year longitudinal study, we found that the intelligence quotient of male hypertensive patients taking ACE inhibitors declined more rapidly than others taking non-ACE-inhibitors. In addition, the serum of AD patients showed a decrease in Aβ42-to-Aβ40-converting activity compared with age-matched normal serum. Using human amyloid precursor protein (hAPP) transgenic mice, we found that a clinical dose of an ACE inhibitor was sufficient to increase brain amyloid deposition. We further generated hAPP/ACE(+/-) mice and found that a decrease in the ACE level promoted Aβ42 deposition and increased the number of apoptotic neurons. These results suggest that inhibition of ACE activity is a risk factor for impaired human cognitive ability and for triggering the onset of AD.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-372 A&beta毒性に対する海馬神経ネットワークの恒常性維持機構 Hiroki Ishikawa（石川裕貴），Tetsuya Hori（堀哲也），Naoto Saitoh（齋藤直人）同志社大院生命医科学 Alzheimer's disease (AD) is characterized by the accumulation of the amyloid-β (Aβ) peptide within the extracellular space of the brain. Aβ may exert its neurotoxic effects via multiple mechanisms and in particular through degradation of excitatory synaptic transmission associated with impaired synaptic plasticity. However, homeostatic plasticity might assure neuronal network stability against Aβ accumulation because AD has long precritical period in spite of the progressive accumulation. We focus on the presymptomatic mechanism which must be the sources of the ideas for the prevention medicine of AD. In this study, we applied 10 nM Aβ42 to cultured hippocampal neurons at 20 day in vitro (DIV) and examined chronic toxicity of Aβ42 on the neural network activity from 20 DIV to 28 DIV. First, we analyzed the pyramidal cells firing rate by calcium imaging using CaMKIIα-GCaMP6f. The Firing rate was maintained at 36°C in spite of the presence of Aβ42, whereas it was transiently 24 DIV reduced by Aβ42 at 28°C. These results might indicate that Aβ42 has toxic effect to the firing rate, but homeostatic plasticity recovers from the damage under the cultured condition. Second, we focused on the number of synapses and expression level of voltage-gated Na channel 1.6 (Nav1.6). The number of excitatory synapses increased by 50% at 24 DIV. The result might suggest that neural network attempted to cancel the Aβ42 toxicity by the amount of the excitatory synapse. On the other hand, the expression level of Nav1.6 at axon initial segments didn't change. It might suggest that Aβ42 has no significant effect on action potential generation. Considering that the firing rate was not changed at 36°C, it was expected that Aβ42 caused serious damage to synaptic transmission in the initial phase. Third, we purified total RNA from cultured hippocampal neurons at 24 DIV, and performed microarray analysis. As a result, we identified the upregulation of oxidative phosphorylation-related genes. Their genes might try to recover Aβ42-induced mitochondrial dysfunction. In summary, our results indicate that neural network resists Aβ toxicity by increasing the number of excitatory synapse and upregulation of mitochondrial function to keep the network activity.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-373 マウス骨髄由来細胞からのTGF-β1の分泌によるミクログリアのAβ貪食の促進 Kazuyuki Takata（高田和幸）¹，Eriko Kuroda（黒田絵莉子）²，Shohei Kawanishi（河西翔平）²，Fumitaka Ueno（植野文貴）²，Kaneyasu Nishimura（西村周泰）¹，Yuki Toda（戸田侑紀）²，Yoshihisa Kitamura（北村佳久）³，Shun Shimohama（下濱俊）⁴，Eishi Ashihara（芦原英司）² ¹京都薬大統合薬科 ²京都薬大病態生理 ³立命館大薬薬効解析 ⁴札幌医大医神経内科 Amyloid-β (Aβ) accumulation in brains is a critical trigger for the development of Alzheimer's disease (AD). Recently, we reported that intrahippocampal transplantation of mouse bone marrow-derived cells stimulated with colony-stimulated factor-1 (CSF-1) ameliorates brain amyloid pathology and cognitive impairment in a mouse model of AD (J Alzheimers Dis, 64, 563-585, 2018). This previous study implies the cell therapeutic strategy against AD. However, it remains unknown how transplanted bone marrow-derived cells interact with residential brain cells including microglia. In this study, we found that the conditioned medium from mouse bone marrow-derived cells increased Aβ phagocytosis by primary-cultured mouse microglia. We therefore analyzed the cytokines in the conditioned medium by enzyme-linked immunosorbent assay-based cytokine array and found that transforming growth factor-β1 (TGF-β1) was exclusively and richly present in the conditioned medium. In fact, bone marrow-derived cells and microglia expressed TGF-β1 mRNA and TGF-β receptor type 1 (TGF-βR1) protein, respectively. We finally treated microglia with recombinant mouse TGF-β1 protein and confirmed the promotion of Aβ phagocytosis and phosphorylation of Smad2 and Smad3 in microglia. The promotion of microglial Aβ phagocytosis with recombinant TGF-β1 protein was suppressed by TGF-βR1 inhibitor, SB525334. Thus, it is suggested that bone marrow-derived cells secrete high levels of TGF-β1, which promote microglial Aβ phagocytosis through TGF-βR1. Together with our previous report, bone marrow-derived cells may ameliorate brain amyloid pathology both by the Aβ phagocytosis by themselves and the secretion of TGF-β1 to promote Aβ phagocytosis by residential microglia. Hence, transplantation of bone marrow-derived cells stimulated with CSF-1 represents a promising strategy for cell therapy against AD.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-374 アルツハイマー病患者脳とAPP-KIマウス由来ミクログリアにおける遺伝子発現解析 Akira Sobue（祖父江顕）¹，Okiru Komine（小峯起）¹，Fumito Endo（遠藤史人）¹，Shigeo Murayama（村山繁雄）²，Takashi Saito（斉藤貴志）³，Takaomi Saido（西道隆臣）³，Koji Yamanaka（山中宏二）¹ ¹名古屋大環境医　病態神経科学 ²東京都健康長寿医療センター ³理研CBS 神経老化制御 Alzheimer's disease (AD) is the most common form of dementia, characterized by accumulation of amyloid β (Aβ) and phosphorylated Tau. On the other hand, glial cells such as astrocytes and microglia migrate and recognize Aβ deposits in the brains of patients with AD and animal models of the disorder. Especially, microglia, the resident phagocytes of the central nervous system, play a critical role in the clearance of Aβ. Activated microglia can be classified as M1 (pro-inflammatory) and M2 (anti-inflammatory), based on the molecular profiles of released cytokines. Recent research results suggest that neuroinflammation mediated by those glial cells is considered to play an important role in the progression of AD. However, the extent to which these events contribute to the Aβ pathologies remains unclear. In this study, we confirmed the changes in microglia in App^{NL-G-F/NL-G-F} knock-in (APP-KI) mice, which reproduce typical Aβ pathology in an age-dependent manner. The APP-KI mice express mutant APP at the level similar to an endogenous gene with appropriate cell-type and temporal specificity, therefore they avoid potential artifacts introduced by APP overexpression. To better understand the molecular mechanisms of Aβ pathologies and neuroinflammations, we performed next-generation sequence using RNA isolated from AD patients' precuneus, which is selectively vulnerable to early amyloid deposition, and isolated-microglia from 4, 8 and 12-month-old APP-KI mice by using magnetic-activated cell sorting. Some genes were commonly altered in microglia of APP-KI mice and AD patients' precuneus. These results suggest a strong correlation between Aβ pathology and microglial activities, and controlling microglia activation may lead to an effective treatment for AD.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-375 上肢運動機能を用いた認知症のスクリーニング手法 Atsuko Miyazaki（宮崎敦子）¹，Takashi Okuyama（奥山卓）²，Hayato Mori（森隼人）¹，Kazuhisa Sato（佐藤和久）³，Yuichi Miura（三浦祐一）⁴，Masahiko Ichiki（市来真彦）⁵，Shinichiro Nakamura（中村振一郎）¹，Rui Nouchi（野内類）^1,6 ¹理研　科技ハブ連携本部 ²神戸大保健リハビリ (非常勤講師） ³株式会社ケア２１ ⁴社会福祉法人天佑 ⁵東京医大精神科 ⁶東北大加齢研認知健康科学研究 We propose a method to screen the degree of dementia with the z-axis height of arm movement during drum playing. Currently, dementia is screened by neuropsychological test. However, often some patients refuse to take the test when they realize it is a dementia test from the questions asked. In addition, many questions cannot be scored for participants with visual and hearing impairments, and participants who do not have dementia may obtain insufficient scores. There is another issue to be considered in the screening of dementia. Previous studies report that hand motor function, hand grip strength in particular, is an important predicator of cognitive dysfunction (Clouston et al., 2013, Fritz et al., 2017, Taekema et al., 2010, Alencar et al., 2012, Jang & Kim, 2015). However, patients with dementia are unable to understand and follow instructions due to apraxia, thus disabling the proper measurement of hand grip strength. Several kinds of apraxia are seen from early stages in patients with various types of dementia (Chandra, et al.,2015). Drum playing is a skilled exercise which requires rhythm reaction movement, and it can be performed by patients with apraxia caused by dementia. We conducted a drum playing program at an elderly nursing home and examined the relationship between upper limb function of the dominant hand and cognitive functions. The height of the participant's arm while beating the drum with a mallet (drum stick) was measured using a 6-axis gyro sensor attached to the participant's wrist. The participants were asked to continue playing the drum for 20 minutes. The average values of the z-axis per second were calculated, and the mean score was 14.73 degree (SD=10.32). Multiple linear regression was also calculated to predict the total neuropsychological score (MMSE and FAB) based on the z-axis per second, sex, and age. A significant regression equation was found (F (3,12) = 4.490, p=0.025, R=0.727). The more severe the dementia, the lower was the mallet raised. In addition, grip strength and the movement range of the flexion of the shoulder joint were measured, and the results showed a strong relationship between these values and dementia score. Based on these findings, we propose the measurement of upper limb movements as a new inexpensive method for evaluating the cognitive function of patients with dementia including those with severe symptoms while treating them humanely and with dignity.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-376 AAVベクターの血清型およびプロモーターによる海馬遺伝子導入効率比較 Yoshihide Sehara（瀬原吉英）¹，Kuniko Shimazaki（島崎久仁子）²，Masashi Urabe（卜部匡司）¹，Kensuke Kawai（川合謙介）²，Hiroaki Mizukami（水上浩明）¹ ¹自治医大分子病態治療研究セ遺伝子治療研究部 ²自治医大医脳外 [Objective] Adeno-associated virus (AAV) vector has now become one of the most useful tools for gene transduction especially in the field of neuroscience because of its safety and efficiency. In this study, we used two different viral capsids of AAV including AAV5 and AAVrh10 with the three different promoters including cytomegalovirus (CMV), chicken β-actin promoter with CMV immediate-early enhancer (CAG), and synapsinI (SynI) to compare the transduction efficiency in different capsids and promoters. [Methods] We stereotaxically injected 1×10¹⁰ vg of AAV5 or AAVrh10 targeting the right side of hippocampus of 4-week-old male gerbils, carrying green fluorescent protein (GFP) driven by one of the above-mentioned promoters. Three weeks later, the gerbils were sacrificed and the brains were cryosected to quantify the areas of GFP signals in the hippocampus (n = 3, each). [Results] The widest GFP signal was observed in the rh10-CMV and rh10-CAG groups (rh10-CMV: 7.1 ± 1.1 mm², rh10-CAG: 6.0 ± 0.5 mm², p < 0.001, for both, compared with the control). The second widest expression was observed in the rh10-SynI and 5-CMV groups (rh10-SynI: 4.2 ± 0.2 mm², 5-CMV 4.2 ± 0.2 mm², p < 0.05, for both, compared with the control). The smallest expression was observed in the 5-SynI group (0.2 ± 0.2 mm², not significant, compared with the control). [Conclusion] Generally, most of the combinations of viral capsids and promoters will be appropriate for AAV-mediated gene transfer. However, we should note that some combinations can fail in transducing our target gene.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-377 Investigations of the early cognitive, behavioral and physiological effects of regular ingestion of natto in the rat Eric P. Wiertelak（Wiertelak Eric P.），Andrew Smith（Smith Andrew），Abigail Methrin（Methrin Abigail） Macalester College, Saint Paul, U.S.A. Natto, a part of the diet in Asia and Japan for many centuries, contributes protein, carbohydrates and fats and many important nutrients. The fermentation that produces natto also produces probiotics supporting healthy microbiomes in humans. The fermentation is dependent on Bacillus subtilis natto, which produces nattokinase, an extracellular enzyme that has both thrombolytic and pro-fibrinolysis effects, of interest in treating cardiovascular disease. Nattokinase is marketed as a nutritional supplement argued to improve circulation. In 2009, Hsu et al (J. Agric. Food Chem. 2009, 57, 503-508) exposed A-beta40, insulin, and prion peptide fibrils, each associated with significant disease processes, to nattokinase and observed degradation of each in vitro. These observations led us to begin a lifespan study of the impact of ingestion of natto on key cognitive, behavioral and physiological aspects of aging in rat. At the conclusion of all phases (3) of the study, all animals' brains will be histologically examined for the prevalence of beta-amyloid plaque. We present here results of phase 1 (early) studies in this program of research. Weanling male and female rat pups were divided into two groups, one provided natto twice weekly across their lifespan, and the other receiving only the standard laboratory rat chow all animals received. In study 1, all rats were assessed for navigational learning in the Morris water maze. Briefly, animals were given ten trials following pretraining to navigate to a hidden platform. Results indicated that young male and female rats ingesting natto regularly were not developmentally different from untreated rats as latencies to find the platform were not significantly different between groups. In study 2, all rats were assessed for anxiety-related behavior using the hole-board test. Briefly, animals were given four trials in the apparatus and assessed for head pokes into a 16-hole array in the floor, and for grooming and rearing behaviors. Results indicated that young male and female rats ingesting natto regularly do not exhibit elevated levels of anxiety-related behaviors compared to controls. Together, phase 1 results support the contention that regular ingestion of natto does not significantly affect juvenile rat learning and behaviors, and further, that any potential between-group differences in phase 2 and phase 3 of the study relate to ingestion of natto across the lifespan.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-378 ゲノム編集技術を用いたアルツハイマー病遺伝子変異のマウス内在性ゲノムへの導入 Kaori Sato（佐藤香織）^1,2，Kenichi Nagata（永田健一）^1,3，Hiroki Sasaguri（笹栗弘貴）¹，Toshio Ohshima（大島登志男）²，Takaomi C. Saido（西道隆臣）¹ ¹理化学研究所脳神経科学研究センター神経老化制御研究チーム ²早稲田大学大学院先進理工学研究科生命医科学専攻分子脳神経科学研究室 ³大阪大学　医学系研究科認知症プレシジョン医療開発学寄付講座 Alzheimer's disease (AD) is the most common cause of dementia in the world. Some AD cases with inherited form of the disease carry mutations in the presenilin proteins (PSEN1 and PSEN2) or in the amyloid precursor protein. PSEN1 is the most frequent causal gene in inherited form of AD, in which more than 250 mutations are identified. However, the functional consequences of each mutation still remain uncovered in vivo. Here we introduced deletion of exon 9 (delta E9) mutation into the mouse Psen1 gene utilizing CRISPR/Cas9 technology in order to explore the physiological functions of exon 9 deficiency in Psen1 protein in vivo. We first designed two sgRNAs on intron 8 and intron 9, respectively to delete 885 bp including exon 9. Both sgRNAs along with SaCas9 mRNA were injected into the cytoplasm of mouse zygotes via microinjection. CRISPR/Cas-mediated genome editing resulted in the efficient generation of the 22 mutant mice (33.8%) which harbored exon 9 deletion of 65 neonates in F0 generation. Unexpectedly, one mouse (1.5%) showed complete deletion of exon 9 with high level of mosaicism (90.1 %) for additional exon 8 deletion. Deletion of exon 8 and exon 9 in Psen1 mRNA of this mouse was also confirmed by RT-PCR. Although it has been reported that some of Psen1 knock-in mice with point mutations exhibited embryonic lethality in a homozygous state, the delta E8-9 mouse was viable and displayed no developmental defects except for tail deformity. These results indicate that CRISPR/Cas system allow the efficient generation of mutant animal models with disease-associated mutations including large deletion, and is highly useful to elucidate functional consequences of the mutations in vivo.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-379 ヒューマニン誘導体による野生型マウス海馬領域のアセチルコリン量の変化 Natsumi Ikegawa（池川夏実），Minetaka Murasaki（村上峰宇），Takako Niikura（新倉貴子）上智大理工情報理工 Humanin(HN) is an endogenous 24-residue peptide, which was first identified as a neuroprotective factor against Alzheimer's disease. HN is secreted from cells, and acts on two types of receptors, G protein-coupled formyl peptide receptor-like (FPRL)-1/2 and an trimeric receptor complex composed of CNTFR, WSX1, and gp130. HN's neuroprotective function is mainly mediated by the trimeric receptor complex. S14G-HN, HN with a substitution of glycine for serine 14 , is a highly potent HN derivative and suppressed cognitive impairment in Alzheimer's disease model mice. S14G-HN also ameliorates memory impairment caused by muscarinic receptor antagonists and GABA receptor activator in normal mice. However, it is still unclear how HN modulates cognitive activities. To understand the mechanism of S14G-HN-induced improvement in cognitive function, we examined the acetylcholine level in the normal mouse brain under the free moving condition. We placed a microdialysis probe in hippocampal region and obtained small molecule sample from interstitial liquid every 30 minutes after intraperitoneal injection of S14G-HN or vehicle. The amount of acetylcholine in the sample was measured by high performance liquid chromatography. Vehicle injection did not cause significant change in the acetylcholine level. On the other hand, S14G-HN increased the level of acetylcholine after 30 min of administration and the level was maintained for 180 min. No significant difference was found in the walking distance between two groups, indicating that the effect of S14G-HN on the acetylcholine level was independent from the locomotor activity. These results suggest that the increase in the amount of acetylcholine in the hippocampal area contributes to the function of HN on cognitive improvement. Using the same method, we also observed an increasing trend of catecholamine levels by S14G-HN treatment in the mouse hippocampal region, suggesting that the effect of S14G-HN is not specific to acetylcholine.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-380 Pin1遺伝子欠損マウスは空間認知機能の異常があり前頭側頭葉が減少を示す。 Hirokazu Ohtaki（大滝博和）¹，Keisuke Kiriyama（桐山恵介）¹，Kenjiro Ono（小野賢二郎）²，Katsuhiko Takahashi（高橋勝彦）³，Takafumi Uchida（内田隆史）⁴，Kazuho Honda（本田一穂）¹ ¹昭和大医顕微解剖 ²昭和大医神経内科 ³星薬大生化 ⁴東北大院農分子酵素 Pin1 is a ubiquitous peptidyl-prolyl cis/trans isomerase (PPIase) and has been shown to be necessary for cell growth and apoptosis. While pin1 deficient (-/-) mice were suggested to contribute to Alzheimer's disease, the relation between behavior and pin1 deficient has not been clarified in detail. Pin1 homozygous gene deficient (-/-) and the wild-type mice were obtained as a littermate by breeding from the heterozygous gene deficient (+/-) mice and housed under standard condition with ad libitum access to standard food and water. Battery of behavioral tests were performed at 12 month old. The tests were involved in Y-maze, open-field spontaneously motor activity, new objective recognition and Morris water maze to examine the recognition, learning and memory function. Another group of animals was examined light/dark transition and resident-intruder test to determine the anxiety-like behavior and social stress behavior at 13 month. The animals were also measured T2-weight MRI (ICON 1T, Burker) coronal images in the brain every 3 month for 15 month, and semi-quantified the area of coronal plate by manually tracing with Osirix software. No significantly differences were recognized in the body weight, Y-maze score, and open-field spontaneously motor activity between Pin1 (-/-) mice and the wild-type (+/+) mice. However, the Pin1 (-/-) mice showed significant difference with wild-type in the new objective recognition and Morris water maze. The Pin1 (-/-) mice less approached to new object at day 2 in the objective recognition test. The mice also took significantly long time to reach the platform at day3 in the Morris water maze. Although there were no differences to the total sojourn time in light/dark room for 3 days (6-trials), the Pin1 (-/-) mice significantly frequent passing number of gate after 3rd trial. Moreover, the Pin1 (-/-) mice were less aggressiveness against the intruder to compare with the wild type mice. The brain volume in the Pin1 (-/-) KO mice was significantly smaller than the wild-type mice, especially in the frontotemporal plane. These results suggest that Pin1 (-/-) mice impaired spatial cognitive function with frontotemporal atrophy.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-381 介入前の前頭葉機能・容積は高齢者に対する体操の効果を予測する：御浜-紀宝スキャンプロジェクト Kenichi Tabei（田部井賢一）^1,2,7，Masayuki Satoh（佐藤正之）¹，Jun-ichi Ogawa（小川純一）³，Tomoko Tokita（時田智子）⁴，Noriko Nakaguchi（中口紀子）⁵，Koji Nakao（仲尾貢二）^6,9，Hirotaka Kida（木田博隆）^1,8，Hidekazu Tomimoto（冨本秀和）² ¹三重大院医認知症医療学 ²三重大院医神経病態内科学 ³ヤマハ音楽振興会 ⁴御浜町役場健康福祉課 ⁵紀宝町役場福祉課 ⁶紀南病院脳神経外科 ⁷(現)産業技術大学院大学産業技術研究科 ⁸(現)みんなのライフサポートクリニック大網 ⁹(現)塩川病院脳神経外科 Background: Non-pharmacological interventions, especially physical exercise has positive effects on cognitive function in healthy elderly people. In older adults, physical exercise with music induces greater positive effects on visuospatial function (Satoh et al., 2014) and leads to more extensive neuroanatomical changes (Tabei et al., 2017) than exercise alone. However, the neuropsychological factors influencing interventions of physical exercise, as well as the neural basis of its efficacy, remain unknown. We aimed to determine whether neuropsychological deficits and brain atrophy could predict the efficacy of physical exercise interventions. Methods: One-hundred twelve participants of healthy elderly people were monitored for 1 year; 51 underwent an intervention involving physical exercise with music, and 61 performed the physical exercise without music. Participants with an increased MMSE score of 2 points or more were included in the improvement subgroup, while the remaining participants were included in the no-improvement subgroup. Results: The no-improvement subgroup performed worse than the improvement subgroup on the word fluency and visuospatial test at baseline. In the no-improvement subgroup, voxel-based morphometric analysis at baseline revealed more extensive gray matter decrease in the left inferior frontal gyrus and medial frontal gyrus. Conclusions: Our findings suggest that some characteristics of pre-intervention cognitive dysfunction and regional brain atrophy may aid clinicians in determining the physical exercise efficacy in healthy elderly people.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-382 Impaired spatial representation in hippocampal-entorhinal circuit of Knock-in Alzheimer Model Heechul Jun（Jun Heechul）¹，Shogo Soma（Soma Shogo）¹，Ananya Dasgupta（Dasgupta Ananya）¹，Kei Igarashi（Igarashi Kei）^1,2,3 ¹Department of Anatomy and Neurobiology, University of California Irvine ²Center for Learning and Neurobiology, University of California Irvine ³Institute for Memory Impairments and Neurological Disorders, University of California Irvine Alzheimer's disease (AD) is a progressive neurological disorder that debilitates our mind and memory. AD patients show impairments in multiple dimensions of memory including spatial memory and navigation, which causes wandering behavioral symptoms. Currently, AD affects 50 million people worldwide and it is expected to reach 131.5 million in 2050. . Despite significant advances made in uncovering molecular and cellular mechanisms behind AD pathology, we still lack the cure to properly treat the disease. A limited number of studies have been performed so far to investigate the changes that occur in the brain circuits of AD. By understanding what type of neuronal activities are lost and demonstrating the relationship between the dysfunctional neural network and cognitive deficits, we may be able to develop novel therapies targeted to reactivate these activities in AD patients. Our lab has been investigating the impairment of brain activity in the AD mouse model using electrophysiological recording methods. We are focusing on the circuit comprised of the entorhinal cortex (EC) and the hippocampus. Neurons in the EC receive input from multiple cortical regions and send projections to the hippocampus. CA1 cells in the hippocampus send their axons back to the EC, thus, forming the EC-hippocampal loop circuit. This connection between the two brain regions is involved in memory formation, and retrieval and damage to this circuit results in memory impairment. Histological and imaging studies in AD patients and animal models have shown that the EC is a primary site of atrophy and activity loss in the early phases of AD. However, it is still unclear what type of activity is lost in the EC of AD patients, or even in AD mouse models. Using a novel amyloid precursor protein (APP) knock-in mouse model (Saito et al., Nat Neurosci 2014), we found that grid cells, a cell type harboring spatial memory-related activity in the medial entorhinal cortex (MEC), are impaired in APP knock-in mice. Place cells, another spatially-tuned neurons in the hippocampus, are also impaired. Our results privide the first evidence to demonstrate that the entorhinal-hippocampal spatial representation is impaired in the AD mouse model. Circuit mechanisms of how the entorhinal-hippocampal spatial representation is impaired will be discussed.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-383 コーヒー豆の焙煎成分によるBACE1発現抑制効果の解析 Hiroomi Tamura（田村悦臣）¹，Kazuya Fukuyama（福山和也）¹，Yosuke Nakazawa（中澤洋介）¹，Megumi Tago（多胡めぐみ）¹，Toshiharu Suzuki（鈴木利治）² ¹慶應義塾大学 ²北海道大学 Scope: Epidemiological studies have shown that coffee consumption may be associated with a lower risk of developing several neurological disorders, including Alzheimer's disease (AD) and Parkinson's disease. Caffeine is thought as a prominent candidate component underlying the preventive effects of coffee; however, the contribution of other constituents is unclear. To clarify this issue, we analyzed the effect of roasting coffee beans on β-secretase (BACE1) expression in human neuroblastoma SH-SY5Y cells and mice. Methods and results: Coffee (2%) reduced Aβ accumulation in culture medium to 80% of control levels after 24 h. Accordingly, BACE1 expression was decreased to 70% of control levels at 12 h. Experiments using cycloheximide and MG-132, a proteasome inhibitor, revealed that coffee enhanced BACE1 degradation through activation of proteasomal activity. Furthermore, coffee activates cAMP-dependent protein kinase, and consequently, phosphorylation of a serine residue of 19S proteasome subunit PSMD11. Pyrocatechol, a product produced from chlorogenic acid during roasting, also reduced BACE1 expression by activation of proteasomal activity. Furthermore, pyrocatechol reduced Aβ production in SH-SY5Y cells. In addition, 60%(v/v) coffee administration to normal mice for 7 weeks significantly reduced hippocampal BACE1 protein expression. Pyrocatechol administration also exhibited the same effect on hippocampal BACE1 expression in mice. Conclusion: Coffee components emerged during the roasting process induce proteasome activity, thereby enhancing BACE1 degradation in cells, leading to repression of Aβ production. Our data suggest that the roasting process may be crucial for the protective effects of coffee consumption in AD.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-384 Tau Phosphorylation at AT8 Pathological Site during Brain Development Tuerde Dilina（Dilina Tuerde）^1,2，Kotaro Furusawa（古澤孝太郎）²，Toshiyuki Takasugi（高杉俊之）²，Taeko Kimura（木村妙子）²，Shinsuke Ishigaki（石垣診祐）¹，Kanae Ando（安藤香奈絵）²，Shin-ichi Hisanaga（久永眞市）²，Gen Sobue（祖父江元）¹ ¹名古屋大学大学院医学系研究科 ²首都大学東京大学院理工学研究科 Tau is a microtubule (MT) associated protein (MAP), stabilizing MTs in axon of neurons and then supporting neuronal network in healthy brains. In contrast, in Alzheimer`s disease (AD) brains, tau is abnormally phosphorylated more than 40 sites, and aggregated into neurofibrillary tangles (NFTs) in neurons undergoing degeneration. Among many pathological phosphorylation sites, phosphorylation at the AT8 site has been most frequently used for diagnosis of AD. The AT8 site comprises two phosphorylation sites at Ser202 and Thr205, although some literatures demonstrate it as a consequence of triple phosphorylation at Ser199, Ser202 and Thr205. However, it is not completely understood yet how the AT8 reactivity is generated in AD brain and how it contributes to AD development. Tau hyperphosphorylation at specific residues occurs not only in AD brains but also in fetal and early postnatal brains. We found that AT8 is one of highly phosphorylated sites in fetal and neonatal stages of mouse, and it suddenly disappear during 2 to 3 weeks after birth when neuronal circuit is established. Moreover, hypothyroidism delayed tau dephosphorylation at the AT8 site specifically about 3 days. These results indicate direct relationship between neuronal development and AT8 site phosphorylation. We think it important to understand the role of AT8 phosphorylation in neuronal maturation at a molecular level. Here, we examined the effect of overexpression of wild-type human tau or AT8 site mutants, both unphosphorylatable Ala and phosphomimetic Glu in developing neurons. Our study would shed light on a physiological role of the AT8 phosphorylation in tau and also provide useful information about a therapeutic target for the treatment of AD.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-385 アルツハイマー病モデルであるAppノックインマウスにおける認知機能障害はアミロイドβ斑の形成とグリオーシスに起因する Yasufumi Sakakibara（榊原泰史）¹，Michiko Sekiya（関谷倫子）¹，Takashi Saito（斉藤貴志）²，Takaomi C Saido（西道隆臣）²，Koichi M Iijima（飯島浩一）¹ ¹国立長寿医療研究センター　アルツハイマー病研究部 ²理研CBS 神経老化制御 Knock-in (KI) mouse models of Alzheimer's disease (AD) that endogenously overproduce amyloid-β (Aβ) without non-physiological overexpression of amyloid precursor protein (APP) provide important insights into the pathogenic mechanisms of AD. Previously, we reported that App^NL-G-F mice, which harbor three familial AD mutations (Swedish, Beyreuther/Iberian, and Arctic) exhibited emotional alterations before the onset of definitive cognitive deficits. To determine whether these mice exhibit deficits in learning and memory at more advanced ages, we compared the Morris water maze performance of App^NL-G-F and App^NL mice, which harbor only the Swedish mutation, with that of wild-type (WT) C57BL/6J mice at the age of 24 months. To correlate cognitive deficits and neuroinflammation, we also examined Aβ plaque formation and reactive gliosis in these mice. In the Morris water maze, a spatial task, 24-month-old App^{NL-G-F/NL-G-F} mice exhibited significantly poorer spatial learning than WT mice during the hidden training sessions, but similarly to WT mice during the visible training sessions. Not surprisingly, App^{NL-G-F/NL-G-F} mice also exhibited spatial memory deficits both 1 and 7 day after the last training session. By contrast, 24-month-old App^NL/NL mice had intact spatial learning and memory relative to WT mice. Immunohistochemical analyses revealed that 24-month-old App^{NL-G-F/NL-G-F} mice developed massive Aβ plaques and reactive gliosis (microgliosis and astrocytosis) throughout the brain, including the cortex and hippocampus. By contrast, we observed no detectable brain pathology in App^NL/NL mice despite overproduction of human Aβ40 and Aβ42 in their brains. Our results indicate that Aβ plaque formation, followed by sustained neuroinflammation, is necessary for the induction of definitive cognitive deficits in App-KI mouse models of AD. Our data also indicate that introduction of the Swedish mutation alone in endogenous APP is not sufficient to produce either AD-related brain pathology or cognitive deficits in mice.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-386 オートファジーの誘導はエクソソームの分泌量の低下を介して細胞内Aβを蓄積させる Shingo Koinuma（鯉沼真吾），Nobuyuki Kimura（木村展之）国立長寿医療セ研究所アルツハイマー Alzheimer's disease (AD) is one of the representative neurodegenerative diseases causing dementia. Since characteristic lesions called senile plaques composed by β-amyloid protein (Aβ) are observed before onset, abnormal accumulation of Aβ would be the key factor for AD pathogenesis. Especially, intraneuronal accumulation of Aβ precedes extracellular Aβ deposition, suggesting that intracellular Aβ would be much important. Several studies showed that autophagy mediates Aβ degradation via autophagic clearance by lysosome, and autophagy-related genes were downregulated in AD patient brains. These findings suggest that autophagy induction would be beneficial to prevent the progression of Aβ pathology. On the other hand, we have previously shown that aging induces endocytic disturbance in brain and that endocytic disturbance disrupts autophagic clearance. Hence, in the present study, we aim to clarify whether autophagy induction can ameliorate Aβ accumulation even when endocytic disturbance occurs. In this study, we treated mouse neuroblastoma Neuro2a cells with chloroquine, which can strongly induce endocytic disturbance by blocking endosomal/lysosomal trafficking pathway. We also used rapamycin to induce autophagy in Neuro2a cells. In Neuro2a cells treated with chloroquine, the amounts of intracellular Aβ were increased. Noteworthy, rapamycin treatment significantly enhanced intracellular accumulation of Aβ as compared to chloroquine treatment alone. In contrast, the amounts of extracellular Aβ strongly decreased with combined treatment of chloroquine and rapamycin. Since Aβ is released extracellularly via exosome, we analyzed exosome secretion in Neuro2a cells with chloroquine and/or rapamycin treatments. Evidently, autophagy induction decreased exosome secretion, and siRNA-induced suppression of autophagy rescued intracellular accumulation of Aβ by upregulation of exosome secretion. Taken together, these findings suggest that autophagy induction would be not beneficial for aged brain, in which age-related endocytic disturbance occurs, and the downregulation of autophagy may rescue intracellular accumulation of Aβ via exosome secretion.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-388 ジヒドロパーオキシドの変異copper-zinc superoxide dismutase 1細胞内凝集体に対する神経保護効果 Tomoyuki Ueda（上田智之）¹，Yuta Asaka（浅香雄太）¹，Masatoshi Inden（位田雅俊）¹，Hisaka Kurita（栗田尚佳）¹，Wakako Tanaka（田中稚子）²，eiji yamaguchi（山口英士）²，akichika itoh（伊藤彰近）²，Isao Hozumi（保住功）¹ ¹岐阜薬大・薬・薬物治療 ²岐阜薬大・薬・合成薬品製造学 Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive muscle weakness, paralysis, and death. Approximately 80% of ALS patients die within 3-5 years after onset of these symptoms. Almost cases are classified as sporadic ALS, whereas about 10% are familial ALS. Although its neuropathology is well understood, currently, effective treatments are unavailable. The mechanism of ALS involves the aggregation and accumulation of several mutant proteins, including mutant copper-zinc superoxide dismutase 1 (SOD1), TAR DNA binding protein 43 kDa (TDP-43) and fused in sarcoma (FUS) proteins. Previous reports have shown that excessive oxidative stress, associated with mitochondrial dysfunction, disturbing degradation system and mutant protein accumulation, contributes to ALS pathology. Edaravone is free radical scavenger and currently approved ALS drug. The present study focuses on the promotion of SOD1 misfolding and the accumulation of aggregates by oxidative stress. Having recently synthesized novel organic gem-dihydroperoxides (DHPs) with high anti-oxidant activity, we now examined whether DHPs reduce the mutant SOD1-induced intracellular aggregates and neurotoxicity involved in oxidative stress. We found that, among DHPs, 12AC2O significantly inhibited mutant SOD1-induced cell death and reduced the intracellular mutant SOD1 aggregates. Moreover, immunofluorescence staining with redox-sensitive dyes showed that 12AC2O reduced the excessive level of intracellular mutant SOD1-induced reactive oxygen species (ROS). Additionally, ESR analysis showed that 12AC2O exerts a direct scavenging effect against the hydroxyl radical and the superoxide anion. These results suggest that 12AC2O may be useful for the prevention of or for the slowing of the progression of ALS, by mitigating mutant SOD1-mediated toxicity.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-389 蛋白分解異常による新規筋萎縮性側索硬化症の発症機構 Yasuaki Watanabe（渡辺靖章）^1,2，Tadashi Nakagawa（中川直）¹，Naoki Suzuki（鈴木直輝）²，Hitoshi Warita（割田仁）²，Keiko Nakayama（中山啓子）¹，Masashi Aoki（青木正志）² ¹東北大学大学院医学系研究科附属創生応用医学研究センター細胞増殖制御分野 ²東北大学大学院医学系研究科神経内科学分野 Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of the human motor system characterized by generalized progressive muscle weakness and atrophy. About 5-10% of ALS is familial, and several novel causal or risk genes have been recently discovered. Two genes, NEK1 and C21orf2 were reported as novel ALS-associated genes in 2016. NEK1 is a serine/threonine kinase acting in multiple biological processes, whereas function of C21ORF2 has not been described. Intriguingly, NEK1 and C21orf2 interact with each other, and both of them are demonstrated to be mutated in some ciliopathies, suggesting they play a crucial role cooperatively in ALS patients. Our purpose was deciphering the interactive functions between C21ORF2 and NEK1, and the effect on them caused by V58L mutation of C21orf2 which is observed in ALS patients. We investigated protein stability of C21orf2 in HEK293T cells. We found C21orf2 was a novel substrate of E3 ligase SCF (Skp1-Cul1-FBXO3) complex in which FBXO3 acts as an adaptor protein. Poly-ubiquitylated C21orf2 was degraded by proteasome. NEK1 phosphorylated C21orf2, leaded to increase the C21orf2 protein by inhibiting its ubiquitylation. V58L mutant of C21orf2 is not ubiquitylated and was stable compared with wild type. On the other hand, we noticed C21orf2 stabilized NEK1 protein vice versa. In C21orf2-disrupted mIMCD3 cells, we observed reduction of the amount of endogenous NEK1 protein. We also confirmed that NEK1 protein level increases after C21ORF2 overexpression in SH-SY5Y cells. We concluded that NEK1 regulates its protein amount by a positive autoregulation system in which NEK1 and C21orf2 stabilize each other. The increase of V58L-C21orf2 protein by non-ubiquitylation results in excessive stabilization of NEK1. In addition, we observed that the C21orf2 knocked-in mutation by CRISPR/Cas9 system induced not only increase of C21orf2 protein also increase of NEK1. The motor neurons which were induced to differentiate from the C21orf2 mutated ES cells tend to extend shorter neurites compared with wild-type. In ALS patients who suffer the C21orf2 rare coding variants, C21orf2 and NEK1 overexpression is suggested to be one of the keys of ALS pathogenesis.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-390 ALS患者の末梢血中の神経由来細胞外小胞miRNAの解析 Masataka Katsu（嘉津政隆）¹，Yuka Hama（浜結香）²，Jun Utsumi（内海潤）²，Ken Takashina（高科謙）¹，Hiroshi Yasumatsu（安松浩）¹，Fumiaki Mori（森文秋）³，Koichi Wakabayashi（若林孝一）³，Mikio Shoji（東海林幹夫）⁴，Hidenao Sasaki（佐々木秀直）² ¹田辺三菱製薬神経科学創薬ユニット ²北海道大院神経内科 ³弘前大院医脳神経病理 ⁴弘前大医院神経内科 Circulating microRNAs (miRNAs) in peripheral blood have been extensively investigated as biomarkers for early diagnosis and monitoring of disease progression. However, their cellular origin as well as their link to the pathophysiology of neurodegenerative disease, remains mostly unknown. Accumulating evidence suggests that extracellular vesicles (EVs) released by neurons are detected in the blood stream and intravesicular expression levels of pathogenic proteins, synaptic proteins and lysosomal proteins are altered in neurological disorder conditions. In the present study, we isolated neuron-derived EVs in plasma by immunoaffinity purification and comprehensively analyzed intravesicular miRNA expression profiles using a microarray technology. To investigate the relationship of miRNA expression between plasma neuron-derived EVs and brain tissue, we compared miRNA expression profiles of plasma neuron-derived EVs with those of formalin-fixed paraffin-embedded (FFPE) samples from the motor cortex. In plasma neuron-derived EVs, a total of 30 miRNAs were found to be differentially regulated in amyotrophic lateral sclerosis (ALS) patients relative to healthy controls. Moreover, of a total of 322 miRNAs detected in plasma neuron-derived EVs, 299 (92.9%) miRNAs were found to be expressed in brain FFPE samples. Gene ontology analysis of miRNA target genes revealed that biological processes implicated in both differentially up-regulated and down-regulated miRNAs were both involved in synaptic vesicle-related pathways. Especially, 4 miRNAs in plasma neuron-derived EVs seemed to be regulated in the similar manner as those in brain FFPE samples from ALS patients. The target genes for the 4 miRNAs were partly overlapped in STX1B (syntaxin 1B), RAB3B (RAB3B, member RAS oncogene family), and UNC13A (unc-13 homolog A) genes. The UNC13A gene has been reported to be associate with increased odds ratio of sporadic ALS in multiple genome-wide association studies. Taken together, our data suggest that miRNAs extracted from neuron-derived EVs in plasma reflect alterations of miRNA expression in brain as potential biomarkers of ALS.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-391 TARDBP のDNAメチル化はTDP-43の自己調節機構関連スプライシングに影響する Yuka Koike（小池佑佳）¹，Akihiro Sugai（須貝章弘）¹，Akio Yokoseki（横関明男）¹，Tomohiko Ishihara（石原智彦）¹，Norikazu Hara（原範和）²，Junko Ito（伊藤絢子）³，Takeshi Ikeuchi（池内健）²，Akiyoshi Kakita（柿田明美）³，Osamu Onodera（小野寺理）¹ ¹新潟大脳研神経内科 ²新潟大脳研遺伝子機能解析学分野 ³新潟大脳研病理学分野 Introduction: Amyotrophic lateral sclerosis (ALS) is characterized by the accumulation of TAR DNA binding protein of 43 kDa (TDP-43) in motor neurons. Increased expression of TARDBP mRNA may underlie the pathogenesis of ALS. However, the factors affecting the amount of TARDBP mRNA in ALS have not been elucidated. The expression level of TARDBP mRNA is auto-regulated via alternative splicing of its 3'UTR. DNA methylation has gained attention as a mechanism for regulating alternative splicing. We hypothesized that the methylation status of the TARDBP 3'UTR affects the autoregulation-relevant splicing. Here, we investigated whether the methylation state of the TARDBP 3'UTR affects alternative splicing in human cultured cells. Moreover, we investigated the DNA methylation status of TARDBP 3 'UTR in post-mortem human brain tissues. Methods: HEK293T cells were transfected with dCas9-TET1 guide RNA vector designed to specifically demethylate the fifteen CpG sites (No.1-15) of TARDBP 3'UTR. The transfected cells were analyzed using bisulfite amplicon sequencing and RT-PCR. Frozen human brain tissues (motor cortex, occipital cortex, and cerebellum) from eight patients without neurological diseases were analyzed by bisulfite amplicon sequencing and droplet digital PCR. Results: The fifteen CpG sites in TARDBP 3 'UTR were originally highly methylated. The dCas9-TET1 guide RNA vector remarkably demethylated six consecutive CpG sites (No.10-15) in TARDBP 3'UTR. The cells transfected this vector reduced alternative splicing and increased the expression levels of TARDBP mRNA. In human motor cortex, the percentage of methylation at these six CpG sites (No.10-15) tended to show a positive correlation with the efficiency of alternative splicing. DNA methylation status of these CpG sites in TARDBP 3'UTR was clearly different depending on the brain regions. The percentages of DNA methylation were higher in the cerebellum than in the motor cortex and in the occipital cortex. Moreover, only in the motor cortex, the DNA methylation percentages were inversely correlated with the age at autopsy. Discussion: We demonstrated that site-specific DNA methylation affects alternative splicing which regulates the expression level of TARDBP. DNA methylation status of TARDBP 3'UTR was different in cell types and aging. To clarify the pathological significance of DNA methylation in ALS, we should analyze the brain tissues from sporadic ALS patients.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-392 TDP-43の多量体化が、その細胞内局在の決定に重要である Kotaro Oiwa（大岩康太郎）^1,2，Seiji Watanabe（渡邊征爾）²，Akira Sobue（祖父江顕）²，Okiru Komine（小峯起）²，Masahisa Katsuno（勝野雅央）¹，Koji Yamanaka（山中宏二）² ¹名古屋大医神経内科 ²名古屋大環境医病態神経科学 Mislocalization of TAR DNA-binding protein 43 (TDP-43) is the pathological hallmark of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). TDP-43 is an RNA binding protein that predominantly localizes to nuclei. However, in ALS/FTLD patients, TDP-43 mislocalizes into cytoplasm and forms aggregates with aberrant phosphorylation. Various studies have pointed out that the mislocalization of TDP-43 is deeply involved in ALS/FTLD pathomechanism(s), but it is still unclear how TDP-43 escapes from nuclei to cytoplasm under the pathological condition in ALS/FTLD. Here, we investigated a role of TDP-43 oligomerization in its intracellular localization. TDP-43 has a classical nuclear localization signal (NLS), and localized to nuclei dependent on its NLS in mouse neuroblastoma Neuro2a (N2a) cells. Intriguingly, TDP-43 variant devoid of its NLS (TDP-dNLS) was also localized to nuclei in N2a cells stably expressing TDP-dNLS but not with overexpression of TDP-dNLS. This suggests that heterodimer formation of TDP-dNLS and endogenous TDP-43 facilitates nuclear translocation of TDP-dNLS. Consistent with this finding, we also found that TDP-43 variant lacking its extreme N-terminal domain (TDP-dN10), which is important for dimerization of TDP-43, did not completely affect cytoplasmic localization of TDP-dNLS. On the other hand, another dimerization-deficient TDP-43 variant, TDP-N6M, carrying 6 amino acid substitutions in its N-terminal domain (E14A/E17A/E21A/Q34A/R52A/R55A), showed prominent cytoplasmic distribution in N2a cells. Moreover, TDP-N6M had a higher propensity to form phosphorylated aggregates compared to the wild type or TDP-dNLS. These results suggest that TDP-43 monomers are prone to translocate from nuclei to cytoplasm and to form aberrantly phosphorylated aggregates. Our findings suggest that aberrant oligomerization of TDP-43 may play a key role in the pathogenesis of TDP-43-linked ALS/FTLD.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-393 TDP-43凝集における後期エンドソームの関わり Yuhei Takahashi（高橋裕平），Yohei Iguchi（井口洋平），Masahisa Katsuno（勝野雅央）名古屋大医神経内科 [Background] Cytoplasmic TDP-43 aggregate formation in neurons and glial cells is a common pathological feature of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) and thought to be a cause of the neurodegeneration. Endosome is a key machinery of membrane trafficking system in eukaryote cells and involved in autophagy-lysosome pathway. Although several endosome-related genes such as CHMP2B, Alsin and TMEM106B are known to be a disease causative gene or a genetic modifier for ALS/FTLD, the role of endosome in the formation of TDP-43 aggregates has been hardly investigated. [METHODS] Neuro2a cells were transfected with wild-type (WT), constitutively active (CA), or dominant negative (DN) form of Rab7 together with aggregate form of TDP-43 (mtTDP), in which nuclear localization signal and RNA-recognition motif are mutated, using Lipofectamine 3000. The transfected cells were cultured for 48 hours, and subjected to immunofluorescence (IF), or time lapse analyses (TL), Western blotting (WB) of sequential extractions using Triton X-100 and Sarkosyl. [Result] IF and TL showed that multiple cytoplasmic mtTDP inclusions fused well with each other and formed large aggregations in the cells with mtTDP and either WT or SA form of Rab7, but mtTDP inclusions rarely fused and sparsely distributed in the cells expressing DN form of Rab7. In addition, insoluble fractions of mtTDP were substantially decreased in the cells with DN Rab7, compared to the cells with either WT or SA Rab7. Live image with LysoTracker revealed that lysosomes did not integrate mtTDP inclusions. [Discussion] Rab7 is intracellular GTPase involved in endosome fusion, which is supposed to promote protein degradation in lysosome. DN form of Rab7, however, decreased the insoluble fraction of mtTDP. A possible reason for this discrepancy is that TDP-43 is not degraded by autophagy-lysosome pathway unless proteasome function is inhibited as shown by previous works and our current data. Further investigation is needed to elucidate these phenomena. [Conclusion] This work indicates that the late endosome function is involved in TDP-43 aggregate formation, and could underlie in the pathogenesis of ALS/FTLD.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-394 AAV投与を介したCRISPR/Cas9ゲノム編集によるDRPLA治療 Taisuke Kato（加藤泰介）¹，Yuka Koike（小池佑佳）²，Sachiko Hirokawa（廣川祥子）¹，Kenta Kobayashi（小林憲太）³，Shoji Tsuji（辻省次）⁴，Osamu Onodera（小野寺理）^1,2 ¹新潟大学脳研究所分子神経疾患資源解析学分野 ²新潟大学脳研究所神経内科学分野 ³生理学研究所ウィルスベクター開発室 ⁴東京大学医学部付属病院 Polyglutamine diseases are caused by the expansion of CAG repeats in the causative genes. The expanded polyglutamine stretch confers toxic functions leading to neurodegeneration. Dentatorubural-pallidoluysian atrophy (DRPLA) is caused by the expansion of CAG repeat in Atrophin-1 (ATN1) gene. The reduction of the expression of ATN1 can be a therapeutic strategy for DRPLA. We had confirmed that non-allele specific gene-silencing approach is adaptable for gene-therapy of DRPLA by producing mosaic transgenic mice harbouring human ATN1 gene with expanded CAG-repeat. As a next step, we developed AAV vector to edit human ATN1 gene in central nervous system. We produced AAV-serotype 9, DJ, DJ/8 and PHP.eB and compared each editing efficiency. AAV9, DJ and DJ/8 injected intracerebroventricularly at PND2 induced relatively high editing efficiency in the brain region around ventricle (e.g. cortex, hippocampus or striatum). In contrast, almost no editing was obtained in distal parts of ventricle when using these AAV serotype (e.g. thalamus, midbrain, cerebellum or medulla oblongata). PHP.eB injected intravenously at PND35 induced extensive editing throughout the brain. Now, using the PHP.eB serotype, we have evaluated the therapeutic effects of the genome editing-mediated gene-silencing by AAV administration after manifestation of disease-related phenotypes. Although currently available data are limited, we have obtained the data supporting efficacy of the AAV-treatment including extending survival rate, reduction of brain-weight loss and alleviation of ataxia phenotypes. These results indicate feasibility of the strategy with genome editing-mediated gene-silencing in vivo for therapy of DRPLA.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-395 Bmp4の選択的発現抑制により筋萎縮性側索硬化症モデルラットの疾患進行が抑制される Tomomi Shijo（四條友望），Hitoshi Warita（割田仁），Kensuke Ikeda（池田謙輔），Naoki Suzuki（鈴木直輝），Tetsuya Akiyama（秋山徹也），Hiroya Ono（小野洋也），Shio Mitsuzawa（光澤志緒），Ayumi Nishiyama（西山亜由美），Rumiko Izumi（井泉瑠美子），Masashi Aoki（青木正志）東北大学神経内科 Amyotrophic lateral sclerosis (ALS) is an adult-onset, intractable neurodegenerative syndrome with systemic loss of motor neurons accompanied by marked activation of astrocytes and microglia, which are considered to accelerate the neurodegeneration. We have previously reported that bone morphogenetic protein (BMP) 4, one of the extracellular molecules that promote astrocytic activation, and its receptor (BMP receptor 1A, BMPR1A) are up-regulated in reactive astrocytes of an ALS rodent model. Moreover, noggin, a physiological antagonist of BMPs, can attenuate both gliosis and neuroinflammation, and eventually extend survival of the ALS model. To prove that the BMP4 is an accelerator in the disease progression of ALS, we tried selective knockdown of Bmp4 by an antisense oligonucleotide (ASO) in vivo. Adult female transgenic rats overexpressing a familial ALS-linked His46Arg mutant human Cu/Zn superoxide dismutase gene (SOD1) at early-symptomatic stage were used as ALS model rats (n = 22), as was in our previous study. A Vivo-Morpholino ASO targeted Rattus norvegicus Bmp4 mRNA, negative control (NC) -ASO, or physiological saline (vehicle) were intrathecally administered for 3 days using osmotic pumps. Fourteen days later, the lumbar spinal cords were harvested and examined by real-time reverse transcription PCR, immunoblotting, and immunohistochemistry. Survival, body weights, and motor functions were also analyzed in the 3 groups. We confirmed that both BMP4 protein and Bmp4 mRNA expression were down-regulated in spinal cords of Bmp4-ASO administered group compared with those of other groups. Furthermore, the selective Bmp4 knockdown by itself significantly attenuated astrocytic activation, and extended survival of ALS model rats. We have revealed that among BMPs, the astrocytic BMP4 plays a key role in enhancing neuroinflammation at the site of neurodegeneration, which exacerbates the disease progression in this ALS rodent model. The BMP4 and its downstream signaling may become a novel therapeutic target in ALS.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-396 パーキンソン病の前駆期モデルにおいて、GBAのハプロ不全は脂質代謝変化を伴いアルファシヌクレイン病理を加速させる Masashi Ikuno（生野真嗣）¹，Hodaka Yamakado（山門穂高）¹，Hisako Akiyama（秋山央子）²，Laxmi Kumar Parajuli（Parajuli Kumar Laxmi）³，Katsutoshi Taguchi（田口勝敏）⁴，Junko Hara（原淳子）¹，Yusuke Hatanaka（畑中悠佑）¹，Katsumi Higaki（檜垣克美）⁵，Masaki Tanaka（田中雅樹）⁴，Masato Koike（小池正人）³，Yoshio Hirabayashi（平林義雄）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理化学研究所脳科学総合研究センター ³順天堂大学大学院神経機能構造学講座（医学部神経生物学・形態学講座） ⁴京都府立医科大学　生体構造科学／解剖学教室（生体構造科学部門） ⁵鳥取大学生命機能研究支援センター Parkinson's disease (PD) is characterized by dopaminergic (DA) cell loss and the accumulation of pathological alpha synuclein (asyn), but its precise pathomechanism remains unclear, and no appropriate animal model has yet been established. Recent studies have shown that a heterozygous mutation of glucocerebrosidase (gba) is one of the most important genetic risk factors in PD. To create mouse model for PD, we crossed asyn Bacterial Artificial Chromosome transgenic mice with gba heterozygous knockout mice. These double-mutant (dm) mice express human asyn in a physiological manner through its native promoter and showed an increase in phosphorylated asyn in the regions vulnerable to PD, such as the olfactory bulb and dorsal motor nucleus of the vagus nerve. Only dm mice showed a significant reduction in DA cells in the substantia nigra pars compacta, suggesting these animals were suitable for a prodromal model of PD. Next, we investigated the in vivo mechanism by which GBA insufficiency accelerates PD pathology, focusing on lipid metabolism. Dm mice showed an increased level of glucosylsphingosine without any noticeable accumulation of glucosylceramide, a direct substrate of GBA. In addition, the overexpression of asyn resulted in decreased GBA activity in mice, while dm mice tended to show an even further decreased level of GBA activity. In conclusion, we created a novel prodromal mouse model to study the disease pathogenesis and develop novel therapeutics for PD and also revealed the mechanism by which heterozygous gba deficiency contributes to PD through abnormal lipid metabolism under conditions of an altered asyn expression in vivo.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-397 Parkin活性化剤の探索 Kahori Shiba（柴佳保里）¹，Tsuyoshi Inoshita（井下強）¹，Yuzuru Imai（今居譲）²，Nobutaka Hattori（服部信孝）³ ¹順天堂大学大学院　医学研究科　多発性硬化症および神経難病治療・研究講座 ²順天堂大学大学院　医学研究科パーキンソン病病態研究解明講 ³順天堂大学大学院　医学研究科　神経学講座 Mutations of genes encoding Parkin and PINK1 cause autosomal recessive early-onset Parkinson's disease (PD), in which selective degeneration of the midbrain dopaminergic neurons occurs. Drosophila and mammalian cell biological studies have revealed that Parkin maintains the survival of dopaminergic neurons through the correction of damaged mitochondrial pool. The reduction of ΔΨm due to mitochondrial damages leads to PINK1 accumulation and activation on the mitochondrial outermembrane, preventing the ΔΨm-dependent import of PINK1 to the internal compartment of mitochondria. Activated PINK1 phosphorylates ubiquitin ligase (E3) Parkin and Ubiquitin, whereby latent Parkin in the cytosol is activated and localized on the mitochondrial outermembrane, ubiquitinating mitochondrial proteins such as Mitofusin and Miro. The ubiquitination of mitochondrial proteins promotes the mitochondrial recruitment of autophagy regulators and receptors. Collaterally, the ubiquitination and subsequent degradation of Mitofusin and Miro promotes mitochondrial fragmentation and suppresses mitochondrial motility, respectively, and facilitates autophagic removal of damaged mitochondria. Although physiological roles of PINK1-Parkin-mediated mitophagy or mitochondrial quality control is not fully understood, some of in vivo evidence suggests that defects of the PINK1-Parkin signals affect the myocardial and skeletal functions, the maintenance of pancreatic β cells and tumorigenesis. On the one hand, our Drosophila study indicates constitutive activation of Parkin is harmful to tissues. Thus, an appropriate control of PINK1-Parkin signals by drugs may be an efficacious treatment of a variety of diseases associated with mitochondrial dysfunctions. In this study, we developed a cell-based high-throughput screening (HTS) system to search compounds to activate the PINK1-Parkin signaling, by which two unique compounds were identified and characterized. Both compounds successfully stimulate Parkin mitochondrial translocation in dopaminergic neurons differentiated from iPS cells without obvious reduction of ΔΨm and cell toxicity. In Drosophila PINK1 model, both compounds improved the motor defects caused by reduced PINK1 activity and resultant Parkin inactivation. Thus, these compounds may be promising drug candidates for diseases associated with mitochondrial damages.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-398 次世代シークエンサを用いたパーキンソン病患者における）GBA遺伝子変異のスクリーニング Yuhei Kanaya（金谷雄平），Takahiro Fukuda（福田崇博），Koudai Kume（久米広大），Yukiko Matsuda（松田由喜子），Ryosuke Ohsawa（大澤亮介），Hiroyuki Morino（森野豊之），Hideshi Kawakami（川上秀史）広島大学　原爆放射線医科学研究所　分子疫学分野 Parkinson's disease (PD) is a progressive neurodegenerative disease with neuronal cell loss in the substantia nigra and aggregation of α synuclein. Symptoms of PD include resting tremor, muscle rigidity, bradykinesia. The estimated prevalence rate is about 150/100,000 population in Japan and the 5-10% of which have a family history of this disorder. The LRRK2, PRKN and SCNA genes have been identified as causative genes for familial PD.The GBA gene is known as a causative gene of Gaucher disease (GD), but it is also known as a susceptible gene of PD. Mutations in GBA causes a reduction or complete loss of glucocerebrosidase activity which leads to accumulation of glucosylceramide in visceral organs, causing a various clinical features including thrombocytopenia, anemia, and hepatosplenomegaly. Some of the cases show central nervous system disorders. Moreover, excess amount of glucosylceramide causes structural changes in α synuclein. That presumably results in aggregation of α synuclein in neuronal cells and develops PD.The frequency of mutations in GBA among PD patients has been reported several studies so far. Thus far, there has been only one study that reports the frequency of mutations among Japanese PD patients. Using next-generation sequencing(NGS), we screened all of exons for GBA mutations and compare the frequency of mutations in PD patients with the previous studies. We screened the GBA gene in 624 PD patients using NGS. Called variants are submitted to four prediction tools (SIFT, PolyPhen-2, Mutation Taster and CADD) for pathogenicity. The variants that are positive in more than one prediction tools or the variants that give CADD scores of 20 or more are considered pathogenic. Mutations that are regarded as pathogenic are confirmed by Sanger sequencing. Overall, 10 different mutations out of 18 PD patients (2.9%) were identified in 624 PD patients. Nine of the mutations are already known while one of the mutation is novel. 20 base deletion and 2 base insertion are found in the new mutation. From the result of our study, the frequency is lower than that of previous studies. One possibility is that this may simply represent the regional differences. Another possibility is that our criteria of narrowing mutations are strict. Further research to elucidate the pathogenesis of GBA-PD will lead to the development of disease-modifying therapy in GBA-PD.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-399 酸化ストレスが関与する神経疾患の新規治療薬の開発 Yoshihisa Koyama（小山佳久）¹，Yuki Kobayashi（小林悠輝）²，Makoto Kondo（近藤誠）¹，Noriyoshi Usui（臼井紀好）^1,3，Hikaru Kobayashi（小林光）²，Shoichi Shimada（島田昌一）¹ ¹大阪大院医神経細胞生物学 ²大阪大産業研半導体材料プロセス研究分野 ³大阪大院医共同研究実習センター Oxidative stress caused by reactive oxygen species such as hydroxyl radicals generated by cellular metabolic respiration is deeply involved in the onset and progression of various brain diseases such as ischemic cerebrovascular disease, neurodegenerative diseases, mental disorders, but treatment methods have not been fully established. A newly developed Silicon-based agent has made it possible to continuously generate large amounts of hydrogen molecules having antioxidant activity in vivo. Using this new method, we tried to establish the therapeutic effect in various neurological diseases associated with oxidative stress. We prepared two neurological disease model mice, dopaminergic neuron-specific neurotoxin, 6-OHDA-induced Parkinson 's disease model mice (PD model) and hemisectioned spinal cord injury model mice (SCI model), and fed these model mice with Silicon-based agent-contained diet (Si group) or normal diet (Control group). As the results, dopaminergic neuronal loss and impairment in motor coordination in the PD models of Si group were significantly reduced as compared to those of Control groups, and the recovery of the hind limb motor function in the SCI models of Si group was significantly accelerated. These findings suggested that hydrogen molecules generated by Silicon-based agent were effective for the prevention of neurological diseases involving oxidative stress. Our Silicon-based agent will be innovative approach to therapy for oxidative stress disease including neurodegenerative and neuroinflammatory diseases.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-400 酸化ストレスの増加とモトプシンの欠損はパルブアルブミン介在ニューロンを減少させる Shiori Miyata（宮田栞），Taiki Kashio（樫尾泰樹），Shinichi Mitsui（三井真一）群馬大院保健 Mutation in 14 genes causes neuronal ceroid lipofuscinoses (NCLs), which is characterized by the accumulation of autofluorescence material and shows various symptoms such as psychomotor retardation. The knockout (KO) of ceroid lipofuscinosis neuronal-1 (CLN1) gene causes increased oxidative stress in the mouse brain and the enhanced expression of serpina1, which inhibits an extracellular protease, motopsin/neurotrypsin. Motopsin, secreted from neuronal cells in various brain regions, cleaves a heparansulfate proteoglycan, agrin, and produces a 22 kDa fragment, which is essential for synaptic formation. The inhibition of motopsin activity by serpina1 in CLN1 KO mice may attribute to the synaptic dysfunctions and probably participates in the reduction of parvalbumin interneurons in hippocampus, although we have not observed such symptoms in motopsin KO mice. Thus, we investigated whether oxidative stress in addition to the lack of motopsin gene leads to synaptic dysfunctions and cognitive impairments using motopsin KO mice. Six mg/kg of doxorubicin (DOX), an anti-cancer drug, or saline was intravenously injected in motopsin KO and wild-type mice every 8 days for 4 weeks. To evaluate cognitive functions, 3 days after the last injection, mice were assessed on an open field test, followed by a novel object recognition test (NOR) and a passive avoidance test (PAT) on 4 consecutive days. Brain were prepared after the PAT and stained by anti-parvalbumin antibody. Parvalbumin-positive cells were counted in hippocampus and cingulate cortex (CG). Regardless of genotype, DOX injection significantly reduced the locomotor activity in an open field test and the duration to investigate objects in a NOR. In a PAT, there was no significant main effects or interaction between groups or genes, suggesting DOX injection in motopsin KO mice did not affect the learning and memory in this test. DOX injection significantly reduced the hippocampal interneurons in CA3, not CA1 or CG. Interestingly, post hoc analysis revealed motopsin deficiency exacerbated the reduction of CA3 interneurons by DOX injection. DOX is a chemotherapeutic agent and increases oxidative stress in the brain, which causes cognitive impairments in some cancer patients. Our results do not support increased oxidative stress for 4 weeks as well as motopsin deficiency induces cognitive deficits. However, such genetic and environmental condition may contribute to the loss of the CA3 interneurons, like CLN1 KO mice.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-401 軽微なHSD17B4変異によって生じる緩徐進行性脊髄小脳変性症 Hiroyuki Morino（森野豊之）¹，Yukiko Matsuda（松田由喜子）¹，Ryosuke Miyamoto（宮本亮介）²，Hideshi Kawakami（川上秀史）¹ ¹広島大原医研分子疫学 ²徳島大院医歯薬臨床神経科学 [Introduction] Spinocerebellar ataxias (SCAs) are clinically and genetically heterogeneous diseases, classified as autosomal dominant, autosomal recessive, X-linked, and mitochondrial. In order to identify a causative gene, we performed genomic analysis of two families with autosomal recessive SCA. [Methods] We identified a causative gene and mutation using exome sequencing and homozygosity mapping. The mutation was confirmed by Sanger sequencing. The pathogenicity of the mutation was evaluated using CADD, and compared with previously reported mutations. Changes in expression levels of mRNA and protein due to the mutation were analyzed by RT-qPCR and western blotting using patient-derived fibroblasts. We also performed western blotting under non-denaturing condition, called native PAGE, to evaluate whether the dimerization of the gene product was changed. [Results] The causative mutation was a homozygous mutation located in the HSD17B4 gene encoding the peroxisomal D-bifunctional protein (DBP). The patients developed cerebellar ataxia after 30 years of age, and the subsequent progression was also slow. Most patients with DBP deficiency develop their symptoms in the neonatal period. In our cases, clinical features were very mild compared to previously reported cases. The expression level of mRNA was not different from that of normal, but the expression level of protein was decreased. Dimerization of DBP was also reduced. The CADD score of the identified mutation was lower than that of mutations reported so far. [Conclusions] We presented cases harboring the mutation in HSD17B4 gene and showing mild SCA clinically. DBP deficiency is a severe disease caused by homozygous mutations of HSD17B4, but it was revealed that the phenotype shows slowly progressive SCA when the functional loss of the protein due to the mutation is very mild.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-402 SCA42モデルマウスにおける小脳プルキンエ細胞の変性 Yukiko Matsuda（松田由喜子）¹，Hiroyuki Morino（森野豊之）¹，Takashi Kurashige（倉重毅志）²，Hisako Nakayama（中山寿子）³，Toshinori Matsuoka（松岡利典）³，Yusuke Sotomaru（外丸祐介）⁴，Kouichi Hashimoto（橋本浩一）³，Hideshi Kawakami（川上秀史）¹ ¹広島大学原医研分子疫学 ²独立行政法人国立病院機構呉医療センター臨床研究部 ³広島大院医歯薬神経生理 ⁴広島大自然科学研究支援開発センター生命科学実験部門 We analyzed two Japanese families with autosomal dominant spinocerebellar ataxia (SCA) using linkage analysis and exome sequencing. We have reported that CACNA1G is a causative gene of SCA, named as SCA42. The SCA42 patients exhibited mild progressive ataxia and cerebellar atrophy. CACNA1G encodes the T-type calcium channel (Cav3.1). The identified mutation p.R1715H is located in the voltage sensor of Cav3.1. Electrophysiological analyses revealed that the membrane potential dependency of the mutant Cav3.1 transfected in HEK293T cells shifted toward a positive potential. We generated the knock-in (KI) mice harboring the same mutation using CRISPR/Cas9 technology. We conducted behavioral analyses including rotarod tests and beam-walking tests. Motor performances were initially normal, but the mice began to show motor dysfunction in both rotarod tests and beam-walking tests around one year after birth. HE staining showed that the cerebellar Purkinje cells (PCs) were also degenerated at the same old. Furthermore, we analyzed KI mice at a much older age. The two-year-old KI mice exhibited tremor-like movement similar to some SCA42 patients. To observe cerebellar atrophy, we measured the size of cerebellum at the age of two years. However, the difference of the size between wild type and KI mice were not detected. To assess the apoptotic cell death of PCs, we stained the cerebellar sections using TUNEL staining kit. TUNEL positive PCs were not detected. These results indicated that the KI mice showed ataxia and involuntary movements similar to SCA42 patients, although macroscopic observations of cerebellar atrophy were negative at an older age. Those behavioral impairments could be caused by dysfunction of the cerebellar PCs. We will search the histological changes at an earlier stage before PCs degeneration to investigate the mechanism of neuronal degeneration in SCA42 KI mice.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-403 脊髄小脳変性症における眼と手の協調運動について – 眼球運動位置の正確性との関連 Satomi Terada-Inomata（寺田さとみ）¹，Shin-ichi Tokushige（徳重真一）²，Shun-ichi Matsuda（松田俊一）³，Masashi Hamada（濱田雅）⁴，Yoshikazu Ugawa（宇川義一）⁵，Shoji Tsuji（辻省次）^4,6，Yasuo Terao（寺尾安生）¹ ¹杏林大医細胞生理 ²杏林大病院　神経内科 ³NTT東日本病院　神経内科 ⁴東大病院　神経内科 ⁵福島県立医大　神経再生医療学　神経内科 ⁶国際医療福祉大　大学院　医学部 Introduction: Gaze and hand movements coordinates in daily motor actions (eye-hand coordination), for which the cerebellum is implicated. We investigated how cerebellar dysfunction affects this coordination in patients with spinocerebellar ataxia (SCA) by recording eye and finger movements simultaneously during reaching tasks. Methods: The subjects were 8 SCA patients with pure cerebellar symptoms (SCA6 and SCA31) and 11 age-matched normal controls (NC). In the visually guided reaching (VGR) task, the subjects placed their index finger on the fixation point at the center of the touch panel and slid their finger toward the target as it jumped to a peripheral location, 10 or 20 degrees from the center. In the memory guided reaching (MGR) task, while subjects placed their finger at the central fixation point, a cue was briefly presented at a peripheral location. Subjects slid their finger toward the remembered target location when the fixation point disappeared thereafter. Results: In both SCA patients and NC, the gaze preceded the finger to the target in VGR, whereas in MGR eye movements preceding the finger movement were often lacking or decreased. Spatial distribution of the eye and finger final positions was more variable in SCA patients than in NC and frequently undershot the target in SCA patients, especially for MGR. For VGR but not in MGR, there was a significant correlation between the final position of the gaze and the accuracy of the finger in both groups (NC: r = 0.69, p < 0.001 (10 degrees), r = 0.83, p < 0.001 (20 degrees) SCA: r = 0.62, p < 0.001(10 degrees), r = 0.52, p < 0.001 (20 degrees). The total time for the finger to reach the target and finger movement time was significantly prolonged in SCA patients in both tasks (VGR: p = 0.04, p = 0.02 (20 degrees) MGR: p = 0.002, p = 0.003 (20 degrees)), whereas the time by which the gaze preceded the finger was comparable. The final position of the gaze did not correlate with the total reaching time nor with the finger movement time in both groups, whereas the interval between the eye and finger movement was correlated with it only in NC. Conclusion: Gaze accuracy correlated with finger accuracy in VGR but no in MGR in both groups, but the variability of both effectors was larger in SCD. Higher gaze accuracy also led the finger to follow the gaze with a shorter interval in NC, but not in SCA patients.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-404 小脳神経系におけるゴルジ体酸性環境の重要性 Yu-shin Sou（曽高友深）¹，Soichiro Kakuta（角田宗一郎）²，Yuji Kamikubo（上窪裕二）³，Kazue Niisato（新里和恵）³，Takashi Sakurai（櫻井隆）³，Kenji Sakimura（崎村建司）⁵，Yusuke Maeda（前田祐輔）⁴，Taroh Kinoshita（木下タロウ）⁴，Yasuo Uchiyama（内山安男）²，Masato Koike（小池正人）¹ ¹順天大院　医神経生物・形態 ²順天大院　医神経疾患病態 ³順天大院　医細胞・分子薬理 ⁴大阪大微生物病研 ⁵新潟大脳研基礎神経科学細胞神経生物 The Golgi apparatus plays an indispensable role in posttranslational modification and transport of proteins to their target destinations. It is well-established that the Golgi requires an acidic luminal pH for optimal functioning. The Golgi pH regulator (GPHR) is an anion channel essential for Golgi luminal acidification. The loss of GPHR in cultured cells results in increased Golgi luminal pH, which in turn causes impaired proper Golgi function and morphology. However, the functional and morphological abnormalities at the neuronal circuit level due to perturbations in Golgi pH are not yet fully understood. In addition, morphological alteration of the Golgi apparatus is associated with several neurodegenerative diseases including Parkinson's disease, Alzheimer's disease and amyotrophic lateral sclerosis. Here, we used anatomical and physiological approaches to characterize morphological and functional abnormalities of neuronal circuit in GPHR conditional knockout mice. Purkinje cells (PCs) from the mutant mice exhibited vesiculation and fragmentation of the Golgi apparatus followed by axonal degeneration and progressive cell loss. Morphological analysis provided unequivocal evidence for the disruption of basket cell (BC) terminals around PC soma. Furthermore, electrophysiological recordings showed selective loss of large amplitude responses, suggesting BC terminal disassembly. In addition, innervation pattern of mutant PCs was altered such that climbing fiber (CF) terminals abnormally synapse on the somatic spines of mutant PCs in matured cerebellum. Taken together, our results describe an essential role of Golgi luminal acidification in maintaining proper neuronal morphology and neuronal circuitry.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-405 パーキンソン病関連遺伝子はsmall GTPase, Arl8の動態を介してαシヌクレインのターンオーバーを制御する Tsuyoshi Inoshita（井下強）¹，Jun Yi Liu（Liu Jun Yi）¹，Daisuke Taniguchi（谷口大祐）¹，Changxu Cui（Cui Changxu）²，Masashi Takanshi（高梨雅史）¹，Yuzuru Imai（今居譲）²，Nobutaka Hattori（服部信孝）^1,2 ¹順天堂大医神経学講座 ²順天堂大医パーキンソン病講座 Parts of Parkinson´s disease (PD) causative or risk genes play roles in the intracellular vesicle transport, which includes endocytosis of synaptic vesicles (SVs). We systematically analyzed the consequence of the loss of these PD-associated genes, LRRK2, VPS35, Auxilin, Synaptojanin 1, INPP5F, RME-8, VPS13C and Rab7L1 and of the combined loss-of-function (LOF) alleles to reveal the possible common pathological pathways in Drosophila. Screening using electrophysiological recordings, SV morphology and organelle distribution at synapses in the larval neuromuscular junctions revealed that the combination of LRRK2 and Auxilin LOF alleles exacerbated synaptic phenotypes, which include increased number of multi-vesicular bodies as well as multi-lamellar structures and accumulation of a small GTPase Arl-8. These phenotypes are signs of dysregulation of endosomal-lysosomal pathway at synapses. Furthermore, α -Synuclein incorporated from synapses were colocalized with Arl-8 puncta. Consistent with the finding, Arl-8b but not Arl-8a was colocalized with Lewy bodies and glial cytoplasmic inclusions, which are neuronal inclusions of α-Synuclein in PD and multiple system atrophy, respectively. These results suggest that dysregulation of Arl-8 dynamics by PD genes at synapses is involved in the transmission of pathogenic α-Synuclein via neural circuits.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-406 In vitroにおけるalpha-synuclein細胞外分泌機構の検討 Hideki Hayakawa（早川英規）¹，Kousuke Baba（馬場孝輔）¹，Chi-Jing Choong（チュンチジン）¹，Kensuke Ikenaka（池中建介）¹，Toshihide Takeuchi（武内敏秀）^1,2，Yoshitaka Nagai（義隆永井）^1,2，Hideki Mochizuki（望月秀樹）¹ ¹大阪大学大学院医学研究科　神経内科学 ²大阪大学大学院医学系研究科神経難病認知症探索治療学 Objective: Alpha-synuclein (α-syn) is the major component of Lewy body (LB), the pathological hallmark in Parkinson's disease (PD), and its genetic mutations cause familial forms of PD. Although α-syn G51D mutation exhibits the most severe clinical symptoms among the PD-linked mutations, its low aggregation propensity lacks correlation with its severe clinical symptoms. Previously, we showed G51D α-syn fibrils induce widespread α-syn pathology, progressive neuronal loss, and motor impairment in mice. However, the detailed mechanism of α-syn secretion from cells and its propagation remains largely unknown. Therefore, we investigated whether neuronal α-syn is secreted by exocytosis. Methods: We established a stable human dopaminergic SH-SY5Y cell line overexpressing human wild-type α-syn and performed assessment of α-syn secretion level. The secreted extracellular α-syn was then collected and added to the neuronal cells and its uptake by cells was examined. Results: A small fraction of extracellularly secreted α-syn was found to be in association with exosomes. α-syn fibril-transfected cells showed an increase in exosomes and extracellular α-syn. The uptake of secreted α-syn by neuronal cells was observed. Conclusion: The increased secretion of exosome carrying α-syn by G51D α-syn fibrils-transfected cells, which may facilitate cell-to-cell spread of α-syn, suggests greater propagation properties of G51D mutated α-syn.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-407 変異型α-シヌクレインフィブリルによるマウス脳でのレビー病理の形成 Kousuke Baba（馬場孝輔）¹，Hideki Hayakawa（早川英規）¹，Kensuke Ikenaka（池中建介）¹，Chi-Jing Choong（鐘其静）¹，Hiroshi Tsuda（津田浩史）¹，Masato Koike（小池正人）²，Seiichi Nagano（長野清一）¹，Takeshi Ikeuchi（池内健）³，Masato Hasegawa（長谷川成人）⁴，Yoshitaka Nagai（永井義隆）⁵，Hideki Mochizuki（望月秀樹）¹ ¹大阪大学大学院医学系研究科　神経内科学 ²順天堂大学大学院医学研究科　神経生物学・形態学 ³新潟大学脳研究所　生命科学リソース研究センター / バイオリソース研究部門　遺伝子機能解析学 ⁴東京都医学総合研　認知症・高次脳機能研究分野 ⁵大阪大学大学院医学系研究科　神経難病認知症探索治療学 Parkinson's disease (PD) is a neurodegenerative disease characterized by movement disorder and its pathological features include progressive neuronal loss in the substantia nigra and Lewy body (LB) formation. α-synuclein (α-syn) is the main component of LB, and its genetic mutation causes familial PD. Among these mutations, G51D mutation exhibits most severe clinical symptoms. However, in vitro studies showed low propensity for α-syn aggregation with G51D mutation. To determine the mechanisms associated with severe neurotoxicity of α-syn G51D mutation, we used a mouse model generated by G51D α-syn fibrils injection into the mouse SN. Analysis of the structure of G51D α-syn fibrils revealed higher β-sheet contents than wild-type α-syn fibrils. The injection of G51D α-syn fibrils into the substantia nigra of wild-type mice induced robust phosphorylated α-syn inclusions in inoculation side at 12 weeks, which further spread to the contralateral side at 24 weeks. Moreover, G51D α-syn fibrils injection resulted in strong induction of reactive astrocyte compared with wild-type α-syn fibrils injection. Mice injected with G51D α-syn fibrils exhibited progressive neuronal loss in the substantia nigra accompanied with mitochondrial abnormalities and motor impairment. However, Mice injected wild-type α-syn fibrls did not exhibited nigral neuronal loss and motor impairment. Our studies showed that G51D α-syn fibrils cause very severe α-syn pathology including neuronal cell death compared to WT α-syn fibrils. These findings indicate that the structural difference of G51D α-syn fibrils plays an important role in the rapidly developed and more severe neurotoxicity of G51D mutation-linked PD.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-408 特発性基底核石灰化症 (IBGC) におけるSLC20A2変異の機能解析 Kazuya Nishii（西飯和哉），Ritsuko Shimogawa（下川梨津子），Hisaka Kurita（栗田尚佳），Masatoshi Inden（位田雅俊），Isao Hozumi（保住功）岐阜薬大・薬・薬物治療 Idiopathic basal ganglia calcification (IBGC) is a rare intractable disease characterized by abnormal deposits of minerals including mostly calcium in the basal ganglia, thalamus and cerebellum. SLC20A2 encoding phosphate transporter PiT-2 was identified in 2012 as the first causative gene of IBGC and occupied approximately 40% in familial IBGC. PiT-2 is a sodium dependent phosphate transporter of type III belonging to the SLC family and involved in the maintenance of inorganic phosphate (Pi) homeostasis. Previous reports have shown that variants of SLC20A2 in IBGC patients caused dysfunction of Pi transport activity in PiT-2 and disrupted Pi homeostasis. The Prodomain region located at the N-terminus and C-terminus of PiT-2 plays an important role in phosphate uptake function. Many SLC20A2 variants have reported so far, but there is no effective cure for IBGC. In this study, we identified four novel SLC20A2 missense variants from IBGC patients and patient's families. Then, we constructed PiT-2 stable Chinese hamster ovary (CHO) cell lines with these variants using the Flp-in system (Flp-in CHO cells) and analyze their functions to clarify the mechanism of IBGC. While Pi transport activity was significantly decreased to null levels in the Flp-in CHO cells with SLC20A2 variants present in Prodomain (variant A, B and D), that in the Flp-in CHO cell with a variant present outside Prodomain (variant C) was maintained at 27.8% of the reference level. Interestingly, the variant C had been discovered by chance in healthy members in an IBGC family. This suggests that preservation of some Pi transport activity may avoid the onset of IBGC. In addition, we studied the virus receptor function and membrane translocation to investigate the process of PiT-2 to cell surface. As the result, we confirmed four variants PiT-2 could be translocated into the cell membrane in the same way as PiT-2-Wild type. In conclusion, we reported four novel variants in SLC20A2 and showed that PiT-2 dysfunction caused by variants would lead to the pathogenesis of IBGC. An interesting case reveals that partial increase in Pi uptake may suppress the onset of IBGC.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-409 特発性基底核石灰化症 (IBGC) 患者由来のiPS細胞から分化させた内皮細胞におけるSLC20A2変異によるリン酸輸送能の障害 Tomohiko Masaka（真坂智彦），Shinichiro Sekine（関根信一郎），kazuya Nishii（西飯和哉），Hisaka Kurita（栗田尚佳），Masatoshi Inden（位田雅俊），Isao Hozumi（保住功）岐阜薬大・薬・薬物治療 Idiopathic Basal Ganglia Calcification (IBGC) is a rare neuropsychiatric illness also known as Fahr's disease or Primary Familial Brain Calcification (PFBC). It is characterized by bilateral calcifications mainly in the basal ganglia, and in the cerebellum, brain stem, and subcortical white matter. At present, there is no effective cure and treatment for IBGC. SLC20A2 encoding phosphate transporter PiT-2 was identified in 2012 as the first causative gene of IBGC and SLC20A2 variants account for approximately 40% of familial IBGC cases in Japan. To date, the effect of variants on SLC20A2 protein function remains unclear, although variants associated with IBGC have been predicted to be unable to increase intracellular Pi uptake, and leading to disruption of intracellular Pi homeostasis based on the in silico analysis using PolyPhen2. We have reported novel SLC20A2 variants in the Japanese population and established an induced pluripotent stem cells (iPSCs) from an IBGC patient carrying a SLC20A2 variant. To investigate the effect of these SLC20A2 variants identified in our previous study, we used Chinese hamster ovary (CHO) cells expressing these variant proteins using the Flp-In system (Flp-In CHO cells), and showed that variant SLC20A2 proteins significantly disrupt the Pi transport activity in Flp-In CHO cells. We considered that endothelial cells (ECs) represent important target cells for elucidating the pathology of IBGC in drug discovery research. So, we differentiated IBGC patient-derived iPSCs into ECs and found same levels of expression of endothelial marker and function into ECs compared with control iPSCs. However, the Pi transport activity of IBGC patient-derived iPS-ECs was significantly decreased compared with that of control iPS-ECs without changing the gene expression of the other SLC 20 family members. We confirmed that SLC20A2 variants cause the loss of function of the Pi transport activity in both Flp-In CHO cells and disease-specific iPSCs. This is the first report to show an in vitro model of iPSCs in IBGC with patient-identified SLC20A2 variants. These useful tools will help in elucidating the disease mechanism and can be used for search drug candidates for IBGC.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-410 培養ニューロン細胞質TDP-43凝集体形成を抑制する分子の解析 Kazuhiko Watabe（渡部和彦）^1,2，Yoichiro Kato（加藤陽一郎）²，Miho Sakuma（佐久間美帆）³，Makiko Murata（村田麻喜子）¹，Motoko Kawaguchi-Niida（新井田（川口）素子）²，Akiyoshi Kakita（柿田明美）⁴，Noriyuki Shibata（柴田亮行）² ¹杏林大学保健学部臨床検査技術学科神経病理学 ²東京女子医大院医病理病態神経科学 ³東京女子医大医学部医学科 ⁴新潟大脳研病理学分野 TAR DNA-binding protein 43 (TDP-43) is a main constituent of cytoplasmic aggregates in neuronal and glial cells in cases of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). We have previously demonstrated neuronal cytoplasmic aggregate formation induced by recombinant adenoviruses expressing human wild type (WT) and C-terminal fragment (CTF) TDP-43 under the condition of proteasome inhibition. These TDP-43 aggregate formation was markedly suppressed by co-infection of an adenovirus expressing heat shock transcription factor 1 (HSF1), a master regulator of heat shock response. In the present study, we performed cDNA microarray analysis to identify candidate molecules locating downstream of HSF1 that counteract TDP-43 aggregate formation. The 1464R rat neural stem cells were differentiated into neurons in the presence of retinoic acid, and co-infected with adenoviruses expressing DsRed-tagged human WT and CTF TDP-43, and EGFP-tagged human HSF1 in the presence of proteasome inhibitor MG-132. Two days in vitro, total RNAs were isolated and cDNA microarray analysis of TDP-43 aggregate formation was performed. We found 64 genes that are up-regulated more than 2-fold by HSF1 transduction, and 393 genes that are up-regulated more than 2-fold in association with aggregate inhibition by HSF1 transduction. We then PCR-cloned candidate genes into EGFP vector and checked their effects on TDP-43 aggregate formation by fluorescence microscopy and western blot analysis. Some of these genes were constructed into adenoviral vectors for further analysis. Our preliminary data showed suppressive effects of some of candidate genes on cytoplasmic TDP-43 aggregate formation.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-411 JNKによるリン酸化を介したSARM1のNAD⁺分解活性とミトコンドリア機能制御 Hitoshi Murata（村田等），Ken-ichi Yamamoto（山本健一），Rie Kinoshita（木下理恵），Masakiyo Sakaguchi（阪口政清）岡山大院医歯薬細胞生物 Mitochondrial dysfunction is a key pathological feature of many different types of neurodegenerative disease. Sterile alpha and Toll/interleukin receptor motif-containing protein 1 (SARM1) has been attracting much attention as an important molecule for inducing axonal degeneration and neuronal cell death by causing loss of NAD⁺ (NADH). However, it has remained unclear what exactly regulates the SARM1 activity. Here, we show that NAD⁺ cleavage activity of SARM1 is regulated by its own phosphorylation at serine 548. The phosphorylation of SARM1 was mediated by c-jun N-terminal kinase (JNK) under oxidative stress conditions, resulting in inhibition of mitochondrial respiration concomitant with enhanced activity of NAD⁺ cleavage. Nonphosphorylatable mutation of Ser548 or treatment with a JNK inhibitor decreased SARM1 activity and suppressed ATP reduction under oxidative stress conditions. Furthermore, neuronal cells derived from a familial Parkinson's disease (PD) patient showed a congenitally increased level of SARM1 phosphorylation compared with that in neuronal cells from a healthy person and were highly sensitive to oxidative stress. The SARM1 phosphorylation also increased in the midbrain of paraquat-treated mice, an in vivo PD model. These results indicate that JNK-mediated phosphorylation of SARM1 at Ser548 is a regulator of SARM1 leading to inhibition of mitochondrial respiration. These findings suggest that an abnormal regulation of SARM1 phosphorylation is involved in the pathogenesis of PD and possibly other neurodegenerative diseases.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-412 C9orf72関連筋萎縮性側索硬化症/前頭側頭型認知症モデルショウジョウバエにおける異常伸長リピートRNAとジペプチドリピートタンパク質の病態での役割 Morio Ueyama（上山盛夫）¹，Taro Ishiguro（石黒太郎）^2,3,4，Takuya Konno（今野卓哉）⁵，Akihide Koyama（小山哲秀）⁵，Kinya Ishikawa（石川欣也）³，Keiji Wada（和田圭司）²，Osamu Onodera（小野寺理）⁵，Yoshitaka Nagai（永井義隆）^1,2 ¹大阪大院医神経難病治療学 ²国立精神・神経セ神経研疾病４ ³東京医歯大脳神経病態学 ⁴Dept. Neurosci, Mayo Clinic, FL, USA ⁵新潟大脳研神経内科 An expanded G4C2 repeat within the C9orf72 gene has been found to be the most common genetic mutation for amyotrophic lateral sclerosis/frontotemporal dementia (C9-ALS/FTD). In the brain of C9-ALS/FTD patients, the expanded repeat RNA transcribed from mutated C9orf72 gene leads to the formation of RNA aggregates, namely RNA foci. The expanded repeat RNA is also translated into dipeptide repeat (DPR) proteins, such as poly(GR), poly(GA), and poly(GP), by the repeat associated non-ATG (RAN) translation. Although these RNA foci and/or DPR proteins are thought to be involved in the neurodegeneration, the molecular mechanisms leading to ALS/FTD pathogenesis remain unclear. To dissect the pathological roles of expanded repeat RNA and DPR proteins caused by an expanded G4C2 repeat, we designed G4C2 repeat constructs without or with a stop codon upstream of the repeat sequences and were expressed in Drosophila under the control of the GAL4-UAS system. We found that flies expressing the expanded repeat without a stop codon in the eye showed severe rough eye phenotype with decreased pigmentation and eye size, while flies expressing the expanded repeat with a stop codon exhibited almost normal eyes. Moreover, flies expressing the expanded repeat without a stop codon in neurons displayed motor dysfunction, while flies expressing the expanded repeat with a stop codon in neurons showed normal motor function. RNA fluorescence in situ hybridization analyses demonstrated that flies expressing the expanded repeat without or with a stop codon had similar RNA foci formation in their salivary glands. Surprisingly, immunohistochemical analyses showed that DPR proteins were only faintly detectable in flies expressing the expanded repeat without a stop codon, although they were expressed in flies expressing the expanded repeat with a stop codon. These results indicate the DPR proteins generated from the expanded G4C2 repeat RNA by the RAN translation mainly contribute to the neurodegeneration in our model flies.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-413 12型先天性大脳白質形成不全症の責任遺伝子産物VPS11の疾患変異解析 Naoto Matsumoto（松本直人），Junji Yamauchi（山内淳司）東京薬科大学・生命科学部・分子神経科学研究室 Hypomyelinating leukodystrophies (HLDs) are inherited dysmyelinating and demyelinating diseases. HLDs are rare diseases, which occur in one every 200,000 to 500,000 people. They are clinically characterized by severe symptoms including nystagmus, spastic quadriplegia, ataxia, and developmental delay. They are commonly due to abnormal myelination in the central nervous system (CNS). Oligodendroglial cells (oligodendrocytes) differentiate to form myelin sheaths and wrap neuronal axons, achieving saltatory conduction and protect axons. So far, 17 HLD-associated genes have been identified; however, it remains unclear how and why their responsible genes cause diseases. It is now difficult to clarify therapeutic targets and drugs based on disease-related molecular mechanisms. Vacuolar protein sorting-associated protein 11 (VPS11) is a common, core component of homotypic fusion and protein sorting (HOPS) and class C core vacuole/endosome tethering (CORVET) protein complexes, which are involved in membrane trafficking. VPS11 also functions as fusion of organelles such as lysosomes and endosomes. The purpose of this study is to elucidate cellular and molecular pathological mechanism(s) underlying HLD12-associated C846G mutation of VPS11. The wild type VPS11 proteins are localized in lysosomes and organelles around lysosomes, being consistent with the reported results. In contrast, the C846G mutant proteins exhibit punctate structures throughout cytoplasm where lysosome-related phosphorylating signaling of S6 and 4E-BP1 through p70 S6 kinase is decreased. We have collected oligodendrocyte progenitor cell line FBD-102b stably harboring the C846G mutant constructs. Following the induction of differentiation, cells harboring the C846G mutant constructs greatly decrease the numbers of multiple processes and myelin web-like structures, comparing to those of parental cells. HLD12-associated VPS11 mutants form aggregates throughout cytoplasm, decreasing oligodendroglial differentiation probably by inhibiting p70 S6 kinase signaling.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-414 AMPA受容体サブユニット特異的RNAアプタマーによるALSモデルマウス（AR2）におけるALS病態の改善効果 Megumi Akamatsu（赤松恵）^1,2，Takenari Yamashita（山下雄也）^1,3，Sayaka Teramoto（寺本さやか）^1,4，Zhen Huang（Huang Zhen）⁵，Tatsushi Toda（戸田達史）²，Li Niu（Niu Li）⁵，Shin Kwak（郭伸）^1,2,4 ¹東京大院医神経病理 ²東京大院医神経内科 ³東京医大院医病態生理 ⁴東京医大院医神経分子病態 ⁵Dept Chem, Univ at Albany, State Univ of New York, Albany, NY, USA Background: In the motor neurons of the vast majority of sporadic ALS patients, the expression of an RNA editing enzyme called ADAR2 is reduced. This results in abnormal expression of glutamine/arginine (Q/R) site-unedited GluA2, a subunit of the AMPA receptor, which ultimately induces death of motor neurons in the conditional ADAR2 knockout (AR2) mice through the Ca²⁺-permeable AMPA receptor-mediated mechanism. Therefore, blocking the exaggerated Ca²⁺ influx through abnormal Ca²⁺-permeable AMPA receptors is a potential therapeutic strategy for developing drugs for sporadic ALS. We previously reported that perampanel, a selective non-competitive AMPA receptor antagonist, robustly prevented the progressive motor dysfunctions and loss of motor neurons with TDP-43 pathology in ADAR2^flox/flox/VAChT.Cre (AR2) mice (Akamatsu et al, Sci Rep. 2016). However, a significant sedative effect of perampanel would limit its dosage should it be used clinically. RNA aptamers are a new group of potential ALS drug candidates, and have the advantage of high potency, high selectivity and low immunogenicity compared to small molecule antagonists. An RNA aptamer (FN1040) specifically blocks AMPA receptors, with selectivity to GluA1 and GluA2. Objective: To investigate whether FN1040 is a potential ALS drug with low sedative effects. Methods: ADAR2 cKO mice (ADAR2^flox/flox/VAChT-Cre.Fast; AR2) were used. FN1040 was delivered to the mouse cerebroventricule through an indwelling catheter connected to a subcutaneously mounted Alzet osmotic pump. After confirming the in vivo stability of FN1040, efficacy on death of motor neurons was tested with rotarod for motor function. and morphological observation of the spinal cord immunostained for ChAT and TDP-43. Results: The local concentration of FN1040 in the brain stem and the spinal cord was determined using ³²P-labeled FN1040. A long-term infusion of FN1040 significantly blocked the progression of motor dysfunction and the death of motor neurons with normalization of TDP-43 subcellular localization. It's worth noting that FN1040-treated mice did not exhibit overt sedation that is an inevitable adverse effect of AMPA receptor antagonists. Conclusion: Our preliminary data have shown a robust neuroprotective effect on this ALS mouse model and suggest that use of AMPA receptor-specific RNA aptamers is potentially a new therapeutic approach for ALS.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-415 先天性大脳白質変性症の疾患変異をもつAIMP1は神経分化を強く阻害する Yu Takeuchi（竹内裕），Junji Ymauchi（山内淳司）東京薬科大学・生命科学部・分子神経科学研究室 Cells in the central nervous system (CNS) are primarily composed of neuronal cells, some glial cells, and other types of cells such as endothelial cells. In CNS glial cells, oligodendroglial cells (oligodendrocytes) wrap myelin sheaths around neuronal axons. Myelin sheaths are differentiated oligodendrocyte plasma membranes. Hypomyelinating leukodystrophies (HLDs) are a group of hereditary CNS neurodegenerative diseases, including demyelinating phenotypes and dying axons, at a rate of 1 in 200,000 to 500,000 people. Advanced techniques including genomic sequencing not only have clarified much more responsible mutated genes than previously expected but also have identified the potential relationship for unexpected primary disease-affecting, pathological cells other than oligodendrocytes. Despite increasing identification of responsible genes and cells, less is known about why and how mutated gene products cause such diseases. In this study, we have investigated the effects of HLD3-associated mutation of aminoacyl tRNA synthase complex-interacting multifunctional protein 1 (AIMP1) on biochemical and cell biological outputs. HLD3-associated mutant proteins (frameshift type 292CA [2 base (CA) deletion at the 292 nucleotide position] and nonsense mutation type Q39X) of AIMP1 are mainly localized in the lysosome where mutated proteins are accumulated. In contrast, wild type proteins are localized throughout cytoplasm, being consistent with the established results. Also, decreased signaling in the lysosome-related mammalian target of rapamycin (mTOR) pathway is observed in cells expressing either 292CA or Q39X. While neuronal cell line N1E-115 exhibits differentiated phenotypes such as elongation of long neurite-like processes with expression of differentiation marker growth associated protein 43 (GAP43), cells harboring the 292CA or Q39X mutant constructs fails to exhibit those ones.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-416 神経系特異的SQSTM1の欠損はALSマウスモデルの発症を早期化させる Shun Mitsui（三井駿）¹，Masahito Ishiyama（石山政仁）¹，Kento Shimakura（島倉健人）¹，Kai Sato（佐藤海）¹，Suzuka Ono（小野鈴花）¹，Asako Otomo（大友麻子）^1,2，Tetsuro Ishii（石井哲郎）³，Toru Yanagawa（柳川徹）³，Masashi Aoki（青木正志）⁴，Shinji Hadano（秦野伸二）¹ ¹東海大医分生 ²東海大マイクロナノ ³筑波大医 ⁴東北大院神内 Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by a selective loss of upper and lower motor neurons. Several studies have demonstrated that mutations in the SQSTM1 gene are linked to ALS. SQSTM1 encodes SQSTM1/p62 that regulates not only autophagy via the association with LC3 and ubiquitinated proteins but also the Keap1-Nrf2 antioxidative stress pathway by interacting with Keap1. Recently, we have demonstrated that systemic depletion of SQSTM1 exacerbates disease phenotypes in a SOD1^H46R transgenic (tg) ALS mouse model, suggesting a neuroprotective role of SQSTM1 in vivo. However, the effect of neural cell-specific knockout (KO) of Sqstm1 in SOD1^H46R mice remains unknown. To address this issue, we newly generated a neural cell-specific SQSTM1 deficient ALS mouse model (Nes;SQSTM1^f/f;SOD1^H46R) by crossing Nes;SQSTM1^f/+ tg mice to SQSTM1^f/+;SOD1^H46R tg mice, and analyzed their survival and pathology. Western blot analysis revealed that expression of SQSTM1 in the brain and spinal cord of Nes;SQSTM1^f/f mice was undetectable, while those in the skeletal muscle and liver were comparable to that of wild-type animal, confirming the nervous system-specific KO of Sqstm1. Lifespan and growth curve analyses demonstrated that nervous system-specific SQSTM1 deficiency caused earlier disease onset and shorter lifespan in SOD1^H46R mice. Immunohistochemical analyses of the lumbar spinal cord revealed that ubiquitin-positive aggregates (Ubs) were present in MAP2-positive neurons in Nes;SQSTM1^f/f;SOD1^H46R mice at end-stage, whereas Ubs were mainly observed in extracellular spaces and/or neuropils in SQSTM1^f/f;SOD1^H46R mice. These results suggest that neural cell-specific SQSTM1 deficiency exacerbates disease symptoms.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-417 遺伝性の白質脳症変異をもつC11orf73は、オリゴデンドロサイト分化を阻害する Kohei Hattori（服部耕平），Junji Yamauchi（山内淳司）東京薬科大学・生命科学部・分子神経科学研究室 During development of the central nervous system (CNS), oligodendroglial cells (oligodendrocytes) wrap neuronal axons with myelin sheaths, which are their differentiated plasma membranes. Genetic hypomyelinating leukoencephalopathies are a heterogeneous group of the CNS disorders with white matter involvement and are often characterized by hypomyelinating phenotypes with periventricular cysts and vermian atrophy. The homozygous disease-related variant (Cys4-to-Ser [C4S]) of the c11orf73 (also called hikeshi) gene is known to be associated with infantile hypomyelinating leukoencephalopathy. Despite a rapidly increasing identification of the genes associated with leukoencephalopathies, the effects of their mutations on protein products and cellular behaviors still remain to be established. Herein we show that in mouse oligodendroglial FBD-102b cells, the C4S mutant proteins of C11orf73 are mislocalized in the lysosome where they can be aggregated. Expression of the C4S mutants leads to localization into the lysosome but not into the endoplasmic reticulum (ER) and the Golgi body whereas the wild types are localized in the nuclei and the cytoplasm. While parental FBD-102b cells exhibit morphological differentiation, being characteristic of myelin web-like structures, cells harboring the C4S mutants fail to exhibit it. Signaling through lysosome-related kinases is downregulated in cells harboring the C4S mutants. Together, leukoencephalopathy-associated C11orf73 mutant is mislocalized in the lysosome to inhibit morphological differentiation, presenting the part of potential cellular phathological mechanisms underlying hypomyelinating leukoencephalopathies.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-418 D-cysteineは脊髄小脳失調症原因タンパク質を発現する初代培養小脳プルキンエ細胞の樹状突起発達低下を改善する Takahiro Seki（関貴弘）¹，Masahiro Sato（佐藤正寛）¹，Ayumu Konno（今野歩）²，Hirokazu Hirai（平井宏和）²，Yuki Kurauchi（倉内祐樹）¹，Hiroshi Katsuki（香月博志）¹ ¹熊本大院生命科学・薬薬物活性 ²群馬大院医脳神経再生 Spinocerebellar ataxia (SCA) is a group of autosomal dominant neurodegenerative diseases, which is characterized by the progressive motor dysfunction and cerebellar atrophy. SCA is classified into SCA1-47 by the differences in the loci of causal genes. We have previously demonstrated that the dendritic development of primary cultured cerebellar Purkinje cells (PCs) is frequently impaired by the expression of several SCA-causing proteins. We consider that this impairment is one of the common phenotypes of in vitro SCA models and that the amelioration of the dendritic development would be a molecular target for SCA treatment. Recently, we have revealed that D-cysteine significantly enhances the dendritic development of primary cultured PCs via the production of hydrogen sulfide. In the present study, we investigated the potential of D-cysteine to ameliorate the dendritic development of cerebellar PCs expressing several SCA-causing proteins. Cerebellar primary cultures were prepared from E16 embryos of Wistar rats. Wild-type and SCA mutant ataxin-1 (SCA1-causing protein) and transmembrane protein 240 (TMEM240, SCA21-causing protein) were expressed to cultured PCs by using adeno-associated viral vectors. D-cysteine (0.2 mM) was treated from 21 to 28 days in vitro. Dendritic development of PCs was evaluated by the immunostaining with anti-calbindin antibody. As we have previously demonstrated, mutant ataxin-1 and mutant TMEM240 significantly impaired the dendritic development of cultured PCs. Although D-cysteine (0.2 mM) did not affect the dendritic development of PCs expressing wild-type ataxin-1 and TMEM240, D-cysteine significantly improved the dendritic development of PCs expressing mutant ataxin-1 and TMEM240. These findings suggest that D-cysteine ameliorates the dendritic development of two different in vitro SCA models and that D-cysteine has a therapeutic potential against several types of SCAs.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-419 トランスジェニックマウス脳内における微小管結合型および遊離型タウの定量 Ayaka Hagita（萩田彩香）同志社大生命医科学 Neurofibrillary tangles (NFTs), a neuropathological hallmark of tauopathies including Alzheimer's disease, are insoluble inclusions composed of abnormally phosphorylated tau. The abnormal tau purified from NFTs lost its physiological functions, microtubule-binding and -stabilization. So, it can be considered that the liberation of tau from microtubule is a crucial step for the pathogenesis of tauopathies. The tau pathologies are reproduced in the brains of tau transgenic mice. However, free-tau that does not bind on microtubules is found in the brains of wild-type mice that never show tau pathologies. This result implies that there is key difference between the wild-type and the transgenic mice in microtubule unbound tau. Microtubules are cytoskeleton abundant in neurons, and are needed for the maintenance of neuronal structure and intracellular transport. Because the microtubules are known to be vulnerable against low temperature, cations, and energy failure, it seems to be difficult to analyze biochemically mimicking the physiological free/microtubule state in the brain. Here we tried to develop the fraction procedure to separate MT-bound and -unbound tau from mice brains and examined the behavior of tau expressed in the brains of wild-type and PS19 tau transgenic mice. We verified and modified the procedure for the fraction of free tubulins and microtubules, that consists of the centrifugation in the taxol-containing buffer, previously reported. Quantifications of the tau and tubulins fractioned into free tubulin fraction and microtubule fractions by western blotting showed that the majority of tau expressed in wild-type mice brains were bound onto microtubules but small fraction of it was left in free tubulins fraction. However, the proportions of exogenously expressed tau recovered in free tubulins fraction were significantly higher compare to that of endogenous mouse tau even in pre-symptomatic period of PS19 mice. Unexpectedly, proportions of tubulins found in free and microtubule fractions were not affected by the expression of exogenous tau. We also found that the tau-inclusions in PS19 mice consist of human tau. These suggest that the abnormal tau which can lead to form tau inclusions is inherently impaired in its microtubule binding ability. We are currently working in the identification of the biochemical characters of microtubule-unbound tau in wild-type and PS19 mice.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-420 Autophagy impairment and unfolded protein response (UPR) induction in 1-methyl-4-phenylpyridinium (MPP⁺) cytotoxicity: potential involvements in Parkinson's disease Ding-I Yang（Yang Ding-I），Zheng-Jie Gong（Gong Zheng-Jie），Yun-Chung Chen（Chen Yun-Chung） National Yang-Ming University Parkinson's disease (PD) is the second most common neurodegenerative disease in the world. Despite the shared pathological feature of dopaminergic neuronal loss in the substantia nigra pars compacta, the etiology of PD is still unknown. Mitochondrial dysfunction and lysosomal damage have all been considered to play a role in the pathogenesis of PD. 1-Methyl-4-phenylpyridinium (MPP⁺) has been proved to be a PD-causing agent in human since 1980s. It is believed that MPP⁺ inhibits complex I in the electron transport chain, thereby causing mitochondrial dysfunction and cell death in the dopamine-secreting cells. Unfolded protein responses (UPR) may affect neuronal survival under the stressful conditions of endoplasmic reticulum (ER) stress. Impairment of autophagy, which is a self-eating process to eradicate dysfunctional components including damaged mitochondria, may also increase the unfolded protein load in the ER lumen with induction of UPR. However, whether MPP⁺ may affect autophagy and UPR to impact on neuronal viability have not been fully investigated. Using PC12 cells, a dopamine-secreting cell line derived from rat pheochromocytoma, we tested whether autophagy and UPR are involved in MPP⁺ cytotoxicity in PC12. We found that MPP⁺ at 1 mM resulted in an approximately 30% decrease in survival within 24 hours without activation of caspase-3, suggesting caspase-independent cell death. Consistent with its role as a mitochondrial inhibitor, MPP⁺ decreased basal oxygen consumption, ATP production, and maximal respiration based on the studies using Seahorse XF24. MPP⁺ impaired autophagy possibly at omegasome elongation step. Furthermore, MPP⁺ also induced expression of ATF4, ATF6, and CHOP without significantly affecting XBP1s in PC12 cells. These results indicated that MPP⁺ impairs autophagy and differentially induces UPR-related pathways, suggesting autophagy and UPR are both involved in MPP⁺ toxicity. Whether autophagy impairment is causally linked to UPR induction, together contributing to cell death, induced by MPP⁺ is currently under investigation.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-421 死細胞上のアミノリン脂質はミクログリア受容体TREM2からシグナルを伝達する Nobuhisa Iwata（岩田修永）¹，Takashi Saito（斉藤隆）²，Shigekazu Nagata（長田重一）³，Marco Colonna（Colonna Marco）⁴，Keiro Shirotani（城谷圭朗）¹ ¹長崎大院・ゲノム創薬学 ²理化学研究所・免疫シグナル ³大阪大学・免疫学フロンティア研究センター ⁴ワシントン大学 Variants of triggering receptor expressed on myeloid cells 2 (TREM2) are associated with an increased incidence of Alzheimer's disease, as well as other neurodegenerative disorders. Understanding how the variants confers neurodegenerative risk promises to provide insights into how microglia, brain-resident myeloid cells, contributes to pathogenesis and progression of the disorders. In this study we developed highly sensitive reporter cell model to monitor signal transduction via TREM2 receptor. The reporter cells expressing TREM2 transduced the signal upon treatment of apoptotic Neuro2a cells while those expressing TREM1 or parent reporter cells did not. The specificity of the signal transduction was also demonstrated as the TREM2 specific antibody inhibited the signal. This TREM2-dependent signal was completely inhibited by milk fat globule-EGF factor 8 (MFGE8), aminophospholipid receptor which is expressed by phagocytes, suggesting that the signal is mediated by phosphatidylserine and phosphatidylethanolamine on apoptotic cells, which are not exposed on the intact cell surface, but become exposed upon apoptosis. We also showed that signal-transducing TREM2 ligands different from aminophospholipids might be present in membrane fractions of 5-month-old mouse cerebral cortex. These results may suggest that TREM2 regulates microglial function by transducing intracellular signals from aminophospholipids on apoptotic cells, as well as unidentified ligands in the membranes of the cerebral cortex.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-422 脆弱X症候群モデルショウジョウバエのシナプス形態および行動の欠陥への薬理学的応用 Sayuri Shiojima（塩島さゆり），Yutaro Matsumoto（松本悠太郎），Junji Yamauchi（山内淳司），Takako Morimoto（森本高子）東京薬科大院生命科学分子神経科学 Recently, we established an experimental system of prepulse inhibition (PPI) in Drosophila larvae (Matsumoto et al., 2018). This is the first description of PPI in Drosophila. PPI is a phenomenon in which the startle response to the sound stimulus (pulse) is greatly suppressed by a precedent weak stimulus (prepulse). It is one of psychophysiological indicators and used for the diagnosis of psychiatric disorders such as schizophrenia. Since Drosophila has long history as an experimental animal and various genetic and molecular tools can be applied, our PPI system in Drosophila is very useful to study the mechanisms of mental disorders and search for drugs potentially effective for these diseases. We have shown that PPI was lost in larvae of fragile X syndrome (FXS) fly model. FXS is known as a mental disorder and caused by mutation in the fmr1 gene. Fmr1 protein (FMRP) is an RNA binding protein and various phenotypes have been reported in FXS model. One of these phenotypes is the enhancement of metabotropic glutamate receptor (mGluR) signaling. In this study, we aimed to investigate the influence of candidate drugs on several phenotypes of FXS. First, we focused on the drugs affecting mGluR signal and investigated effects on the synaptic morphology of larval neuromuscular junctions (NMJs). When ACPD, an agonist of Group I/II of mGluR, was fed to wild type larvae, the total area of synaptic boutons was significantly increased as compared with control larvae, which were not fed with ACPD. On the other hand, DHPG, an agonist of mGluR Group I, did not affect synaptic morphology, which was consistent with the knowledge that Drosophila has only mGluR Group II but not Group I. Zhang et al., (2001) described that the synaptic terminals of FXS larval NMJs enlarged and has an excessive number of synaptic boutons compared to that in the wild type. Taken together, our results suggest that excess activation of mGluR Group II may affect synaptic morphology in Drosophila FXS. Next, in order to investigate candidate drugs that may be useful to treat FXS, we investigated effects of several drugs, metformin and chlorpromazine on loss of PPI in FXS. Metformin has been reported to be a potential FXS therapeutic chemical and chlorpromazine is a typical antipsychotic drug. Currently, we feed FXS larvae with these drugs and investigate whether PPI is observed or not and will discuss effects of these drugs.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-423 化学物質誘発性自閉症モデルラットの行動異常とそのメカニズム Rie Matsufusa（松房利恵）¹，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 Multiple environmental agents, including medical drugs, have been associated with an increased risk of autism and mental disorders. In autism, some structural abnormalities in the cerebellum have been reported. Especially, a reduction in size and number of Purkinje cells is revealed in the postmortem human studies. We have reported the development of Valproate (VPA)-treated autistic animal models, in which excess development of Purkinje cells and excess folds in some cerebellar lobule were observed. Additionally, VPA-administrated mature rats showed ADHD-like behavior. They had less impairment of social interaction and much curiosity. In this study, we report the behavior and cerebellar development of Trichostatin (TSA)- or Chlorpyrifos (CPF)-administrated rats and compare the difference between TSA-, CPF- and VPA-ad. animals. CPF is one the well-known autism inducer, and TSA is a HDAC inhibitor in same group of VPA. An observed animal was placed on the open-field circle (80 cm in diameter) with an overhead camera, and observed 10 minutes. All tracking data was analyzed with Tracker Video Analysis and Modeling Tool produced by D. Brown. Chemical-administrated or vehicle male rats from 6 to 8 weeks were tested their individual behavior, social interaction to same- or bigger-size strange rats and target interaction to well-known or new objects. In the individual behavior, both CPF and TSA animals moved less than VPA and control animals. In the social interaction to bigger-size stranger, the behavior of CPF animals showed similar to VPA animals, whereas TSA animals showed fear than VPA one. In target behavior to objects, in turn, CPF animals show nervous, while VPA animals got excited. We suggest TSA-administrated animals would show some similar behavior to VPA-animals due to similar neuronal background, while CPF-administrated animals would have different neuronal impairing points and behavior.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-424 胎児期葉酸欠乏マウスにおける認識・記憶能の低下について Misato Yoshikawa（吉川弥里），Hiroaki Aso（阿蘓寛明），Masahiko Watanabe（渡辺雅彦），Katsuya Suemaru（末丸克矢）就実大薬公衆衛生 Folate is important for normal fetal development and growth. Animal experiments and human studies have suggested that fetal folic acid exposure influence postnatal brain development and reduce the risk of psychiatric disorder. And prenatal folate deficiency is related to neural tube defects. However, the association between prenatal folate deficiency and developmental disorder such as autism remains unknown enough. In this study, we investigated whether prenatal folate deficiency is associated with the offspring's behavior related to developmental disorder. ICR mice were fed a control diet of chow (2 mg folate/kg diet) or a folate deficiency diet of chow (0.3 mg folate/kg diet) from embryonic day 1 (E1) to delivery. The serum folate concentration was decreased by 33% after 3 weeks of dietary folate restriction. We evaluated the function of sociability and memory/recognition in male offspring mice at age 7-10 week. The preference for social novelty, but not sociability, was impaired in the male offspring of prenatal folate deficiency compared with the male control offspring in the three-chambered social test. Additionally, male offspring of prenatal folate deficiency decreased the function of spatial memory in Morris water maze test and object recognition memory in novel object recognition test. Next, we performed histological analysis by Hematoxylin-Eosin staining. The size of hippocampus didn't change in the male offspring of prenatal folate deficiency. These results may suggest that prenatal folate deficiency affects function of memory and recognition without the histological change of hippocampus.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-425 遺伝子補充は、Cyclin-dependent kinase-like 5 (CDKL5)欠損マウスのシナプス・行動学的表現型をレスキューするか？ Teruyuki Tanaka（田中輝幸）¹，Elizabeth Zheng（Zheng Elizabeth）¹，Shin-ichi Muramatsu（村松慎一）²，Masashi Mizuguchi（水口雅）¹ ¹東京大院医発達医科学 ²自治医大医神経内科 Loss-of-function (LOF) mutations in the Cyclin-dependent kinase-like 5 (CDKL5) gene cause severe neurodevelopmental disorders accompanied by early-onset intractable epilepsies, i.e. CDKL5 deficiency disorder (CDD). Previously, we have generated Cdkl5 KO mice, and demonstrated postsynaptic overaccumulation of GluN2B-containing N-methyl-D-aspartate (NMDA) receptors in the hippocampus, significant hyperexcitability to NMDA, significantly enhanced anxiety-like behaviors, and impaired acquisition and retention of spatial reference memory. But the questions on how CDKL5 regulates postsynaptic localization of the GluN2B and emotional and cognitional behaviors are unanswered. To answer these questions, we sought to test whether the gene replacement restores the synaptic and behavioral phenotypes in Cdkl5 KO mice. We have generated an adeno-associated virus (AAV) vector in which the brain isoform of the human CDKL5 gene is expressed under the synapsin I promoter. We are examining the optimal age, administration route, and gene dosage of the AAV vector for effective transduction to cortical neurons. Detailed analyses of synapses and behaviors of the transduced mice will follow. The outcome of this study should clarify whether the CDKL5 kinase activity by itself is sufficient for the normal function of the postsynaptic NMDA receptors or behaviors, or any precondition is necessary for the CDKL5 activity to regulate these functions. These data should demonstrate the effect and scope of the gene replacement therapy for the CDD.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-426 Cell-autonomous roles of RBFOX3 in the brain function Hsien-Sung Huang（Huang Hsien-Sung） National Taiwan University RBFOX3 (RNA binding protein, fox-1 homolog (C. elegans) 3), an RNA alternative splicing regulator, is exclusively expressed in neurons. The other well-known alias for RBFOX3 is NeuN which is widely used in the neuroscience community as a marker for postmitotic neurons. Despite its popularity, the functional roles of RBFOX3 are still unclear in the brain. Importantly, dysfunctional RBFOX3 is associated with cognitive impairments and epilepsy; however, the pathogenesis of these disorders is not well-studied. In our previous studies, we used a Rbfox3 knockout mouse to recapitulate symptoms from persons with dysfunctional RBFOX3. It provides a causal relationship between deficits of RBFOX3 and its related human brain diseases. Our Rbfox3 knockout mice demonstrated decreased anxiety-related behaviors, increased seizure susceptibility, and decreased learning ability. We also showed that deletion of Rbfox3 contributes to dysfunctional postnatal neurogenesis and synaptogenesis. Furthermore, we displayed that RBFOX3 is dispensable for visual function, which is an important piece of evidence to interpret the behavioral results from Rbfox3 knockout mice. Currently, we are interested in investigating the cell-autonomous roles of RBFOX3 in the brain function with Rbfox3 conditional knockout mice. Our preliminary data showed that RBFOX3 works in a cell-type-specific manner in the brain. Our work will give us a significant insight regarding how RNA alternative splicing operates in a cell-autonomous manner within our brains.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-427 遺伝子 Slc52a3の欠損はマウス胎仔大脳皮質の発達に影響する。 Congyun Jin（金叢芸）^1,2，Atsushi Yonezawa（米澤淳）^1,2，Hiroki Yoshimatsu（吉松宏樹）^1,2，Satoshi Imai（今井哲司）¹，Madoka Koyanagi（小柳円花）^1,2，Kaori Yamanishi（山西香里）^1,2，Shunsaku Nakagawa（中川俊作）¹，Tomohiro Omura（大村友博）¹，Takayuki Nakagawa（中川貴之）¹，Kazuo Matsubara（松原和夫）¹ ¹京都大学医学部附属病院薬剤部 ²京都大学大学院薬学研究科 Background: Riboflavin transporter 3 (RFVT3), encoded by the SLC52A3 gene, plays a role in the homeostasis of riboflavin. It is highly expressed in the small intestine, kidney, testis and placenta, and its mutations are linked to a rare neurological disorder, Brown-Vialetto-Van Laere syndrome. Our previous study demonstrated that Slc52a3 knockout ( Slc52a3-/-) mice exhibits neonatal lethality, riboflavin deficiency and metabolic disorder. Here, we investigated the inference of Slc52a3 gene disruption to the development of brain using Slc52a3-/- mice. Methods: Slc52a3-/- mice were obtained from Knockout Mouse Project repository. Brains were dissected at embryonic day 13.5 (E13.5) and immediately after delivery (P0). Riboflavin concentrations were measured by HPLC. Morphological changes in the whole brain at P0 were observed by histochemistry. Expressions of neuron and intermediate neural progenitors (INPs) makers (Tuj1 and Tbr2) were evaluated by immunohistochemical analysis. Results: Brain riboflavin levels were decreased in Slc52a3-/- mice compared with WT mice. Histological and immunohistochemical analysis of brains in Slc52a3-/- newborn pups showed hypoplasia and global reduction of cerebral cortex. The cortical layers of Slc52a3-/- pups were thinner. Evaluation of Slc52a3-/- embryos at E13.5 indicated that Tuj1-positive neurons and Tbr2-positive INPs were significantly decreased, compared with WT mice. Conclusions: Rfvt3 can be important for the embryonic development of cerebral cortex in mice.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-428 母胎免疫活性化による発達障害モデルマウスの行動とドーパミン回路の解析 Gen Murakami（村上元）¹，Takayuki Murakoshi（村越隆之）² ¹埼玉医科大学医学部教養教育 ²埼玉医科大学医学部生化学教室 Developmental disorders such as attention-deficit hyperactivity disorder (ADHD), autism spectrum disorder (ASD) and schizophrenia remain a serious public health problem worldwide. Although many researchers have investigated the pathogenesis using animal models, the mechanism has not been elucidated and further understanding is important. Maternal immune activation (MIA) is one of the most frequently used animal models for the study of developmental disorders. This model mimics the environmental risk factor of immune activation during the fetal period by treating pregnant animals with immune activating agents such as poly I:C. In this model, the timing and doses of the treatments and different rodent species result in different behavioral consequences in adults. Therefore, it is important to identify the behavioral phenotypes according to the MIA treatment paradigm particularly used in the experiment to relate the phenotypes to the underlying mechanisms. In this study, we treated fetuses at GD9 with poly I:C (5 mg/kg body weight), and the poly I:C-treated male C57BL/6 mice were analyzed for locomotor activity, social behaviors and pre-pulse inhibition in adults, which are characteristically impaired in ADHD, ASD and schizophrenia, respectively. To screen the genes related to the pathogenesis, we will perform RNA-seq experiment for comprehensive analysis of gene expression in brain regions related to the dopamine system of the poly I:C-treated mice because dysfunction of the dopamine system is common in these disorders. To examine the detailed effect of MIA on the dopamine system, we will analyze expression of dopamine-related genes such as dopamine receptor D1/D2, dopamine transporter and tyrosine hydroxylase in the same brain regions by qPCR. These results will provide a novel landmark particularly in the dopamine system in the pathogenesis of developmental disorders.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-429 ピロロキノリンキノンは生後10日間マウスの頬ひげを切除することにより生じる触覚認知の機能及び社会性行動発達の異常を改善する。 Hitomi Soumiya（宗宮仁美），Hiromi Araiso（荒磯ひろみ），Ayumi Godai（五代あゆみ），Shoei Furukawa（古川昭栄），Hidefumi Fukumitsu（福光秀文）岐阜薬大・分子生物 Abnormalities in tactile perception, such as sensory defensiveness, are common features in autism spectrum disorder (ASD). The deficits in tactile perception might associated to impairment of social communication skills in ASD. However, the influence of tactile perception deficits on the development of social behaviors remains uncertain, as do the effects on neuronal circuits related to the emotional regulation of social interactions. In neonatal rodents, whiskers are the most important tactile apparatus, so bilateral whisker trimming is used as a model of early tactile deprivation. To address the influence of tactile deprivation on adult behavior, we performed bilateral whisker trimming in mice for 10 days after birth (BWT10 mice) and examined social behaviors, tactile discrimination, and c-Fos expression, a marker of neural activation, in adults after full whisker regrowth. Adult BWT10 mice exhibited deficient in the social behavior, whisker-dependent tactile perception. Based on the c-Fos expression pattern analysis, hyperactivity was found in BWT10 amygdala circuits for processing fear/anxiety-related responses to height stress but not in circuits for processing reward stimuli during whisker-dependent cued learning. Pyrroloquinoline quinone (PQQ) is an essential nutrient and is known to play important roles in central nervous system via modification of glutamatergic NMDAR activity. Next, we examined the effect of neonatal PQQ administration on the behavior abnormalities of BWT10 mice. The results indicated that the PQQ treatment significantly prevent abnormal behavioral development of BWT10 mice, as detected in gap crossing, social interaction and social-dominance tube tests. In addition, PQQ also improve mal-development of emotional system, judging from c-Fos expression pattern induced by psychological elevated-platform stress. These data suggest that PQQ might be one of promising candidates for the therapeutic drug of neurodevelopmental disorders.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-430 小児神経発達障害で同定されたCACNA1G変異におけるチャネル機能の検討 Misako Kunii（國井美紗子）¹，Hiroshi Doi（土井宏）¹，Shunta Hashiguchi（橋口俊太）¹，Masaki Okubo（大久保正紀）¹，Hideyuki Takeuchi（竹内英之）¹，Toyojiro Matsuishi（松石豊次郎）²，Yasunari Sakai（酒井康成）³，Mizue Iai（井合瑞江）⁴，Taro Ishikawa（石川太郎）⁵，Noriko Miyake（三宅紀子）⁶，Hirotomo Saitsu（才津浩智）⁷，Naomichi Matsumoto（松本直通）⁶ ¹横浜市立大学医学部神経内科学・脳卒中医学 ²聖マリア病院小児総合研究センター・レット症候群研究センター ³九州大学大学院医学研究院成長発達医学 ⁴神奈川県立こども医療センター神経内科 ⁵東京慈恵会医科大学薬理学講座 ⁶横浜市立大学大学院医学研究科遺伝学 ⁷浜松医科大学医学部医化学講座 Variants of CACNA1G which encodes Ca_V3.1 have been reported to be associated with various neurological disorders. In this study, we performed exome analysis on genomic DNAs from 348 patients with neurodevelopmental disorders. As results, we found three patients with de novo CACNA1G mutations. The effects of identified Ca_V3.1 mutations on channel functions were studied by the whole-cell patch clamp technique using HEK293T cells overexpressing wild type or mutant channels. Two patients diagnosed as Rett syndrome and West syndrome were found to have known pathological CACNA1G mutations: c.2881G>A [p.A961T] and c.4591A>G, [p.M1531V], respectively. One patient diagnosed as Lennox-Gastaut syndrome was revealed to have an unreported heterozygous mutation: c.3817A>T, [p.I1273P]. While changes of electrophysiological properties of known mutations were almost same as those described in the previous report, the clinical symptoms and MRI findings were ununiform. It is suggested that the clinical spectrum of the diseases caused by CACNA1G mutations are heterogeneous even in the same mutation. The patient with novel p.I1273P mutation showed developmental delay, intractable seizure and mild cerebral atrophy in MRI. Electrophysiological study demonstrated that p.I1273P mutation did not change the T-type Ca²⁺ current though multiple in silico algorithms predicted p.I1273P as disease causing. The pathological significance of I1273P mutation remains uncertain although we speculate that p.I1273P mutation may cause subtle Ca_V3.1 functional changes undetectable by our patch clamp methods. In conclusion, we report three neurodevelopmental disorders patients with CACNA1G mutation including one unreported variant. Functional study revealed that two of three mutations changed the T-type Ca²⁺ currents. Our study would contribute to understand the genotype-phenotype correlations of the patients with CACNA1G mutations.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-431 時計遺伝子Per3の大脳皮質形成における機能解明 Mariko Noda（野田万理子），Hidenori Tabata（田畑秀典），Hidenori Ito（伊東秀記），Koh-ichi Nagata（永田浩一）愛知県心身障害者コロニー発達障害研 Period gene (Per) is the first identified circadian clock component in Drosophila. Mammalian Per family has three homologs (Per1, 2, and 3). While Per1 and Per2 expression profiles have been clarified in the developing mouse cortex, Per3 distribution pattern is largely unknown. Given that circadian dysregulation is involved in the pathogenesis of several neuropsychiatric diseases, clock genes may participate in brain function and development. We thus focused on and examined the role of Per3 in the development of the mouse cerebral cortex. We first have performed in situ hybridization to determine the Per3 mRNA expression profile in the developing mouse brain. Per3 was expressed in the ventricular/subventricular zones and cortical plate (CP) in E16 mouse cortex, and showed time-dependent increase until P10. We then performed acute knockdown of Per3 with in utero electroporation and found abnormal positioning of cortical neurons, which was rescued by RNAi-resistant Per3. Since the abnormal positioning of Per3-deficient neurons may be due to reduced migration velocity, we have performed time-lapse imaging of migrating cortical excitatory neurons in the intermediate zone (IZ) and CP. As the result, Per3-deficient cells were stranded at the IZ/CP boundary and their migration velocity was reduced compared with control cells. Per3-deficient cells also showed impaired axon extension and dendritic arbor formation in vivo. Taken together, Per3 was found to play a pivotal role in corticogenesis via regulation of excitatory neuron migration and synaptic network formation. Loss-of-function of PER3 was likely to be related to neuropsychiatric disease etiology.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-432 バルプロ酸誘導自閉症モデル雌マウスにおける社会性障害行動を検出できる行動パラダイムの確立 Chiharu Tsuji（辻知陽）¹，Pinyue Fu（傅品悦）¹，Zing Zhong（钟静）¹，Takahiro Tsuji（辻隆宏）^1,2 ¹金沢大学子どものこころの発達研究センター ²福井大学 The characteristic symptoms of Autism spectrum disorder (ASD) are deficits in the social skills, disruption in the speech and nonverbal communication, and the repetitive behaviors. Currently, due to the higher prevalence of ASD in males, little attention has been paid to the female symptoms. However, the gender roles of female in the society may hide the behavioral deficits, and so the number of females with ASD could be estimated much more. Therefore, it is necessary to extend our study to examine the female ASD symptoms. Valproic acid (VPA) is a widely used drug to treat epilepsy, migraine and bipolar disorder, but it is shown to increases the risks of malformation, ASD, and mental retardation in children, when used during pregnancy. Recently, many studies have shown the prenatally exposed VPA mice display ASD-like behaviors such as hyper locomotion in a novel environment, anxiety-like behaviors, deficits in social interaction, and recognition memory impairment. However, most of the behavioral analysis was performed in males and not in females. Here, we focused to establish the social behavioral paradigm in the VPA female mice. We demonstrated that the examination of the interaction-induced ultrasonic vocalization and maternal behaviors may be useful experimental paradigm to investigate the female social responsiveness.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-433 ニューレキシン遺伝子のSNP(rs858932)は健常成人において白質体積とSQスコアに関連がある Yuka Shiota（塩田友果）¹，Izumi Matsudaira（松平泉）¹，Hikaru Takeuchi（竹内光）²，Chiaki Ono（小野千晶）³，Hiroaki Tomita（富田博秋）³，Ryuta Kawashima（川島隆太）^2,4，Yasuyuki Taki（瀧靖之）^1,2,4,5 ¹東北大院機能画像医学研究 ²東北大加齢研認知機能発達 ³東北大精神医学 ⁴東北大スマートエイジングリサーチセンター ⁵東北メディカルメガバンク機構 Neurexin-1 (NRXN1) gene encodes the neurexin-1α and neurexin-1βproteins that function as pre-synaptic neural adhesion molecules. The neurexin superfamily gene variants has been recognized as associated with Autism Spectrum Disorder (ASD). rs858932 (C/G) is a single nucleotide polymorphism (SNP) of NRXN1, and the G-allele carrier heterozygotes had increased white matter volume of frontal lobe compared to `CC' genotype (Voineskos et al., 2011). However, it is unclear whether this SNP correlates with Empathy Quotient (EQ) - Systemizing Quotient (SQ) score and brain structure. Therefore, the aim of this study was to investigate the relationship among the alleic status of rs858932, EQ-SQ score and brain structure of the area associated with ASD in healthy adults. The participants were 312 healthy Japanese adults (age range: 18-27 years). EQ/SQ score was assessed using the EQ/SQ -Japanese version. We isolated DNA from saliva and genotyped the NRXN1 rs858932 using TaqMan analysis. We collected brain 3D T1-weighted images of MPRAGE protocol using 3T Phillips Intra Achieva scanner. Preprocessing of the images and statistical analyses were performed using SPM12. We employed ANCOVA to assess the gray and white matter volume differences between subjects with `CC' genotype and `GC' genotype by adjusting for age, sex, and total brain volume. The statistical significance was set at p<0.05. We found a significant correlation between SQ score of subjects with `GC' genotype and white matter volume of the bilateral pallidum, basal forebrain, and accumbens (MNI coordinates=[17, 8, -3], cluster-level: k=1728, p=0.001) (MNI coordinates=[-9, -3, -15], cluster-level: k=1316, p=0.003). Subjects with `GC' genotype who have increased white matter volume of the bilateral basal ganglia pathway had a higher Systemize score than `CC' genotype (F=5.65, p=0.018). Individuals with ASD had significantly larger pallidum and lateral ventricle volumes compared to healthy volunteers (Turner et al., 2016). In addition, the increase in white matter volume may be due to an increase in the number of synapses per neuron and myelination (Taki et al., 2013), so it is possible that there is an acceleration of myelination in `GC' genotype. Therefore, NRXN1 polymorphisms would confer an intermediate phenotype related to ASD, and this indicates the SNP may confer the phenotype of autistic traits. Our findings in healthy adults would contribute to understanding the mechanisms of ASD.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-434 C57BL6/Jマウスにおいて他者の存在が自発運動を促進する Kaichi Yoshizaki（吉崎嘉一），Masato Asai（浅井真人），Atsuo Nakayama（中山敦雄）愛知県心身障害者コロニー発達障害研究所 Autism spectrum disorder (ASD) is a developmental disorder characterized by severe impairment in socialization and in particular, "lack of spontaneous seeking to share enjoyment, interests, or achievements with other people" and "lack of social or emotional reciprocity". Recent study revealed that susceptibility to the presence of others was impaired in ASD, i.e. deficits in social facilitation. Social facilitation is referred to social facilitation refers to the increase in frequency and intensity of an individual's behavior due to the mere presence of other individuals. Here we developed a special breeding cage with running wheel and examined the effect of observer on voluntary exercise of highly social C57BL6/J and poorly social BALB/c mice. Wheel-running performance of C57BL6/J mice was significantly increased in the presence of observer, compared to that in the absence of observer. In contrast, the presence of observer did not affect the wheel-running performance of BALB/c mice. Moreover, observed experience did not impact subsequent wheel-running performance of either C57BL6/J or BALB/c mouse, i.e. no effect on observational learning. These studies suggest that highly social C57BL6/J mice, but not poorly social BALB/c mice, exhibited social facilitation on wheel-running performance. Our system for social facilitation would provide a useful tool to examine pathophysiology behind communication and socialization deficits in ASD.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-435 加速度センサを用いた自閉症モデル霊長類の生体リズム機構の解明 Madoka Nakamura（中村月香）^1,2，Akiko Nakagami（中神明子）^1,3，Keiko Nakagaki（中垣慶子）¹，Nobuyuki Kawai（川合伸幸）²，Noritaka Ichinohe（一戸紀孝）¹ ¹国立精神・神経セ神経研微細構造 ²名古屋大院情報 ³日本女子大人間社会心理 Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder characterized by core impairments associated with social interactions and repetitive behaviors. Recent human studies have shown that ASD is accompanied by abnormalities in daily activity, sleep patterns and endocrine function as well as social communication skills. To investigate whether similar aberrations in biological rhythms can also be detected in a non-human primate model of ASD, we used the common marmoset model induced by prenatal exposure to valproic acid (VPA) since our previous studies indicated that the VPA marmosets showed ASD-like behaviors. Home-cage activity and sleep-wake pattern of adult marmosets individually housed under a 12h light/dark cycle were measured using an unobtrusive collar-worn actigraphy device (Actiwatch Mini; Camtech, Cambridge, UK) for three weeks. The data collected from the first two weeks were excluded from the analysis to avoid confounding effects from their unfamiliarity of the apparatus. The VPA marmosets demonstrated significantly higher activity in the light period than a VPA-unexposed (UE) group. Although there was no significant difference in sleep latency between the two groups, the duration of immobility during the dark period was considerably longer in the VPA marmosets than in the UE controls. Besides, there was a negative correlation between the light and dark period activity levels in the VPA group, revealing that the activities of the VPA marmosets are influenced by their activities on a previous day while there was no carry-over in the activities of the UE group. These results indicate that the VPA marmosets may have ASD-like phenotypes including abnormal biological rhythms, suggesting that our primate model of ASD is one of the clinically useful animal models contributing to the understanding of neurodevelopmental symptoms associated with ASD.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-436 Ca2+-dependent activator protein for secretion 2 (CAPS2)ノックアウトマウスはオキシトシン分泌の低下と社会行動の障害を示す Shuhei Fujima（藤間秀平）¹，Ryosuke Yamaga（山鹿亮祐）¹，Haruka Minami（南春花）¹，Rina Maniwa（真庭梨奈）¹，Yo Shinoda（篠田陽）²，Manabu Abe（阿部学）³，Kenji Sakimura（﨑村建司）³，Yoshitake Sano（佐野良威）¹，Teiichi Furuichi（古市貞一）¹ ¹東京理科大学理工学部応用生物科学科 ²東京薬科大学薬学部 ³新潟大学脳研究所 CAPS2 is a protein that promotes the release of peptidergic neurotransmitters, peptide hormones and monoamine neurotransmitters via exocytosis of dense-core vesicles. CAPS2 knockout (KO) mice cause abnormalities in synaptic development and function as well as social behavior. There are some reports showing a possible association of CAPS2 with autism spectrum disorder (ASD) due to finding of expression of a rare alternative splicing variant lacking axonal release function as well as of copy number variation and single nucleotide variation in some patients with ASD. However, it is yet unknown how CAPS2-mediated release mechanism of transmitters and hormones contributes to ASD. Interestingly, CAPS2 is expressed in oxytocin (OXT)-producing neurons in the hypothalamic paraventricular nucleus (PVN) of mice. OXT is known to be a social neuropeptide/peptide hormone whose administration could improve symptoms of ASD. From this point of view, we got an idea that CAPS2 regulates OXT release, thereby being involved in social behavior. To address this question, we compared serum OXT levels between control and CAPS2 KO mice by OXT ELISA. As a result, CAPS2 KO mice showed decreased serum OXT levels compared to those of control mice. We also analyzed OXT immunoreactivity in the posterior lobe of the pituitary (PP), in which OXT is released from OXT terminals to blood vessels, in CAPS2 KO and OXT neuron-specific conditional knockout (cKO) mice. The results showed the accumulated levels of OXT in the PP of both CAPS2 KO and cKO mice compared to control mice. Taken together, we suggest that the CAPS2-mediated OXT releasing mechanism is associated with OXT-related social behavior.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-437 オキシトシン受容体発現を正確に再現できるレポーターマウス・Creドライバーマウス Yukiko U. Inoue（井上-上野由紀子）¹，Ryosuke Kaneko（金子涼輔）²，Yuki Morimoto（森本由起）¹，Takayoshi Inoue（井上高良）¹ ¹国立精神・神経セ神経研疾病６ ²群馬大院医 Oxytocin (Oxt), a natural brain peptide produced within the hypothalamus, plays an important role in regulating social behaviors. It is also expected as a potential therapeutic for social deficits in autism spectrum disorders. Oxytocin receptor (Oxtr) is abundantly expressed in the brain and its relationship with social behaviors have also been extensively studied in mouse brains. However, multiple independent tools have been used to visualize Oxtr expressions and the resulted expression patterns are not fully consistent with each other. First, commercially available antibodies don't have enough specificities to Oxtr protein, and detection of Oxtr mRNA by in situ hybridization has been reported to be difficult. Secondly, several reporter lines and Cre recombinase driver lines have been generated by BAC transgenesis or ES cell-based gene targeting, yet they have somewhat different expression profiles probably because of the different integrity and context for their reporter/driver expressions in each line. Here, we employ CRISPR/Cas9 genome editing technology to generate novel knock-in mouse lines to precisely recapitulate the endogenous Oxtr expression profiles. We establish Oxtr-PAtag-T2A-tdTomato reporter line and Oxtr-PAtag-T2A-iCre driver line, in which fluorescent reporter or codon-improved Cre recombinase is bicistronically expressed within the Oxtr-expressing cells depending on T2A peptide cleavage. Furthermore, in order to directly determine the endogenous Oxtr localizations, PA-tag is fused to C-terminus of the receptor protein. By using these knock-in lines, we carefully analyze the co-localization of tdTomato/iCre with PA-tag in the brain tissues to obtain a precise Oxtr expression atlas. As the results, in addition to the previously reported brain regions where Oxtr expressions have been detected, we confirmed the receptor expressions in the other regions such as the nucleus accumbens. It is noteworthy that Oxtr-PAtag-T2A-tdTomato reporter line can be maintained in the homozygous state without the laborious genotyping procedure and the tdTomato fluorescence can immediately illuminate brain tissues without immunostaining. Our knock-in mouse lines could thus serve useful tools to investigate roles of Oxt-Oxtr system in the social brain.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-438 自閉スペクトラム症者におけるクロスモーダル干渉効果に対するラバーハンド提示の効果 Makoto Wada（和田真）^1,2，Hanako Ikeda（池田華子）¹，Shinichiro Kumagaya（熊谷晋一郎）³ ¹国立障害者リハビリテーションセンター研究所脳機能系障害研究部 ²静岡大学情報学部 ³東京大学先端科学技術研究センター Participants sometimes perceive successive sensory signals as a flow ("" apparent motion "" ), and multisensory signals interact each other at specific situations ("" cross-modal dynamic capture "", CDC) (Soto-Faraco et al., 2002). Tactile temporal order judgment (TOJ) at moderately short stimulus onset asynchronies (SOAs) is affected by visual distracters that elicit ""apparent motion"" (Shibuya et al., 2007). Furthermore, presentation of a rubber hand enhances this effect (Wada & Ide, 2016). In this study, we examined the effect of rubber hand presentation on CDC in individuals with autism spectrum disorder (ASD) when they performed TOJ. Because they sometimes have atypical characters in sensory processing (Foss-Feig et al., 2010). In the present study, successive two tactile stimuli were delivered to the index and ring fingers of a participant's right hand, which was always hidden in an opaque box. On the box, a rubber hand was placed in a forward or inverted direction (Forward / Inverted conditions). At the timing of the tactile stimulus delivery, visual stimuli, which were provided by lighting LEDs on fingers of the rubber hand, were simultaneously delivered with congruent or incongruent orders to the tactile stimuli with a same SOA, and participants who had diagnosis of ASD (n = 11) were required to judge temporal orders of the tactile stimuli regardless of the visual distracters. When incongruent visual stimuli were delivered, judgment was notably reversed at the moderately SOAs (200-300ms) in both the Forward and Inverted conditions. There was no significant difference between the both conditions. With regards to the degree of the reversals, there were large individual differences, compared with responses of typically developing participants in the previous study (Wada & Ide, 2016). In contrast, the participants judged quite precisely with congruent visual stimuli. Present results suggest that large individual differences in the CDC effects exist in individuals with ASD, and the presentation of the rubber hand in a forward direction had different effects on them. Large individual difference in the task may reflect atypical sensory processing in each ASD individuals.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-439 バルプロ酸誘発性自閉症モデルマーモセットにおけるシナプス構造と機能の生後発達 Satoshi Watanabe（渡辺恵）¹，Tohru Kurotani（黒谷亨）²，Tomofumi Oga（小賀智文）¹，Keiko Nakagaki（中垣慶子）¹，Jun Noguchi（野口潤）¹，Noritaka Ichinohe（一戸紀孝）^1,2 ¹国立精神・神経セ神経研微細構造 ²理研CBS 高次脳機能分子解析 Autism spectrum disorder (ASD) is characterized by impaired social interaction and communication, restricted interests and repetitive behaviors. Cortical neurons from ASD patients have abnormalities in the number and structure of dendritic spines. How the synaptic pathology develops during postnatal development, or how it is related to neural circuit functions has been poorly understood. To clarify these issues, we used the common marmoset, which shows similar cortical structure and development to humans. We have reported an ASD model marmoset with exposure to valproic acid (VPA) in utero (Yasue et al. Behav. Brain Res. 2015). These marmosets show abnormalities in complex social behavior as seen in human ASD patients. We made acute slices of the prefrontal cortex (areas 8 and 9) of VPA-exposed and unexposed neonates and infants (3 and 6 months), and performed electrophysiological and structural analyses in layer 3 pyramidal neurons. In unexposed animals, the density of dendritic spines initially increased toward 3 months, and then decreased toward 6 months, showing synaptic pruning after 3 months. In VPA-exposed neonates, the spine density was lower than in UE neonates. In VPA-exposed infants at 6 months, in contrast, the spine density was higher than in unexposed infants. Consistently with these data, the frequency of miniature EPSCs tended to be lower in VPA-exposed neonates and higher in VPA-exposed infants at 6 months. We also recorded EPSCs and IPSCs evoked by stimulation of layer 4, and revealed that the E/I ratio was reduced in VPA animals at 3 months. These results suggest that synaptic structure and function are modulated in VPA animals in a time-dependent manner during postnatal development, and these modulations may be related to development of ASD in infants.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-440 15q11-13重複自閉症モデル動物の前頭前皮質における興奮性/抑制性バランス異常に対するセロトニン受容体の役割 Fumihito Saitow（斎藤文仁）¹，Toru Takumi（内匠透）²，Hidenori Suzuki（鈴木秀典）¹ ¹日本医科大学　薬理学 ²理化学研究所CBS Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by three symptoms: deficits in social communication, impaired language and communication, and stereotyped and repetitive behaviors. A chromosome 15q11-13 paternal duplication (15q dup) mouse corresponding to cytogenetically frequent CNV in ASD have core symptoms. Prior to this study, we found that 15q dup mice showed a hypoactive state in the dorsal raphe nucleus (DRN) and a hyposerotonergic state in the brain. Also, we found excitatory/inhibitory (E/I) balance of synaptic transmission at the somatosensory (S1) cortex was shifted to excitatory state in 15q dup mice. Together with the serotonin (5-HT) restoration using SSRI treatment at the postnatal period, hypo-neuronal activity in the DRN and E/I imbalance in the S1 cortex were ameliorated. In this study, we explored the roles of 5-HT-mediated modulation in the prefrontal cortex (PFC) using 15q dup mice. Using whole cell recording from layer 5 (L5) neurons in acute adult brain slices of the PFC (~3 month-aged), we first found 5-HT elicited significantly larger 5-HT_1A receptor-mediated outward currents in 15q dup mice than in WT. In contrast, baclofen-induced GABA_B receptor-mediated outward currents were not significantly different between genotypes. Further, the E/I ratio at the PFC L5 neurons were higher in 15q dup mice, consistent with our previous study observed in the S1 cortex. We found that two mechanisms underlying the imbalance in the PFC network. One is differences in synaptic inputs of both glutamatergic and GABAergic transmissions at fast-spiking interneurons (FSINs). FSINs in 15q dup mice shifted to inhibitory state. Second is difference in level of 5-HT₂ receptors (5-HT₂Rs) activation in FSINs. The E/I ratio in WT was largely elevated under the blocking of 5-HT₂Rs, while the same treatment had small impact on the E/I ratio in 15q dup. These results suggested that the level of 5-HT₂R-activation in the FSINs contributed to maintaining inhibitory tone at the PFC network. Taken together, the levels of both 5-HT_1A and 5-HT₂ receptors expressions and their activities at the PFC are important for regulating the optimal balance of the synaptic transmission in the PFC. Therefore, this study proposes that interventions of 5-HT-related neuronal modulation can be considered as a promising strategy for ASD treatment.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-441 自閉症様なNSF欠損マウスは異常なシナプス可塑性を示す Min-Jue Xie（謝敏かく）^1,2,3，Keiko Iwata（岩田圭子）^1,2,3，Yasuyuki Ishikawa（石川保幸）⁴，Yugo Fukazawa（深澤有吾）^1,2,5，Hideo Matsuzaki（松﨑秀夫）^1,2,3 ¹福井大学　子どものこころの発達研究センター　脳機能発達研究部門 ²福井大学　ライフサイエンスイノベーションセンター ³大阪大学大学院　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 ⁴前橋工科大・システム生体工学 ⁵福井大学大学院　医学系研究科　脳形態機能学領域 Autism, characterized by profound impairment in social interactions and communicative skills, is the most common neurodevelopmental disorder. Many studies on the mechanisms of autism have focused on the serotonergic system, but its underlying molecular mechanisms remain controversial. In our previous report, we reidentified N-ethylmaleimide-sensitive factor (NSF) as new serotonin transporter (SERT) binding protein. In this study, we generated the NSF^+/- mice and investigated their behavioral phenotypes. As previous report has already shown that NSF is necessary for AMPA receptors location in the synapse, we examined AMPA receptors location in the synapse of the NSF^+/- mice by using freeze-fractured replica-immunolabeling study at first, and revealed the significant decrease in postsynaptic expression of AMPA receptors in CA1 of the hippocampus. Interesting, we also found the membrane expression of SERT half-reduced in the raphe of the NSF^+/- mice. Furthermore, to examine synaptic functions we measured evoked field EPSPs in the hippocampal slice. The long-term depression (LTD) impaired in the CA1 of the NSF^+/- mice, but was normal long-term potentiation(LTP). Then, we assessed the autistic like behaviors using the three-chambered task and ultrasonic vocalizations analysis. We found that the social interaction and communication reduced in the NSF^+/- mice, compared with the wild mice. It is suggested that NSF is regulators of synaptic plasticity and membrane protein trafficking, might be related to the pathophysiology of autistic behavior.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-442 Auditory brainstem development of sound-stimulated juvenile rats: a preliminary study of autism model rats Lucila Emiko Tsugiyama（Tsugiyama Lucila Emiko）¹，Michiru Ida-Eto（Ida-Eto Michiru）¹，Takeshi Ohkawara（Ohkawara Takeshi）¹，Yuichi Noro（Noro Yuichi）²，Masaaki Narita（Narita Masaaki）¹ ¹Dept. of Dev. Regen. Med., Mie Univ., Mie, Japan ²Dept. of Physics Eng., Mie, Japan [INTRODUCTION] Autism Spectrum Disorder (ASD) is a neurodevelopmental condition, characterized by impairments in social interaction, communication, repetitive behavior and sensory abnormalities, such as auditory hypersensitivity. However, the mechanisms of ASD and auditory hypersensitivity has not been elucidated yet. We have established autism model rat using prenatal thalidomide exposure. Previously, we found anatomical difference of auditory brainstem between autism model rats and control rat¹. We also found increased neuronal activity in the auditory brainstem of autism model rats on postnatal day (P) 50 after sound stimulus². In this study, we used P14 and P21 infantile rats to know whether similar results would be obtained or not. [MEHTODS] Autism model rats using prenatal exposure to thalidomide were made as described previously. On P14 and 21, control rats were placed into a sound-attenuated box and exposed to 30 minutes of no sound stimulation, followed by one hour of sound stimulation of 16 kHz (62 dB) or one hour of no stimulation at all. Immediately after, the rats were anesthetized, perfused and the brain was dissected out. The brainstem was sectioned at a 50 µm thickness. Immunohistochemistry was performed using c-Fos antibody, a marker of neuronal activity. [RESULTS] The P21 sound stimulated animals showed c-Fos expression in the auditory brainstem, which was consistent to adult rats (P50). Although the P14 animals responded to the auditory stimulus, c-Fos expression in the auditory brainstem of these animals seemed to be inconsistent, compared to P21 animals. [CONCLUSION] These results indicate immaturity of hearing mechanisms and still under development of the hearing accuracy of P14 rats when compared to P21 sound stimulated rats. We are now conducting auditory brainstem development experiments in autism model rats. [REFERENCES] 1.Ida-Eto M, Hara N, Ohkawara T, Narita M. Mechanism of auditory hypersensitivity in human autism using autism model rats. 2017, Pediatr. Int. 59: 404-7. 2.Tsugiyama LE, Ida-Eto M, Ohkawara T, Noro Y, Narita M. Altered neuronal activity in the auditory brainstem following sound stimulation in autism model rats. (manuscript in submission).

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-443 Identifying schizophrenia genes through a forward genetic screen in mice Liqin Cao（Cao Liqin）¹，Daniela Klewe-Nebenius（Klewe-Nebenius Daniela）¹，Greg Asher（Asher Greg）¹，Chika Miyoshi（Miyoshi Chika）¹，Takeshi Sakurai（Sakurai Takeshi）¹，Hiromasa Funato（Funato Hiromasa）¹，Masashi Yanagisawa（Yanagisawa Masashi）¹，Qinghua Liu（Liu Qinghua）^1,2,3 ¹International Institute for Integrative Sleep Medicine(WPI-IIIS), University of Tsukuba ²University of Texas Southwestern Medical Center, USA ³National Institute of Biological Sciences, China Schizophrenia is a severe psychiatric disorder characterized by disturbances of thoughts, perceptions, emotions and cognition. Twin and adoption studies have showed that schizophrenia is highly heritable, and disease risk results from the interplay of diverse genetic variants as well as environmental factors. It affects around 1% of the general population worldwide, and is among the most disabling medical disorders. The disease is devastating to the affected individuals and enormously costly to society. However, given the etiologic complexity the mechanisms of molecular basis and neural circuits of schizophrenia remain largely unclear. Schizophrenia is often associated with emotional response deficits for fear stimuli. We developed a forward genetic screen, using a novel predator odor-evoked innate fear assay, to identify ethylnitrosourea (ENU)-mutagenized mice with abnormal fear behaviors in response to innate fear stimuli. By single nucleotide polymorphism (SNP) mapping and whole-genome sequencing, we identified the causative mutation of a dominant "schizophrenia-like" mutant pedigree, named Psycho. The Psychom/+ mice exhibited hyperactivity, risk-taking behavior, diminished motivation, and impaired cognition that correspond to the clinical presentation of schizophrenia in humans. Psychom/+ mice may serve as an excellent model to understand the pathophysiology of schizophrenia and develop novel treatments for psychiatric disorders.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-444 統合失調症モデルマウスのシナプス病態の形態学的および機能的解析 Kisho Obi（小尾紀翔）¹，Fukutoshi Shirai（白井福寿）^1,2，Mari Umezawa（梅沢茉里）¹，Norimitsu Suzuki（鈴木紀光）¹，Akiko Hayashi-Takagi（林（高木）朗子）¹ ¹群馬大体調節研脳病態制御分野 ²東京大院医構造生理 Various lines of evidence have suggested that synaptopathy is involved in schizophrenia. However, it is unknown whether synaptopathy is an underlying mechanism of disease or a secondary consequence. Thus, we performed a longitudinal in vivo 2-photon imaging analysis of the brain of a schizophrenia model (DISC1 knockdown mice) and found that this model exhibited a decrease in the density of dendritic spines. Furthermore, we also found a significantly greater number of large dendritic spines in the model mice. The presence of the large spines in the schizophrenia mice model mirrors findings from another schizophrenia mice model, calcineurin knockout mice. It is well-known that there is a strong correlation between spine head size and its synaptic efficacy, whereby the large spines can generate a larger synaptic current. This led us to hypothesize that large spines can non-linearly affect the dendritic computation, causally resulting in subsequent behavioral alterations. To test this hypothesis, we use a multi-scale analysis that consists of an uncaging-evoked single spine EPSC measurement and Ca2+ imaging to visualize the synaptic input, dendritic event, action potential, and behavioral manifestations. First, we present our conditional KD model in which DISC1 shRNA together with a neuronal morphological marker is induced by the treatment of tamoxifen for late adolescence specific gene regulation. And we found DISC1 conditional KD does not reduce spine density, but reproducibly induced the huge spines. And our result of Y maze spontaneous alternation test suggests that the huge spines in the model disrupt the working memory. In addition, by applying 2-photon glutamate uncaging, we performed simultaneous recording of dendritic Ca2+ event and somatic EPSPs in response to the activation of large spines and other spines. We found that the stimulation of large spines including huge spines evoked action potentials. This result implies that the huge spines have a non-linear effect on the generation of action potential. Now, with use of in vivo optical and in silico manipulation of the spines in the model animals, we are trying to causally examine what kind of synaptic pathology would underlie the pathology of disorders.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-445 難治性統合失調症患者由来ARHGAP10遺伝子変異マウスにおけるメタンフェタミン誘発性認知機能障害に対するファスジルの効果 Jingzhu Liao（廖婧竹）¹，Taku Nagai（永井拓）¹，Daisuke Mori（森大輔）²，Bolati Wulae（吾拉尔波拉提）¹，Kazuhiro Hada（羽田和弘）¹，Toshitaka Nabeshima（鍋島俊隆）^3,4，Norio Ozaki（尾崎紀夫）²，Kiyofumi Yamada（山田清文）¹ ¹名古屋大学大学院医学系研究科 ²名古屋大院医精神医学 ³藤田医科大学 ⁴藍野大学 ARHGAP10, a member of Rho GTPase-activating protein (RhoGAP) superfamily which contributes to neuronal development, polarization and function. A mouse model of schizophrenia with both a deletion-type copy-number variation (CNV) and a single-nucleotide variation (SNV) in the RhoGAP domain of ARHGAP10 gene (ARHGAP10 mutant mice) has been developed. In recent years, touchscreen-based cognitive tasks have been developed for rodents to provide a better translational approach across species for further understanding of the cognitive impairments observed in various neuropsychiatric disorders and for testing potential pharmacological interventions. Methamphetamine (METH) is one kind of highly addictive drug which induces cognitive deficit in human and rodents. Furthermore, it is well known that METH potentiates schizophrenia symptoms. In this study, we investigated the performance of ARHGAP10 mutant mice with or without MEHT pretreatment in the touchscreen-based visual discrimination task. Mice were initially trained to discriminate between a pair of stimuli (marble and fan, above 80% accuracy for 2 consecutive days). On the testing day, mice were injected with either saline or METH (0.3 or 1.0 mg/kg) 30 min before the test. In order to examine the effect of a Rho kinase inhibitor fasudil, mice were administrated saline or fasudil (3, 10 or 20 mg/kg) 25 min after the METH treatment. Cognitive function of saline-treated ARHGAP10 mutant were comparable to wild-type mice. However, low dose of METH (0.3 mg/kg) significantly reduced the accuracy of visual discrimination in ARHGAP10 mutant mice, but not wild-type mice. METH-treatment at a dose of 1.0 mg/kg induced cognitive impairment in both wild-type and ARHGAP10 mutant mice. METH (0.3 mg/kg)-induced cognitive impairment in ARHGAP10 mutant mice was ameliorated by fasudil in a dose-dependent manner. We demonstrated by using the translatable visual discrimination task that cognitive function in ARHGAP10 mutant mice is highly vulnerable to acute METH treatment, and this impairment could be rescued by fasudil.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-446 覚醒ラットにおける音長変化を使った聴覚ミスマッチ反応；周波数依存性とガンマオシレーション効果 Hiroyoshi Inaba（稲葉洋芳）¹，Hidekazu Sotoyama（外山英和）¹，Itaru Narihara（成原格）¹，Hisaaki Namba（難波寿明）¹，Eiichi Jodo（浄土英一）²，Satoshi Eifuku（永福智志）²，Hirooki Yabe（矢部博興）³，Hiroyuki Nawa（那波宏之）¹ ¹新潟大脳研基礎神経科学分子神経生物 ²福島県立医大医システム神経科学 ³福島県立医大医神経精神医学 The brain function that detects deviance in the acoustic environment in humans can be evaluated with auditory mismatch negativity (MMN). MMN is triggered in response to rare deviations from the regular pattern of standard sounds and calculated as a subtracted value of the event-related potential (ERP) for standard sounds from that for deviant sounds. The reduction in MMN, particularly in sound duration MMN, is often seen in patients with schizophrenia. To date, the neurophysiological underpinnings and neural circuits responsible for the MMN production are poorly understood. Monitoring ERPs from electrocorticography (ECoG) electrodes placed on the auditory and frontal cortices of free-moving rats, we characterized the effectors and components of duration MMN in the oddball paradigm consisting of standard tones (50 ms) and deviant tones (150 ms) of 3, 12, or 50 kHz. To control the effects of the absolute duration of these stimulants and that of the standard-deviant sequence, we adopt the equal probability paradigm; the non-standard tones (50 ms) and non-deviant tones (150 ms) were equally given in a random order and pseudo MMN was calculated as a subtracted value of ERP for non-standards from that for non-deviants. In comparison between ERPs for deviants and standards (either 3, 12, or 50 kHz), significant MMN-like potentials were commonly detected 150-200 ms after stimulus onset. In comparison between MMN and pseudo MMN, there were significant differences at 150-200 ms, irrespective of the tone frequencies. In contrast, we failed to detect any significant MMN-like potentials from the electrode of the frontal cortex with any tone frequency. Individual ERPs were subjected to time-frequency analysis and the results were compared between the oddball and equal probability paradigms. Wavelet analysis revealed that the deviant ERPs showed significantly higher induced-power in the high-gamma range (60-100 Hz) than that of the non-deviant ERPs at the same time period (100-150 ms after stimulus onset) in all tone frequencies. These results suggest that, like human duration MMN, duration MMN-like potentials can be detected from the electrode place on the primary auditory cortex of awake rats. The production of the duration MMN appeared to involve the gamma oscillations induced by the tone-duration difference.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-447 栄養環境依存的な新規統合失調症モデルマウスより得られた新たな中間表現型 Shinobu Hirai（平井志伸）¹，Makoto Arai（新井誠）²，Hideki Mira（三輪秀樹）³，Yasuto Kunii（國井泰人）^4,5，Haruo Okado（岡戸晴生）¹，Mizuki Hino（日野瑞城）⁴，Nagaoka Atsuko（長岡敦子）⁴，Hirooki Yabe（矢部博興）⁴ ¹東京都医学総合研神経細胞分化PT ²東京都医学総合研統合失調症PT ³国立精神神経センター　分子精神薬理 ⁴福島県立医科大学　医学部神経精神医学講座 ⁵福島県立医科大学　会津医療センター　精神医学講座 Objective: In modern times, the number of patients with psychiatric disorders has increased drastically concurrently with the time when a sugar refining technology was established. It is less clear that the influence of sugar on psychological disorders during development with the experimental proof. In this study, we tried to uncover the effect of sugar on the brain development, using mice which have a genetic risk factor. We picked up Glyoxylase1 (Glo1) as a genetic risk factor for schizophrenia. Although Glo1 mutations are associated with schizophrenia, same mutations are found in healthy people with a lower probability than patients. Therefore, we thought that an additional environmental risk factor could be required to cause schizophrenia combined with Glo1 mutations. Methods: We prepared two types of diets that contain increased carbohydrates of the same quantity and consist of different kind of carbohydrate. One contains starch which is polymers of glucose. Another contains sugar which is the disaccharides consisting of glucose and fructose. We fed each diet to young mice for short duration and then performed various behavior tests, in vivo microdialysis and electroencephalogram (EEG). After that, we sacrificed them for an immunohistological analysis. Results: Glo1 deficiency significantly worsens the phenotypes observed in Sugar fed mice. Sugar fed Glo1 hetero mice are accompanied with the defects of PV positive neurons. Dysfunction of PV neurons is a famous intermediate phenotype with the most causal relationship to the onset of schizophrenia. The abnormality of PV neurons causes characteristic symptoms in schizophrenia and these phenotypes are also observed in our created model mice. We also found a novel schizophrenia phenotype. Glo1 is an enzyme that inhibits creating an advanced glycated endproducts (AGEs). Excess sugar intake caused an accumulation of AGEs in and outside of the cerebral vascular endothelial cells independent of genotypes. However, injured endothelial cell marker was observed only in the sugar fed glo1 heterozygote mice. Moreover, same phenotype was detected in the postmortem brain of schizophrenia patients. Conclusions: We found that the combination of genetic vulnerability and excess sugar becomes a novel model mouse for schizophrenia with detailed scientific proof. We also found a novel schizophrenic phenotype, which is an injured vascular endothelial cells. It could expand the selections of therapeutic medicines.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-448 The Effects of Acute Dopamine Depletion on Resting-State Functional Connectivity and Striatal Glutamate in Healthy Humans Fernando Caravaggio（Caravaggio Fernando），Yusuke Iwata（Iwata Yusuke），Alex Barnett（Barnett Alex），Shinichiro Nakajima（Nakajima Shinichiro），Carol Borlido（Borlido Carol），Eric Plitman（Plitman Eric），Yukiko Mihashi（Mihashi Yukiko），Julia Kim（Kim Julia），Philip Gerretsen（Gerretsen Philip），Ariel Graff-Guerrero（Graff-Guerrero Ariel） Research Imaging Centre, Centre for Addiction and Mental Health, Toronto, Canada Understanding the interplay between the neurotransmitters dopamine and glutamate in the striatum has become the highlight of several theories of neuropsychiatric illnesses, such as schizophrenia. However, it is unclear how acute dopamine depletion in healthy humans may affect the functional connectivity of the striatum and glutamate concentrations therein. Sixteen healthy participants (8 female, age [mean ± SD] = 27.4 ± 9.3) participated in the study. Participants provided both resting-state MRI (rsMRI) and 1H-MRS scans (3T GE Discovery MR750; 8-channel head coil) before and after acute dopamine depletion. Dopamine depletion was induced by oral metyrosine intake over 25 hours (64mg/kg over 24 hours). Metyrosine administration resulted in increased self-reported fatigue (t(15)=-3.66, p=0.002) and reduced vigor (t(15)=3.95, p=0.001). Compared to baseline, dopamine depletion reduced functional connectivity between the caudate and medial prefrontal cortex, as well as between the ventral tegmental area and hippocampus. However, acute dopamine depletion did not alter striatal glutamate concentrations measured with 1H-MRS. Acute dopamine depletion in healthy humans may reduce the functional connectivity between the dorsal striatum and prefrontal cortex. This work has important implications for theorizing about neurochemical alterations in neuropsychiatric diseases and, in turn, our understanding of the functioning of the basal ganglia in humans.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-449 RNA干渉法を利用した脳内リーリンシグナルを増強させる手法の開発 Tumi Tsuneura（常浦祐未）¹，Kanako Kitagawa（北川佳奈子）¹，Saeko Takase（高瀬冴子）¹，Masahito Sawahata（澤幡雅仁）¹，Norimichi Ito（伊藤教道）¹，Takao Kohno（河野孝夫）²，Mitsuharu Hattori（服部光治）²，Taku Nagai（永井拓）¹，Kiyofumi Yamada（山田清文）¹ ¹名古屋大院医医療薬学・附属病院薬剤部 ²名古屋市大薬病態生化学 Reelin is a secreted protein that is important for development of brain structures and regulating brain function. Previous research suggests that decreased Reelin expression causes brain dysfunction, which may lead to mental and neuronal disorders such as schizophrenia and Alzheimer's disease. It has been reported that microinjection of Reelin protein into the brain enhances cognitive ability, synaptic plasticity and dendritic spine density in adult mice. Although Reelin microinjection could be effective to treat neuropsychological disorders, alternative methods may also be applicable to stimulate Reelin signaling in clinical application. A disintegrin and metalloproteinase with thrombospondin motifs-3 (ADAMTS-3) has been identified as a protease that inactivates Reelin. To establish novel approach to stimulate Reelin signaling, we developed a gene silencing method for adamts-3. In order to accomplish specific knockdown of ADAMTS-3 using RNA interference (RNAi), we prepared lentiviral vector expressing shRNA targets adamts-3 (lenti-shADAMTS-3). Primary cultured cortical neurons were obtained from embryos of C57BL/6J mice. We checked knockdown efficiency of the shRNA in primary cultured neurons by real-time RT-PCR analysis. For measurement of Reelin signaling activation, we determined the Reelin content in conditioned medium and intercellular Dab1 expression in primary cultured neurons by western blotting. In primary cultured cortical neurons, the treatment with lentivirus-mediated RNAi reduced the expression level of adamts-3 mRNA. Knockdown efficiency of adamts-3 was around 84%. In cultured medium of primary cortical neuron treated with lenti-shADAMTS-3, Reelin degradation was suppressed, and the ratio of full length Reelin was increased. Lenti-shADAMTS-3 treatment also decreased the expression level of intercellular Dab1 suggesting that Reelin signaling can be activated by ADAMTS-3 knockdown. We developed lentivirus-mediated RNAi of adamts-3 knockdown in primary cultured cortical neurons. But further experiments are required to analyze functional activity of these tools in vivo.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-450 RNA-seq analysis of stratum reveals sex-differential gene expression in AKT1-deficient mouse model of schizophrenia Tzu-Ping Lee（Lee Tzu-Ping）¹，Bin Xu（Xu Bin）²，Wen-Sung Lai（Lai Wen-Sung）^1,3,4 ¹Department of Psychology, National Taiwan Unitversity ²Department of Psychiatry, Columbia University Irving Medical Center, U.S.A. ³Graduate Institute of Brain and Mind Sciences, National Taiwan University, Taipei, Taiwan ⁴Neurobiology and Cognitive Science Center, National Taiwan University, Taipei, Taiwan Schizophrenia is a severe mental disorder with a strong genetic component. Sexual differences in schizophrenia are widely reported, which are displayed in the prevalence of disease, age of onset, symptoms, and responses to antipsychotics. In general, female patients have later age of onset and less severe symptoms. Yet, the underlying molecular mechanism of sex difference remains elusive. Accumulating evidence from human genetic studies suggests that AKT1 (protein kinase B α), a key signaling kinase downstream of dopamine receptor D₂, is associated with schizophrenia in several ethnic groups and it is one of the susceptibility genes which contribute to the pathogenesis of the disorder. Intriguingly, AKT1-deficient mice exhibited some characteristics of schizophrenia and sex differences were also reported. A better understanding of the mechanisms of sexual differences may provide new ways of treatment and/or insights to the disorder. To further understand the molecular mechanism behind the sexual differences in Akt1 heterozygous mutant mice (Akt1^+/-) and their wild-type littermate controls (WT), RNA-sequencing technique was used in this study. Transcriptomic expression of striatum tissues from male and female early adult mice were analyzed. Differential expression analysis was conducted by DESeq2. We found that a total of 92 genes had significant effects of interaction. We further analyzed the simple main effect of sex at both genotypes (male and female: WT vs. Akt1^+/-) and found that there are 40 significant genes for males and 19 for females. For gene set enrichment analysis, our Enrichr gene ontology analysis did not reveal any neuronal-related ontology, therefore further individual gene search was conducted. For dopamine regulation, Slc3a1 and Cadm2 were differentially expressed in male Akt1^+/- mice compared to WT. For AKT1 signaling transduction, Kcnmb4, Map4k5, Bmi1, and Hipk3 were differentially express in male Akt1^+/- mice. For female mice, Tspan7, known to involve in dopamine receptor D₂ internalization, was down regulated in Akt1^+/- mice. Genes and gene lists reported by this study could be candidates for the mechanism of sexual differences behind AKT1-deficient mouse model of schizophrenia. Further analyses are still in progress and further experiments are needed to confirm the roles of each gene.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-451 新生仔期のEGF投与は内側前頭前皮質の高頻度発火型GABA神経に対して成熟期により著明となる機能異常をもたらす Hisaaki Namba（難波寿明），Hiroyuki Nawa（那波宏之）新潟大脳研分子神経生物学 Rodent animals, exposed with epidermal growth factor (EGF) as neonate, are so far established as animal models for schizophrenia. These animals exhibit impaired development of neurochemical phenotypes of GABAergic neurons in frontal cortex during neonatal period. In adulthood, phase coherence of gamma bands of sensory evoked-electroencephalogram in frontal cortex are diminished, which suggests some functional impairments of GABAergic neurons. We so far reported immunoreactivity of parvalbumin, a neurochemical marker of fast-spiking (FS) GABAergic neurons, is diminished in prelimbic region (PrL) of medial prefrontal cortex (mPFC) and anterior cingulate (ACC) in adult EGF-treated mice. In this study, we analyzed firing property of FS neurons of PrL in adult mice (> postnatal days 70). As a result, pre-treatment with EGF significantly diminished the number of spikes evoked by higher depolarizing current injection (0.6-0.8 nA), which reveals a dysfunction of sustained spike generation at higher frequency of more than 100Hz. In parallel, the amplitude of spike after-hyperpolarization of each action potential as well as protein expression of voltage-gated potassium channel, Kv3.1b, were significantly attenuated. These alterations in firing properties of fast spiking neurons and in channel protein were not detected in other regions of frontal cortex, such as ACC and primary motor area. We also analyzed FS firing properties in PrL of pre-adolescent mice at postnatal days 28-34. As a result, only moderate alterations were detected in the firing properties in this developmental stage. Thus, pre-treatment with EGF as neonate markedly affected FS properties in mPFC during post-adolescent period.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-452 多発家系患者のiPS分化神経細胞を用いた統合失調症の分子病態解析 Kana Yamamoto（山本果奈）¹，Toshihiko Kuriu（栗生俊彦）²，Takanobu Nakazawa（中澤敬信）^1,3，Kensuke Matsumura（松村憲佑）¹，Kazuki Nagayasu（永安一樹）^1,4，Masayuki Baba（馬場優志）¹，Hidenaga Yamamori（山森英長）^5,6，Yuka Yasuda（安田由華）^5,7，Michiko Fujimoto（藤本美智子）^5,8，Yoshinori Tsurusaki（鶴崎美徳）^9,10，Naomichi Matsumoto（松本直通）⁹，Kazuhiro Takuma（田熊一敞）^3,11，Ryota Hashimoto（橋本亮太）^5,12，Hitoshi Hashimoto（橋本均）^1,11,13,14 ¹大阪大院薬 ²大阪医大・研究支援セ ³阪大・院歯・薬理 ⁴京大・院薬・生体機能解析 ⁵国立精神・神経医療研究セ・精神保健研・精神疾患病態 ⁶大阪病院 ⁷生きる・育む・輝くメンタルクリニック ⁸阪大・院医・精神医学 ⁹横浜市大・院医・遺伝学 ¹⁰神奈川県立こども医療セ ¹¹阪大・院連合小児・子どものこころセ ¹²大阪大 ¹³阪大・データビリティフロンティア機構・バイオサイエンス ¹⁴阪大・先導的学際研究機構・超次元ライフイメージング研究 Schizophrenia is a severe neuropsychiatric disorder with a prevalence rate of approximately 1% in the world wide, characterized by delusions, hallucinations, impaired cognitive function, emotional blunting and incoherent behavior. While many antipsychotic drugs have been demonstrated as effective in the treatment of schizophrenia, a substantial number of patients with schizophrenia are partially or fully unresponsive to the treatment; schizophrenia is a significant medical and social problem and drug development and treatment strategies based on the molecular mechanisms of the etiology and pathology are imperative. Currently, the molecular defects in neurons leading to the onset and progression of this disease remain largely unclear, partly because of the limited accessibility to live neurons from patients with schizophrenia. Here, we analyzed the neural function of neurons from induced pluripotent stem cell (iPSC) lines derived from patients from a family with multiple occurrence of schizophrenia (familial schizophrenia). We established iPSC lines from 2 patients with familial schizophrenia and 2 unaffected healthy controls and differentiated these iPSC lines into neurons by standard methods. We firstly evaluated synaptic transmission using whole-cell patch-clamp recordings from these neurons and observed that the amplitude of miniature excitatory postsynaptic currents was increased in neurons from the patients. We then performed transcriptome analysis of these neurons and found that the expression levels of several synaptic proteins were altered in neurons from the patients. The present results suggest that the increased excitatory neurotransmission and the differential expression patterns of synaptic proteins may be involved in the molecular pathophysiology of schizophrenia in the patients. As we previously identified common genetic mutations in these patients with familial schizophrenia, we are currently analyzing the causal relationship between the abnormal neural functions and the genomic mutations.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-453 22q11.2欠失症候群モデルマウスにおける視覚誘発電位の評価 Ryo Saito（齋藤遼）¹，Michinori Koebis（古戎道典）¹，Yuki Sugaya（菅谷佑樹）^2,3，Masanobu Kano（狩野方伸）^2,3，Atsu Aiba（饗場篤）¹ ¹東京大院医疾患生命工学動物資源 ²東京大院医神経生理学 ³東京大・WPI-IRCN Impairments of visual sensory processing represent one of the most frequent neurophysiological disturbances in individuals with schizophrenia. The reduced amplitude of visual evoked potential (VEP), especially of the P1 component has been recurrently observed in patients with schizophrenia, suggesting this deficit as a diagnostic marker for the disease. Therefore, Assessment of change in VEP could be a clue to evaluate the validity of schizophrenia animal models. The 22q11.2 deletion syndrome (22q11.2DS) is estimated to affect up to 1 in 4000 live birth. The 22q11.2DS has been shown to increase a risk of developing a variety of psychiatric and developmental disorders including schizophrenia, intellectual disability, autism spectrum disorder and attention deficient hyperactive disorder (ADHD). In particular, the risk of schizophrenia due to this deletion (odds ratio: 16.3-44.2) is higher than any other single genetic variations that have been associated with schizophrenia to date. Although most (~90%) of the 22q11.2DS patients have a 3.0-Mb deletion, mouse models that have been generated so far mimic a partial 1.5-Mb mutation found in a minor population of 22q11.2DS patients. Forty-six protein cording genes are located in the 3.0-Mb region and 29 of them are included in the 1.5-Mb region. A deletion containing the remaining 17 genes has never been introduced to model animals. Thus, it is yet unknown whether these 17 genes contribute to psychiatric symptoms of 22q11.2DS. Recently, we have generated a new mouse model that reproduced the 3.0-Mb deletion of 22q11.2DS (hereinafter, referred to Del(3.0Mb)/+) by a genome editing technique using CRISPR/Cas9 system. In this study, we investigated VEP of Del(3.0Mb)/+ mice in order to evaluate their validity as a model of schizophrenia. We recorded local field potentials from the primary visual cortex (V1) of awake mice while 500-ms visual stimuli were repeatedly presented. We observed generally smaller VEP in Del(3.0Mb)/+ than in WT mice. In particular, the amplitude of the P1 and N2 components were significantly smaller in Del(3.0Mb)/+ mice. These changes in VEP in Del(3.0Mb)/+ mice were consistent with those observed in patients with schizophrenia. Therefore, our results support the validity of Del(3.0Mb)/+ mice as a schizophrenia animal model and suggest that our mouse model should be a useful resource to study pathophysiology of schizophrenia associated with 22q11.2DS.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-454 更年期障害における精神神経症状を改善する加味逍遥散の作用機序解明 Shoko Shimizu（清水尚子），Yugo Ishino（石野雄吾），Masaya Tohyama（遠山正彌），Shingo Miyata（宮田信吾）近畿大東洋医学研究所 Current literature indicates an increase in the prevalence of depression in women going through menopause. In addition to a decline in circulating ovarian hormones, environmental and social stress are considered to contribute as additional factors in the development of depressive behavior in women. Kamishoyosan (KSS) is a traditional Japanese Kampo medicine, and it is composed of specified mixtures of 10 crude compounds from plant, animal, and mineral sources. KSS has been clinically used for neuropsychiatric symptoms such as depression, irritation, and anxiety in menopausal women. However, the precise mechanism by which KSS ameliorates these symptoms remains unknown. In this study, we aimed to determine whether the administration of KSS modulated menopausal depressive behavior and elucidate the underlying action mechanism of KSS. We assessed whether the administration of KSS modulated stress-induced depression-like behavior by assessing the immobility time of chronically stressed and ovariectomized female mice in the forced swimming test. Furthermore, we examined whether the administration of KSS in the same animals affected expression levels of the serotonin transporter and the serotonin 1A receptor as key proteins in the serotonergic nervous system. It is widely recognized that the neurotransmitter serotonin plays an important role in modulating mood, and dysfunctions in serotonergic neurotransmission are associated with various neuropsychiatric disorders. In addition, previous studies have shown that the ovarian hormone estrogen modulates numerous factors regulating serotonin synthesis and serotonin levels in the central nervous system. KSS is a mixture of several natural ingredients, and it is important to determine which components of KSS are effective for menopausal neuropsychiatric symptoms during stress. The results might suggest one or more compounds as alternatives to current antidepressant drugs.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-455 幼若期における慢性社会的敗北ストレスは小脳キヌレニンと盲腸内細菌叢に影響する Atsushi Toyoda（豊田淳）^1,2，Natsuki Ohsawa（大澤夏樹）¹，Alato Okuno（奥野海良人）³，Takamitsu Tsukahara（塚原隆充）⁴ ¹茨城大農食生命 ²東京農工大院連合農 ³つくば国際大医療保健 ⁴栄養病理研 Depression in young generation is one of the serious health problems especially in advanced societies. In order to study depression in children and young people, several stress models of animals have been developed and characterized. In this study, we have developed a mouse model of childhood depression using the paradigm which juvenile mice are exposed to subchronic and mild social defeat stress (sCSDS), and analyzed kynurenine which is one of the critical metabolites of inflammation, and cecal microbiota in this model. Twenty four C57BL/6J (B6) male mice (3 weeks, CLEA Japan, Tokyo, Japan) were purchased and habituated for 1 week. For developing the juvenile sCSDS mice, B6 mice (n=12) were placed into a home cage of an aggressive ICR male mouse (Japan SLC, Shizuoka, Japan) and were allowed to be attacked by the ICR mice for 5 min. After that, the mice were kept in the same cage but separated by an acrylic board with small holes for 24 hours to allow sensory contact. The procedure was then repeated with another aggressive ICR mouse for 10 consecutive days but the attacking time was reduced by 0.5 min/day from days 2 to 10. After 10 days of sCSDS, the mice were subjected to social interaction and nest-building tests. Finally, kynurenine and tryptophan levels in prefrontal cortex, hippocampus, and cerebellum were determined by HPLC. Furthermore, the metagenome analysis was carried out to reveal microbiome in the cecum. All B6 mice were fed semi-purified diet (AIN-93G, Oriental Yeast, Tokyo, Japan). sCSDS suppressed body weight gain compared with control, although it did not impact food intake. The juvenile sCSDS mice showed several deficits in social behavior such as reduced interaction time in the social interaction test. Also, nest building behavior was partially inhibited by sCSDS. Kynurenine level of cerebellum was increased by sCSDS, although those of prefrontal cortex and hippocampus were not changed. Cecal microbiota was drastically changed by sCSDS, whereas diversity values (Chao1 and Shannon index) was not affected between the groups. Phylum Proteobacteria might be associated their behavior, because the abundance was 7-times higher in sCSDS mice rather than in healthy control (p<0.05). These results suggested that sCSDS in juvenile mice induces deficits of social and nesting behaviors following increased cerebellar kynurenine and changed cecal microbiota.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-456 内側前頭前野内レゾルビンE1投与はBDNFとVEGFの遊離およびmTORC1活性化を介してリポポリサッカライド誘発うつ病様行動を抑制する Satoshi Deyama（出山諭司）¹，Kohei Ishimura（石村航平）²，Hayato Fukuda（福田隼）^2,3，Satoshi Shuto（周東智）²，Masabumi Minami（南雅文）⁴，Katsuyuki Kaneda（金田勝幸）¹ ¹金沢大院医薬保健薬理 ²北海道大院薬創薬有機化学 ³長崎大院医歯薬薬品製造化学 ⁴北海道大院薬薬理 Resolvin E1 (RvE1) is one of the bioactive lipid mediators generated from eicosapentaenoic acid. We have recently reported that the intracerebroventricular infusion of RvE1 produces antidepressant effects in a murine lipopolysaccharide (LPS)-induced depression model and these antidepressant effects are completely blocked by systemic injection of a mechanistic target of rapamaycin complex 1 (mTORC1) inhibitor rapamycin (Deyama et al., Psychopharmacology, 2018). We have also demonstrated that infusion of RvE1 (50 pg/side) into the medial prefrontal cortex (mPFC) exerts antidepressant effects. Although these findings indicate the crucial role of the mPFC in the antidepressant actions of RvE1, the underlying mechanisms remain unknown. Previous studies have shown that activity-dependent release of brain-derived neurotrophic factor (BDNF) and downstream mTORC1 signaling in the mPFC are essential for the rapid and sustained antidepressant actions of ketamine, an NMDA receptor antagonist. Our recent studies show a similar requirement for vascular endothelial growth factor (VEGF) (Deyama et al., Am J Psychiatry, in press) and the pivotal role of BDNF-VEGF interplay in the antidepressant and neurotrophic effects (Deyama et al., Biol Psychiatry, in press). Thus, here we examined the roles of BDNF/VEGF release and mTORC1 activation in the antidepressant actions of intra-mPFC RvE1 infusion in LPS-induced depression model mice using the tail suspension and forced swim tests. The results demonstrate that the antidepressant effects of intra-mPFC RvE1 infusion are completely blocked by co-infusion of a BDNF neutralizing antibody (nAb), a VEGF nAb or rapamycin. We also demonstrate that the antidepressant effects of intra-mPFC BDNF or VEGF infusion are blocked by co-infusion of rapamycin. Together, the current results indicate that BDNF/VEGF release and subsequent activation of mTORC1 in the mPFC are required for the antidepressant actions of RvE1.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-457 新規うつ病治療薬の探索 Shingo Miyata（宮田信吾）¹，Shoko Shimizu（清水尚子）¹，Yugo Ishino（石野雄吾）¹，Masaya Tohyama（遠山正彌）^1,2 ¹近畿大東医分子脳科学 ²大阪府立病院機構 Major depressive disorder (MDD) is probably the oldest and still one of the most frequently diagnosed psychiatric illnesses. MDD is one of the leading causes of disturbances in emotional, cognitive, autonomic, and endocrine functions, affecting nearly 7% of the population in Japan. According to the large amount of information on MDD that has been accumulated during recent years, patients with MDD show an enhanced biologic stress-response mechanism, especially a hyperactive hypothalamic-pituitary-adrenal (HPA) axis and high levels of circulating cortisol. Although dysregulation of the HPA axis by chronic stress is indicative of MDD, the molecular mechanisms and functional changes in the brain underlying depression are largely unknown. In the previous study, we have developed an animal model of depression by exposing mice to chronic stress. These mice showed depression-like symptoms including chronically elevated plasma levels of corticosterone. We previously showed oligodendrocyte (OL)-specific activation of the serum/glucocorticoid-regulated kinase (SGK)1 cascade, increased expression of axon-myelin adhesion molecules, and elaboration of the oligodendrocytic arbor in the corpus callosum of chronically stressed mice. In the current study, we demonstrate that the nodes and paranodes of Ranvier in the corpus callosum were narrower in these mice. Chronic stress also led to diffuse redistribution of Caspr and Kv 1.1 and decreased the activity in white matter, suggesting a link between morphological changes in OLs and inhibition of axonal activity. OL primary cultures subjected to chronic stress resulted in SGK1 activation and inhibited the transcription of metabotropic glutamate receptors (mGluRs). Furthermore, the cAMP level and membrane potential of OLs were reduced by chronic stress exposure. We showed by diffusion tensor imaging that the corpus callosum of patients with MDD exhibited reduced fractional anisotropy, reflecting compromised white matter integrity possibly caused by axonal damage. Our findings suggest that chronic stress disrupts the organization of the nodes of Ranvier by suppressing mGluR activation in OLs, and that specific white matter abnormalities are closely associated with MDD onset. In this study, we investigate the molecular mechanisms underlying the pathogenesis of MDD by using transcriptomics and metabolomics data.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-458 慢性社会的挫折ストレスモデルマウスにおいて乳酸産生阻害剤の投与は不安様行動を悪化させる Hideo Hagihara（萩原英雄），Hirotaka Shoji（昌子浩孝），Yoshihiro Takamiya（高宮義博），Tsuyoshi Miyakawa（宮川剛）藤田医大総医研システム医科学 Exogenous treatment with lactate has been found to have antidepressant effects through studies on animal models of stress, such as corticosterone treatment model and social defeat stress model, while increased lactate levels have been found in the brain of patients with depression and anxiety disorders, and the corresponding animal models. Here, we investigated the role of endogenous lactate production in response to stress on anxiety and depression-like behavior in a mouse model of social defeat stress. We first confirmed that lactate levels in the brain increase immediately after a single exposure to defeat stress, and then attenuated these levels by prior treatment with sodium oxamate (OX), a lactate dehydrogenase inhibitor that blocks lactate production. We then conducted a chronic experiment in which mice experienced daily defeat stress for ten days, and thereafter, were processed for behavioral tests. Mice were treated with OX or vehicle before each defeat session. There were no differences in susceptibility to the repeated defeat stress, as assessed by social avoidance test, between defeated mice with OX and vehicle treatment. However, we found that anxiety-like behavior, as assessed by the open field test and light-dark transition test was increased in OX-treated, defeated mice compared to vehicle-treated, defeated mice. Treatment with OX did not affect depression-like behavior, as assessed by the Porsolt forced swim test and tail suspension test, in defeated mice. Following the behavioral tests, whole brains were harvested and processed for lactate measurement. Results showed that when compared with control mice, chronic defeat stress increased brain lactate levels in vehicle-treated mice and to a large extent in OX-treated mice, when focused on the mice with susceptibility to stress. Mice chronically treated with only OX did not show such enhanced anxiety-like behavior or increased lactate levels in the brain. The results suggest that repeated inhibition of acute lactate production in response to defeat stress leads to its sustained elevation and enhanced anxiety-like behavior in chronic phase.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-459 近交系BALB/cおよびC57BL/6マウスにおける産後のうつ様行動の増加 Hirotaka Shoji（昌子浩孝），Tsuyoshi Miyakawa（宮川剛）藤田医科大総医研システム医科学 Pregnancy and lactation are characterized by various changes in the endocrine system, brain, and behavior of females. These changes are potential risk factors for the development of postpartum depression. A valid animal model of postpartum depression is needed to understand the neurobiological basis of the depressive state of females. To explore a potential mouse model of postpartum depression, we assessed anxiety- and depression-like behaviors in four inbred strains (C57BL/6J, C57BL/6JJcl, BALB/cAnNCrlCrlj, and BALB/cAJcl) of female mice. For each strain, three different reproductive state groups (nulliparous, non-lactating primiparous, and lactating primiparous females) were used. In the non-lactating female group, pups were removed from the primiparous females after parturition to examine the influences of physical interaction with pups on the behavior of females during the postpartum period. The results indicate that BALB/c females showed decreased locomotor activity, increased anxiety-like behavior, and increased depression-related behavior compared with C57BL/6 females. Additionally, there were some behavioral differences between the two BALB/c substrains and between the two C57BL/6 substrains. Moreover, pregnancy- and lactation-dependent behavioral differences were found in some strains of mice. In the BALB/cAJcl strain, lactating females showed decreased distance traveled and reduced sucrose preference compared with the other reproductive state groups, while non-lactating and lactating females showed increased immobility in the forced swim test compared with the nulliparous females. Increased immobility in the forced swim test in lactating females was also observed in the C57BL/6J strain, while lactating females of the C57BL/6JJcl strain showed a decreased sucrose preference. Our data show that female mice of the commonly used mouse strains BALB/c and C57BL/6 differ with regards to anxiety-like and depression-related behaviors. The substrains of mice are also not behaviorally equivalent. These observations suggest genetic contributions in the regulation of the behaviors of female mice. Furthermore, this study suggests that lactating females of the inbred strains BALB/cAJcl and C57BL/6 are potential models of postpartum depression, and thus these female mice could be used to further study genetic and neurobiological mechanisms underlying the development of postpartum depression.		7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-460 歯状回ドパミンD1受容体はSSRIの抗うつ作用発現に重要な役割をもつ Takahide Shuto（首藤隆秀），Mahomi Kuroiwa（黒岩真帆美），Naoki Sotogaku（外角直樹），Yukie Kawahara（河原幸江），Yoshinori Ohnishi（大西克典），Yuuki Hanada（花田雄樹），Akinori Nishi（西昭徳）久留米大医薬理 Major depression is a psychiatric disorder with high lifetime prevalence. Selective serotonin reuptake inhibitors (SSRIs) are commonly used for treatment of depression. However, current pharmacological treatment of depression is insufficient, and development of improved treatments especially for treatment-resistant depression is desired. Recently, chronic administration of SSRI is reported to increase the expression of dopamine D1 receptors in mature granule cells of the hippocampal dentate gyrus, but the mechanisms underlying involvement of D1 receptors of SSRIs to improve symptoms of depression are not fully understood. In this study, we investigated the role of D1 receptors in the dentate gyrus in antidepressant actions of SSRIs. Mice were treated with fluoxetine, an SSRI, chronically by subcutaneous implantation of matrix-driven delivery pellets (15 mg/kg/day, 14 days). The fluoxetine-induced changes in gene and protein expression were analyzed by quantitative real-time PCR and Western blot analysis, respectively. In addition, depression-like behaviors were evaluated with the novelty-suppressed feeding test (NSFT) and tail suspension test (TST) in mice subjected to chronic restraint stress. Chronic treatment with fluoxetine induced the expression of D1 receptor mRNA and protein in the dentate gyrus, but not other subtypes of dopamine receptor mRNA. The overexpression of D1 receptors in the dentate gyrus reduced the feeding latency in the NSFT and the immobility time in the TST. Regular restraint stress (2 hr/day, 14 days) increased the feeding latency in the NSFT, and chronic treatment with fluoxetine reduced the increased feeding latency. However, in mice subjected to severe restraint stress (4 hr/day, 28 days), chronic treatment with fluoxetine failed to reduce the stress-induced increase in feeding latency in the NSFT and immobility time in the TST. Chronic co-administration of a dopamine D1 receptor agonist, R(+)-SKF81297 (1.5 mg/kg/day, i.p. for 5 days), with fluoxetine resulted in the reversal of the depression-like behaviors. These results suggest that the increase in D1 receptor signaling in the dentate gyrus is essential for the antidepressant actions, and activation of dopamine D1 receptors in the dentate gyrus enhances therapeutic actions of SSRI under SSRI-resistant stress conditions.

7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-461 マウス反復社会的敗北ストレスが報酬獲得における"やる気"に及ぼす影響 Nozomi Endo（遠藤のぞみ）¹，Nami Somayama（杣山奈実）¹，Takashi Komori（小森崇史）²，Mayumi Nishi（西真弓）¹ ¹奈良県立医科大学第一解剖学 ²奈良県立医科大学精神医学講座 Stress is a major risk factor for the development of psychiatric disorders such as depression, anxiety, and post-traumatic stress disorder. Repeated social defeat stress (RSDS) is commonly used as an ethologically relevant stressor in rodents. Recent numerous studies have shown that the influences of RSDS on depressive- and anxiety-like behaviors in mice. In this study, we examined effects of RSDS on effort-based decision-making behavior related to reward acquisition in mice by using our own behavioral test. In our protocol, C57BL/6N mice were exposed to physical contact from aggressive ICR mice for 10 min a day in consecutive 10days. Then we performed social avoidance test and selected RSDS mice that exhibited 40% or more avoidance ratio to ICR mice in subsequent behavior test. Control mice had never been exposed to the physical attack of the ICR mice. Then, all mice were food restricted to approximately 85-90% of free-feeding weight before the behavior test. In the effort-based decision-making test, milk chocolate (20 mg) as a high-reward and food pellet (25 mg) as a low-reward were placed in each side of the experimental box. A high wall was set upped as a high-cost in front of chocolate reward, and a low wall was set upped as a low-cost in front of the food pellet reward. Mice were allowed to freely select either high-reward/high-cost or low-reward/low-cost option, and mice have to overcome the higher wall to earn the high-reward. Control mice mostly selected high-reward/high-cost option (approximately 90 % of choice for milk chocolate). In contrast, RSDS mice were divided into two groups; RSDS mice that chose the comparable percent of high-reward/high-cost option as with control mice and RSDS mice that did not absolutely choose high-reward/high-cost option. Importantly, even RSDS mice that only chose low-reward/low-cost option remained ability to overcome the high wall. These findings suggest RSDS produces abnormal reward-cost ratios in effort-based decision-making in mice which causes individual differences in susceptibility to the same stress. We are now analyzing the neural mechanism of the effects of RSDS on effort-based decision-making behavior.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-462 Intestinal dysbacteriosis mediates the reference memory deficit induced by anaesthesia and surgery in aged mice Xiyao Gu（Gu Xiyao），Weifeng Yu（Yu Weifeng），Diansan Su（Su Diansan） School of Medicine, Shanghai Jiaotong Univ Background: Postoperative cognitive dysfunction (POCD) is associated with increased morbidity and mortality and has become a major concern of patients and caregivers. POCD is most common in older patients, but it can occur in younger patients as well. Previous studies demonstrated that the gut microbiome affects cognitive function and behaviour, and perioperative factors, including the operation itself, antibiotics, opioids, or acid-inducing drugs, affect the gut microbiome. Thus, we hypothesised that intestinal dysbacteriosis caused by anaesthesia and surgery induces POCD. Methods: Tibial fracture internal fixation was performed in 18-month-old C57BL/6 mice under isoflurane anaesthesia to establish the POCD model. The Morris water maze was used to measure reference memory after anaesthesia and surgery. We used high-throughput sequencing of 16S rRNA from faecal samples to investigate changes in the abundance of intestinal bacteria after anaesthesia and surgery. To confirm the role of the gut microbiome in POCD, we pretreated mice with compound antibiotics or mixed probiotics (VSL#3). Results: Anaesthesia and surgery impaired reference memory and induced intestinal dysbacteriosis in aged mice. The 16S rRNA sequencing data revealed 37 genera (18 families) of bacteria that changed in abundance after anaesthesia and surgery. Pretreating mice with compound antibiotics or mixed probiotics (VSL#3) prevented the learning and memory deficits induced by anaesthesia and surgery. Conclusions: Our data suggest that the deficits in reference memory induced by anaesthesia and surgery are mediated by intestinal dysbacteriosis. Keywords: gut microbiome, intestinal dysbacteriosis, postoperative cognitive dysfunction

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-463 先天性高ビリルビン血症は前部帯状回皮質におけるドーパミン伝達異常を引き起こす Keiko Tsuchie（土江景子）¹，Toshiko Tsumori（津森登志子）²，Shoko Miura（三浦章子）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）^3,4 ¹島根大医精神医学 ²県立広島大保健福祉看護 ³島根大医免疫精神神経学 ⁴株式会社　RESVO There are many reports on Jaundice or neonatal hyperbilirubinemia induced psychotic symptoms although the molecular mechanisms are not yet well understood. We previously reported that the Gunn rat, which is a congenital animal model of hyperbilirubinemia, has abnormal behavior similar to schizophrenia and autism. Further, we reported that the administration of antipsychotics such as dopamine receptor antagonists alleviated behavior abnormalities in Gunn rats (Tsuchie et al. 2013). The abnormality of the dopaminergic system is related to the abnormal behavior. Here we found dopaminergic dysregulation in the anterior cingulate cortex (ACC) of the Gunn rat. The amounts of dopamine and its metabolites, 3, 4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the 4 brain regions (frontal cortex, ACC, striatum, hippocampus) of the Gunn rats were measured by high performance liquid chromatography. There were significantly higher dopamine, DOPAC and HVA in the ACC region of the Gunn rats when compared to the control rats. On the other hand, the ratio of metaborates to dopamine found no significant difference between the 4 brain regions of the Gunn rats and of the control rats. Dopamine in the ACC region seems to play a key role in governing cognition and associated behaviors. Our results suggest that there are dopaminergic system abnormalities in the Gunn rats' ACC regions. This result could explain why the Gunn rats have abnormal behavior and why after being treated with dopamine receptor antagonists, their abnormal behavior improved.		7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-464 Phf24遺伝子の欠失ラットにおけるけいれん感受性の亢進 Masaki Kato（加藤将貴）¹，Naofumi Kunisawa（國澤直史）¹，Saki Shimizu（清水佐紀）¹，Kazuma Kawakita（河北和馬）¹，Shohei Kawaji（川路翔平）¹，Higor A. Iha（伊波 A. イーゴル）¹，Takashi Kuramoto（庫本高志）^2,3，Tomoji Mashimo（真下知士）^2,4，Tadao Serikawa（芹川忠夫）^1,2，Yukihiro Ohno（大野行弘）¹ ¹大阪薬大薬品作用解析学 ²京都大実験動物施設 ³東京農大動物科学動物栄養学 ⁴大阪大医実験動物学 We previously demonstrated that Phf24, also known as G_αi-interacting protein (GINIP) interacting with GABA_B receptors, was markedly down-regulated in Noda epileptic rat (NER) (Kuramoto et al., Behav. Genet., 47, 609, 2017). To clarify the role of Phf24 in regulating seizure susceptibility, we created the novel animal model, Phf24-knockout (KO) rats, and analysed their behavioral phenotypes. Seizure susceptibility of Phf24-KO rats was assessed using chemically- and electrically-induced convulsion tests. Intensity and incidence of seizures induced by pentylenetetrazole (PTZ, 40 mg/kg) were significantly increased in Phf24-KO rats than in control (F344) rats. Phf24-KO rats also showed higher sensitivity to electrical shock-induced seizures. In addition, PTZ-induced kindling (30 mg/kg/day, 10 days) was significantly facilitated by the Phf24-KO. Furthermore, to explore the excited brain regions following generalized tonic-clonic seizure (GTCS), we performed immunohistochemical analysis of c-Fos protein expression, a biological marker of neural excitation, after electrical shock-induced seizures. Phf24-KO rats showed a significantly higher c-Fos protein expression than control animals in the cerebral cortex, amygdala, hippocampus and thalamus. These results suggest that Phf24 plays a crucial role in controlling the susceptibility to epileptic seizures, which is probably involved in epileptogenicity of NER. Moreover, it seems possible that Phf24 negatively regulates the neural excitation in the cerebral cortex, amygdala, hippocampus and thalamus.

7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-465 β-N-methylamino-L-alanine (BMAA)は非神経株化細胞の細胞周期進行を抑制する Shun Hamada（濱田俊）¹，Saki Okamoto（岡本彩希）¹，Shigeyuki Esumi（江角重行）²，Kayoko Hamaguchi-Hamada（濱田香世子）¹ ¹福岡女子大国際文理 ²熊本大院生命科学・医脳回路構造 β-N-methylamino-L-alanine (BMAA), a natural non-proteinaceous amino acid, is a neurotoxin produced by a wide range of cyanobacteria living in various environments. BMAA becomes concentrated through the food chain, and high concentrations of BMAA have been detected in aquatic animals at high trophic levels, such as mussels, oysters, and fish. BMAA is a candidate environmental risk factor for neurodegenerative diseases such as amyotrophic lateral sclerosis and Parkinson-dementia complex. Although BMAA is known to exhibit weak neuronal excitotoxicity via glutamate receptors, the underlying mechanism of toxicity has yet to be fully elucidated. To examine the glutamate receptor-independent toxicity of BMAA, we investigated the effects of BMAA in non-neuronal cell lines. BMAA potently suppressed the cell cycle progression of NIH3T3 cells at the G1/S checkpoint without inducing plasma membrane damage, apoptosis, or overproduction of reactive oxygen species, which were previously reported for neurons and neuroblastoma cells treated with BMAA. We found no evidence that activation of glutamate receptors was involved in the suppression of the G1/S transition by BMAA. Our results indicate that BMAA affects basic cellular functions, such as the division of non-neuronal cells, through glutamate receptor-independent mechanisms. As non-neuronal cell lines can easily be analyzed for metabolic changes and functional alterations, the present results provide a new perspective to understanding the mechanisms by which BMAA affects cellular function.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-467 視床下部から視床背内側核・外側手綱核へ伝播するてんかん性放電を模した光遺伝学的神経刺激による作業記憶の障害 Hisao Aimi（會見比佐夫）¹，Masaki Sonoda（園田真樹）^1,2，Haruo Toda（戸田春男）¹，Keisuke Kawasaki（川嵜圭祐）¹，Reiko Meguro（目黒玲子）³，Haruo Okado（岡戸晴生）⁴，Shigeki Kameyama（亀山茂樹）⁵，Tetsuya Yamamoto（山本哲哉）²，Isao Hasegawa（長谷川功）¹ ¹新潟大学医学部生理学第一教室 ²横浜市立大学医学部脳神経外科学教室 ³新潟大学大学院　保健学研究科 ⁴東京都医学総合研究所神経細胞分化ブロジェクト ⁵国立病院機構西新潟中央病院機能神経外科 Hypothalamic hamartoma (HH) is a rare congenital brain lesion. Whereas one hallmark of HH syndrome is intractable gelastic seizures (GS), about half of HH patients exhibit cognitive impairments and behavioral disorders, referred to as epileptic encephalopathy. The pathophysiological mechanisms underlying cognitive deficits associated with HH still remain unclear. Subtraction ictal SPECT coregistered to MRI (SISCOM) and EEG-fMRI that detect blood-oxygen-level dependent changes related to interictal discharges suggest increased activity in the medio-dorsal thalamus (MD)/ lateral habenula (LHb) as a key nucleus of epileptic circuits of GS with HH. We hypothesized that epileptic discharges from HH to MD/LHb deteriorates cognitive functions related to the prefrontal cortex, which has rich connections from MD/LHb. To test the hypothesis, we induced pathway-specific abnormal neural activity mimicking epileptic discharges from HH by using an optogenetic approach with channelrhodpsin2 (ChR2) in rats, and examined changes in the performance of a working memory task, the most representative behavioral test of the prefrontal functions. Specifically, we injected pCAG-ChR2-GFP incorporating adeno-associated virus serotype 9 (AAV9) into the lateral hypothalamus area (LHA) and histologically observed GFP expression at the injected area and ipsilateral MD/LHb. We then photostimulated the MD/LHb while recording neuronal activity from LHA, MD/LHb and bilateral prelimbic cortex. Head-fixed rats learned, within 3 months, to perform a working memory task in which the rats were required to pull the appropriate side of spout-lever with their forelimb after 0-9 seconds following presentation of a visual cue. We optogenetically generated the abnormal neural activity in MD/ LHb in different periods of the working memory task. We found that the optogenetically induced abnormal neural activity in the instruction period, not in the delay period, of the working memory task selectively impaired the performance of the tasks. These findings support our hypothesis and suggest that epileptic discharges propagated from HH to MD/ LHb contribute to the cognitive symptoms observed in epileptic encephalopathy associated with HH.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-468 Effects of Dominant/Subordinate Social Status on Thermal-induced Nociception and Correlation with Concentrations of Catecholamine in Prefrontal Cortex and Hippocampus Soomaayeh Heysieattalab（Heysieattalab Soomaayeh）¹，Fatemeh Bagheri（Bagheri Fatemeh）²，Emad Khalilzadeh（Khalilzadeh Emad）³，Leila Sadeghi（Sadeghi Leila）⁴，Mahdi Dolatyari（Dolatyari Mahdi）³，Mohamad Ali Nazari（Nazari Mohamad Ali）¹ ¹Division of Cognitive Neuroscience, University of Tabriz,Tabriz,Iran ²Department of Psychology, Faculty of Education and Psychology, University of Tabriz, Tabriz, Iran ³Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran ⁴Subdivision of Biochemistry, Department of Animal Biology, Faculty of Natural Science, University of Tabriz, Tabriz, Iran Social dominance is a natural and widespread phenomenon extending to most social species, including humans and animals. However, hierarchical social status greatly influence access to limited resources, reproductive opportunities and health. Laboratory and clinical studies have revealed that social interactions between pairs of conspecifics can affect the response of individuals to noxious external stimuli. Pain is an important sensorial modality with an elevated degree of complexity and subjectivity that involves not only the transduction of noxious stimuli, but also cognitive and emotional features. There has been growing interest in the relationship between pain perception and social hierarchies with a focus on exploration of mechanism(s) and mediators of pain in particular and such studies in rodents addressing the link between the pain and social hierarchical are likely to be relevant in humans. The goal of the current study was to examine the relationship between dominant/subordinate social status and the perception of pain with the concentration of Catecholamine in prefrontal cortex (PFC) and hippocampus on these processes. For this purpose, we used the well-known tube dominance test to determine social dominance in animals, and also hot plate for evaluating thermal pain model effects. Catecholamine levels were determined in rat brain regions by the enzymatic method described by Bernt and Bergmeyer. Our results showed that dominant rats significantly decreased latency to hot-plate test as compared to subordinate rats. However, Catecholamine levels in hippocampal dominant rats was significantly higher than subordinate rats but no significant differences were found in Catecholamine levels in the PFC region. It seems that social dominance increased the perception of pain and this effect may exert through increase in Catecholamine concentration in the hippocampus.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-469 Girdin/ccsc88a KOマウスに発症したてんかん発作 Machiko Iida（飯田真智子）¹，Motoki Tanaka（田中基樹）¹，Yuka Mizutani（水谷友香）¹，Kaichi Yoshizaki（吉崎嘉一）¹，Yoshihito Tokita（時田義人）¹，Naoya Asai（浅井直也）²，Masahide Takahashi（高橋雅英）²，Masato Asai（浅井真人）¹ ¹愛知県心身障害者コロニー発達障害研究所 ²名古屋大学大学院医学系研究科病理病態学講座腫瘍病理学分野 Epilepsy is one of the most common chronic brain diseases. Approximately 50 million people are estimated to suffer from epilepsy in the world. According to a recent epidemiological study, epileptic seizures in more than 30% of patients become refractory. Therefore, there is an urgent need for a better understanding of the mechanisms of epileptic seizure onset and for the development of innovative therapies for epileptic seizures. In order to achieve these objectives, a monogenic animal model with reproducible epileptic seizures would be valuable; however, such animal models are very limited. Girdin/ccdc88a is an actin-binding protein and a binding partner of Akt/PKB. Our group recently developed Girdin germline global knockout (KO) mice and nestin-lineage conditional KO mice. We found that both lines spontaneously develop generalized tonic-clonic seizures (GTCSs) with 100% genetic penetrance. GTCSs are generated almost every day with a high frequency (5-25 times per day) throughout their life (3 weeks to 2 years of age). Histological analyses showed granule cell dispersion (GCD) in the dentate gyrus and hippocampal sclerosis (HS) with GFAP-positive astrocytes in global Girdin KO mice. Results of temporal analyses of the GCD and HS will also be provided in the presentation. Electroencephalograms (EEGs) obtained in nestin-lineage conditional KO mice were examined. High-amplitude waves were continuously detected for 24 hours and the largest amplitude was recorded at an electrode just above the hippocampus. High-amplitude and high-frequency EEGs (bursts) were synchronously observed with a GTCS. Considering the lesions (GCD and HS) in the hippocampus, the GTCSs in Girdin KO mice may originate from the hippocampus. GCD, HS and hippocampus-originating GTCSs found in Girdin KO mice are characteristic of mesial-temporal lobe epilepsy (MTLE). We also reported that a human Girdin/ccdc88a loss-of-function mutation in patients caused a PEHO-like neurodevelopmental phenotype including early onset tonic-clonic seizures (Nahorski et al., 2016 Brain). Together with the unique characteristics in the Girdin KO mouse, which is a monogenic animal model, the complete penetrance and high frequency of GTCSs, and the non-requirement of chemical induction for epileptic seizures, the Girdin KO mouse is a potentially valuable animal model for investigating the mechanisms of MTLE onset and for the development of innovative therapies for MTLE.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-470 線虫神経回路の信号伝達におけるギャップ結合の機能のシミュレーションによる解析 Kazumi Sakata（坂田和実），Tokumitsu Wakabayashi（若林篤光），Tarou Ogurusu（小栗栖太郎）岩手大理工化学生命 Caenorhabditis elegans has 302 neurons and all neural connections were revealed with electron microscopy. The nematode shows various responses to the variety of environmental stimuli with its small neural network. Even though all connections were revealed, properties of almost connections, being excitatory or inhibitory, were not determined. To determine the properties of the neural connection and to reveal the mechanism which processes environmental signal to actuate body muscles for locomotion, computer simulation was performed. In the neural network model, which was a part of the neural network of the nematode and of which neurons were involved in the behavior responding to the various chemical stimuli, there were two kinds of connection, chemical synapse and gap junction. Connection weight of each connection was determined by genetic algorithm. At first, the gap junction was implemented as a pair of two connections of which directions were opposite and their connection weight were implemented as they always had an identical value. In the simulation with this connection model, the connection weights of the gap junctions were close to zero. Second, connection weight of each direction of gap junction was varied independently. With this model, the weights of both directions of the gap junction were distributed around value of zero. Difference of the weights of two directions were also distributed around zero. Two connection weights obtained under the second condition did not show correlation. It means that two connection weights of two directions between two neurons consisting of gap junction could be non-identical.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-471 深層ニューラルネットワークにおける単一細胞の両耳間時間差チューニング Takuya Koumura（上村卓也），Hiroki Terashima（寺島裕貴），Shigeto Furukawa（古川茂人） NTTコミュニケーション科学基礎研 Sound usually reaches the ear nearer to the sound source faster than it does to the other ear. The difference in the timing of a sound's arrival at the two ears is called the interaural time difference (ITD). Since the ITD depends on the azimuth of the sound source relative to the listener, it is utilized by animals as an important binaural cue for sound localization. Moreover, binaural hearing is also advantageous for sound detection and identification (Kidd, 1995, J. Acoust. Soc. Am.; Dubno, 2008, J. Speech Lang. Hear. Res.), presumably due to the presence of implicit information on sound location. Some neurons in the auditory system respond selectively to sounds with specific ITDs (Grothe et al., 2010, Physiol. Rev.). ITD tuning is considered to be one of the neural mechanisms of binaural hearing, including binaural sound localization, detection, and identification. However, it is still unknown why ITD tuning has emerged in the auditory system. For example, a question that arises is ""does ITD tuning emerge only from binaural sound detection and/or identification without explicit sound localization?"" This pilot study attempted to answer the above question by taking a modeling approach. Specifically, we trained a computational model of the auditory system for binaural sound detection and identification, and calculated the ITD tuning in the trained model. We used a deep neural network (DNN) as a model. The DNN is a machine learning model for sound recognition. Since it processes a sound through its cascaded layers for recognition, it is suitable for modeling the auditory system, which processes a sound for recognition through cascaded brain regions. We trained a DNN to detect and identify acoustic events from binaural natural sounds (Mesaros, et al., 2016, EUSIPCO). Then, we analyzed the trained DNN using neurophysiological methods (Koumura, et al., 2018, bioRxiv). We recorded single-unit activities in the DNN that arose in response to tones with various ITDs. Some units in the DNN exhibited asymmetric ITD tuning with respect to zero ITD. On the other hand, the ITD tunings in a DNN trained on diotic sounds were more symmetrical, indicating insensitivity to the sign of the ITD. These results suggest that ITD tuning emerged from optimization to binaural natural sound detection and identification even without explicit optimization to localization.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-472 脳構造に着想を得た初期化手法のRNNとLSTMへの適用 Tomohiro Fusauchi（房内智弘）¹，Toshikazu Samura（佐村俊和）^1,2 ¹山口大院創成科学研究科 ²玉川大脳研 The performance of a neural network is affected by initial constraint on network structure (e.g. Glorot and Bengio, 2010, He et al., 2015). A neural network is inspired from the mechanisms of information processing in the brain. It is expected that the computational performance of a neural network is improved by the introduction of brain structures. On the other hand, the brain is known to be robust to failure. The partial lesions of the brain network degrade functions instead of total functional failure (e.g. Moser et al., 1995). In this study, we introduced brain anatomical structures (neuron type, partial connection, small world structure) into all-to-all connected RNN and Long Short-Term Memory (LSTM) network as an initial structure. We evaluated the performance of the RNNs in various tasks and their robustness against the partial failure of a network after training through BPTT. We replaced synaptic weights of inputs or outputs (input destruction, output destruction) of a part of neurons by zeros to evaluate the robustness. The performance of all-to-all connected RNN improves by the introduction of neuron type. In the network, neurons were connected by excitatory and inhibitory connections at the same ratio as the brain. The introduction of partial connection or small world structure or both did not improve the performance of a RNN. However, the introduction of partial connection reduced the effect of partial failure. Although All-to-all connected RNNs severely degraded their performance by the failure, partially connected RNNs degraded them moderately. There is a trade-off between performance and robustness in RNNs with the introduction of brain structure. Moreover, we found that two types of destruction cause different degradation in performance even though the same number of connections was destructed. On the other hand, the introduction of neuron type also improved the performance of all-to-all connected LSTM network. The difference in degradation between the two types of destruction was smaller in the LSTM network compared to RNN. However, the performance was apparently degraded. Consequently, the introduction of excitatory and inhibitory connections as an initial constraint improves the performance of all-to-all RNNs and LSTM network. The other types of initial constraint may be needed to improve robustness against partial failure.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-473 Brain hierarchy score: ヒト視覚野に似た階層性を持つ深層ニューラルネットワークはどれか？ Soma Nonaka（野中聡馬）¹，Shuntaro C. Aoki（青木 C. 俊太郎）²，Kei Majima（間島慶）³，Yukiyasu Kamitani（神谷之康）^2,3 ¹京都大総合人間 ²ATR脳情報研究所 ³京都大院情報 Multistage hierarchical processing in both deep neural networks (DNNs) and human brain is considered to play a vital role in achieving high-performance visual recognition. A previous study from our group has shown that, by analyzing brain activity patterns obtained with functional magnetic resonance imaging (fMRI), low-/high-layer DNN features are well decoded from low-/high-level visual areas, respectively (Horikawa & Kamitani, 2017). This suggests that there is a hierarchical correspondence (homology) of the internal representations between DNN and the human brain. While one of the representative DNNs, AlexNet, was used in this study, a variety of DNNs with different architectures have been proposed in recent years, and which types of DNN are similar to the human brain is of great interest. A recent study quantified the similarity of the representation to a specific brain area such as V4, and compared it across several types of DNN (Schrimpf et al, 2018). However, no study compared the degree of the hierarchical correspondence to the human visual hierarchy across DNNs. In this study, we compared 13 DNNs in terms of the hierarchical correspondence to the human brain. Using fMRI responses to 1250 natural images in five subjects, we performed decoding analysis where individual units in each layer of the 13 DNNs were predicted from seven human visual areas. We found that some DNNs showed clear hierarchical correspondence to the human visual hierarchy up to middle- or high-level visual areas, while others exhibited almost no hierarchical correspondence even though they performed object recognition at a near-human level. To quantitatively evaluate this hierarchical correspondence, we developed an index called "brain hierarchy score (BH score)" and compared it across the 13 DNNs. DNNs with relatively small numbers of layers showed high BH scores and those with more than 50 layers did not. This result is consistent with the view that recently proposed DNNs with many layers are too complex as a model of the human visual system (Kubilius et al., 2017). DNNs with recurrent or skip connections showed low BH scores, suggesting that there is a large gap between such connections in DNNs and those in the human brain. Our results provide a new insight into the similarity and the difference between DNNs and the human brain.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-474 スパースモデリングに基づく神経回路の構造推定 Ren Masahiro（政廣蓮），Toshiaki Omori（大森敏明）神戸大学大学院工学研究科電気電子工学専攻 In recent years, tremendous advances have been made in measurement technology of neuroscience. In particular, imaging techniques such as voltage imaging and calcium imaging enable us to observe neuronal responses from multiple neurons simultaneously in a more precise manner. In order to deepen understanding of neural systems, it is important to develop a data-driven method for extracting a latent dynamics of neural network underlying observable imaging data. In this study, we propose a data-driven approach for estimating neural network structure. In order to achieve this, we consider the following three properties. The first property is that each neuron obeys nonlinear dynamics. By reflecting neuronal nonlinear dynamics, we derive nonlinear state space model of neural network dynamics consisting of conductance-based neuron models. The second property is that neural networks have sparsity in their synaptic connections. The number of synaptic connections is assumed to be relatively small for the number of neurons. The third property is that neurons obey Dale's principle; each neuron can emit either excitatory transmitters or inhibitory transmitters. In order to develop a data-driven approach reflecting these three properties, we construct a statistical data-driven method based on sequential Monte Carlo (SMC) method and Group LASSO. The SMC realizes extraction of nonlinear dynamics from observable data, whereas the Group LASSO realizes estimation of the neural network structure based on prior information of both sparsity in synaptic connections and Dale's principle. To show effectiveness of the proposed method, we estimate neural network structure using simulated observable data. Using only noisy membrane potentials from multiple neurons, we find that the proposed method successfully extract neural network structure. Furthermore, reconstructing potentials from estimated network structures by existing methods and proposed method, we show that the proposed method is superior to the existing methods in structure estimation of neural networks.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-475 マウスのテトロード単一ユニット記録データにおける自動スパイクソーティング Sumire Matsumoto（松本すみ礼）^1,2，Keiichi Morikuni（保國惠一）³，Momo Matsuda（松田萌望）⁴，Kotaro Sakamoto（坂本航太郎）¹，Kaoru Ohyama（大山薫）²，Tetsuya Sakurai（櫻井鉄也）³，Kaspar Vogt（Vogt Kaspar）² ¹筑波大学　グローバル教育院　ヒューマンバイオロジー学位プログラム ²筑波大学国際統合睡眠医科学研究機構 ³筑波大学システム情報系情報工学域 ⁴筑波大学システム情報工学研究科コンピュータサイエンス専攻 Single spike sorting for extracellular recording data is a necessary technique for investigation of neural activity in brain, yet remains challenging. Although several algorithms have been proposed (Chung et al., 2017; Dhawale et al., 2017), a fully automated sorting system has not been well established, and sorting is still mostly done manually requiring substantial effort and time. There are several obstacles to automation; drifting of clusters and a high level of background noise. Moreover, automated algorithms were so far mainly applied to rat recordings because rats represent one of the most popular experimental animal models in extracellular recording. However, it is also crucial to develop automated sorting for mouse recording data, since mice with their shorter generation time and genetic tractability offer key advantages as experimental models. We thus use the mixture of drifting t-distributions (MoDT) (Shan et al., 2017) for 24 hour tetrode recording data from freely behaving mice. This spike sorting mechanism is based on an expectation-maximization (EM) algorithm with the t-distribution which has heavier tails than the Gaussian distribution. There are three modifications to the basic EM model; first, replacing the multivariate Gaussian distribution to the multivariate t-distribution, second, breaking up the time series into T time frames (1 minute per frame in our case), third, penalizing large drifts of the clusters over consecutive series. We investigated applying this clustering algorithm to our data in a hierarchical manner and obtained some stable single clusters per tetrode throughout 24 hours. The clusters were evaluated using the isolation distance, L-ratio and interspike interval. We are planning to apply the algorithm to more data and evaluate its performance in various experimental situation or gene modified animals in the future.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-476 様々な認知機能間の関係性を大規模fMRIデータベース分析とネットワーク理論から明らかにする Hiroki Kurashige（倉重宏樹）^1,2，Yuichi Yamashita（山下祐一）²，Rieko Osu（大須理英子）³，Yohei Otaka（大高洋平）^4,5，Takashi Hanakawa（花川隆）⁶，Manabu Honda（本田学）²，Hideaki Kawabata（川畑秀明）⁷ ¹東海大創造科学技術研究機構 ²国立精神・神経セ神経研 ³早稲田大人間科学学術院 ⁴藤田保衛大医 ⁵東京湾岸リハビリテーション病院 ⁶国立精神・神経セ脳病態統合イメージングセ ⁷慶應大文心理 In the present study, we propose an approach to reveal relationships among dozens of cognitive functions based on the functional brain mapping and network analysis. According to the previous study, from the fMRI database we first reconstructed 109 pseudo-activation maps (cognitive function maps; CFM), each of which are corresponding to 109 cognitive functions such as ""emotion"", ""attention"", ""episodic memory"", and so on. Then, to survey a whole picture of relationships among the cognitive functions, we mapped those onto a two-dimensional space where the relevant functions were located close to each other on the basis of the resting-state functional connectivity among CFMs. Moreover, we tried to conduct conceptual analysis of cognitive functions using clustering of voxels in each CFM based on the functional connectivity from the voxels to the other 108 CFMs. As a result, a CFM for each cognitive function was subdivided to several parts, each of which are strongly associated with some CFMs for subset of the other cognitive functions, which brought in sub-concepts (i.e. sub-functions) of the cognitive function. In addition, we conducted whole-brain parcellation using spatially constrained spectral clustering based on the connectivity from each voxel in the brain to the 109 CFMs, resulting in the ~200 parcels, each of which is featured with degrees of associations with the 109 cognitive functions. Applying pseudo-heat diffusion analysis to the network consisting of these parcels as nodes, we revealed that memory-related parcels interact with functionally most diverse parcels. Moreover, using the clique percolation method, we showed that vision-, motion-, and language-related subnetworks have most dense intra-subnetwork connectivity. In addition, we investigated functional homogeneity within each network community identified with the clustering analysis and found the high inhomogeneity in the cerebellar community. Based on the present method, revealing relationships among cognitive functions is expected to suggest natures of each cognitive function and neural mechanisms to work such a function out. This will provide hypotheses to test from both neural and behavioral views to the neuroscience community. In addition, knowledge of the relationships among cognitive functions will give the AI community guidelines and inspirations to develop novel algorithms. Finally, it will bring us a view of the brain as a unity of cognitive functions.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-477 Framework for predicting probability from brain activity Shirin Vafaei（Vafaei Shirin）^1,2，Kei Majima（Majima Kei）¹，Hiroaki Yamane（Yamane Hiroaki）¹，Yukiyasu Kamitani（Kamitani Yukiyasu）^1,2 ¹Grad Informatics, Kyoto Univ, Kyoto ²ATR Computational Neuroscience Lab, Kyoto, Japan Brain decoding with machine learning methods has provided a powerful framework for extracting information encoded in population neural activity. Multivariate regression methods such as variants of linear regression are used when a continuous variable is predicted. However, variables we want to predict in brain decoding analysis sometimes take the form of probability, the variable with special properties that separates it from other continuous variables. For example, the performance in a cognitive task (e.g., attention task and working memory task) has been predicted from functional magnetic resonance imaging (fMRI) activity in previous studies, and the task performance was measured by the proportion of correct answers (i.e., accuracy). Also, impression scores for images/videos/products (e.g. valence, arousal) were often defined by what proportion of raters feel a specific impression in subjective judgements. While these probabilities were predicted by applying standard regression methods, this is not appropriate because most of the regression methods do not take into account the fact that the variable to be predicted is probability. To treat this in an appropriate manner in brain decoding analysis, we introduced probit regression, a generalized linear model whose output is assumed to be probability, as a regression model and trained it using the Kullback–Leibler divergence as the cost function, which is known as a suitable distance measure for probability distributions. We also introduced sparse regularization into probit regression (sparse probit regression) to prevent overfitting. In both simulation and real fMRI data analysis, sparse probit regression better predicted probabilities than probit regression without sparse regularization, indicating that sparseness leads to better decoding performance. Sparse probit regression also outperformed linear regression with the same type of sparse regularization, indicating the advantage of our appropriate treatment of the probability. Our results suggest that our framework with sparse probit regression provides an effective method for predicting variables of interest quantified as probability, such as task performance, emotion/ impression scores, and confidence, from brain activity.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-478 神経細胞の誕生日を利用した分類と操作を可能にする神経細胞の誕生日タグづけ法 Tatsumi Hirata（平田たつみ）国立遺伝研脳機能 During development, neurons are generated over a protracted time window. Their differentiation timing, which is often referred to as the ""neuronal birthdate°, has immense impacts on phenotypes of differentiated neurons. Directly or indirectly, it can determine their molecular, morphological or physiological properties, and influences their axon trajectories and connection patterns. This is the reason why neuronal birthdating has been so widely used traditionally for classification of neurons in various nervous systems. However, the current methods using nucleotide analogues have obvious limitations; they are purely histological and can only label cell nuclei after fixation. To overcome the limitations, we developed ""birthdate tag mouse lines°, in which tamoxifen-inducible Cre recombinase, Cre-ER^T2 is only transiently expressed in newly born neurons under enhancers of neuronal differentiation genes such as neurogenins. In the mice, administration of tamoxifen at a certain developmental stage induces loxP recombination only in the neurons several hours after the final mitosis. Subsequently, the birthdate tagged neurons can be studied and manipulated later for various purposes in combination with different reporter or effector genes. In this meeting, I will introduce four independent birthdate tag lines that were established using enhancers of different genes, and discuss potential applications of these mouse lines. The new mouse resources will be useful for many researchers working in neuroscience fields.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-479 オプトジェネティクスに向けた一価カチオン選択性チャネルロドプシンの機能解析 Shunta Shigemura（重村竣太）¹，Shoko Hososhima（細島頌子）¹，Hideki Kandori（神取秀樹）¹，Satoshi Tsunoda（角田聡）^1,2 ¹名古屋工大院工 ²JSTさきがけ Channelrhodopsin is 7-transmembrane protein, functioning as a light-gated passive ion channel [1, 2]. Optogenetics enables to control biological functions simply by light after expressing light-sensitive proteins into living tissues [3, 4]. Gt_CCR4 is a novel light-gated cation channelrhodopsin from a cryptophyte algae Guillardia theta [5]. Surprisingly, Guillardia theta encodes four genes of cation channel rhodopsins in the genome (Gt_CCR1~4) [5-7]. Gt_CCR4 has close sequence homology to bacteriorhodopsin of H+ pumping rhodopsin (BR) rather than the well-known channelrhodopsin-2 from Chlamydomonas reinhardtii (Cr_ChR2), in which several critical amino acids are not conserved. Thus, the channel gating and ion transport mechanism of Gt_CCR4 could be different from Cr_ChR2. Actually Gt_CCR4 exhibits large photocurrent without large inactivation and desensitization, serving as a new optogenetics tool. We performed electrophysiological measurements to characterize Gt_CCR4 and Cr_ChR2 in ND7/23 cells and primary cultured cortical neurons. The channel properties such as ion selectivity, light sensitivity, kinetics were compared. We further explored a putative channel pore region on a basis of ion permeation properties. As the result, Gt_CCR4 shows high sodium selectivity and low calcium and proton selectivity than Cr_ChR2. Gt_CCR4 shows high light sensitivity than Cr_ChR2. Thus, Gt_CCR4 serves as an alternative optogenetics tool, especially when large Na+ change is required while pH and/or calcium change should be minimized. [1] Nagel et al. 2003, PNAS 100, 13940-13945 [2] Kato et al. 2012, Nature. 482(7385), 369-74 [3] Boyden et al. 2005, Nat. Neurosci. 8, 1263-1268 [4] Ishizuka et al. 2006, Neurosci Res. 54, 85-94 [5] Yamauchi et al. 2017, Biophys. Physicobiol. 14, 57-66 [6] Sineshchekov et al. 2016, Biophys. J. 110(11), 2302-2304 [7] Sineshchekov et al. 2017, PNAS 114(45), E9512-E9519

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-480 複合体特異的なAMPA受容体光不活化法の開発 Kiwamu Takemoto（竹本研），Takuya Takahashi（高橋琢哉）横浜市大医生理 Acute inactivation of synaptic neurotransmitter receptors in vivo with spatio-temporal precision should be a powerful tool to understand their roles in cognitive functions. Excitatory and inhibitory neurotransmitter receptors usually form protein complex by various combinations with subunits and express their spatio-temporal function. For example, AMPA type glutamate receptors (AMPA-Rs), which are well known to be important glutamate receptors for learning, are composed of variable combinations of four subunits, GluA1-4. Among them, GluA1 homomer, GluA1/2 heteromer and GluA2/3 heteromer were known to be expressed in adult brain. AMPA-Rs with GluA1 subunits require plasticity-inducing protocols and NMDA-Rs activation to be driven into synapses and serve to enhance transmission. In contrast, GluA2/3 heteromer complex continuously replace synaptic receptors in a manner that maintains transmission (Shi, S-H. et al. Cell 2001, Takahashi, T. et al. Science 2003 etc.). In addition, subunit combinations affect the functions of ion channel. Complexes lacking GluA2 subunit shows calcium permeability and high single channel conductance (Dingledine R et al. Pharmacol Rev 1999, Coombs ID et al. J. Neurosci. 2012). These observations support the idea that AMPA-Rs complexes should have different physiological functions in vivo. To elucidate their complex-specific functions in vivo, we have developed an optical technology for acute inactivation of synaptic GluA1 homomeric receptors in vivo by chromophore assisted light inactivation (Takemoto et al. Nat. Biotechnol. 2017). This technology enabled us to elucidate their physiological functions of contextual fear learning in hippocampus with complex specific manner. In this presantation, we will discuss our new CALI technology for other AMPA-Rs complex.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-481 ゼブラフィッシュにおける逆行性トレーシングとカルシウムイメージングを目指した水疱性口炎ウイルスベクターの開発 Hisaya Kakinuma（柿沼久哉），Toshiyuki Shiraki（白木利幸），Hitoshi Okamoto（岡本仁）理研CBS 意思決定回路動態 An increasing number of comparative neuroanatomical and behavioral analyses have revealed that the basic structures and functions of brain are conserved between teleosts and mammals. However, compared to mammalian model animal, there have been little reports on utilization of virus vectors for studying neural circuits of the zebrafish brain. Previously we applied in vivo anterograde tracing mediated by vesicular stomatitis virus (VSV) to the adult zebrafish dorsal pallium, and found that the central part of the dorsal pallium (Dc) neurons project to the dorsal nucleus of the ventral telencephalic area (Vd), the dorsal entopeduncular nucleus (dEN) and the preglomerular nucleus (PG). And comparison of the tectal afferents mediated by VSV and lipophilic tracer revealed that the Dc neurons directly projected to the middle layer of the tectum and indirectly projected to the deep layers via dEN neurons. In this study, we attempted to confirm the pallio-tectal projection by VSV(RABV-G)-mediated retrograde tracing. A VSV(RABV-G) encodes rabies virus glycoprotein (RABV-G) instead of its own glycoprotein VSV-G and can transmit retrogradely. Injection of a recombinant VSV(RABV-G), rVSV(RABV-G) into the tectum showed that retrogradely labeled neurons were found on the surface of the dorsal pallium ipsilaterally, however, not in the Dc and the dEN. In some cases, radial glia-like neural stem cells on the surface of the dorsal pallium were also retrogradely labeled. These data suggested that surperficial neurons on the dorsal pallium projected to tectum ipsilaterally not via Dc or dEN neurons. And it also suggested that VSV(RABV-G) might have a different cellular tropism from native VSV due to its glycoprotein. Next, we took advantage of low toxicity of VSV and applied it to in vivo calcium imaging of adult zebrafish brain. rVSV-G-CaMP7 was injected to the dorsal pallium and the infected fishes were trained with an active avoidance paradigm. 24 hrs after last training, we measured fluorescence changes in response to LED stimulation as the CS presentation. In learner fish, neural activities in the telencephalon and tectum ipsilaterally were detected upon the LED stimulation as shown in our previous study with transgenic fish. These data indicated that VSV-mediated calcium imaging could be applicable to monitoring neural activities in vivo. Now we are trying to apply optogenetics with VSV harboring opsins to manipulate neural activity in adult zebrafish.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-482 細胞内局所的活性酸素発生によるアクチンフィラメントの操作 Tetsuya Ishimoto（石本哲也），Hisashi Mori（森寿）富山大学　院医薬 Actin cytoskeleton is known to define the morphology of peripheral structures of the cells including neuronal spine. Artificial manipulation of actin filament could modulate cellular activity such as neuronal motility, neurite outgrowth and synaptic transmission. In this study, we attempted to establish a new method to manipulate actin filament by targeted generation of reactive oxygen species on actin filament. Firefly luciferase which emits green light was fused with KillerRed protein which is known to generate reactive oxygen species when excited by green light. In this fusion protein, treatment with luciferin, a substrate of luciferase, leads to generate reactive oxygen species by bioluminescence resonance energy transfer. The fusion protein tagged with F-actin targeting peptide was expressed in HEK293T cells and stimulated with luciferin. We observed dramatically increased actin filament under fluorescent microscopy 24 h after luciferin treatment. The increase in actin filament was confirmed by biochemical separation analysis using ultracentrifuge. On the other hand, no increase in actin polymerization was seen in the HEK293T that expressed luciferase-KillerRed fusion protein without F-actin targeting peptide. These results indicate targeted, not global, generation of reactive oxygen species on actin filament can modulate the activity of actin or actin-binding proteins and induce polymerization. Since the actin filament induced by this method was co-localized with cofilin, the filament is likely a structure called cofilin-actin rod. As cofilin-actin rod is seen in the neurons of Alzheimer's disease patient, our method may be used to mimic pathology of Alzheimer's disease in vitro and in vivo. Furthermore, there are many proteins whose activities are regulated by oxidization and reduction. Our method can be a general technique to manipulate the activity of cellular proteins which are sensitive to reactive oxygen species.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-483 抑制性ニューロン特異的プロモーターの開発と解析 Chiaki Hoshino（星野千秋）¹，Ayumu Konno（今野歩）¹，Ryosuke Kaneko（金子涼輔）²，Hirokazu Hirai（平井宏和）¹ ¹群馬大院医脳神経再生医学 ²群馬大院医生物資源セ Viral vectors with a cell type-specific promoter allow us to express a transgene in a specific cell population. Although various cell type-specific promoters are currently available, promoters specific to inhibitory neurons still have not been developed sufficiently. Here, we aimed to develop a new inhibitory neuron-specific promoter. We cloned possible promoter regions upstream of genes expressing specifically in GABAergic neurons, which include VGAT, GAD65, GAD67. Then, lentiviral vectors expressing GFP under the control of above candidate promoters were produced. Those lentiviral vectors were injected into mouse cerebral cortex to examine the cell type-specific expression profiles. We found that all candidate promoters have strong promoter activity in excitatory neurons. However, GAD65 and GAD67 promoters have some promoter activities also in inhibitory neurons. Then, using AAV9 vectors, we further examined those GAD65 and GAD67 promoters in mouse cerebral cortex. The results showed the GAD65 promoter having higher specificity to inhibitory neurons than the GAD67 promoter. After AAV9-based analyses of shortened versions of the GAD65 promoter, we found that a region of approximately 2.5 kb, GAD65(de1) served as an inhibitory neuron-specific promoter. The specificity and transduction efficiency of the GAD65(de1) promoter in other brain areas were examined by AAV-PHP.B, which shows a great blood brain barrier (BBB) permeability in mice (Deverman et al, 2016). Intravenous injection of PHP.B expressing GFP under the control of the GAD65(de1) promoter resulted in inhibitory neuron-specific GFP expression in the hindbrain (although no transduction in Purkinje cells) as well as the forebrain. Since the Dlx enhancer shows the forebrain specific transduction, the GAD65(de1) promoter would be a useful piece in neuroscience research.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-484 アデノ随伴ウイルスベクターの超簡易精製法 Ayumu Konno（今野歩），Hirokazu Hirai（平井宏和）群馬大院医脳神経再生医学 Adeno-associated virus (AAV) vectors are widely used in neuroscience research, since they can infect and express a transgene efficiently in both neurons and glial cells in the central nervous system (CNS) of living animals. However, a density gradient centrifugation step, which is needed for purification of AAV particles, costs substantially and takes time. Here, we report a super simplified method (SSM) for a purification of AAV-PHP.eB, a capsid variant of AAV9 having a great permeability to the blood brain barrier (BBB) in mice (Chan et al., Nat Neurosci 2017). The SSM does not need an ultracentrifuge, and requires only conventional machines normally equipped in a culture room, i.e., a bench for cell culture (clean bench or safety cabinet) and a centrifuge for collecting cells. In the SSM, the 293T cells were grown on 15 cm dish, followed by transfection with an expression vector (pAAV/CBh GFP WPRE), rep/cap plasmid (pAAV-PHP.eB), and pHelper. One day after transfection, the medium was changed to FBS-deficient D-MEM. AAV particles were released from 293T cells to the medium due to the undernutrition. Six days after the incubation, the medium was collected, and AAV particles were concentrated and formulated in PBS with Vivaspin 20 centrifugal concentrator (MWCO 100 kDa). This solution can be used directly as viral solution. The yield of AAV vectors from one 15 cm dish was 1.97 × 10¹² ± 9.2 × 10¹⁰ vg/ml. The AAV vectors obtained by the SSM was significantly less, in terms of the purity, than AAV vectors conventionally obtained using a ultracentrifuge. However, the brain-transduction efficiency by intravenous infusion of AAV-PHP.eB obtained by the SSM was almost comparable to that by the ultracentrifuge-purified one. Thus, the SSM, which saves costs and time greatly, is useful for obtaining high titer AAV vectors available for in vivo administration.		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-485 クリスパー活性化によるアルツハイマー病モデル細胞の作製とその利用 Keiichi Inoue（井上敬一）^1,2，Luis MA Oliveira（Oliveira MA Luis）¹，Asa Abeliovich（Abeliovich Asa）¹ ¹Dept Pathol, Columbia Univ, New York, USA ²新潟大学院医歯学機能制御学 A challenge to the study of human neurodegenerative diseases, such as Alzheimer's disease (AD), is the limited availability of facile cell-based disease models. Mutations in presenilin (PSEN) 1 and 2, which encode components of the gamma-secretase complex, cause familial Alzheimer's disease (FAD). It is hypothesized that altered gamma-secretase-mediated processing of the amyloid precursor protein (APP) to the Abeta42 fragment, which is accumulated in diseased brain, may be pathogenic. However, the alteration in the processing has not been fully presented in the FAD patient-derived skin fibroblast cultures. Here we describe an in vitro fibroblast model system that enables the facile analysis of neurological disease mechanisms in non-neuronal patient cells using CRISPR transcriptional activation of endogenous disease-relevant genes. CRISPR activation of APP and/or BACE1 increases beta-CTF, the substrate for gamma-secretase, and extracellular Abeta levels (Abeta42, Abeta40, and Abeta38) in skin fibroblast cultures. In FAD patient-derived fibroblasts, CRISPR activation of APP and/or BACE1 unmasked an occult processivity defect in downstream gamma-secretase-mediated carboxypeptidase cleavage of APP (the lowered ratio of Abeta38 per Abeta42 in FAD fibroblasts), ultimately leading to higher toxic Abeta42 levels. These data suggest that, selectively in neurons, relatively high levels of BACE1 activity lead to substrate pressure on FAD-mutant gamma-secretase complexes, promoting CNS Abeta42 accumulation. Our results introduce an additional platform for analysis of neurological disease. We have further characterized the AD-associated genes in the context of gamma-secretase processivity using our CRISPR-fibroblast system.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-486 Sensitive and stable quantitation of endogenous oxytocin in mice using reduction/alkylation approach for ELISA Stanislav M. Cherepanov（Cherepanov Stanislav M.），Maria Gerasimenko（Gerasimenko Maria），Teruko Yuhi（Yuhi Teruko），Shigeru Yokoyama（Yokoyama Shigeru），Haruhiro Higashida（Higashida Haruhiro） Kanazawa University Oxytocin (OT) is a nonapeptide essential for the social brain. Several studies have reported a strong relationship between central and peripheral levels of OT as well as the ability of plasma OT to be a neurological biomarker of autism, anxiety and other mental disorders. There are several proteomic tools to measure OT in peripheral fluids, such are radioimmunoassays (RIA), immunosorbent assays (ELISA) and mass spectrometry (MS). However, specificity and selectivity of these methods are criticized. High matrix interference of plasma OT with albumins in data of direct determination is doubtful and overestimated. An established way to solve this problem – is a measurement of only the free OT fraction, employing solid phase extraction (SPE). However, SPE required a significant amount of plasma (1-5 ml) which made it difficult to use in basic or translational research. Additionally, several studies indicated large variations of measured OT levels after SPE or even absence of detectable levels of OT. Recently, Brandtzaeg group discovered reduction/alkylation followed by plasma precipitation (RAPP) methods that enable to save a sufficiently high amount of OT released from plasma proteins as well as provide a reduction of matrix interference. Importantly, the required volume of plasma is just 100 μl. This method was implemented for the MS platform. However, we aim to adopt this approach for ELISA. First, we evaluated 8 commercially available ELISA kits using a set of diluted plasma samples and samples after SPE from mice plasma pool. Data indicated a high variation of obtained results. The absence of correlation between diluted plasma and SPE plasma also was notable. After that, we have performed RAPP prior ELISA measurement, by 2 selected kits with most stable previous results. Obtained results slightly but significantly lower from results of diluted plasma samples, and demonstrated low variation between two assays. Protein precipitation only leads to a dramatic loss of OT from the sample. This data indicated essential roles of RAPP to release OT from binding with plasma albumins. Finally, we validated RAPP sample treatment in comparison of plasma pools of ICR and CD38KO mice, a model with disrupted OT release from the paraventricular nucleus. Obtained data confirmed lower levels of OT in CD38KO plasma. Overall data indicated RAPP prior ELISA – potentially a good alternative of SPE to obtain stable and true-positive levels of OT in plasma		7月25日（木）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-487 LINuSによる蛍光タンパク質の核内移行を利用した脳内における青色光の伝達マッピング Norihiro Kimizuka（君塚悟洋）¹，Ayumu Inutsuka（犬束歩）²，Hisashi Yakabu（屋嘉部久志）³，Natsuki Takanohashi（高野橋菜月）³，Tatsushi Onaka（尾仲達史）² ¹自治医大院神経脳生理 ²自治医大医神経脳生理 ³自治医大医 Optical manipulations are widely used to analyze neuronal functions in vivo. Blue light is frequently used to activate channelrhodopsins or LOV domains although its absorption and scattering is higher compared with longer wavelength light. High spatial resolution of optical manipulation is easily achieved in vitro, while light is unevenly scattered and absorbed dependent on many factors in the tissue. However, it is difficult to spatially measure a blue light transmission area in vivo. Here, we propose a genetic method to visualize the blue light transmission in the brain using light-induced nuclear translocation of fluorescent proteins using LOV domain. Light-inducible nuclear localization signal (LINuS) consists of LOV2 domain fused with NLS. LINuS can be used as a transporter unit for light-induced nuclear translocation. We confirmed that blue light illumination induced reversible translocation of NES-tdTomato-LINuS from the cytosol to the nucleus with in 30 minutes in HEK293 cells. By employing PHP.eb capsid, which can penetrate the blood brain barrier, a retro-orbital sinus injection of adeno-associated virus vectors induced ubiquitous expression of NES-tdTomato-LINuS in the brain. We confirmed that 30-minutes transcranial blue light illumination with a LED pad induced nuclear translocation of NES-tdTomato-LINuS in pyramidal neurons in the cortex and the granule cells in the dentate gyrus. We also found that mice exposed to blue light at the shaved abdominal area exhibited a substantial increase in nuclear translocation in the surface area of the liver. These results provide a simple way to get useful information on light transmission in the tissue without any transgenic animals or skillful procedures.

7月26日（金）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-488 KANPHOS (Kinase-Associated Phospho-Signaling) Platform - 包括的リン酸化DBプラットフォーム - Keisuke Kuroda（黒田啓介）¹，Mutsuki Amano（天野睦紀）¹，Junichiro Yoshimoto（吉本潤一郎）²，Taku Nagai（永井拓）³，Takayuki Kannon（観音隆幸）⁴，Tomoki Nishioka（西岡朋生）¹，Shiro Usui（臼井支朗）⁵，Kozo Kaibuchi（貝淵弘三）¹ ¹名古屋大院医神経情報薬理 ²奈良先端科学技術大学院大学情報科学領域数理情報学研究室 ³名古屋大院医医療薬学 ⁴金沢大学医薬保健研究域　医学系革新ゲノム情報学分野 ⁵(独)理化学研究所脳神経科学研究センター神経情報基盤開発ユニット Protein phosphorylation is a major and essential post-translational modification in eukaryotic cells that plays a critical role in various cellular processes. While recent advances in mass spectrometry based proteomics allowed us to identify approximately 200,000 phosphorylation sites, it is not fully understood which sites are phosphorylated by a specific kinase and which extracellular stimuli regulate the protein phosphorylation via intracellular signaling cascades. Recently, we have developed an in vitro approach termed the kinase-interacting substrate screening (KISS) method and an in vivo approach termed kinase-oriented substrate screening (KIOSS) method. Using KIOSS method, we analyzed the phosphorylation signals downstream of dopamine in mouse striatal slices, and found that about 100 proteins including ion channels and transcription factors were phosphorylated probably by PKA or MAPK. Here, we present an on-line database system which provides the phosphorylation signals identified by our KISS and KIOSS methods as well as those previously reported in the literature. The database system and its web portal, named KANPHOS (Kinase-Associated PHOspho-Signaling), were built based on the Next Generation XooNIps. All data are controlled for quality via review and curation by our professional staffs. In the portal site, we can search for the data of interest in three ways: 1) Search for substrates phosphorylated by a specific kinase; 2) Search for kinases phosphorylating a specific protein; and 3) Search for kinases and their target substrates by a specific signaling pathway. Each substrate is linked with external databases such as Uniprot KB (proteomics database), HGNC DB (human genomics database), and Allen Brain Atlas, enabling us to easily predict unknown functions of the protein phosphorylation. As an application of the database, we also demonstrate how to retrieve proteins and pathways in striatal medium-sized spiny neurons modulated by extracellular dopaminergic stimulation.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-489 方位分布関数と内包率を利用した白質神経線維束クラスタリング新手法の検討 Koma Suzuki（鈴木康真），Shiho Okuhata（奥畑志帆），Tetsuo Kobayashi（小林哲生）京都大院工 Introduction: Recently, clustering white matter nerve fiber bundles from the reconstructed streamlines using the probabilistic tracking has been intensively studied. Selecting and removing false positive streamlines that are generated at a certain rate in probabilistic algorithms is one of major challenges. In a previous study, we reported an automatic fiber clustering method based on fiber orientation distribution (FOD) [1]. However, over-removal of streamlines and the individual differences of the number of survived streamlines among subjects have been remained as limitations. Here, we purpose a new method that extracts anatomically relevant fiber bundles to overcome these limitations. Methods: 3T-MRI scanner (Siemens, Verio) was used to acquire dMRIs from 6 healthy adults (24.33 ± 0.58 years old, 5 males). The tract ROIs were created from JHU ICBM tracts atlas [2]. The proposed method reconstructed streamlines using inverse normalized tract ROIs as seeds [2]. The representative FOD vector in each tract was defined by averaging FOD vectors of all the voxels of the ROIs. Following three attributes were calculated for each reconstructed streamlines; 1) intension ratio, 2) mean and 3) SD FOD peak angle from the representative FOD vector. The intension ratio is the proportion of the number of steps of the reconstructed streamline within the tract ROI. FOD peak angles from the representative vector were calculated for each step to yield mean and SD for each streamline. After the Euclidean distance between each point and the ideal point was measured based on the three attributes in each fiber bundle, streamlines with greater distance were removed. Results & Discussion: The proposed method could successfully cluster fiber bundles with effective number of streamlines with less individual differences among subjects. In particular, the variation in each fiber bundle was suppressed to 10% or less, while sufficient number of streamlines was secured in the main 14 fiber bundles, such as the cortico-spinal tract. These results demonstrate the feasibility of the proposed method for clustering fiber bundles. Acknowledgements: This work was supported by AMED (JPdm0207009) and was conducted using the MRI scanner at Kokoro Research Center, Kyoto University. We thank to Dr. Ryusuke Nakai for his assistance in data acquisition. [1] K. Ida, et al., IEICE Technical report, 117(360), pp.29-34, 2017. [2] K. Hua, et al, NeuroImage, 39(1), pp.336-347. 2008.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-490 イソフルラン麻酔はマーモセット大脳皮質-視床間機能的結合を変化させる-覚醒下安静時fMRIとの比較 Yuki Hori（堀祐樹）¹，David J. Schaeffer（Schaeffer J. David）¹，Kyle M. Gilbert（Gilbert M. Kyle）¹，Joseph S. Gati（Gati S. Joseph）¹，Ravi S. Menon（Menon S. Ravi）¹，Stefan Everling（Everling Stefan）^1,2 ¹Robarts Research Institute, Univ of Western Ontario, London, Canada ²Dept Phys and Pharm, Univ of Western Ontario, London, Canada The common marmoset (Callithrix jacchus), a New World Primate, is becoming increasingly popular as an additional non-human primate model to assess functional brain networks using resting-state functional magnetic resonance imaging (RS-fMRI). RS-fMRI studies in animals often use anesthetic agents to avoid the effects of motion, physiological stress and training requirements, despite clear evidence that anesthetics affect brain activation, in particular of the thalamus (Alkire et al, 2008). In marmosets, however, there is only one report detailing the effects of anesthesia on functional connectivity (FC) (Liu et al., 2013) which focused on the effects of propofol anesthesia on the BOLD signal caused by somatosensory stimulation. As such, it remains unclear how anesthetic agents affect FC of brain networks in marmosets. Here, we investigated the effects of isoflurane on functional networks in marmosets by comparing RS-fMRI under a fully awake state with that under 1.5% isoflurane anesthesia. We had two primary objectives: The first was to elucidate the global changes of functional networks caused by anesthesia in marmosets. The second was to assess which thalamic FC were modulated by anesthetics. Our main results showed that: (1) isoflurane globally decreased FC in resting-state networks, but the structure of the networks was preserved except for the default mode network; (2) isoflurane modulated frontal-thalamic FC, while it did not modulate intra-thalamic FC. These findings bridge the gap between resting-state FC studies under anesthesia and awake conditions and provide insights into the effects of isoflurane on functional brain networks in primates. Support was provided by the Canadian Institutes of Health Research (FRN 148365, FRN353372) and the Canada First Research Excellence Fund to BrainsCAN.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-491 広域電顕を用いた老化マウス大脳皮質の微細構造学的解析 Mitsuyo Maeda（前田光代）^1,2，Mitsuo Suga（須賀三雄）¹，Motohiro Nakamura（中村元弘）³，Katsuyuki Suzuki（鈴木克之）¹，Yuji Hasebe（長谷部祐治）¹，Naoki Kikuchi（菊池真樹）¹，Yuuki Yamaguchi（山口祐樹）¹，Asami Eguchi（江口麻美）^1,2，Satoshi Kume（久米慧嗣）^1,2，Rie Matsuzaki（松崎理恵）²，Yosky Kataoka（片岡洋祐）^1,2 ¹国立研究開発法人理化学研究所理研-JEOL連携センター　マルチモダル微細構造解析連携ユニット ²バイオシステムダイナミクス研究理研センター、細胞機能イメージング ³日本電子株式会社 In November 2014, RIKEN-JEOL Collaboration Center was established. Multi-Modal Microstructure Analysis Unit in the research center has been developing technologies with scanning electron microscopy (SEM) for new medical and biological applications. We have developed a SEM system realizing serial and automatic capturing of thousands to tens of 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. Using such a newly-developed electron microscopic technique, we acquired large-scale electron microscopic images (0.25 mm2) of cerebral cortex including all cortical layers (layers I-VI) from mice at various ages. Segmentation of cell nuclei and cytoplasmic organelles, such as mitochondria in neurons and glial cells were performed, and morphological changes in aging were quantitatively evaluated. In this meeting, we will demonstrate the increase in area of heterochromatin in each nucleus of neurons and glial progenitor cells in aging. Further, the amount of lysosomes in the cytoplasm increased in microglia. The large-scale electron microscopic technique will bring about morphological big data (Micro-Morphomics), and realize quantitative analysis of biological tissue microstructure.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-492 近赤外発光を利用した生体マウス脳内における活動依存的なBDNF遺伝子発現誘導の可視化 Mamoru Fukuchi（福地守）¹，Ryohei Saito（齊藤亮平）²，Shojiro Maki（牧昌次郎）²，Nami Hagiwara（萩原なみ）¹，Satoru Mitazaki（三反崎聖）¹，Hisashi Mori（森寿）³ ¹高崎健康福祉大薬分子神経科学 ²電気通信大情報理工学域 ³富山大院医薬分子神経科学 We previously generated a novel transgenic mouse strain, termed BDNF-Luciferase transgenic (BDNF-Luc Tg) mouse, to visualize changes in BDNF gene expression in living mice, using a luciferase as an imaging probe. We successfully detected bioluminescence signals from the Tg mice after the administration of D-luciferin, a substrate for luciferase. However, it was quite difficult to visualize changes in BDNF gene expression in the brain region of the Tg mice, because the bioluminescence light producing D-luciferin and luciferase poorly penetrates biological tissues. We here used novel substrates for luciferase, TokeOni and seMpai, water-soluble substrates producing highly penetrable near-infrared bioluminescence light. Comparing with D-luciferin, we clearly detected the bioluminescence signal from the brain region of the Tg mice after the administration of TokeOni, but not seMpai, probably due to the difference in permeability of blood-brain-barrier. Using TokeOni, we could also visualize kainic acid- or light stimulation-induced BDNF gene expression in the brain region of the Tg mice. Taken together, TokeOni would be a beneficial substrate for luciferase to visualize changes in BDNF gene expression in the brain region.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-493 神経細胞への有害作用を検出するための培養神経細胞を用いたシナプスの状態、樹状突起長、神経細胞数のハイコンテントイメージング解析 Kenji Hanamura（花村健次）¹，Anggraeini Puspitasari（プスピタサリアングラエイニ）²，Noriko Koganezawa（小金澤紀子）¹，Yuko Sekino（関野祐子）³，Tomoaki Shirao（白尾智明）¹ ¹群馬大院医神経薬理 ²群馬大未来先端 ³東京大学院薬ヒト細胞創薬学 To detect adverse effects of toxic substances on neurons, we quantitated neuron number, dendrite length and synaptic status of cultured neurons. An actin-binding protein drebrin accumulated in the postsynaptic sites of glutamatergic synapses was used as a marker to detect synaptic status. In addition, a tubulin-binding protein, MAP2 was used to visualize neuronal cell body and dendrites. Our previous studies have shown that activation of N-methyl-D-aspartic acid-type glutamate (NMDA) receptors, the application of amyloid beta or the X-irradiation decrease the amount of drebrin in postsynaptic sites of glutamatergic synapses. To use this method for high-throughput analysis, we applied the drebrin-based evaluation of synapses and MAP2-based quantification of dendrite length and neuron number to high-content imaging analysis using microplates. In this study, we showed that glutamate treatment for 10 min significantly reduced drebrin cluster density of 21-days-in-vitro (DIV) neurons in a dose-dependent manner. We also confirmed that exposure of 1 Gy X-irradiation to 1-DIV neurons reduces neuron number, dendrite length and drebrin cluster density in the neurons at 21-DIV. In addition, our analysis could efficiently detect staurosporine-induced neuronal cell death in mature neurons. 24 hours exposure of 0.3 and 1.0 μM staurosporine to 21-DIV neurons significantly reduced neuron number. These results suggest that our high-content imaging analysis for detecting synaptic status using drebrin clusters along dendrites, dendrite length and neuron number is useful for analyzing the effects of various toxic substances.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-494 CUBIC-Cloud: 組織透明化による全脳全細胞解析フレームワーク Tomoyuki Mano（真野智之）^1,2，Etsuo Susaki（洲崎悦生）^3,4,5，Ken Murata（村田健）^6,7，Chika Shimizu（清水知佳）⁵，Kazuhiro Kon（昆一弘）³，Hiroaki Ono（小野宏晃）³，Kazunari Miyamichi（宮道和成）^6,7，Kazushige Touhara（東原和成）^2,6,7，Hiroki R Ueda（上田 R 泰）^1,2,3,5 ¹東京大院情報理工学系研究科システム情報学専攻 ²東京大学ニューロインテリジェンス国際研究機 ³東京大院医システムズ薬理学 ⁴科学技術振興機構さきがけ ⁵理化学研究所生命システム研究センター合成生物学研究グループ ⁶東京大院農学生命科学 ⁷科学技術振興機構ERATO東原化学感覚シグナルプロジェクト In the past decade, tissue clearing has established itself as an innovative technology in neuroscience as high-performance clearing protocols and advanced imaging methods emerged, allowing researchers to investigate the entire neural system with single cell resolution. Less explored challenges in this field, however, is how to effectively share and utilize whole-brain data obtained by individual studies, and to derive collective insights beyond single research project. To that end, we designed and implemented CUBIC-Cloud, a computational framework to analyze, visualize and share whole-brain data. CUBIC-Cloud offers an open source library which contains all essential image analysis routines (cell counting, registration, plotting, etc) in one seamless and user-friendly Python package (CloudMap). In addition, we created an web-browser-based interactive brain viewer (CloudEye), with which researchers can visualize millions of cells downloaded from online cloud server. To demonstrate the capability of CUBIC-Cloud, we first investigated whole-brain distribution of popular cell types, including PV, Sst, ChAT, Th and Iba1 expressing cells. Secondly, we quantified neuronal activity change under pharmacological perturbation by imaging immediate early genes and identified distinct brain regions affected by the drugs. Thirdly, we profiled brain-wide monosynaptic connection by using Rabies virus tracers and successfully detected extremely rare cell populations which would otherwise be overlooked with conventional, slice-based methods. Together, we established a general computational framework designed for tissue clearing studies, which will accelerate systems-level identification of cellular cluster and circuits. All the data analyzed in this study is going to be freely available at https://cubic-cloud.org (website under construction).		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-495 ２次元表面プラズモン共鳴観察による神経細胞への分化過程のモニタリング Hiroaki Shinohara（篠原寛明）¹，Shota Yashiki（屋敷尚汰）²，Masaki Fujii（藤井正貴）²，Minoru Suga（須加実）¹ ¹富山大学大学院理工学研究部(工学) ²富山大学大学院理工学教育部 In this paper, new method for monitoring of differentiation stage from stem cell to neuronal cell by two dimensional surface plasmon resonance (2D-SPR) observation is introduced. Mouse embryonal carcinoma cell line, P19 cell was used as a model stem cell. P19 cells were differentiated to neuronal cells by retinoic acid treatment and adhered onto a gold sensor chip. The 2D-SPR imaging of the individual cell was performed with a SPR imager made by NTT-AT, Japan. Neurite outgrowth was very active and SPR signal vibration was clearly measured at the cell at 6th day from differentiation start. The neurite network was completed and SPR signal vibration was weakened at 12th day. In our study, it was considered that the vibration of SPR signal may be related the development of tublin and actin filament in differentiation stage. Furthermore, the glutamate receptor expression at the cell membrane was also investigated from SPR response upon AMPA stimulation. The SPR signal at cell region significantly increased upon AMPA stimulation. On the other hand, no signal change was observed at undifferentiated cell and a slight increase of SPR signal was observed at 12th day cell. These results also indicated the difference of the differentiation stage between 6th day cell and 12th day cell. In conclusion, we would like to say our 2D-SPR observation method is very useful for identification of the differentiation stage from stem cell to immature and mature neuronal cell.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-496 再構成可能な迷路は記憶学習研究のための繰り返し精度の高い再現性を有するテストを提供する Susumu Takahashi（高橋晋），Satoshi Hoshino（星野諭），Kaoru Ide（井出薫）同志社大院脳科学研究科 Several tests for learning and memory are routinely conducted in a variety of shapes of maze such as the T maze, Plus maze, and Radial arm maze. The shape of conventional mazes cannot be easily changed in the same room. Visual-based virtual reality produces unlimited shapes of the maze. However, the hippocampal place coding dramatically remaps between different mazes in both virtual and real environments. Thus the evidence observed in the maze tests cannot be simply utilized to understand the learning and memory in a complementary way. Here, to address this issue, we develop a reconfigurable maze system to flexibly assemble and disassemble parts for configuring a variety of shapes of the maze to produce the existing mazes even in the same room and to control the sensor and actuator for task scheduling in them. It consists of interlockable parts including runways, feeders, movable walls, shut-off sensors, and treadmills. Using the maze system, we could rapidly configure the existing mazes: T maze, W maze, Figure-8 maze, Plus maze and Radial arm maze in an enclosure, which demonstrates the reproducibility of the reconfigurable maze. To ask whether the interlockable parts internally or externally affect animal behaviors in the maze, we simultaneously monitored rat's behavioral trajectory and hippocampal CA1 place cell activity. Results suggested that the interlocking mechanisms do not impair the navigational behavior and hippocampal place coding. Furthermore, to demonstrate the repeatability, the maze was morphing a square to a cruciform. As expected, the place field of hippocampal place cells remapped across morphing the shape of the maze. The results suggest that our system has flexibility, consistency, repeatability, and reproducibility from the internal and external viewpoints. Therefore, we propose that it facilitates examination into the neuronal underpinning of learning and memory.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-497 脳波による認知機能評価システムの開発 Ryohei P Hasegawa（長谷川良平），Yoshiko Nakamura（中村美子）産総研ニューロテクノロジー研究グループ A brain-machine(BMI) provides a direct link between the brain and external devices. The cognitive type of BMIs especially utilizes the neurotechnologies about the neural decoding of internal decision on single trials. We have been developing such a cognitive BMI-based communication aid ""Neurocommunicator"" (NC) for people with severe motor disabilities. The NC utilizes an event-related potential (ERP), the cognitive component of the EEG, as a ""switch"" for the user to select messages (pictograms). One of the benefits of this system is the compact headgear with a wireless EEG recorder that localizes the electrode positions around the top of the head, around which the maximum ERP response is observed. The other is the high-speed and high-accuracy decoding algorithm that predicts user's internal choice based upon the pattern recognition of the single ERP waveforms. However, there remains a problem to be solved for its practical realization; the appropriate compatibility check for user candidates should be introduced in terms of cognitive ability because of the risk of the cognitive impairment by the Alzheimer's disease as well as by the disuse syndrome. Here, we studied the feasibility of the NC as a convenient system for the cognitive assessment system in order to detect subtle symptoms of mild cognitive impairment. As a first step, we examined the effect of the involvement of bottom-up and top-down attention on the ERP response pattern. The former was tested by the oddball target task with a single kind of frequent non-target (the popular paradigm for ERP studies), and the latter was tested by the target selection task with multiple kinds of non-targets (the model paradigm for our daily life situations). We supposed that the task became more difficult in this order. We recorded the EEG data from 40 normal adult subjects during the target only task, the oddball target task, and the target selection task. Although the ERP waveforms systematically changed, it was difficult to evaluate how strongly they responded. Therefore, we focused on a single index, the accuracy of the decoding of the target, which was originally the indicator of the spec of the proposed BMI system. The average of the accuracy of all subjects gradually became lower along with the task difficulty. These results suggest that the accuracy to decode the target by the NC might be a good ""biomarker"" of the individual's cognitive state.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-498 歪みを最小化しながら神経生体信号を抽出する非線形フィルターの実装と性能評価 Toshihide Tabata（田端俊英）¹，Rihito Nagamatsu（永松利一）¹，Masayoshi Abe（阿部匡良）¹，Kazuki Kitamura（北村和希）¹，Tomoki Matsui（松井友希）¹，Ryo Takemura（竹村諒）¹，Hakushun Sakairi（坂入伯駿）²，Yuji Kamikubo（上窪裕二）²，Takashi Sakurai（櫻井隆）² ¹富山大院理工（工）生体情報工 ²順天堂大医薬理 It is difficult to analyze a neurobiological signal masked by relatively large background noise. To improve the signal-to-noise ratio, one may employ low-pass filter, moving average, or another smoothing technique. However, signal processing using such a technique may slow down and/or shrink the rapidly changing components of neurobiological signals. An alternative technique is a non-linear filter (NLF) originally developed by Chung and Kennedy. In this study, we have implemented the NFL as an application of Igor Pro (WaveMetrics), a widely used commercial software to process massive sequential data. The NLF expresses the value of the data at the given time point by the weighted average of multiple time windows with different durations around the time point. The relative weights for the shorter windows to the longer windows are increased when the value greatly changes around the time point. The NLF application could extract a train of pulses from the background white noise without reducing the amplitudes of the pulses or obscuring the timing of the pulse onsets and offsets. The NLF application had no obvious phase delay regardless of input signal frequency and a wide frequency-response as compared with low-pass filter and moving average processing with similar extents of noise reduction. Furthermore, the NFL application could efficiently smooth real biological signals including fluorescence resonance energy transfer time trajectories indicating the molecular interaction of type-1 metabotropic glutamate receptor subunits and single-channel recording traces indicating the gating of type-4 hyperpolarization-activated cyclic nucleotide-gated channel. The NFL application would be a powerful tool to facilitate analysis of neurobiological signals.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-499 成体大脳新皮質に存在する神経幹細胞前駆細胞の培養法の確立 Koji Ohira（大平耕司），Yukari Manabe（間部由佳理），Honoka Shibata（柴田歩乃佳），Saki Okamoto（岡本彩希）武庫女大食物栄養 Currently, studies have reported that neural stem cells (NSCs)/neural progenitor cells (NPCs) produce new neurons in a few regions of the adult mammalian brain, such as the subventricular zone (SVZ), the subgranular zone (SGZ), and the cerebral cortex. The molecular and cellular mechanisms and functions of adult neurogenesis in the SVZ and SGZ are becoming clear. In contrast, neither the underlying mechanism nor functions of cortical adult neurogenesis is understood. Establishment of culture system for cortical NSCs/NPCs makes it possible to reveal the mechanisms of proliferation and differentiation of cortical adult neurogenesis. In the way of developing the culture method of cortical NSCs/NPCs, the enzymatic treatment time to dissociate NSCs/NPCs from cortical tissues is particularly important. When the treatment time is too long, the cells weaken, and conversely if the treatment time is short, the number of cells that can be isolated decreases. In this study, using the neurosphere assay, we conducted the time course analysis for the enzymatic dissociation of NSCs/NPCs from cortical tissues of mice. The cortical tissues (3 day-, 1 week, 2 week-, and 4 weeks-old) were treated by the protease papain for 10, 20, 30, and 60 minutes with oxygen bubbling, and then cell fractions containing NSCs/NPCs were taken by percoll density gradient centrifugation. The cells obtained were incubated for 1 week, being added growth factors (epidermal growth factor, basic fibroblast growth factors, and brain-derived neurotrophic factor) every 2 days. We found that some neurospheres were formed in all the ages and treatment periods. At ages 3 days and 1 week, neurospheres were well formed with enzyme treatment time for 20 minutes. On the other hand, for cultures obtained from tissues of 2 weeks or more, a longer enzyme treatment time was required to form neurospheres. In almost all culture conditions, neurospheres with diameters approximately 100 microns were formed. We are now trying to form neurospheres from older mice.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-500 マニュピレータ一体型小型tetrodeアレイの開発 Yoshiya Mori（森理也），Hiroyuki Ito（伊藤浩之）京都産業大学情報理工学部 Information processing in the brain involves coordinated activities of a large neuronal population. Stable and simultaneous recording of multiple neuronal activities is a fundamental technology needed for understanding of population coding and developing decoding algorithm of the BMI and BCI. For a long-term recording from behaving animals, chronically implanted extracellular probes have been widely used (such as Utah probes and Silicon Probes). Spatial positions of the conventional recording probes are fixed once implanted in the brain and additional surgery is required to record from another neuronal population. To overcome this problem, we developed an array of electrode manipulators integrated with a miniature motor drive. We adopted the tetrode (Thomas Recordings), in which four fine wires are enclosed in a glass fiber, because of its high spike isolation quality that enables a large scale population recording. Each manipulator drives a single tetrode fiber to the longitudinal direction by miniature piezo linear motor (NEW SCALE, SQUIGGLE RV) in a spatial resolution of less than 0.5 μm. Each motor is controlled independently through the cable connected with the computer and the travel range is more than 7 mm. The body dimension of each integrated drive is 4 x 4 x 30 mm. In the recording chamber we arrange four tetrodes with a tight spacing that enables the penetrations of four tetrodes within an area of 500 x 500 μm on the cortical surface. Most parts constituting the drive can be easily fabricated by the conventional 3D printer. To test the performance of this electrode array, we have implanted the drives to two Rats (primary visual cortex or somatosensory cortex). Neuronal activities have been recorded for more than 6 months. We are trying to start neuron operant conditioning experiment by using this recording system.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-501 インクジェット印刷による超薄・柔軟皮質脳波(ECoG)電極の開発 Takumi Kishi（岸巧）¹，Shinji Takeoka（武岡真司）¹，Hiroyuki Ohta（太田宏之）²，Toshinori Fujie（藤枝俊宣）^3,4,5 ¹早稲田大学先進理工学部 ²防衛医科大学校　生理学講座 ³東京工業大学　生命理工学院 ⁴早稲田大学ナノ・ライフ創新研究機構 ⁵JSTさきがけ ECoG electrodes are expected for long-term monitoring of brain activity without mechanical mismatch between electrodes and brain surface. In this study, we developed an ultra-thin and flexible ECoG electrode consisting of inkjet-printed electrodes on a polymeric ultra-thin film (referred to as "nanosheet"). The nanosheet was made of poly(d,l-lactic acid) (PDLLA) with the thickness of 302 nm, on which electrode pads (400 μm square, 4 channels) and conductive wiring (100 μm width) were inkjet-printed using poly(3,4-ethylenedioxythiophene): polystyrene sulfonate (PEDOT:PSS) and Au nanoinks. Then, as an insulating layer, an SU-8 film with thickness of 3.1 μm was prepared by spincoating and micropatterned by standard UV-photolithography, which was transferred to the electrode pads. Total thickness and bending stiffness of a resulting nanosheet-based ECoG electrodes (i.e., nanosheet, electrode pads and SU-8 film) was 3.7 μm and 5.1 nN m, respectively, that were ca. 27 times thinner and more flexible than conventional ECoG electrodes (thickness: ca. 100 μm, bending stiffness: >>100 nN m). Finally, the nanosheet-based ECoG electrode was placed on a hippocampus slice of channel rhodopsin-2(ChR2) expressed transgenic mice. Under the blue-light stimulation (wavelength: 470 nm), local field potential (LFP) was recorded selectively from the region of an electrode close to the neuron cells. The present nanosheet-based ECoG electrode will be useful for conformable attachment to the brain tissues and mapping out the neuronal activity.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-502 脳深部からのホールセルパッチクランプ記録法 Yu Sato（佐藤由宇），Yuji Ikegaya（池谷裕二）東京大院薬薬品作用 Neurons transmit information to other neurons via electrochemical signals. Several electrophysiological methods have developed to investigate activities of neurons. One method for recording from a single neuron is the patch-clamp technique. In this technique, a glass pipette is tightly sealed with the membrane surface of a neuron (gigaseal), and then the plasma membranes are broken and are integrated with pipettes (whole-cell mode). The whole-cell recordings make it possible to clamp the voltages or currents of a single neuron, by which the dynamics in the inputs or outputs of the targeted neuron are monitored. The patch-clamp technique is applied to not only ex vivo brain samples but also in vivo brain surface regions, such as the cerebral cortex and the hippocampus. However, there are a few reports using in vivo whole-cell recording from deep brain regions, mainly because it is extremely difficult to form gigaseal due to impurities on the pipette tips. Here we developed a new method to attain whole-cell recordings to deep regions in in vivo mouse brains. We used double tubes; one is a guide cannulae for patch-clamp pipettes, and the other is a tissue-boring stick. Thereby, we were able to easily achieve in vivo whole-cell recordings from deep brain regions, because the path lengths for which the pipettes needed to go through the brain tissue parenchyma were minimized. In this poster, we will report some examples for recordings from neurons of the basolateral amygdala, which locates about 4 mm deep from the brain surface.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-503 3D quantitative synthetic MRIによるヒト脳の皮質厚および脳部分容積測定：3Tでの再現性および従来法との比較 Shohei Fujita（藤田翔平）^1,2，Akifumi Hagiwara（萩原彰文）^1,2，Masaaki Hori（堀正明）¹，Issei Fukunaga（福永一星）¹，Christina Andica（Andica Christina）¹，Tomoko Maekawa（前川朋子）^1,2，Ryusuke Irie（入江隆介）^1,2，Koji Kamagata（鎌形康司）¹，Shigeki Aoki（青木茂樹）¹ ¹順天堂大学医学部　放射線診断学講座 ²東京大学大学院　医学系研究科　放射線医学講座 Previous quantitative synthetic MRI of the brain has been solely performed in 2D. We propose an application of the 3D-QALAS (3D-quantification using an interleaved Look-Locker acquisition sequence with T2 preparation pulse) sequence for simultaneous acquisition of relaxometry parameters as well as for obtaining volumetric information in high-resolution 3D. Here, we evaluate the feasibility of the recently developed sequence 3D-QALAS, for brain cortical thickness and volumetric analysis. For each of the twenty-one healthy volunteers (35.6 ± 13.8 years), 3D T1-weighted fast spoiled gradient recalled echo (FSPGR) sequence was performed once, and 3D-QALAS sequence was performed twice on a 3 T scanner. Widely used free-software FreeSurfer and FIRST were used to measure cortical thickness and volume of subcortical structures, respectively. Agreement with FSPGR and scan-rescan repeatability were evaluated for 3D-QALAS. Percent relative difference compared with FSPGR in cortical thickness of the whole cortex was 3.1%, and 89% of the regional areas showed less than 10% relative difference in cortical thickness. The mean ICC across all regions was 0.65, and 74% of the structures showed substantial to almost perfect agreement. For volumes of subcortical structures, the median percent relative differences were lower than 10% across all subcortical structures, except for the accumbens area, and all structures showed ICCs of substantial to almost perfect agreement. For the scan-rescan test, the percent relative difference in cortical thickness of the whole cortex was 2.3%, and 97% of the regional areas showed less than 10% relative difference in cortical thickness. The mean ICC across all regions was 0.73, and 80% showed substantial to almost perfect agreement. For volumes of subcortical structures, relative differences were less than 10% across all subcortical structures except for the accumbens area, and all structures showed ICCs of substantial to almost perfect agreement. In conclusion, 3D-QALAS could be reliably used for measuring cortical thickness and subcortical volumes in most brain regions.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-504 多細胞スパイクデータからシナプス結合を推定する技術の開発 Ryota Kobayashi（小林亮太）^1,2，Shuhei Kurita（栗田修平）³，Katsunori Kitano（北野勝則）⁴，Kenji Mizuseki（水関健司）⁵，Barry J Richmond（Richmond J Barry）⁶，Shigeru Shinomoto（篠本滋）⁷ ¹国立情報学研究所 ²総研大院複合科学情報学 ³京都大院情報 ⁴立命館大情報理工知能情報 ⁵大阪市大院医 ⁶NIH ⁷京都大院理物理 Advances in experimental techniques have enables us to record spikes from more neurons for longer durations. Indeed, the number of that can be simultaneously recorded neurons has been doubled in every seven years (Stevenson and Kording, 2011). Such recordings allow us to infer the structure of neural circuits, i.e., the synaptic connectivity between neurons. Unfortunately, it is notoriously difficult to estimate interneuronal connectivity because neuronal activity is highly irregular and the firing activity of surrounding neurons fluctuates greatly. Cross-correlation (CC) (Perkel et al., 1967) has become the standard tool for estimating interneuronal connectivity. However, CC is vulnerable to such fluctuations in neuronal activity. Here we propose a novel method, termed CC-GLM, for estimating synaptic connectivity as postsynaptic potentials (PSP) between neurons from the spike trains of multiple neurons. It is notable that this method is able to absorb fluctuations in data by combining CC with the generalized linear model (GLM). To evaluate its performance, we applied CC-GLM to simulated data of 1,000 Hodgkin-Huxley neurons with AMPA, NMDA, and GABA model synapses. The results showed that CC-GLM detected the majority of strong connections, i.e., 80% (100%) of excitatory (inhibitory) connections with PSP greater than 1 mV. Further, it outperformed conventional methods for estimating neural connectivity, including the standard cross correlation method (Perkel et al., 1967) and jittering method (Amarasingham et al., 2012). Lastly, we applied CC-GLM to spike trains recorded from rat hippocampal CA1 neurons while the rats explored a square field (Mizuseki et al., 2009). The connections estimated by CC-GLM were consistent with those inferred by experts from other physiological evidence.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-505 マウスてんかん発作検出のための画像解析と機械学習 Masato Asai（浅井真人）¹，Machiko Iida（飯田真智子）¹，Motoki Tanaka（田中基樹）¹，Yuka Mizutani（水谷友香）¹，Kaichi Yoshizaki（吉崎嘉一）¹，Tsuyoshi Takagi（高木豪）¹，Yoshihito Tokita（時田義人）¹，Naoya Asai（浅井直也）²，Masahide Takahashi（高橋雅英）² ¹愛知県心身障害者コロニー発達障害研究所 ²名古屋大学大学院医学系研究科病理病態学講座腫瘍病理学分野 Rationale We have developed mice generating abundant generalized tonic clonic seizures (GTCSs) with complete genetic penetrance. These mice exhibit epilepsy-related lesions in the hippocampus without the need of epileptogenic induction. These mice faithfully mimic human patients with refractory mesial-temporal lobe epilepsy. The GTCS counts represent overall seizure severity, which can be utilized to determine the presence of the therapeutic effects of anti-epileptic treatments in mice. The GTCS counting currently relies on manual procedures. However, such manual counting in long-term video has problems, including oversights, judgment fluctuation, and technostress in human. Video content analyses (VCA) automates tracking a particular movement of an object, which came to be widely used in recent years for health care. We presently applied VCA on counting GTCSs with the aims of labor reduction, and externalization of seizure judgment. Methods Singly-housed epileptic animal was maintained in a home cage equipped with an overhead network camera, which allows long-term video archiving and the post hoc analyses. Adult Girdin/ccdc88a knockout mice (female n=3, P103-132) were used. An aluminum home cage (CKS Inc.) was combined with a network camera (TS-WLC2), a router, and a network-attached-storage (HDL-XR4.0W/2D). Authors and IVIS Inc. jointly developed two versions (machine-learning and image-analysis) of seizure detection software on a PC (HP Pavilion 15-cx0105TX) using a free and open-source programming language (Python 3.6.4, Anaconda). Human's GTCS counts were considered as reference. Results Machine-learning version properly detected 67 out of 67 human-verified GTCSs, while it falsely judged 936 motions as GTCSs, of which processing time was approximately thirty times as long as actual time. Image analysis version successfully compressed total video time from 14 h 46 m to 1 h 42 m without losing any of GTCS scenes (18 out of 18 human-verified GTCSs), of which processing time was 13 h 13 m, which was 7.8 times as long as actual time. Conclusions Neither machine-learning nor image-analysis software did not realize automatic counting of GTCSs. However, image-analysis successfully shortened the video without losing any of GTCS scenes within a practical processing time, which reduced analyzers' technostress. Further machine power enhancement and algorithm refinements may implement the full-automatic seizure detection in the future.

7月25日（木）10:35～11:55　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-506 効率的モデルラット開発および免疫不全ラットリソースの確立 Kosuke Hattori（服部晃佑）¹，Yoshiki Miyasaka（宮坂佳樹）¹，Yuko Yamauchi（山内祐子）²，Yuko Kotani（小谷祐子）¹，Kanako Shimizu（清水加奈子）¹，Rieko Ando（安藤理恵子）¹，Yoshihiro Uno（夘野善弘）¹，Kazuto Yoshimi（吉見一人）²，Tomoji Mashimo（真下知士）^1,2 ¹大阪大院医附属動物実験施設 ²大阪大院医附属共同研ゲノム編集センター Immunodeficient mice have been utilized as transplant recipients of cancer cell lines, cancer tissues, and human hematopoietic cells. They are useful models for human disease treatment, and research on blood cell differentiation and immune response. On the other hand, rats (Rattus norvegicus) that have a history of more than 150 years as experimental animals, have several advantages such as ease of handling, behavioral experiments, and the size of tissues. However, they haven't been used for preclinical research due to the difficulty of genetic modification. Recently, genome editing technology expands the potential of rat resources. NBRP-Rat as a part of the National BioResource Project in Japan has started since 2002. The main roles of the NBRP-Rat are the collection, preservation, and supply of so many various rat strains. In 2017, Osaka University has joined this project as the allocation organization for preserving and distributing immunodeficient SCID rats. We provide three SCID rat strains such as Il2rg knockout (NBRP Rat No.0883), Rag2 knockout (NBRP Rat No.0894), and Il2rg/Rag2 double knockout (NBRP Rat No.0895). We also the member of Platform of advanced Animal Model Support established by MEXT in Japan since 2016. This platform aims to support researchers who want to generate and to analyze various gene-editing animals. Here we report the outline of `NBRP-immunodeficient rats' and `Support Animal Model Platform', and the basic technologies that we recently established as genome editing technology.		7月26日（金）13:15～14:35　ポスター会場（朱鷺メッセ　1F　展示ホール） PA-507 脳血管内皮細胞の多次元解析による老化に特徴的な亜集団の同定 Takeshi Fukuhara（福原武志），Shu-sei Hamamichi（濱道修生），Nobutaka Hattori（服部信孝）順天堂大学医学部神経学 Blood brain barrier (BBB) in neurovascular unit plays a pivotal role to keep homeostatic condition. We are particularly interested in vascular state under aged or inflammatory condition that are often observed in neurodegenerative diseases. Under both acute and chronic inflammation, it has been considered that trans-endothelial migration (TEM) of immune cells regulate a subsequent trigger of multiple deleterious events. Therefore, it is critical to reveal the precise mechanism of TEM for further development of diagnostic or therapeutic tools against neurodegenerative diseases. Recent technological advantages such as single cell RNA-seq analysis enabled us to refer the molecular basis of heterogeneous vascular cell types, however, it still has been challenging to specify the cell populations that are relevant as diagnostic or therapeutic target in neurodegenerative diseases due to multiple reasons. From the practical point of view in drug delivery, we have been screening antibodies to target inflammatory vasculature. By taking advantages of DT3C-immunotoxin screening system, multiple functional antibodies with internalization capacity were established. Here, we present the functional characterization of monoclonal anti-CD146 antibody (clone 29-1709) that are cross-reactive to mouse and human CD146. Curiously, upon ex vivo stimulation by ionomycin, CD146+ subpopulation was newly generated in mouse brain. Hi-D FACS analysis revealed that this CD146+ subpopulation in the brain lost typical surface markers of endothelial cells. Further, Hi-D FACS technology was employed to analyze the mouse brain in different age, resulting in the identification of young- or aged-specific subpopulation of endothelial cells. To develop theranostic moiety, we formulated immunoliposome encapsulated various ingredients such as ICG or drug. Reproducible results demonstrated a novel immunoliposome conjugated with anti-CD146 MoAb was selectively incorporated in endothelial cells line, bEnd.3 in vitro. We are currently evaluating the applicability of this immunoliposome in mouse model.