Late Breaking Abstract

Late Breaking Abstract

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-L-508 The temporal patterning of neural progenitor cells during complex brain formation Merve Bilgic（Bilgic Merve）^1,2，Wu Quan（Quan Wu）²，Fumio Matsuzaki（Matsuzaki Fumio）^1,2 ¹Graduate School of Biostudies of Kyoto University ²RIKEN Biosystems for Dynamics Research Neural progenitors (NPs) generate the cerebral cortex in a timely-controlled manner, with neurogenesis occurring first, followed by the gliogenesis. As a result, changes in the temporal progression of NPs determine the cellular diversity within the cortex. In the developing cortex, germinal layers where NPs divide are located next to cerebral ventricles (known as Ventricular Zone, VZ and Subventricular Zone, SVZ). During mammalian brain evolution, a novel germinal layer, called outer SVZ (oSVZ), is emerged into the gyrencephalic species, such as primates and carnivores. Nevertheless, in complex brains, it remains largely unknown whether and how the clock in NPs from distinct layers proceed and contribute to the cortical development. Our lab's previous study revealed a set of genes that represents the temporal identity of NPs of mouse cortex (Okamoto et al., 2016), a simple brain model. In this study, we aim to understand how the spatiotemporal identity of NPs progress in developing complex brains. We thus address to in situ hybridization (ISH) method to examine the spatiotemporal pattern of mouse clock markers on ferret brain, our complex brain model. We perform single-cell transcription profiling covering embryonic and post-natal development to distinguish molecular features of NPs derived from different germinal layers. Finally, we investigate the transitions of cell fate in different germinal layers at late-neurogenesis by immunohistochemistry. Our preliminary results suggest that, during embryonic development, VZ and SVZ NPs undergo similar temporal identity changes, while some temporal marker genes are delayed in their expression in the ferret SVZ towards the end of neurogenesis. Moreover, we observe that VZ and SVZ seem to undergo different fate transitions during late-neurogenic stages at early post-natal development: VZ acquire ependyma-like features, while SVZ may maintain late-neurogenic and/or gliogenic properties longer, hence contributing to the complex brain enlargement. The combination of single-cell transcriptome analysis with ISH and immunohistochemistry will thus allow us to determine the temporal progression of NP identity and cell fate changes occurring in VZ and oSVZ. It will be important to distinguish molecular and cellular features of different NPs in the complex brains in order to better understand the complex brain 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-L-509 細胞間相互作用を介した大脳皮質領野形成機構の解析 Rui Iizuka（飯塚瑠伊）¹，Peishan Hou（侯珮珊）¹，Chihiro Nishiyama（西山千尋）²，Carina Hanashima（花嶋かりな）¹ ¹早稲田大院先進理工 ²理研　生命機能科学研究センター The cerebral cortex holds a remarkable capacity to integrate multimodal sensory information and generate coordinated outputs that underlies higher cognitive functions. While the localization of cortical areas responsible for modality-specific information processing has been mapped over a century ago, how each area is assembled to serve its unique function has remained unknown. Neurons of layer 4, lying in between the deep- and upper-layers of the neocortex are the major recipient of sensory inputs relayed through thalamic nuclei. The thickness and number of layer 4 neurons is the most prominent feature between distinct areas, which is enriched in the somatosensory area but virtually absent in the motor area. Despite their functional significance in cortical arealization, the ontogeny of these neurons and how they integrate into biased areas is unknown. To explore this, we established a genetic labeling system using Cre-mediated reporter recombination and tracked temporal cohorts of newborn neurons. Our results indicate that the migration and integration dynamics of neurons differ significantly among cortical regions. In vitro cultivation of these temporal precursors recapitulated in vivo cortical development, suggesting the critical impact of intrinsic regulation in regional identity establishment. Using single cell RNA-sequencing, we further identify potential intrinsic regulators that mediate interaction between precursor cells and extrinsic signals to confer regional migratory behavior. In this presentation, we will further discuss the underlying mechanisms involved in these processes.

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-L-510 発生初期の抗うつ剤SSRI処理によるゼブラフィッシュ脳における作用解析 Tomomi Sato（佐藤智美）^1,2,3，Takeshi Kajihara（梶原健）²，Hiroshi Handa（半田宏）³，Masabumi Nagashima（永島雅文）¹ ¹埼玉医大医　解剖学 ²埼玉医大医　産婦人科 ³東京医大　ナノ粒子 For risk management between maternal mental health and fetal development, it is important to understand effects on embryos exposed to antidepressant, selective serotonin reuptake inhibitor (SSRI). Recently, ingestion of SSRI is reported to increase the risk of autism. However, it remains to be elucidated the role of serotonin transporter (SERT), the target of SSRI during development of the brain. Placenta-derived serotonin is transiently localized in the forebrain of early mouse embryos, suggesting a fundamental role of serotonin in early development of the brain. To understand the role of SERT in early brain development, we investigated the effects of SSRI on brain development using zebrafish embryos as a vertebrate model system. Immunohistochemistry with anti-serotonin antibody showed that serotonergic neurons were developed in the raphe nuclei of zebrafish embryos from 52 hours post fertilization (hpf). Treatment with SSRI from 9 hpf, a stage when the head primordium is developed, generated embryos with a small head and decreased neurons at 54 hpf. We found that SERT was localized in the apical surface of radial glial cells in the ventricular region at 24 hpf. Knockdown of SERT showed a small head and decreased radial glial cells at 30 hpf, suggesting that SERT is involved in development of radial glial cells. However, phospho-histone H3 (pH3)-positive mitotic cells were obviously increased in the brain of SERT-knockdown embryos. At a later stage 3 dpf, we observed that the head was slightly enlarged. Consistently, retinotectal projection areas were significantly expanded, whereas arborization/neuronal densities were significantly decreased in SERT-knockdown embryos. These results suggest that transient knockdown of SERT at early developmental stages causes a decrease of neural stem/progenitor cells in the brain, consequently generates abnormal development of neural circuitry, implicating a possibility that exposure of early embryos to SSRI is a risk factor for autism.		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-L-511 大脳皮質領野形成における放射状グリア細胞の役割 Koichio Irie（入江浩一郎）¹，Chihiro Nishiyama（西山千尋）²，Carina Hanashima（花嶋かりな）^1,3 ¹早稲田大　教育・総合科学学術院　発生生物学研究室 ²理化学研究所生命機能科学研究センター ³早稲田大院先進理工 The mammalian cerebral cortex comprises of diverse types of neurons that exhibit distinct function in transmitting modality-specific information. These neurons are organized into laminar and areal compartments, each of which can be distinguished by molecular, morphological and hodological properties. During development, neocortical neurons are sequentially produced from progenitor cells called radial glial cells, that elongate fiber towards the pial surfaces. Newly generated neurons migrate and are arranged superficial to early-generated neurons. It has been considered that these radial fibers contribute to the formation of layer structures through functioning as a scaffold for migrating newborn neurons. However, whether radial glial fibers play a role in cortical arealization has remained unknown. In this study, to assess whether radial glial fibers contribute to cortical arealization, we investigated the property and morphology of radial glial cells among cortical areas. We found region-specific neuronal migration velocity of migrating neurons. We also demonstrate region-specific morphologies of radial glial fiber and gene expression. Further analysis implied the relation between subplate neurons and direction of radial glial fibers. The region-specific changes in radial glia property may serve as an underlying mechanism for cortical arealization in addition to its known function for lamination in the developing cerebral 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-L-512 神経幹細胞の分化に対するGPR40とFATPの役割 Kenta Watanabe（渡邊健太），Hisami Koito（小糸寿美），Kenji Sugibayashi（杉林堅次），Masanori Katakura（片倉賢紀）城西大学薬学部薬学研究科栄養生理学講座 G protein coupled receptor (GPR) 40 have been identified as receptor of polyunsaturated fatty acids (PUFA), especially docosahexaenoic acid (DHA) and arachidonic acid (ARA) in the brain. However, little has been reported on how changes GPR40 fatty acid transport protein (FATP), a long chain fatty acid transporter, and expression in the brain. In this experiment, expression level of GPR40 have analyzed and measured fatty acid composition in the brain. Furthermore, neural stem cell (NSC) from fetus rat brain were isolated. The role of GPR40 on differentiation of NSC by pharmacological approach to use antagonist and agonist. Cerebral cortex and hippocampus from Wistar male rats were isolated at embryonic stage (E12.5, 14.5, 18.5, 22.5 days), 1 day age, 1 week age, 6.5 weeks age, 3 months age, 6 months age, and 10 months age. Fatty acid composition in each brain were measured by gas chromatography. Tuj-1, a neural marker and GPR40 protein expression level in cortex and hippocampus were quantitated by western blotting. NSCs were cultured from the forebrain isolated from E14.5 days using the neurosphere method. NSCs were added inhibitor and differentiated for 4 days. Then, NSCs were analyzed survival rate and differentiation rate by MTS assay and by immunofluorescence staining with specific neural markers. Although ratio of arachidonic acid (ARA) was decrease, ratio of DHA was increase during embryonic days. Before birth, ARA ratio was 12% and DHA ratio was 9%. In after birth, both ratios were higher than embryonic stage, but significant changes were not observed in fatty acid composition in growth stage. DHA increasing and neuron generation period were overlapped as a result of comparison of fatty acid composition and neural cells marker expression. Therefore, DHA was suggested that significant for generation of the neuron. GPR40 was expressed in cortex and hippocampus in embryonic stage that neurogenesis has been activated. After birth, it changing was similar to GFAP expression level. GPR40 was not expressed in undifferentiated NSCs. GPR40 expression was observed in all types of neuronal cells together with differentiation. Also, GPR40 agonist promoted differentiation in astrocytes as well as neurons. These results suggest that GPR40 is involved not only in neurons but also in astrocyte differentiation.		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-L-513 Cdc7キナーゼ活性化サブユニットASK/Dbf4の脳の発生・機能における役割 Shunsuke Kobayashi（小林駿介）^1,2，Tomohiro Iguchi（井口智仁）¹，Karin Hori（堀かりん）^1,2，Tomio Ono（小野富男）³，Chiaki Maruyama（丸山千秋）⁴，Satoshi Yamazaki（山崎聡志）¹，Hisao Masai（正井久雄）^1,2 ¹東京都医学総合研ゲノム動態プロジェクト ²東京大院新領域創成科学メディカル情報生命専攻 ³東京都医学総合研遺伝子改変室 ⁴東京都医学総合研神経回路形成プロジェクト The Cdc7-ASK kinase complex is evolutionally conserved and plays crucial roles in initiation of DNA replication as well as in other chromosome events including meiotic recombination and repair of DNA damages through trans-lesion DNA synthesis. ASK (activator of S phase kinase) is an activation subunit for mammalian Cdc7 kinase. It was previously reported that ASK of <I>Xenopus</I> is an inhibitor of the canonical Wnt signaling pathway and is required for heart development in a manner independent of its role for Cdc7. ASK has three conserved motifs, motif-N, -M, and -C. Motif-M (unique proline-rich motif) and -C (Z2H2-type Zn-finger) are required for full activation of Cdc7 kinase. Motif-N, a BRCT-like motif, is required for cell proliferation in ES cells, although not essential for Cdc7 kinase activation in vitro. Mammalian ASK has a long C-terminal tail sequence, whose functions are unknown. We previously reported that ASK is essential for growth of ES cells and its depletion leads to arrest of DNA synthesis (Yamashita <I>et al</I>. Genes Cells. 2005), but ASK functions in various organs are largely unknown. To examine the function of ASK in development of various organs, we have produced mice in which the exon 7 and 8 were flanked with loxP site. The expression pattern of ASK in mice brain is quite distinct from that of Cdc7. Therefore, we crossed ASK floxed mice to Nestin-Cre mice that express Cre recombinase during neural stem cell. Mice deficient in ASK in neural stem cells showed weight loss, gait disorder and body convulsion at 8 days after birth, phenotypes similar to that of Cdc7, and died within 22 days. Unlike Cdc7 knockout mice, we noted that female ASK knockout mice were not born, suggesting female-specific embryonic lethality. Our results indicate the roles of ASK in brain development, both overlapping with and distinct from those of Cdc7.

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-L-514 A Human Induced Pluripotent Stem Cell-Derived Tissue Model of a Cerebral Tract Connecting Two Cortical Regions. Siu Yu Angela Chow（Chow Siu Yu Angela）^1,2，Takaaki Kirihara（Kirihara Takaaki）^1,2，Zhongyue Luo（Luo Zhongyue）^1,2，Ryuji Misawa（Misawa Ryuji）^1,2，Jiro Kawada（Kawada Jiro）¹，Shinsuke Shibata（Shibata Shinsuke）⁴，Farad Khoyratee（Khoyratee Farad）^1,3，Carole Anne Vollette（Vollette Carole Anne）^1,3，Valentine Volz（Volz Valentine）¹，Timothee Levi（Levi Timothee）^1,3，Teruo Fujii（Fujii Teruo）¹，Yoshiho Ikeuchi（Ikeuchi Yoshiho）^1,2 ¹Institute of Industrial Science, The University of Tokyo, Tokyo, Japan ²Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Tokyo, Japan ³Laboratoire de l Integration du Materiau au Systeme (IMS), University Bordeaux, Bordeaux INP, CNRS UMR 5218, Talence, France ⁴Electron Microscope Laboratory, Keio University School of Medicine, Tokyo, Japan Cerebral tracts connect separated regions within a brain and serve as fundamental structures that support integrative brain functions. However, understanding the mechanisms of cerebral tract development, macro-circuit formation, and related disorders has been hampered by the lack of an in vitro model. In this poster, we report the generation of a model tissue mimicking a cerebral tract, which consists of a culture of two spheroids with an interconnecting fascicle of axons derived from human induced pluripotent stem (iPS) cells. We used a tissue culture microdevice equipped with a narrow channel with two chambers at its ends. Two cerebral spheroids were placed into the chambers, and they grew axons reciprocally into the channel. The axons spontaneously formed a robust fascicle, and the two spheroids became connected as one continuous tissue through an axon fascicle. Immunostaining and western blotting demonstrated that the fascicle of axons did not include cell bodies or dendrites. Axons assembled into fascicle significantly faster when two spheroids extended axons reciprocally than when one spheroid extended axons unidirectionally. Importantly, the two spheroids were electrically connected and coupled through the axon fascicle with distinct response kinetics. To model the developmental defect of cerebral tract formation including agenesis of corpus callosum (ACC), we knocked down the ACC-related gene L1CAM in neurons in our model tissue using RNAi. Axons from the L1CAM knockdown cells exhibited significantly lower ratio of axons assembled into a bundle than the control cells, suggesting that the axon fascicle formation process in our tissue is relevant to cerebral tract formation in vivo. The cerebral tract model tissue should provide a promising platform to study the mechanisms underlying cerebral tract development and related 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-L-515 高次視覚野は開眼直後にはすでに一次視覚野から機能的に選択的な入力を受けている Toshiki Kato（加藤利樹），Masato Uemura（上村允人），Tomonari Murakami（村上知成），Kenichi Ohki（大木研一）東京大院医統合生理 Multiple cortical areas hierarchically process different visual information, which is achieved by specific connections between lower and higher order areas. In adult mice, there are several higher visual areas (HVAs) which process different visual information (orientation, direction and spatiotemporal frequency tunings) and receive functionally specific inputs from V1. Soon after eye opening (EO, P14), HVAs already show functional differences which are slightly immature. Anatomically, HVAs receives axonal projections from V1 soon after EO. However, it is unknown whether HVAs already receive functionally specific inputs from V1 and this specificity is involved in development of higher visual areas soon after EO. If the functional differences of HVAs are attributed to the specific inputs from V1, specific inputs from V1 to HVAs will be functionally segregated in this stage. To test this hypothesis, we performed in vivo calcium imaging of the axon terminals from V1 to HVAs at several developmental stages(P18&tilde;30). We injected adeno associated virus (AAV) encoding calcium indicator into V1 and observed axon terminals in HVAs soon after EO. To visualize axon terminals specifically form V1, we developed a method which allow us to localize the infection site into small neonatal cortex and effectively observe axonal response during developmental stages. By using this method combining with in vivo wide-field calcium imaging, we examined axonal responses to moving gratings with various spatiotemporal frequencies. We found that axonal responses from V1 were functionally segregated between HVAs soon after EO. The axonal response property was similar to that of neurons in target HVAs. Moreover, these responses became gradually stronger, and tunings became broader across the ages. In summary, HVAs have already received functionally specific inputs from V1 soon after EO and this functionally specific connection between lower and higher order areas may be important for development of higher order areas.

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-L-516 NT-3/TrkCシグナルによる膨大後部皮質モジュールの形成機構 Toshio Miyashita（宮下俊雄），Haruo Hagiwara（萩原治夫）帝京大学医学部解剖学講座 Apical dendritic bundles from pyramidal neurons are a prominent feature of the cortical neuropil but with significant area specializations. A potentially useful model system for investigating dendritic bundles is the rat granular retrosplenial cortex (RSCg). The RSCg has a highly modular organization in layer (L) 1. This modular structure is consisting of prominent apical dendritic bundles from callosally projecting pyramidal neurons in L2. We had investigated the mechanisms of bundle formation, focusing on L1/2 bundles, in the rat RSCg (Miyashita et al., 2010). By using microarrays, we first searched for genes highly and specifically expressed in RSCg L2 before bundle formation on postnatal day 3 (P3). This screening identified neurotrophin-3 (NT-3) as highly and specifically expressed in the RSCg L2 at P3. To test the importance of NT-3 mediated events in bundle formation, we used in utero electroporation to overexpress NT-3 in cortical areas where L2 bundles are normally absent. This intervention in fact successfully induced apical dendritic bundling in the barrel cortex and adjacent regions. The controlled ectopic induction of dendritic bundles identifies a new role for NT-3, but it remains unclear how the NT-3 signals are transduced into the L2 neuron and regulate dendritic bundle formation. Here, we investigate mechanisms of bundle formation via NT-3. We show that the NT-3 receptor TrkC has important roles in bundle formation in the L2 of RSCg. We induce the expression of a dominant negative variant of TrkC (DN-TrkC), which lacks the tyrosine kinase domain, by in utero electroporation in the RSCg. The RSCg dendritic bundle with DN-TrkC presented induced malformation in which the bundle become wider, especially within its distal tufts. In addition, single-cell labeling and further morphological analysis revealed that inhibition of TrkC signaling induced ectopic dendritic branching. These results suggest that NT-3/TrkC signaling shape L1/2 dendritic bundles by regulating the dendritic branching of individual L2 pyramidal neurons in the RSCg.		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-L-517 神経活動パターン依存的な嗅覚回路形成 Ai Nakashima（中嶋藍）¹，Naoki Ihara（伊原尚樹）¹，Yuji Ikegaya（池谷裕二）^1,2，Haruki Takeuchi（竹内春樹）¹ ¹東京大院薬薬品作用 ²脳情報通信融合研究センター Precise neural circuits emerge through the interplay of genetic programming and activity-dependent refinement. The refinement of olfactory map is instructed by olfactory receptors (ORs) expressed in olfactory sensory neurons. OR identity is represented as a combinatorial code of axon-sorting molecules, whose expression is regulated by neural activity. However, how neural activity induces OR-specific expression patterns of axon-sorting molecules remains unclear. Here, we applied genetic approaches to explore the regulatory mechanisms underlying combinatorial expression of these molecules. Using calcium imaging, we revealed that the spatiotemporal patterns of spontaneous neuronal spikes varied among neurons and were uniquely correlated with the expressed ORs. Receptor substitution experiments demonstrated that ORs determine spontaneous activity patterns. Moreover, optogenetically differentiated patterns of neuronal activity induced specific expression of the corresponding axon-sorting molecules and regulated axon targeting. Thus, temporal patterns of spontaneous activity play instructive roles in generating the combinatorial code of axon-sorting molecules during olfactory map 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-L-518 多小脳回症原因遺伝子Gpr56のe1mプロモーター領域はGABA作動性ニューロンの発生に関与している Ayako Murayama（村山綾子）^1,2，Kenichiro Kuwako（桑子賢一郎）^1,3，Junko Okahara（岡原純子）^1,4，Erika Sasaki（佐々木えりか）⁴，Hideyuki Okano（岡野栄之）^1,2 ¹慶應義塾大学医学部生理 ²理研CBS マーモセット神経構造 ³島根大医生理 ⁴実中研 GPR56, a member of the adhesion G protein-coupled receptor family, is expressed in various tissues including the brain. In the nervous system, GPR56 is highly expressed in neural progenitor cells, and also expressed in developing neurons. The human GPR56 gene has multiple presumptive promoters that drive the expression of the full length GPR56 protein with distinct expression patterns. Similar to the coding mutations of the human GPR56 gene that may cause GPR56 dysfunction, the 15-bp homozygous deletion in the cis-regulatory element upstream of the noncoding exon 1 of Gpr56 (Gpr56 e1m) leads to the cerebral cortex malformation called bilateral frontoparietal polymicrogyria (BFPP) and frontal lobe-associated dysfunctions such as epilepsy. To clarify the expression profile of the Gpr56 e1m promoter-driven GPR56 in primate brain, we generated the transgenic common marmoset line, in which EGFP is expressed under the control of the human minimal Gpr56 e1m promoter. In contrast to the total GPR56 protein that is highly enriched in the ventricular zone of the cerebral cortex, Gpr56 e1m-EGFP is mostly expressed in the developing neurons in the fetus. In addition, Gpr56 e1m-EGFP is predominantly expressed in GABAergic neurons, whereas the total GPR56 protein is evenly expressed in GABAergic and glutamatergic neurons, suggesting the GABAergic neuron-preferential activity of the Gpr56 e1m promoter. These results imply GABAergic neuron-related pathogenesis in the cerebral cortex of the patients with the GPR56 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-L-519 Cdc7-ASKキナーゼ複合体の脳の発生における役割 Karin Hori（堀かりん）^1,2，Tomohiro Iguchi（井口智弘）¹，Shunsuke Kobayashi（小林駿介）^1,2，Tomio Ono（小野富男）³，Chiaki Maruyama（丸山千秋）⁴，Satoshi Yamazaki（山崎聡志）¹，Hisao Masai（正井久雄）^1,2 ¹東京都医学総合研究所・ゲノム動態プロジェクト ²東京大学大学院新領域創成科学研究科メディカル情報生命専攻 ³東京都医学総合研究所・遺伝子改変動物室 ⁴東京都医学総合研究所・神経回路形成プロジェクト The Cdc7-Dbf4(ASK) kinase complex is evolutionally conserved and plays crucial roles in initiation of DNA replication as well as in other chromosome events including meiotic recombination and repair of DNA damages through trans-lesion DNA synthesis. We previously reported that Cdc7 knockout mice die in an early embryronic stage (E3.5-6.5), while hypomorphic Cdc7 mutant mice survive but are deficient in germ cell development (Kim et al. EMBO J. 2002, 2003). Conditional knockout (KO) of Cdc7 in embryonic stem cells results in arrests of DNA replication and accumulation of DNA damages, and eventually to p53-dependent cell death. Cdc7 p53 doubleknockout mice survived until viviparous 8.5 days, probably because cell death was partially circumvented. In order to more precisely examine the roles of Cdc7 during development, we have generated mice with Cdc7 flox mutation, and induced Cdc7 KO in various organs and tissues. Combination of Nestin-Cre with the Cdc7 flox generated mice that lose Cdc7 expression in neural stem cells. Unexpectedly, the mice were born without significant phenotype, but exhibited growth retardation after birth and did not survive beyond 3 weeks of age. The mutant mice showed convulsion as well as impaired body movement, and brain layer formation was severely impaired, indicating crucial functions of Cdc7 in neonatal brain development. In contrast, similar knockout of Dbf4/ASK, the activation subunit of Cdc7, in neural stem cells, did not result in neonatal death, suggesting that some of the Cdc7 functions in brain may be independent of the Dbf4/ASK subunit, that has been identified so far. Cdc7, although required for DNA replication in mammalian culture cells, may control brain development in a manner independent of its function in cell proliferation. We are now trying to identify the target(s) of Cdc7 in brain devlopment. We are also searching for a possible brain-specific activation subunit for Cdc7 kinase. Our results also indicate that Cdc7 functions are dispensable for embryonic neural development, which suggests the presence of a pathway(s) that bypass the role of Cdc7 kinase in proliferation/ development of some organs/ tissues at some developmental stages.

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-L-520 成体でのRP58発現抑制は認知機能低下を惹起する Hiroko Shimbo（新保裕子）^1,3，Shinobu Hirai（平井志伸）¹，Tomoko Tanaka（田中智子）¹，Kenji Tanaka（田中謙二）²，Haruo Okado（岡戸晴生）¹ ¹東京都医学総合研究所神経細胞分化 ²慶應義塾大学医学部・精神神経科学 ³神奈川県立こども医療センター臨床研究所 The <I>Rp58</I> (<I>Zbtb18</I>, <I>Znf238</I>, and <I>Zfp238</I>) gene encodes a BTB/POZ zinc finger transcription factor that regulates cell division and neuronal migration during brain development. The phenotype of patients with RP58 deficiency is characterized by intellectual disability, microcephaly, and agenesis of the corpus callosum. Interesting, expression of RP58 is also reduced in the frontal cortex of the aging human brain. Expression of RP58 is thus important for regulating brain development across the lifespan. Here, we used the tetracycline/doxycycline-regulated transgenic mouse line Actin-tTS::Rp58 tetO to study the effect of RP58 on memory in adult animals. The tetracycline-controlled transcriptional silencer (tTS) was inserted upstream of the <I>Rp58</I> translation initiation site. Expression of <I>Rp58</I> can be reversibly regulated by doxycycline (Dox) administration. tTS mediated tetO mice function without Dox as knockout or knockdown phenotype. Actin-tTS::Rp58 tetO mice were fed doxycycline until they were three-weeks-old and behavioral tests were performed at 4-5 months of age. In the object location recognition test (OLT), control mice spent more time exploring an object moved to a novel place. By contrast, Actin-tTS::Rp58 tetO heterozygous mice spent a similar amount of time exploring the object whether it was moved to a novel place or remained in a familiar place. Further, Actin-tTS::Rp58 tetO homozygous mice exposed to the object that remained in a familiar place than the object moved to a novel place. Reduced RP58 expression after three weeks of age resulted in abnormal location memory in the OLT. RP58 is an important factor for brain function in adulthood.		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-L-521 ドーパミンはD1受容体を介してオレキシン神経細胞内カルシウム濃度を長期的に上昇させる Yasutaka Mukai（向井康敬）^1,2,3,4，Hidenori Aizawa（相澤秀紀）⁵，Kenji F Tanaka（田中謙二）⁶，Takeharu Nagai（永井健治）⁷，Akihiro Yamanaka（山中章弘）^1,2,4 ¹名古屋大環境医神経系２ ²名古屋大院医神経性調節 ³日本学術振興会特別研究員DC ⁴科学技術振興機構CREST ⁵広島大院医歯薬保神経生物 ⁶慶應大医精神・神経科学 ⁷大阪大産研生体分子機能科学 Orexin neurons in the hypothalamus have a critical role in the maintenance of wakefulness. Although several neurotransmitters and bioactive substances modulating the activity of orexin neurons was reported, these studies depended on electrophysiological recording which allowed us to record the activity for few minutes. Therefore, the mechanism of sleep/wakefulness state change, which lasts for seconds to hours, is still unclear. We previously reported that dopamine (DA) application immediately inhibited activity of orexin neurons. However, calcium imaging revealed that DA application induced long-lasting intracellular calcium ([Ca<sup>2+</sup>]<sub>i</sub>) increase in orexin neurons for more than an hour after transient inhibition. The experiments using selective antagonists and agonists suggested involvement of dopamine D<sub>1</sub>-like receptor in this long-lasting increase in [Ca<sup>2+</sup>]<sub>i</sub> in orexin neurons. Here we further genetically investigated the receptor involved in this response using <i>in vivo</i> genome editing (CRISPR-Cas9). To achieve this, adeno-associated virus (AAV) co-expressing SaCas9 and sgRNA for dopamine D<sub>1</sub> receptor (D1R) gene was injected into the hypothalamus of transgenic mice in which orexin neurons exclusively expressed Ca<sup>2+</sup> indicator, Yellow Cameleon-Nano50. More than 4 weeks after AAV injection, we made an acute brain slice and monitored [Ca<sup>2+</sup>]<sub>i</sub> after application of DA. As a result, orexin neurons in D1R knocked out (D1R-KO) slices showed diminished long-lasting calcium increase suggested involvement of D1R in this response. This result strongly suggested that the long-lasting increase in [Ca<sup>2+</sup>]<sub>i</sub> after application of DA in orexin neurons was mediated through the D1R. We will study the physiological role of the long-lasting increase in [Ca<sup>2+</sup>]<sub>i</sub> after application of DA in orexin neurons <i>in vivo</i>.

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-L-522 ドーパミンは線虫C. elegansの運動量の性差を生み出す Satoshi Suo（周防諭）¹，Kazuki Harada（原田一貴）²，Shogo Matsuda（松田翔吾）³，Koki Kyo（姜興起）⁴，Min Wang（王旻）¹，Kei Maruyama（丸山敬）¹，Takeo Awaji（淡路健雄）¹，Takashi Tsuboi（坪井貴司）^2,3 ¹埼玉医大　薬理学 ²東京大院総合文化 ³東京大院理 ⁴帯広畜産大学人間科学 Sex differences in behavior allow animals to effectively mate and reproduce. However, the mechanism by which biological sex regulates behavioral states, which underlie the regulation of sex-shared behaviors such as locomotion, is largely unknown. C. elegans hermaphrodites switch between two distinct behavioral states, roaming and dwelling, in which animals have high and low locomotor activity, respectively. Hermaphrodites spend most of the time in the dwelling state and various neuromodulators are involved in this regulation. However, the behavioral states of males have yet been studied. In this study, we analyzed sex differences in the behavioral states and found that males spend less time in the low activity state than hermaphrodites on food. In dopamine-deficient mutants, the dwelling state was increased in males and, as previously reported, decreased in hermaphrodites compared to the wildtype counterparts, suggesting that dopamine generates the sex differences in the behavioral states by having the opposite effects in males and hermaphrodites. We also found that, in males, dopamine reduces the dwelling state by acting in the same pathway as polycystic kidney disease-related genes, which function exclusively in male-specific neurons. In hermaphrodites, octopamine signaling in the sex-shared SIA neurons promotes locomotion and dopamine increases the dwelling state by suppressing the octopamine signaling. Calcium imaging showed that the SIA neurons of hermaphrodites respond to octopamine by evoking calcium oscillation. In contrast, the calcium response to octopamine was severely reduced in the SIA neurons of males. Furthermore, males explored a larger area of the bacteria lawn than hermaphrodites and dopamine was responsible for this sex difference. In addition, dopamine promotes exploration outside of a lawn in males and suppresses it in hermaphrodites: males leave a lawn at a much higher rate than hermaphrodites in the absence of mates and dopamine was also responsible for this difference. These behavioral differences are beneficial for males to find mates and hermaphrodites that can self-fertilize to stay with food. Together, the results demonstrate that dopamine controls behavioral states in the opposite way through sexually dimorphic signaling to promote adaptive behavior for each sex.		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-L-523 ヒストン脱アセチル化酵素の阻害を伴う運動は大脳皮質における神経栄養因子の発現を修飾する Hiroshi Maejima（前島洋）¹，Mika Kitahara（北原美佳）²，Takahiro Inoue（井上貴博）^2,3，Yasuyuki Takamatsu（高松泰行）¹ ¹北海道大院保健機能回復 ²北海道大院保健 ³日本学術振興会 Neurotrophins play importnat roles in neuroplasticity, neurogenesis, and neuroprotection in the central nervous system. Aerobic exercise increases the expression of neurotrophins, particularly brain-derived neurotrophic factor (BDNF) in the brain and positively contributes to motor and cognitive functions. Epigenetic regulation plays a crucial role to regulate gene transcriptions. The activity levels of histone acetyltransferases (HATs) and histone deacetylases (HDACs) regulate histone acetylation and modulate gene transcription. Specifically, HDAC inhibitors including sodium butyrate (NaB) acetylate histones and enhance gene transcription. Thus, researches focus on pharmacological treatments using HDAC inhibitors to improve or maintain motor and cognitive functions in the patients with central nervous system (CNS) disorders. The objective of the present study was to examine the interactive effects of exercise and pharmacological treatment using HDAC inhibitor (HDACi) on the expressions of neurotrophins including NGF, BDNF and NT-4, and neurotrophin receptors in the cerebral cortex. ICR mice were randomly distributed among 4 groups based on two factors of exercise and HDACi, i.e. a control group, an exercise group, a HDACi group, and an exercise plus HDACi group. We administered a HDAC inhibitor, NaB intraperitoneally to the mice (the HDACi group and the exercise plus HDACi group) at a dose of 1.2g/kg, whereas the mice (the exercise group and the exercise plus HDACi group) were exercised on a treadmill for approximately 1 h every day. After four week intervention, the expression of mRNA of neurotrophins (NGF, BDNF and NT-4) and neurotrophin receptors (Trks and p75) were assayed using real time PCR following behavioral tests. All study procedures were approved by the ethics committee for animal research of Hokkaido University in Japan. Although we could not confirm a significant improvement of motor and cognitive functions, HDACi significantly increased the expression of NGF and neurotrophin receptor, p75, specifically in the presence of exercise. Therefore, the present study suggested that HDAC inhibition with repetitive administration of NaB could enhance the exercise-induced expression of neurotrophin and neurotrophin receptor, and positively contribute to neuroplasticity, neurogenesis, and neuroprotection in the cerebral cortex of the patients with CNS 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-L-524 マウス生体脳におけるL-ドパニューロンの分布と神経化学的特性 Shunsuke Nakagawa（中川舜介），Masahiko Watanabe（渡辺雅彦），Kohtaro Konno（今野幸太郎）北海道大院医解剖発生学教室 L-DOPA is synthesized from tyrosine by the action of tyrosine hydroxylase (TH), the rate-limiting enzyme in the catecholamine biosynthesis, and is converted to dopamine by aromatic L-amino acid decarboxylase (AADC). L-DOPA has been thought as a precursor molecule for dopamine, adrenaline, noradrenalin. Nevertheless, there is immunocytochemical evidence for existence of neurons that express TH, but not AADC, suggesting that L-DOPA could also function as a transmitter or modulator. In the present study, we defined TH(+)/AADC(-) neurons as potential L-DOPA neurons, and examined their distribution and neurochemical properties in the adult mouse brain. By double immunofluorescence, TH(+)/AADC(-) neurons were densely populated in the arcuate nucleus, olfactory bulb, A1/C1 region, and A3 region, as reported previously. In addition, we found the ventral part of periaqueductal gray (PAG) to be enriched with TH(+)/AADC(-) neurons. These neurons in the PAG were negative to dopamine transporter (DAT) and vesicular monoamine transporter 2 (VMAT2), but expressed VGluT2 mRNA. In comparison, TH(+)/AADC(+) neurons in hypothalamic A11 region, which is located rostral to the PAG and projects dopaminergic fibers to the spinal cord, expressed VMAT2, but not DAT or VGluT2 mRNA. These results suggest that TH(+)/AADC(-) neurons in the PAG are not endowed with molecular machineries for dopamine synthesis, vesicular filling, or re-uptake, and may co-release glutamate. Furthermore, the lack of VMAT2 implies that L-DOPA might be released from these neurons by non-vesicular mechanisms. Projection targets of TH(+)/AADC(-) neurons in the PAG should be examined in future studies.		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-L-525 ラット脳におけるムスカリン性アセチルコリン受容体M2の局在解析 Takashi Hashimoto（橋本隆）¹，Hiroshi Nagano（永野宏）²，Toshihiro Unno（海野年弘）²，Satoshi Iino（飯野哲）³ ¹愛知医科大・医・薬理 ²岐阜大応用生物獣医・獣医薬理 ³福井大医人体解剖 Muscarinic acetylcholine receptors are family A G-protein-coupled receptors activated by muscarine as the transmitter, and five subtypes (M1 to M5) have been identified. It is known that muscarinic receptors play a significant role in cholinergic responses in the various tissue including CNS, heart, lung, etc. M1 in the hippocampus are important for learning and memory. M5 expressed in ventral tegmental is thought to be involved in reward system. It is reported that, M2 is located on the sinus node and regulates the heart rate, and we revealed that M2 is distributed in the smooth muscle cells and interstitial cells of Cajal in gastrointestinal tract (Iino S et al., 2006, Neuroscience). However, little is known about the localization of M2 in the CNS. In this study, we analyzed the immunohistochemical distribution of M2 in the brain of adult male rats by using specific monoclonal antibody. M2 immunoreactivity detected in the telencephalon including cerebral cortex (layer III and V) and hippocampus (pyramidal and oriens layer). In basal ganglia, we observed robust M2 positive fibers and cell bodies in the horizontal and lateral diagonal band nuclei. Prominent immunoreactivity of M2 was seen in anterodorsal thalamic nucleus, and M2 immunoreactive fibers were also found in lateral hypothalamic area. In the midbrain, strong immunopositive reaction of M2 was observed in superior colliculus and pretectal area. In addition, robust immunostaining for M2 was located in the pontine nuclei and motor nuclei of the trigeminal, facial and hypoglossal nerve.Our histological data show that distribution pattern of M2 protein is surely correlate with that of acetylcholine, but is not always correspond to that of other muscarinic receptors, suggesting its unique functions in the 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-L-526 HCN1 and HCN2 at specific sites of the cerebellar circuit Anne Gunther（Gunther Anne），Thomas Launey（Launey Thomas） RIKEN Center for Brain Science, Saitama, Japan Among the various ion channels that contribute to initiation, coordination and modulation of neuronal signals, hyperpolarization-activated and cyclic nucleotide-gated (HCN) channels play an essential role in the determination of biophysical properties of membranes. HCN channels are activated at negative membrane potentials and their activation can additionally be modulated by direct binding of cAMP. However, while it has been shown that HCN channels affect neuronal excitability throughout the murine CNS, the question how they contribute during neuronal development remains largely unaddressed. Here, we assessed the expression of individual HCN isoforms in the murine cerebellum. The cerebellum is the center of motor learning and motor control and thus plays an important role during development. Additionally, due to its distinct cellular organization and defined pathways of signal relay, the cerebellum is an appealing target for the study of neuronal development. We assessed expression profiles of individual HCN isoforms in cerebellar tissue at different ages from embryo to adult. We found distinct expression patterns of the subunit isoforms HCN1, HCN2 and HCN4 on the RNA level as well as on the protein level, as well as correlation with identified molecular marker proteins of specific cell types. Additionally, we could show specific localization of individual HCN isoforms to distinct, subcellular sites of the cerebellar circuit. For functional studies, we established recombinant adeno-associated viral vectors as efficient tools for the modification of neuronal cell function based on RNAi. Application of RNAi-mediating viral vectors in vivo allows assessment of the functional contribution of individual HCN isoforms to cerebellar processes including neuronal development as well as learning 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-L-527 ミクログリアは末梢神経損傷に伴う視床回路の再構築を制御する Yoshifumi Ueta（植田禎史），Mariko Miyata（宮田麻理子）東京女子医大院医神経生理 Peripheral nerve injury rapidly reorganizes somatotopic maps in the adult brain. Using the mouse whisker sensory pathway, we previously reported that transection of whisker sensory nerve (IONC, infraorbital nerve cut) induces aberrant changes in ascending synaptic inputs from neurons in the principal sensory trigeminal (Pr5) nucleus of the brainstem to neurons in the ventral posteromedial (VPM) nucleus of the thalamus. In the normal mature VPM, neurons in barreloids receive a single ascending fiber input from Pr5 barrelette neurons. In the IONC group, on the other hand, VPM neurons receive multiple fiber inputs from the Pr5 nucleus. These multiple inputs contain both whisker-derived and ectopic axon terminals, which are originated from barrelettes and non-barrelettes, respectively. Thus, IONC reorganizes whisker representation in the VPM nucleus. However, it is unclear what mechanisms regulate this nerve injury-induced thalamic circuit reorganization. Here we show that removal of microglia, which are well-known to associate with formation and refinement of synapses, prevents nerve injury-induced aberrant changes in thalamic circuit and function. IONC induced activation of microglia specifically in the Pr5 barrelettes of nerve-injured pathway. Thus, we examined the involvement of microglia in nerve injury-induced thalamic circuit reorganization by using depletion of microglia. Orally administered plx3397, an inhibitor for colony stimulating factor 1 receptors, removed almost all of microglia from the brain. We found that the depletion of microglia prevented IONC-induced increase of multiple fiber inputs to VPM neurons. We also found that microglia depletion prevented IONC-induced increase of ectopic axon terminals in the VPM nucleus. In addition, local depletion of microglia from the Pr5 nucleus prevented IONC-induced increase of ectopic axon terminals, suggesting that removal of activated microglia in the Pr5 nucleus is sufficient for preventing IONC-induced remodeling of ascending inputs to postsynaptic VPM neurons. We previously showed that IONC induces mechanical hypersensitivity to tactile stimulation. This hypersensitivity was disappeared by the depletion of microglia. Thus, our results demonstrate an important role of microglia for regulating aberrant plasticity changes in thalamic circuit structure and function induced by peripheral nerve 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-L-528 光遺伝学により細胞体活動を抑制した条件下でのスパインの視覚応答記録 Satoru Kondo（根東覚）^1,2，Kohei Kikuta（菊田浩平）²，Kenichi Ohki（大木研一）^1,2 ¹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 ²東京大学大学院医学系研究科統合生理学分野 Understanding how neurons integrate thousands of synaptic inputs is critical to discern cortical information processing. Substantial evidences suggest the importance of spatial arrangement of synaptic inputs onto dendrites for neuronal computation. However, the principle of spatial arrangement and integration mechanisms of inputs remain largely unsolved. Recent advances of functional imaging technique enable recordings of activities of individual spines using various calcium sensors. Due to the low time resolution of calcium signal transient, back propagating action potential (bAP) invades spines and makes the accurate spine analysis difficult. To solve this problem two methods are currently used. (1) Subtract the contaminated bAP signal from the spine signal and estimate the `true' spine signal. (2) Prevent the somatic action potential generation by voltage-clamping the imaged neuron. However, both methods have some technical problems. In this study we developed a new method to overcome this problem by using inhibitory optogenetics. We sparsely co-expressed GCaMP6s and inhibitory optogenetic protein with soma-localized signal in mouse primary visual cortex (V1) by adeno associated virus (AAV) and recorded visually evoked spine signals from excitatory neurons. Without inhibition, spine signals were contaminated with bAP. On the contrary, with inhibition, we successfully suppressed the somatic depolarization and individual spine signal was detected. Then, we recorded the visually-evoked signals from the same spines without inhibition followed by with inhibition and compared them. We found that the bAP-subtracted method does not always estimate the accurate visual response of spines especially when the spine shows the simultaneous response with the soma. Next, to investigate the input-output relationship, we recorded ~1,000 spine responses from individual orientation or direction selective neurons. We found that ~30% of recorded spines were visually responsive and among them ~90% were selective for either orientation or direction. These selectively responded spine were distributed all over the dendrites without apparent clustering of similar selectivity in terms of the distance from the soma or the same dendritic segments.		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-L-529 グリオキサール固定液は脳におけるシナプス分子の検出に有効かつ有用である Kohtarou Konno（今野幸太郎），Miwako Yamasaki（山崎美和子），Masahiko Watanabe（渡辺雅彦）北海道大院医解剖発生 Paraformaldehyde (PFA) is the most commonly used fixative for immunostaining of cells and tissues. Nevertheless, PFA fixation has various problems, as exemplified by loss of antigenicity, tissue shrinkage, and hindered antibody access to antigens, particularly, at postsynaptic sites. Recently, it has been reported that glyoxal, a dialdehyde with OHC-CHO (M.W. 58.04), is a valuable alternative to PFA, a monoaldehyde with HCHO (30.03) for immunostaining by its faster cross-linking ability of proteins and better preservation of cellular morphology (Richter et al, 2018). Here we compared these fixatives in immunohistochemical detection for synaptic molecules in the adult mouse brain. Noteworthily, antigen exposing techniques, such as protease digestion for light microscopic immunohistochemistry (Watanabe et al., 1998) and postembedding methods for electron microscopic one (Landsend et al., 1997), is necessary for ionotropic glutamate receptors and PSD proteins in PFA-fixed tissues, whereas these postsynaptic molecules were readily detectable in glyoxal-fixed tissues without antigen exposing techniques. For example, selective localization of GluD2 at parallel fiber-Purkinje cell synapses was demonstrated in glyoxal-fixed cerebellar tissues by conventional immunofluorescence and preembedding immunoelectron microscopy. Furthermore, the intensity of immunohistochemical signals was intensified for most other synaptic molecules, including VGluT, VIAAT, bassoon, Neurexin, NMDA receptor, AMPA receptor, PSD-95, Gephyrin, and GABAA receptor. Therefore, glyoxal is an effective and useful fixative for immunohistochemical detection of synaptic molecules. It is particularly worth trying this fixative when immunosignals are unsatisfactory in PFA-fixed brain tissues using unfamiliar or first time antibodies.

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-L-530 プロテインキナーゼCγは登上線維のシナプス伝達を制御する Masashi Watanave（渡邊将），Nobutaka Takahashi（髙橋伸卓），Ayumu Konno（今野歩），Hirokazu Hirai（平井宏和）群馬大院医神経生理 Protein kinase Cγ (PKCγ) is expressed exclusively in neurons of the brain and spinal cord. In the cerebellar cortex, only Purkinje cells (PCs) expressed PKCγ. PKCγ-deficient PCs show normal cerebellar long-term depression (LTD), but persistent innervation by multiple climbing fibers (CFs) and impaired motor coordination. We found that viral vector-mediated re-expression of PKCγ in PCs resulted in significant rescue of behavioral defects seen in PKCγ-deficient mice, suggesting that PKCγ had a pivotal role in adult PCs and thereby, regulating the cerebellar function. To clarify a role of PKCγ in mature PCs, we compared the electrophysiological properties of PKCγ-null PCs with those of wild-type (WT) PCs. We found no difference in synaptic transmission from PF to PC and from interneuron to PC between WT and KO mice. In contrast, we found significant decrease in CF-EPSC amplitudes in KO PCs, which was elicited in the presence of EGTA in the internal solution at holding current (Hc) of -10mV, but not at Hc of -70mV (in presence of 0.5 μM NBQX), suggesting the voltage dependency of CF-PC EPSC modulation by PKCγ. Intriguingly, the significant decrease in CF-EPSC amplitudes in KO PCs was not observed when using the internal solution containing 10mM BAPTA, instead of EGTA, or the extracellular solution containing 500 μM TEA, suggesting that modulation of CF-EPSC amplitude by PKCγ was Ca2+ and K+ dependent. Based on these results, we hypothesized that the modulation on CF-EPSC was mediated through activation of large conductance potassium (BK) channels, which was activated by voltage-gated calcium channels (VGCCs). Namely, we assumed that BK channel function was augmented in KO mouse PCs. Enhanced BK current in KO PCs could cause shunting effect and attenuate the excitatory signal during the propagation from CF-PC synapses to cell-body. To test this, we are going to compare CF-PC EPSC amplitude in the presence of iberiotoxin (BK channel blocker) between genotypes, and more directly, to measure the BK current. We are planning to show the results in this poster 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-L-531 AMPA受容体のGluA1ホモマーのN型糖鎖修飾はシナプス可塑性に重要な役割を担う Yoshihiko Wakazono（若園佳彦）¹，Ryosuke Midorikawa（緑川良介）¹，Munal B Kandel（カンデル B ムナール）¹，Shogo Oka（岡昌吾）²，Kogo Takamiya（高宮考悟）¹ ¹宮崎大医機能制御・統合生理 ²京都大院医人間健康科学生化 The intracellular molecular mechanisms underlying the regulation of the AMPA receptor have been dramatically elucidated in the past few decades. In contrast, the functional regulation of the extracellular domain remains unclear. Here, we focused on N-glycosylation of the AMPA receptor in the extracellular domain and tried to clarify their functions by combining molecular biological and electrophysiological techniques. We previously reported that 401 asparagine residues (N401), putative N-glycosylation site, in GluA1 subunit might be a responsive site for drastic changes of glutamate responses of AMPA receptor from desensitization to re-sensitization, and that long-term potentiation (LTP) induction of hippocampal slices prepared from N401Q GluA1-expressing lentivirus vector-injected GluA1 KO mice was impossible to maintain its potentiation. In the present study, we tried to clarify molecular mechanisms underlying the defect of LTP maintenance in N401Q GluA1 mutant. Analysis of miniature EPSC (mEPSC) revealed that the frequency, amplitude and rise times showed no significant change, and that the decay time became significantly faster than that of wild-type GluA1. In addition, when expressed N401Q GluA1 in primary cultured neurons prepared from embryonic GluA1 KO mice brain, the glutamate response did not always show the re-sensitization. These are not consistent with results in N401Q GluA1-expressing HEK cells. To explain this discrepancy, we next examined the involvement of GluA2 subunit, which is possible to form heteromers with GluA1 subunit. Glutamate response in N401Q GluA1-expressing HEK cells reverted the re-sensitization to the desensitization by co-expression with GluA2 subunit. These results suggest that the re-sensitization in N401Q GluA1 homomers might influence to synaptic currents during the LTP induction period, not in the resting state.

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-L-532 小脳におけるシナプス伝達・長期抑圧へのノルアドレナリンの作用の違い Takuma Inoshita（井下拓真），Tomoo Hirano（平野丈夫）京都大院理生物物理 The cerebellum is involved in motor learning and contribution of noradrenaline (NA) to cerebellum-dependent learning paradigms has been reported. On the other hand, long-term depression (LTD) at parallel fiber to Purkinje neuron (PF-PN) synapses in the cerebellum has been regarded as a primary cellular mechanism for motor learning. However, the relationship between NA and LTD at PF-PN synapses has not been clarified. We previously reported that application of NA to the cerebellar flocculus enhanced optokinetic response (OKR) which works to stabilize an image on the retina during movement of visual field (Wakita et al., 2017). The flocculus is known as a regulation center of oculomotor reflexes including OKR, and we also reported that during adaptation of OKR, LTD takes place in the flocculus (Inoshita et al., 2018). Thus, we considered that NA might induce or facilitate LTD, and examined effects of NA on excitatory postsynaptic currents (EPSCs) and LTD at PF-PN synapses in the flocculus by whole-cell patch-clamp recording. NA application did not significantly change EPSC at PF-PC synapses in slice preparations. In order to examine the possibility that NA facilitates LTD induction, we tried to induce LTD by a relatively weak conditioning stimulation after NA application. The weak conditioning stimulation which did not induce LTD in the control solution induced LTD at PF-PN synapses in the NA containing solution. Thus, NA facilitated LTD induction without affecting basal synaptic transmission in the flocculus. Next, we asked whether the NA effects on PF-PN synapses are common in the cerebellum, and examined effects of NA on the EPSC amplitude and LTD in the vermis, another region of the cerebellar cortex. NA application to a slice preparation decreased the amplitude of EPSC at PF-PN synapses, and the weak conditioning stimulation did not induce LTD in the NA containing solution. Thus, NA suppressed PF-PN synaptic transmission, and LTD induction might have been occluded in the vermis. These results indicate that effects of NA on PF-PN synapses are different among cerebellar areas.		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-L-533 シナプス可塑性におけるArc-Drebrin相互作用に関する研究 Yuta Ishizuka（石塚佑太），Clive R. Bramham（Bramham R. Clive） Dept of Biomed, Univ of Bergen, Bergen, Norway Arc (activity-regulated cytoskeleton-associated protein) is involved in multiple forms of synaptic plasticity. In long-term potentiation (LTP), Arc promotes the phosphorylation of cofilin, resulting in the inhibition of its actin-severing activity. In long-term depression (LTD), Arc binds to inactive CaMKIIβ at inactive synapses and triggers endocytosis of AMPA receptors. Because Arc is functioning both in LTP and LTD, Arc is implicated as master regulator of synaptic plasticity. However, little is known about molecular basis of Arc function in synaptic plasticity. Also, the mechanism of Arc in actin-cytoskeletal regulation is unknown. We previously showed that Arc forms a complex with the F-actin binding protein drebrin A in cytoskeletal fraction <I>in vivo</I>. Recently, it has been also shown that Arc regulates intracellular actin dynamics via myosin II and Arc is necessary for the migration of skin-migratory dendritic cells. From several lines of evidence, Arc may regulate actin-cytoskeleton dynamics by cooperating with actin-binding proteins (ABPs) in synaptic plasticity because the translocation of drebrin driven by myosin II is a trigger for actin-cytoskeletal reorganization during LTP, and this translocation may allow some synaptic proteins including newly synthesized Arc to flow into dendritic spines. In this study, we hypothesized that protein-protein interactions (PPIs) among Arc and ABPs are temporally and spatially controlled during LTP and the modification of PPIs underlie actin-cytoskeletal reorganization. To address this hypothesis, cultured neurons were chemically induced LTP as the cellular model of LTP. Co-immunoprecipitation (co-IP) analysis revealed that Arc interacts with several ABPs such as drebrin and myosin IIA/B in basal state and these PPIs were enhanced by LTP stimulation. To clarify these PPIs at synapses, we isolated synaptoneurosome compartment, then performed co-IP and western blotting. We found that chemical LTP increased Arc expression, and promoted these PPIs in synaptoneurosomes. Our results suggest that Arc-drebrin-myosin II interaction may contribute to actin polymerization during LTP because the translocation of drebrin-decorated F-actin is mediated by myosin II activity and also drebrin-decorated F-actin is resistant to cofilin-mediated actin-severing.

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-L-534 背内側コリン作動性ニューロンは発火の停止により皮質線条体シナプス可塑性の方向性を調節する Atsushi Tamura（田村篤史），Julie Chouinard（Chouinard Julie），Kiyoto Kurima（Kurima Kiyoto），Yumiko Akamine（Akamine Yumiko），Jeffery R Wickens（Wickens R Jeffery）沖縄科学技術大院神経生物学研究ユニット The cholinergic interneurons (CINs) of the striatum are crucial for behavioral flexibility. CINs of the dorsomedial striatum (DMS) play a role in strategy switching. However, how CINs modulate the neural circuitry underlying strategy switching is unclear. The glutamatergic afferents from the cerebral cortex to the striatum display activity-dependent plasticity in the corticostriatal synapses, and may be involved in certain types of learning. One hypothesis is that strategy switching may be realized by a modulatory effect of CINs on corticostriatal plasticity. Here, we investigated the effect of CINs on activity-dependent plasticity in the corticostriatal synapses. To control tonically firing of CINs, AAV encoding halorhodopsin (NpHR) was injected into DMS of ChAT-cre mice. AAV injected mice expressed NpHR in CINs and we can optogenetically inactivate CINs firing. We recorded EPSPs induced by electrical stimulation of corpus callosum using ex vivo slice whole-cell recording from spiny projection neurons (SPNs), which are the output neurons of the striatum. Activity dependent synaptic plasticity was induced by high-frequency stimulation under the Mg-free conditions. This conditioning stimuli combined with optogenetically inactivation of CINs during HFS induced the long-term potentiation in some SPNs. However, other group of SPNs showed long-term depression to the same conditioning stimuli. This result might indicate that CIN activity modulate corticostriatal plasticity in different manner between direct and indirect SPNs.		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-L-535 海馬顆粒細胞における抑制入力による応答特性への影響 Naoki Nakajima（中島直樹）¹，Gennosuke Tasaka（田坂源之助）¹，Hirofumi Hayakawa（早川博章）²，Takeshi Aihara（相原威）^1,2 ¹玉川大学大学院工学研究科 ²玉川大学工学部 Spatial information (place) and non-spatial information (mainly odor) are integrated in hippocampal dentate granule cells (GCs). It was reported that spatial information is propagated at theta oscillation (4-8 Hz) to the medial dendrite (MD) of GCs. On the other hands, non-spatial information is propagated at gamma oscillation (20-40 Hz, average 21.0 Hz) to the lateral dendrite (LD) of GCs. However, how those information is integrated is still unknown. To investigate the integration mechanism of two inputs to MD and LD, the frequency responses of MD and LD of GCs were measured using rat hippocampal slices. We applied 5 pulses stimulus at 10-40 Hz to MD or LD. During experiment, to block the synaptic plasticity induced by frequency stimulus, NMDA-receptor antagonist, D-APV, was applied. In addition, GABA-A receptor antagonist, picrotoxin, was applied for controlling inhibitory inputs. As the physiological experimental result, responses were transiently decreased at MD of GCs in both the presence and the absence of inhibitory inputs. On the other hand, successive responses for five inputs were sustained at LD of GCs in the presence of inhibitory inputs. However, when inhibitory inputs were blocked by picrotoxin, these successive responses were not sustained and transiently decreased at LD of GCs. The result suggest that inhibitory inputs may play an important role as a stabilizer for keeping the amplitude of successive response at LD. In addition, computer simulation was performed using NUERO simulator with a multi-compartment model of the GC. This model was fixed by parameter fitting for the physiological data. Theta burst input and random pulses (10-40 Hz, gamma) were applied to MD and LD of the GC model, respectively. As the computational experimental result, the temporal-pattern sensitivity for theta burst inputs applied to MD was not clear. However, when random pulse inputs were additionally applied to LD at 10 Hz, GC activation for theta burst input was increased. Moreover, 20Hz-30Hz random pulse inputs were simultaneously applied to LD, GC activation was more facilitated so that the proportion of the response area for input temporal patterns changed from broad to narrow area. It suggests that temporal pattern sensitivity for theta inputs was tuned and clearly observed in the presence of DD inputs. Our results suggest that processing of spatial information was enhanced depending on the input frequency of non-spatial information in GC.

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-L-536 リハビリテーション促進薬投与後の損傷周囲におけるシナプス構造の変化 Sayaka Kogami（湖上爽）¹，Susumu Jitsuki（實木亨）¹，Yutaka Kawakami（川上裕）¹，Aoi Jitsuki-Takahashi（實木－高橋葵）²，Tomomi Yamanoue（山ノ上友美）¹，Hitoshi Masuyama（増山仁）³，Takeaki Yano（矢野武明）³，Takuya Takahashi（高橋琢哉）¹ ¹横浜市大院医生理 ²東京女子医大生理 ³富士フィルム医薬品・ヘルスケア研究所 Damage to the brain causes severe neurological condition, such as sustained sensory, motor, cognitive dysfunction and compromise work capacity and self-care. No pharmacological intervention that could foster recovery and complement current rehabilitation has yet been established as effective. Restoration of motor impairment after brain damage is considered to be the result of compensative neural plasticity in spared neural circuit, and the Experience-dependent synaptic AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic-acid) receptor (AMPAR) delivery underlies behaviors that require neural plasticity such as learning. We have previously found that a small compound, edonerpic-maleate (also known as T-817MA), facilitated experience-driven synaptic glutamate AMPA receptor delivery in the peri-injured area and resulted in the acceleration of motor function recovery after cortical damage. However, it remains unclear whether the recovered motor function could be maintained over the long term after drug withdrawal. Moreover, if the motor function is maintained, the structural synaptic plasticity in that period is also unknown. To address this issue, we tested forelimb motor performance 2 months after termination of rehabilitative training combined with drug treatment, and quantified three types of dendritic spines (mushroom, stubby, thin) in the peri-injured area of cortex at this period. We found that the recovered motor function was maintained 2 months after termination of the treatment. We also found that only in the edonerpic treated animal, the number of mushroom-type spine was negatively correlated with recovery rate, while the number of thin-type spine was positively correlated with recovery rate. These results suggest that rehabilitation with edonerpic treatment affect dendritic spine morphology for maintaining of recovered motor performance.		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-L-537 海馬顆粒細胞におけるLTP誘導により調節される逆伝播活動電位の解析 Shoya Sugimoto（杉本翔哉）¹，Motoya Kwai（河合元弥）¹，Masashi Kondo（近藤将史）²，Yasuhiro R Tanaka（田中康裕）³，Takeshi Aihara（相原威）^1,4 ¹玉川大院工 ²東京大院医機能生物 ³玉川大脳研 ⁴玉川大 The dentate gyrus plays as an initial integrator of various inputs in the hippocampal circuit. The dentate granule cells receive spatial information (e.g. place) and non-spatial information (e.g. odor) from entrhinal cortex (EC) to medial dendrite (MD) and lateral dendrite (DD) at the molecular layer, respectively. Depending on the precise timing of presynaptic spikes (associated EPSP) and postsynaptic spikes (associated back-propagated action potential, bAP), repetitive spiking induces either Long-term potentiation (LTP) or depression (LTD), known as spike-timing-dependent plasticity (STDP), in hippocampal neurons. The bAP is one of important factors for the STDP induction. In previous studies, we reported that amplitudes and transmission distances of the bAP were modulated by LTP induction along dendrites in hippocampal CA1 pyramidal neurons. The result suggested that association of the context information from CA3 and the other information through a direct pass from EC was facilitated in CA1 pyramidal neurons. However, it is unknown how LTP induction effects amplitudes and transmission distances of the bAP in hippocampal dentate granule cells. Hence, the association mechanism of spatial and non-spatial information is still unclear in dentate gyrus granule cells. In this study, to investigate the influence of synaptic-plasticity induction on the amplitude changes of the bAP in dentate granule cells, we measured amplitude changes of the bAP depending on LTP induction in the hippocampal granule cells, using optical imaging method with voltage sensitive dye. As the result, the bAP transmission distance was spread from MD to DD after LTP induction. Consequently, it is considered that the association of place information to MD and the non-spatial information to DD was facilitated, depending on the bAP modulation caused by the induction of synaptic plasticity in dentate granule cell.

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-L-538 小脳顆粒細胞における軸索起始部の可塑性と選択的スプライシング制御 Takatoshi Iijima（飯島崇利）¹，Satoko Suzuki（鈴木暁子）¹，Hiroshi Kuba（久場博司）²，Yoko Iijima（飯島陽子）¹，Yuji Sato（佐藤悠司）¹ ¹東海大医基礎医分子生命 ²名古屋大院医細胞生理 Neuronal alternative splicing is a powerful mechanism for functional diversification. Multiple of neural genes are subjected to alternative splicing with neuronal activity via Ca2+-dependent signaling pathways. The activity-regulated gene regulation could underlie plasticity-dependent changes in the molecular composition. However, the functional role in activity-regulated alternative splicing programs are just beginning to be uncovered. Here we found that activity-regulated alternative splicing by Rbfox1 causes the molecular changes at the axon initial segment (AIS), a specialized compartment of proximal axons that initiates action potentials, in the cerebellar granule cells (CGCs); Rbfox1 regulates alternative splicing of the transcripts encoding AIS constituents such as Neurofascin and Nav1.6 in response to high K+-induced depolarization. In addition, we revealed that the depolarization diffusely elongated the length of AIS in the CGCs. Notably, the knockdown of Rbfox1 significantly attenuated the high K+-induced elongation of AIS. Both the molecular and structural changes at the AIS by high K+-induced depolarization were late-onset, long-lasting and reversible with order of day through L-type calcium channel and ERK signaling, indicating that the activity-regulated alternative splicing by Rbfox1 couples to structural changes at the AIS in CGCs. This structural change accompanies a decrease in the density of voltage-gated Na+ (Nav) channels at the AIS, thereby which functionally leads to the homeostatic reduction in neuronal excitability. Therefore, this study uncovers the strong link between the activity-regulated alternative splicing and homeostatic plasticity at AIS.		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-L-539 Histamine H<sub>1</sub> receptors on astrocytes regulate aggressive behaviour, circadian rhythm, and wakefulness in mice Aniko Karpati（Karpati Aniko）¹，Takeo Yoshikawa（Yoshikawa Takeo）¹，Fumito Naganuma（Naganuma Fumito）^1,2，Kazuhiko Yanai（Yanai Kazuhiko）¹ ¹Dep. of Pharmacology, Tohoku University, Sendai, Japan ²Division of Pharmacology, Tohoku Medical and Pharmaceutical University, Sendai, Japan The neurotransmitter histamine contributes to various processes, including the sleep-wake cycle, learning, memory, and stress responses. Alterations in brain histamine levels are closely related to central nervous system (CNS) dysfunction and are thought to contribute to neurological disorders, such as Alzheimer's disease and depression. Actions of histamine are mediated through G protein-coupled histamine H<sub>1</sub>-H<sub>4</sub> receptors. Previous studies have reported the expression of H<sub>1</sub>-H<sub>3</sub> receptors in the CNS. Conventional H<sub>1</sub> receptor gene (<I>Hrh1</I>) knockout mice showed significant phenotypic differences compared to controls. For example, exploratory behaviour, aggression, learning, and memory were affected by <I>Hrh1</I> deficiency. Further, pharmacological studies have emphasised the importance of H<sub>1</sub> receptors for wakefulness and cognition in humans. H<sub>1</sub> receptors are abundantly expressed on neurons and astrocytes. Astrocytes are multifunctional cells that critically contribute to CNS physiology, including cerebral blood flow, energy metabolism, ionic homeostasis, and synaptic function. Studies with rodents have shown the importance of astrocyte signalling in complex behaviours, such as memory and circadian rhythm. Furthermore, various neurological diseases, including Alzheimer's disease and epilepsy, are associated with pathological states of astrocytes in rodents and humans. Growing evidence demonstrates functions of astrocytic H<sub>1</sub> receptors in vitro, emphasising the need to selectively assess the impact of neuronal and astrocytic H<sub>1</sub> receptor signalling on behaviour. However, studies selectively investigating the roles of neuronal and astrocytic H<sub>1</sub> receptors in behaviour are lacking. We generated novel astrocyte- and neuron-specific conditional knockout (cKO) mice to address this knowledge gap. cKO mice showed cell-specific reduction of <I>Hrh1</I> expression. H<sub>1</sub> receptors on both cell types played a significant role in anxiety. Astrocytic H<sub>1</sub> receptors were involved in regulating aggressive behaviour, circadian rhythms, and quality of wakefulness, but the sleep-wake cycle remained unaffected. Our results emphasise the importance of neuronal H<sub>1</sub> receptors in recognition memory. In conclusion, this study highlights the novel roles of H<sub>1</sub> receptors on astrocytes and neurons in various brain functions.

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-L-540 Neuron-microglial contacts are required for endotoxin-tolerance-mediated inhibition of PGE2 Chun-Hsien Chu（Chu Chun-Hsien） National Cheng Kung University Endotoxin tolerance (ET), a reduced responsiveness to an endotoxin challenge following a previous encounter with the endotoxin, has been well studied in peripheral innate immune cells such as macrophages/monocytes. However, little is known about ET in the brain. After the immune challenge, the inducible cyclooxygenase (COX-2)/prostaglandin E2 (PGE2) pathway trigger neuroinflammation and neurodegeneration. The present study determined that the regulatory mechanism of PGE2 reduction in brain immune cells, microglia, during ET challenge. Our data showed that expression of cox-2 mRNA at 3 hours and PGE2 at 24 hours was significantly reduced in primary neuron-glial cells with two lipopolysaccharide (LPS) treatments compared to these cells with one LPS treatment, indicated LPS-tolerant microglia produce lower PGE2. To determine if TLR4 is required for the reduction of PGE2 in tolerated microglia, the serum-free medium without LPS binding protein (LBP) was used to culture neuron-glia cells. Our data showed that PGE2 production did not be reduced, even higher, when the cells were encountered with twice LPS in serum-free medium. Conversely, adding LBP back into the medium rescue the LPS-tolerant capacity of microglia on PGE2 production. Additionally, blockage of new protein synthesis by cycloheximide disrupted PGE2 reduction in tolerized microglia. These data indicated that TLR and new protein synthesis are required for tolerized microglia. Next, we determined if interactions among neurons and glia play roles in the reduction of PGE2 under endotoxin tolerance. Our findings showed that the LPS tolerance-mediated PGE2 reduction was disappeared in mixed-glia and microglia-enriched cultures, implying the presence of neurons was necessary for PGE2 reduction under endotoxin tolerance. To determine if soluble factors secreted by neurons can regulate the LPS tolerance-mediated PGE2 reduction, our results showed that pre-incubation of the mixed-glia cells (loss of endotoxin tolerance-mediated PGE2 reduction) with neuron-glial condition medium failed to induce the occurrence of PGE2 reduction under endotoxin tolerance. Interestingly, after incubation with the fixed neurons, microglia with twice LPS treatment produced lower PGE2 than the cells treated with once LPS. Taken together, our study demonstrated that neuronal contacts play essential roles in the formation of PGE2 reduction in tolerized microglia.		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-L-541 脳虚血障害におけるミクログリアVNUTの役割 Yuri Hirayama（平山友里）¹，Ha Pham Ngoc Le（Pham Ngoc Le Ha）³，Itsuko Ishii（石井伊都子）¹，Naohiko Anzai（安西尚彦）²，Schuichi Koizumi（小泉修一）³ ¹千葉大学医学部附属病院薬剤部 ²千葉大院医薬理 ³山梨大院医工薬理 ATP is known to be increased in response to transient brain ischemia, but whether this ATP increase is beneficial or harmful to cerebral ischemic injury is controversial, and a mechanism underlying the ATP increase is unknown. Vesicular nucleotide transporter (VNUT), a transporter responsible for vesicular storage of ATP, is recently an emerging key factor for purinergic chemical transmission in various physiological and pathological phenomena. However, a contribution of VNUT and its physiological consequences to ischemic injury also remain unknown. Here, we show that the ATP exocytosis via VNUT contributes to neuroprotection against cerebral ischemic injury. Using <I>in vivo</I> middle cerebral artery occlusion (MCAO) in mice, we found that transient brain ischemia increased expression of VNUT preferentially in microglia in the ischemic region and VNUT deficiency exacerbated brain infarction following transient MCAO, which indicated neuroprotective potential of microglial VNUT. Extracellular ATP levels were increased during transient ischemic period and returned to the baseline immediately after reperfusion in both WT and VNUT knockout (VNUT-KO) mice. Interestingly, after returning to the baseline, ATP levels were increased again and persisted at high levels until 3 days after MCAO in WT mice, but not in VNUT-KO mice. In order to investigate the mechanism underlying microglial VNUT upregulation following brain ischemia, cytokine array was performed. We focused on IL-6, the cytokine that was significantly increased at 1 day after transient MCAO, and found that when added to cultured microglia, IL-6 was able to induce microglial VNUT mRNA upregulation in <I>in vitro</I> experiment. In addition, IL-6 receptor antibody (IL-6RA) administration exacerbated MCAO-evoked brain infarction in WT mice. However, the infarction in VNUT-KO mice was comparable between control and IL-6RA groups. Taken together, VNUT upregulated in response to ischemia via IL-6-mediated mechanism has a vital role for exocytosis of ATP from microglia and the events induce neuroprotection against ischemic 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-L-542 外側視床下部におけるオレキシン細胞とシナプス近辺星状膠細胞(アストロサイト)についての超構造的分析 Kazue Semba（仙波和惠）¹，Chantalle Briggs（Briggs Chantalle）¹，Sayuri Hatada（畑田小百合）²，Samuel Deurveilher（Deurveilher Samuel）¹，Yoshiyuki Kubota（窪田芳之）² ¹Dept Med Neurosci, Dalhousie Univ, Halifax, NS, Canada ²生理学研究所、大脳神経回路論研究部門 Sleep-wake cycles are regulated by the alternate activation of sleep- and wake-promoting neurons. We recently showed that astrocytes dynamically regulate excitatory transmission to wake-promoting orexin (ORX) neurons and sleep-promoting melanin concentrating hormone (MCH) neurons in the lateral hypothalamus via glutamate transport in a cell type-specific manner and according to sleep history (Briggs et al., 2018, J Neurosci). To obtain a better understanding of the structural basis of astrocytic regulation of synaptic inputs to ORX neurons, we have been conducting 3D reconstruction of ORX neurons and associated perisynaptic astrocytes, using correlative light-electron microscopy (EM). ORX neurons were identified using an orexin-A antibody with laser confocal microscopy. The same neurons were then identified in serial ultrathin sections obtained with automated tape-collecting ultramicrotomy (ATUM) and scanning EM. Depending on the orientation, dendrites could be followed up to 120 um from the soma of ORX neurons. ORX cell bodies and dendrites had very few spines. Synapses were frequently found both on cell bodies and along dendrites. At virtually all of these synapses, astrocytic processes were located near the cleft but with variable distances. Typically, the same astrocytic process approached multiple synapses with the same ORX neuron, as well as multiple synapses with non-ORX dendrites. Direct apposition between different parts of the same, and apparently-different, astrocytic process was very common, and direct apposition was also observed between two astrocytic cell bodies. These astrocytic appositions may represent gap junctions, which are known to occur frequently between astrocytes. Reconstruction of single astrocytes indicated that each astrocyte emits branching processes of complex morphologies that surround various neuronal elements (dendrites, axons), including ORX neurons, while also making contacts among themselves. These preliminary results suggest that astrocytes play an important role in regulating synaptic transmission to ORX as well as other neurons in the lateral hypothalamus to control sleep-wake and other behaviours.		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-L-543 背側前脳アストロサイトにおける慢性的カルシウムイオン上昇の機能解析 Haruna Aikawa（相川治奈），Yuichi Hiraoka（平岡優一），Mizuki Tanicha（谷茶みづき），Kohichi Tanaka（田中光一）東医歯大難治疾患研分子神経科学 Astrocytes are the most abundant glial cell in the central nervous system. It is known that astrocytes have various functions such as forming blood brain barrier, delivering neurotrophic factors, clearance of neurotransmitters, and so on. Until a few decades ago,, astrocytes were considered to be the supportive cells for neurons.. However, for the last couple of decade, researchers found that astrocytes display a form of excitability based on intracellular Ca2+ dynamics and have capabilities to evoke neural firing via releasing gliotransmitters. Recent studies suggest that astrocytes play crucial and independent roles by this Ca2+ dysregulation in the pathogenesis of psychiatric or neurological disorders, such as Alzheimer's disease(AD), epilepsy and Huntington disease. In AD mouse models, astrocytes exhibited a higher frequency of spontaneous Ca2+ oscillations when compared to wild-type mice. In addition, astrocytes spatially associated with amyloid plaques show spontaneous propagating Ca2+ waves that occur independently of neuronal activity, suggesting the involvement of aberrant astrocytic Ca2+ signals in AD pathogenesis. In contrast, a recent study has demonstrated that astrocytic Ca2+ elevations generates synaptic potentiation and enhance memory. In this study, we assess whether astrocyte Ca2+ elevation in dorsal forebrain could be a primary cause of cognitive impairment by using an intersectional chemogenetic tool. We found that chronic astrocyte Ca2+ elevation in dorsal forebrain induced an increase in the number of c-fos positive neurons, resulting in cognitive impairment in object recognition test. These results imply that chronic astrocytic activation may contribute to cognitive impairment.

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-L-544 マイクログリアは海馬スライス培養においてカイニン酸誘導性の神経細胞死を軽減する Tasuku Araki（荒木匡）¹，Yuji Ikegaya（池谷裕二）^1,2，Ryuta Koyama（小山隆太）¹ ¹東京大院薬薬品作用 ²脳情報通信融合研究センター Status epilepticus-induced hippocampal sclerosis is a common pathological change in mesial temporal lobe epilepsy (mTLE) with resistance to anti-epileptic drug. The main symptoms of hippocampal sclerosis include neuronal death, astrogliosis, and activation of microglia. Neuronal death in hippocampal sclerosis gradually progresses and is involved in the aggravation of epilepsy. Thus, clarifying the cellular mechanisms to protect neurons in hippocampal sclerosis will contribute to the treatment of mTLE. Here, mainly using hippocampal slice cultures with or without pharmacological depletion of microglia, we directly examined whether microglia, brain-resident immune cells that could act either neurotoxically or neuroprotectively, accelerate or attenuate neuronal death in kainic acid (KA)-induced hippocampal sclerosis in vitro. KA was treated to hippocampal slice cultures to induce hippocampal sclerosis. Clodronate-containing liposome or PLX3397 was used to deplete microglia in hippocampal slice cultures and its effect on kainic acid-induced neuronal death was immunohistochemically assessed. Treatment of clodronate-containing liposome or PLX3397 depleted microglia from hippocampal slice culture without inducing significant neuronal death. However, the depletion of microglia significantly promoted neuronal death in KA-treated hippocampal slice cultures. Thus, our results suggest that microglia possibly exert neuroprotective effects against KA-induced neuronal death in slice cultures.		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-L-545 頭部外傷後早期の脳ペリサイトPDGFRβ発現量増加は、ミクログリア活性化およびピロカルピンけいれん感受性亢進を誘導する Fuyuko Takata（高田芙友子）¹，Kenta Sakai（坂井研太）¹，Atsushi Yamauchi（山内淳史）²，Shinya Dohgu（道具伸也）¹，Yasufumi Kataoka（片岡泰文）¹ ¹福岡大学薬学部応用薬剤学教室 ²福岡大学薬学部生物薬剤学教室 The coordinated crosstalk among cells in the neurovascular unit (NVU) is critical for finetuning of the brain functions. Traumatic brain injury (TBI) has been well known to produce the secondary damages including late-onset epilepsy and chronic cognitive impairment, months or years after the initial trauma. TBI causes various different reactions in NVU-constituting cell types including pericytes, microglia, astrocytes and neurons. The TBI-dysregulated NVU function is likely to be associated with the secondary brain damages. Thus, to better understand the dynamics of each cell type is important to assume the therapeutic strategy for TBI. In this study, to investigate the spatiotemporal dynamics of NVU cells (pericytes, microglia and astrocytes) in the hippocampus of the injured side (except for the core region of injury), we used mice subjected to controlled cortical impact (CCI) for TBI. Histological analyses of the NVU cells were performed on 1 h and 1, 4, 7 and 28 days after CCI. In addition, we evaluated seizure susceptibility to the sub-threshold dose of pilocarpine on 7, 14, 21 and 28 days after CCI. The PDGFRβ immunoreactivities in pericytes were increased in the ipsilateral hippocampus during a period from 1 h to 28 days after CCI. The expressions of GFAP and Iba1 as a marker for astrocyte and microglia reactivity, respectively, were increased from 4 to 28 postoperative days. The severities of seizure induced by pilocarpine were gradually increased from 14 days after, with a significant increase at 28 days after CCI. These findings suggest that the increased PDGFRβ expression in pericytes precedes glial activation and neuronal alteration after CCI. To determine whether pericytes in response to CCI triggers the induction of glial activation and the increased seizure susceptibility, CCI mice were treated with an inhibitor of PDGFR signaling pathway, imatinib (200 mg/kg p.o., twice a day) for 5 days after injury and then were supplied for pilocarpine seizure test and histological analysis. Imatinib apparently lowered seizure susceptibility to pilocarpine and suppressed microglial activation in the ipsilateral hippocampus at 28 postoperative days. These findings suggest that the increased PDGFRβ expressions in pericytes may drive TBI-induced dysregulation of NVU function. The imatinib administration in the early stage after TBI could be operative for protecting against TBI-induced brain damage in the late stage.

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-L-546 ミクログリアの突起動態はマウス運動学習時にシナプス可塑性を誘導する Ako Ikegami（池上暁湖）¹，Koichiro Haruwaka（春若航一路）^1,2,3，Hiroaki Wake（和氣弘明）^1,2 ¹神戸大院医 ²生理研 ³総研大院 Microglia are sole immune cells in the brain. They possess characteristic morphology and kinetics; long and ramified processes, and are continuously surveying local environment with these processes. During this surveillance, they are known to contact with synapses, and increasing evidence is revealing that microglia are necessary for normal development of neural circuit. However, it is not known that whether and how this microglial process movement control synapses and neural circuit maturation. Here we investigated the change of microglial movement upon motor learning in vivo, which is known to be accompanied with synapse remodeling. We recruited 2 photon microscopy to follow the microglial change in the same mice, which undergoes the 2 weeks lever pulling motor learning, and quantified the microglial process trajectory. In the later phase of learning where the neural circuit matures, microglial process movement gradually converged, indicating they became to contact with more specific places (synapses). Further we have verified that there were higher chances of synaptic formation/elimination where microglia touched more frequently, and this synaptic turnover controlled by microglia was continuously occurring in brain of adult mice, even without any stimulus or learning tasks. Our results have given the direct evidence for microglial modification of synapses with their processes in the mature brain in vivo. Microglia changed their process movement to contact with more specific spines in the later phase of motor learning, and by following the microglia-synapse contact in the same mice, we have found that microglial contact induce synapse remodeling. These indicate that microglia are contributing for synapse remodeling, by controlling the duration and frequency to touch the specific synapses with their processes. Therefore, controlled morphology and kinetics of microglia is essential for maintaining our cognitive function such as learning and even simple daily perception. This could further explain why altered microglia phenotype cause several neurological / psychiatric conditions with synapse abnormalities.		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-L-547 捕食者臭誘発ストレス関連活動には嗅球の内側部と背外側部の両部位とも必要らしい Mutsumi Matsukawa（松川睦）¹，Narumi Katsuyama（勝山成美）²，Masato Imada（今田正人）¹，Shin Aizawa（相澤信）¹，Takaaki Sato（佐藤孝明）³ ¹日大医機能形態生体構造医 ²京都大霊長研高次脳 ³産総研バイオメディカル In the mouse olfactory bulb (OB), odor input from the olfactory epithelium innervate topographically to form odorant maps which are mirror-image arrangement of glomerular clusters with domain organization. It has been still unknown why and what for the mirror-image representation exists in OB. Predator odors, such as 2,5-dihydro-2,4,5-trimethylthiazoline (TMT), have known to induce a stress-like behavior in rodents. The dorsal domain (especially the cluster J in the DII domain) in the dorsolateral wall of OB (dlOB) have shown to be involved in this process and photoactivation on a single posterodorsal glomeruli, which represent a TMT-reactive olfactory receptor Olfr1019, induce immobility responses in mice. However, it remains unclear whether the activities in the medial wall of OB (mOB) are essentially needed to induce the stress-related activities in mice. First, we determined the region of TMT-receptive cells in mOB. Second, we lesioned the mOB and/or dlOB, and counted the number of immunohistochemically identified cells using antibody against c-Fos in the anterior piriform cortex (APC), bed nucleus of the stria terminalis (BST), amygdalo-piriform transition area (AmPir), and posterior piriform cortex (PPC) in mice brain which were removed at 30 min after TMT presentation after a week of surgeon. Third, we assessed the density of neurons which activated following electrical stimulation in mOB and/or dlOB. Although there were no significant differences in the number of activated neurons in BST, AmPir, and PPC wherever we lesioned, less activated neurons were observed in APC following lesion in both of mOB and dlOB. There were significant differences between the stimulated regions in OB. No changes were observed in the density of activated cells in all of brain regions we observed following the stimulation either mOB or dlOB alone. However, the number of c-Fos immunoreactive neurons in APC, BST, and AmPir were significantly increased following electrical stimulation in both of mOB and dlOB simultaneously. It suggests that the simultaneous activation in both of mOB and dlOB is needed to induce the neural activation in APC which causes stress-like behavior in rodents. These observations might be important not only for clarification of the mechanisms of the olfactory information processing but also for suppression of odor induced stress-like 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-L-548 マウス副嗅球相反性シナプス伝達におけるバゾプレッシンの役割：顆粒細胞－僧帽細胞間GABA作動性シナプスに対する作用 Mutsuo Taniguchi（谷口睦男），Yoshihiro Murata（村田芳博），Masahiro Yamaguchi（山口正洋），Hideto Kaba（椛秀人）高知大学医学部生理学講座 Central vasopressin (AVP) facilitates social recognition and modulates numerous complex social behaviors in mammals. Recently, new population of vasopressin neurons were reported to exist in the accessory olfactory bulb (AOB). The AOB has been demonstrated to be a critical site for mating-induced mate recognition (olfactory memory) in female mice. The effect of AVP, however, on the synaptic transmission between dendrites in the AOB of female mice is largely unknown. To address this issue, we previously measured synaptic currents (IPSCs) from mitral cells in the AOB, We have demonstrated that AVP significantly reduced the IPSCs in Mg<SUP>2+</SUP>-free solution. An agonist for V1a receptors, [Phe2, Orn8]-vasotocin mimicked the AVP action on the IPSCs. The suppressive effect of AVP on the IPSCs was diminished by an antagonist for V1a receptors, Manning compound, while an antagonist for V1b receptors, SSR149415 unaffected the effects of it. These results suggest that AVP attenuates reciprocal transmission between mitral and granule cells through AVP V1a receptors. The reciprocal transmission, however, contains both glutamatergic transmission from mitral to granule cells and GABAergic transmission from granule to mitral cells. Thus, it is unclear whether AVP acts on the excitatory and/or the inhibitory transmissions. In the present study, we have given attention to the effect of V1a receptor activation on GABAergic trasmission. AOB slices were prepared from 23- to 35-day-old Balb/c mice. With uUsing the patch-clamp technique in whole-cell configuration, the current responses of mitral cells were recorded in the presence of antagonists for glutamatergic transmission, CNQX (10 μM) and AP5 (50 μM). An extracellular application of vasopressin did not affect the magnitude of the response of mitral cells to GABA (10 μM and 100 μM), suggesting that AVP modulate the synaptic transmission from granule to mitral cells not through a postsynaptic mechanism.		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-L-549 コモンマーモセットにおける頭皮上聴覚誘発電位の無麻酔・無侵襲記録 Kosuke Itoh（伊藤浩介）¹，Haruhiko Iwaoki（岩沖晴彦）²，Naho Konoike（鴻池菜保）²，Hironaka Igarashi（五十嵐博中）¹，Katsuki Nakamura（中村克樹）² ¹新潟大学脳研究所 ²京都大学霊長類研究所 The common marmoset (<I>Callithrix jacchus</I>), a New World monkey, can serve as important animal models for biomedical sciences, including auditory neuroscience. This study recorded scalp cortical auditory evoked potentials (CAEPs) to pure tone stimuli in non-sedated common marmoset monkeys by using a noninvasive method that is essentially identical to those used for humans. Adult common marmosets (<I>n</I> = 4, three males, 5-7 years old) were seated in a monkey chair, and their head movements were restricted by a customized thermoplastic mask placed around their head. Individual electrodes were fixated on their scalp using adhesives and electrode gel. A pure tone stimulus (300 ms duration) was presented using two different stimulus onset asynchronies (SOAs, 330 ms or 1130 ms) while electroencephalograms were recorded using up to sixteen electrodes placed on the scalp and earlobes. When the SOA was 1130 ms, candidate homologues of the major components of human CAEP, i.e., P1, N1, P2, N2, and sustained potential (SP), were clearly identified in the grand-average. These waves were labeled as CjP1, CjN1, CjP2, and CjN2, and CjSP, respectively. (Cj stands for <I>Callithrix jacchus</I>.) Preceding CjP1, an earlier component CjNb was also observed, which likely corresponded to the human middle latency response Nb. In addition, the stimulus also induced a slow oscillatory activity of approximately 10 Hz. The peak latencies of the marmoset CAEP components were substantially shorter than in humans. When the SOA was short (330 ms), the wave morphology changed drastically, and an unambiguous identification of the above CAEP components became difficult; the 10 Hz oscillatory response also diminished. The study represents the first scalp-recorded CAEP in the alert common marmoset.

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-L-550 音刺激に対する瞳孔反応を用いた情動推定 Kengo Shimizu（Shimizu Kengo）¹，Satoshi Nakakoga（中古賀理）¹，Tetsuto Minami（南哲人）^1,2，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 Recently, estimation of emotional states using physiological response has become more important. Previous studies reported that pupillary dilatation response (PDR) was induced when viewing emotionally arousing pictures (Bradley et al., 2008). However, the estimation method by the pupillary response so far is directly linked to the timing of emotion elicitation such as the onset of emotional pictures. Other recent studies have reported that brain activity to deviant sound is enhanced to negative emotions (Tartar et al., 2012). If the similar phenomenon is found in pupillary responses, estimation of emotional states become possible using pupillary responses to auditory stimuli. In this study, we investigated pupillary responses to auditory oddball tasks in each emotion state elicited by emotional pictures (positive, negative, and neutral) for 32 participants. The picture was followed 600 ms later by a beep sound in an auditory oddball paradigm. Pupil diameter was recorded by EyeLink 1000PLUS while presentation of the visual and auditory stimuli. At the end of each trial, the participants categorized (picture and tone combination) with a button press. Our results showed that PDR to beep sound in negative and positive emotional states was more dilated compared to neutral, and PDR to beep sound in negative emotional states was induced earlier compared to other emotional states. In addition, the emotion-dependent PDR was not related to the difference between standard and oddball sounds. Taken together, our study suggested that it is possible to estimate the emotional states from the amplitude and the latency of pupil activity by using beep sounds. This finding has broad implications for emotion estimation using pupillary response.		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-L-551 蝸牛損傷後に起こる下丘内での興奮ー抑制神経活動バランスの異常 Lanlan Ma（馬蘭蘭），Munenori Ono（小野宗範），Nobuo Kato（加藤伸郎）金沢医大院医生理 The cochlear insults often accompany with tinnitus. Previous studies showed that the spontaneous neuronal activities were enhanced in the auditory centers by the cochlear insults. The hyperactivity after the cochlear insults is considered to be induced in order to compensate for the hearing loss, and underlie the tinnitus. However, it remains unclear what mechanism enhance the neuronal activities after the cochlear injury. In the auditory centers, it is well known that the balance of the excitatory and inhibitory processes is critical in shaping the neuronal activities. It is likely that the aberrant activities in the excitatory and inhibitory neuronal network underlie the hyperactivity in the auditory pathway. However, the changes in the excitatory and inhibitory neuronal activities after cochlear insults have not been investigated yet. In this study, we utilized the VGAT-ChR2 mice, in which the inhibitory neurons specifically express the channelrhodopsin2, and identified the excitatory and inhibitory neurons in the inferior colliculus (IC) during in vivo electrical recordings using light stimuli. We recorded the neuronal activities in the normal hearing animals and cochlea-damaged animals. The cochlear injury was induced by the noise exposure (1 octave noise centered at 16 kHz, 116 dB, 1hr). The level of the hearing loss was assessed by recording the auditory brainstem response (ABR), and noise exposure was found to induce a long-lasting ABR threshold shift in the exposed ear. In the normal hearing animals, the inhibitory neurons had higher spontaneous firing level than the excitatory neurons. Interestingly, in the noise exposed animals, the balance between the activities of these cell types changed: the excitatory neurons had significantly higher spontaneous firing rate than the inhibitory neurons. This result suggested that the change of the activity of the excitatory and inhibitory neural circuits in the IC might be relevant to the enhancement of the neuronal activity triggered by the cochlear insults.

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-L-552 弁別不可能な聴覚課題における正解の神経デコーディング Motoshige Sato（佐藤元重），Nobuyoshi Matsumoto（松本信圭），Yuji Ikegaya（池谷裕二）東京大学大学院薬学系研究科薬品作用学教室 There are difficult tasks that humans are not able to solve even they have sufficient information to solve the problems. For example, many Japanese people have difficulty in discriminating between sounds of L and R although the two types of sounds are represented in distinct cell assemblies in the auditory cortices. Therefore, since sensory information is conveyed from sensory organs to each sensory cortex when humans are exposed to sensory stimuli, sensory cortices contain the information required to solve a sensory-discrimination task. Particularly in primary sensory cortices, there is little information loss in the process of sensory information transmission from the sensory organs to the sensory cortices. Therefore, sensory information in a sensory-relevant task may be decoded from neural activity in the primary sensory cortex. We hypothesized that when the human language is presented to rats in an auditory discrimination task, the auditory information (e.g., Spanish or English) can be decoded from neural activity, although rats do not innately discriminate between the phonetic features of spoken English and Spanish languages. To test this hypothesis, we developed a new behavioral task, where we performed extracellular recording of neural activity (i.e., local field potentials (LFPs)) from the rat primary auditory cortex while a rat was given short phrases of Spanish or English sentences. Then, we tried to decode the language given to the rat from the neural activity. Before we presented voices of human languages to rats, as a pilot study, we recorded LFPs from the primary auditory cortex while rats were given pure tones (1500 - 7000 Hz, 500 msec) intervened between silent periods, and found the differences in LFPs between the onsets of the silent and tone periods. We examined whether deep neural network can decode the difference of the pitches and silent periods from LFPs. We found neural network model with a structure that performed one-dimensional convolution in the time axis direction and the electrode channel direction independently, improved decoding performance partially because the spatiotemporal information of LFPs was efficiently used. With this deep learning model, we are planning to feedback the decoded information (i.e. English or Spanish languages) to the brain via electrical stimulation and to enhance the discrimination accuracy, which would enhance animals' intelligence.		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-L-553 情動的手がかりが自閉スペクトラム症者の視覚時間分解能に与える効果 Masakazu Ide（井手正和）¹，Atsumi Takeshi（渥美剛史）^1,2，Reiko Fukatsu（深津玲子）¹，Mrinmoy Chakrabarty（Chakrabarty Mrinmoy）¹ ¹国立リハ研究所・脳機能部 ²杏林大医病態生理 While atypical sensory profiles across auditory, tactile, visual modalities are well known in autism spectrum disorders (ASD), the physiological states that influence sensory sensitivities in ASD are less studied. It is reported that a sizeable proportion of ASD have anxiety as co-morbidity, which in turn is associated with dysfunctions of autonomic nervous system. We investigated if sudden induction of autonomic arousal using disgust facial emotion cues, influenced visual temporal resolution in ASD (n = 9) and typically-developing individuals (TD; n = 13; controls). Participants reported the laterality of the last of two sequential, Gaussian blob flashes. The key manipulation was presenting an irrelevant picture prime (duration = 100 ms), randomly 300-500 ms before the Gaussian blobs. The trials in two separate blocks contained Disgust and Neutral facial emotions randomly intermixed with scrambled images as baseline. Temporal resolution was calculated for each participant by fitting cumulative Gaussian function to the response data. Comparisons were done after adjusting the temporal resolution in each type of facial emotions trials with their respective baseline and thereafter subtracting values of Neutral trials from those of Disgust. Disgust cues improved visual temporal resolution and increased the inter-session temporal resolution variability only in ASD. Individual values of disgust-induced variability in temporal resolution across all participants (n = 22), associated positively with subjective trait anxiety as well as sensory sensitivity / sensation avoiding scores (representing diagnostic sensory-hyperresponsivity). The preliminary results indicate that emotion-induced fluctuations of subjective, internal physiological states may influence temporal sensitivity of visual perception.

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-L-554 自由視中のサル視覚系ニューロン群の活動は繰り返し注視で疎に変化する Yukako Yamane（山根ゆか子）^1,2，Junji Ito（伊藤淳司）³，Cristian Joana（Joana Cristian）³，Pedro E. Maldonado（Maldonado E. Pedro）⁴，Hiroshi Tamura（田村弘）^1,5，Ichiro Fujita（藤田一郎）^1,5，Kenji Doya（銅谷賢治）²，Sonja Gruen（Gruen Sonja）^3,6 ¹大阪大学大学院生命機能研究科 ²沖縄科学技術大学院大学 ³Inst. of Neuroscience and Medicine (INM-6) and Inst. for Advanced Simulation (IAS-6), Juelich Research Centre and JARA Brain Institute I, Juelich, Germany ⁴Dept. of Neuroscience and BNI, Faculty of Medicine, Universidad de Chile, Santiago, Chile ⁵脳情報通信融合研究センター ⁶Theoretical Systems Neurobiology, RWTH Aachen Univ., Aachen, Germany We constantly move our eyes to visually sense their surrounding environment. The visual system has to process successively incoming images after each saccade, while the decision for the next fixation position is made in parallel to this process. In such a natural context, the eye movement pattern is expected to affect sensory processing. For example, the activity can be modulated across the history of fixated objects. To test this possibility, we recorded the spiking activity of multiple neurons in primary visual (V1) and inferotemporal (IT) cortices by multi-probe array electrodes from awake macaque monkeys freely viewing images containing complex visual objects. Each stimulus image was generated by randomly placing 5 objects, which were selected from 20 objects, on a gray background. Under this paradigm, the monkeys mostly fixated on the placed objects. To examine the effect of the fixation history, we classified each fixation based on its order as follows. 1) 1st fixations: 1st fixation on an object after stimulus onset. 2) re-visit fixations: fixation from other objects or from background to a previously fixated object. 3) 2nd+ fixations: successive fixations on the same object after a 1st fixation or a re-visit fixation. We found that approximately 30% of neurons in both V1 and IT showed a lower mean firing rate in later fixations (2nd+ and re-visit fixations) than in 1st fixations. To see how population activity differs between fixation orders, we examined the population activity pattern, defined as a vector composed of firing rates of individual neurons during 0.05-0.15ms (V1) or 0.08-0.18ms (IT) after fixation onset, on different objects and in different fixation orders. Cosine similarity as well as Pearson's correlation was calculated between population activity patterns in the fixations of different pairs of objects. The result showed higher similarity between population activity patterns for different objects in 1st fixations compared to later (2nd+ and re-visit) fixations. This result suggests that populational neural representation of objects are not very well separated in 1st fixations, and become better separated in later (2nd+ and re-visit) fixations, and that the population firing pattern dynamically changes according to the history of fixation so as to better discriminate an individual object by a smaller number of spikes.		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-L-555 一次視覚野の方位判別能はトレンド除去により恒常性を保つ Shu Kyono（京野秀）¹，Yoshiko Maruyama（圓山由子）²，Hiroyuki Ito（伊藤浩之）³，Keiji Miura（三浦佳二）¹ ¹関西学院大学理工学部生命科学科 ²函館工業高等専門学校　生産システム工学科 ³京都産業大学　情報理工学部 The spontaneous neural activities in the primary visual cortex have been reported to show temporal drifts (Okun et al., 2015). However, the mechanism by which the brain can recognize visual stimuli robustly despite the non-stationary baseline drifts is still unclear. In this study, to understand the effects of the trends in baseline activities, we compared the orientation discriminability from 566 neurons in the cat primary visual cortex before and after the trend removal. To see if the trends do harm on population coding, we performed linear discriminant analysis to discriminate orientations based on the neural activities. First, of all the 40 trials for each stimulus, we learned from the former 20 trials and discriminated 16 orientations for the same former 20 trials by using the leave-one-out cross validation. Note that we presented 16 visual stimuli with different orientations pseudo-randomly in each block and we repeated 40 blocks. The classification success rate was mild. Next, when we learned from the former 20 trials and discriminated 16 orientations for the latter 20 trials, the hit rate decreased significantly. Thus, the effect of the baseline drifts was large enough to deteriorate the discriminability. Finally, to clarify how the brain overcomes the non-stationarity, we hypothesized that the brain could remove the trends in the time series of neural activities. By removing the trends in advance, we equated the mean activities for the former 20 trials and latter 20 trials. Then, we found that the hit rate recovered. That is, the cross-validated hit rate for the latter trials learned with the latter trials was not significantly different from the one learned with the former trials, after we removed the trends. This suggests that the brain could achieve robust visual recognition by trend removal.

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-L-556 前頭前皮質からのトップダウン投射は高次視覚野の視覚応答を増強する Chiaki Kasahara（笠原千晶），Tomonari Murakami（村上知成），Kenichi Ohki（大木研一）東京大学医学部統合生理学 The hierarchical cortical network consists of bottom-up and top-down pathways. The bottom-up pathway progressively processes sensory inputs from low-order to high-order areas, and the top-down pathway modifies the bottom-up sensory processing. In the mouse cortex, neurons in the frontal cortex send projections to the visual cortex including the primary (V1) and higher visual areas (HVAs), and this top-down projection affects the visual response of neurons in V1 and improves visual discrimination. However, it still remains unclear whether the top-down projection from the frontal cortex modulates the visual responses of mouse HVAs. In this study, we investigated this question by combining wide-field calcium imaging with optogenetics, and found that the visual responses of HVAs were enhanced by optogenetic activation of the frontal cortex. We injected three types of adeno-associated viruses (AAV9-CaMKII-tTA-WPRE, AAV9-TRE-C1V1-p2a-mcherry-WPRE, and AAV9-TRE-GCamp6f-WPRE) into the mouse cortex at postnatal day 0, so that GCaMP and C1V1 were expressed in excitatory neurons in a large cortical region including the visual and frontal cortices. About 4 weeks after virus injection, we performed wide-field calcium imaging (wavelengths of GCaMP excitation/emission: 480/520 nm) and acquired the visual responses of V1 and HVAs. During the presentation of the drifting gratings with various spatiotemporal frequencies, neurons in the frontal cortex were optogenetically activated by yellow light stimulation (wavelength: 594 nm, pulse duration: 15-25 msec, frequency: 10 Hz, pulse repetition: 10 times). The comparison of the visual response amplitudes between the presence or absence of the light stimulation revealed that the visual responses of not only V1 but also of HVAs were enhanced by optogenetic activation of the frontal cortex. In addition, this enhancement was stronger in the response to rapidly drifting gratings compared with slowly drifting ones. Together, these results show that the top-down projection from the frontal cortex enhances the visual response both of V1 and HVAs, and suggest that this enhancement seems more related to the visual motion processing.		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-L-557 マウス一次視覚野における開眼後の動的機能変化について Fumiaki Kishino（岸野文昭）¹，Masato Uemura（上村允人）¹，Sigrid Traegenap（Traegenap Sigrid）^2,3，Matthias Kaschube（Kaschube Matthias）²，Kenichi Ohki（大木研一）^1,4 ¹東京大院医統合生理 ²Frankfurt Institute for Advanced Studies, Frankfurt, Germany ³The International Max Planck Research School for Neural Circuits, Frankfurt, Germany ⁴International Research Center for Neurointelligence, Univ. of Tokyo, Tokyo, Japan Neuronal function matures during the early stage of life. In mouse primary visual cortex (V1), the distribution of preferred orientation changes after eye-opening. At eye-opening, neurons responsive to cardinal (i.e. vertical and horizontal) orientations outnumber those responsive to oblique ones, and this cardinal bias becomes small in adults. Currently, it is unclear whether this decrease in cardinal bias is due to a shift in the distribution of responsive cells, or due to a change of tuning in individual cells. In order to address this issue, we first conducted acute two-photon calcium imaging at multiple time points and found that the cardinal bias decreases mainly during the first week of eye-opening (P14-21). Next, to examine how this functional change occurs in this period, we pursued the orientation tuning of identical neurons in V1 during the first week of eye-opening every other day using chronic two-photon calcium imaging. We found three types of changes contributing to the decrease in cardinal bias: 1) neurons shifting their preferred orientation from cardinal toward oblique, 2) neurons not responsive to grating stimuli at eye-opening acquiring response to oblique orientations, and 3) neurons responsive to cardinal orientations at eye-opening becoming unresponsive. Also, some neurons showed changes that do not decrease the cardinal bias, and some remained stably selective to the same orientation across all days. We will further investigate functional properties of these neurons, such as response amplitude, selectivity, trial-to-trial variability, and signal and noise correlation, and what factors affect the functional changes of these neurons in the first week of eye-opening.

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-L-558 サル側頭葉ニューロンにおける顔の質感情報表現 Kazuko Hayashi（林和子）¹，Narihisa Matsumoto（松本有央）¹，Keiji Matsuda（松田圭司）¹，Kenichiro Miura（三浦健一郎）²，Shigeru Yamane（山根茂）¹，Shin Matsuo（松尾真）³，Keiji Yanai（柳井啓司）³，Mark A. G. Eldridge（Eldridge A. G. Mark）⁴，Saunders C. Richard（Richard C. Saunders）⁴，Barry J. Richmond（Richmond J. Barry）⁴，Yuji Nagai（永井裕司）⁵，Naohisa Miyakawa（宮川尚久）⁵，Takafumi Minamimoto（南本敬史）⁵，Masato Okada（岡田真人）⁶，Kenji Kawano（河野憲二）¹，Yasuko Sugase-Miyamoto（菅生-宮本康子）¹ ¹産業技術総合研究所 ²国立精神・神経医療研究センター ³電気通信大学 ⁴アメリカ国立衛生研究所 ⁵放射線医学総合研究所 ⁶東京大学 In our social interaction, it is important to detect a face and recognize its individual identity and emotional expressions. This ability is assumed to rely on the workings of the primate temporal lobe where visual characteristics of face parts are processed. We have previously shown temporal processing of facial information in temporal cortex neurons (Sugase et al., 1999; Sugase-Miyamoto et al., 2014). The physical properties of faces, such as texture of skin, twinkle in eyes, and gloss of hair, also provide abundant information associated with age, health conditions and emotional states. It has been reported that the material properties of objects, including gloss and texture, are processed in the posterior inferior temporal cortex of humans and monkeys (Okazawa et al., 2012; Goda et al., 2014). These findings suggest that the temporal cortical areas involved in the representation of material properties partially overlap with the face-responsive regions. To examine how temporal cortex neurons respond to physical properties of faces, we recorded neuronal activities from two monkeys during the presentation of facial images with different skin textures in a fixation task. The original stimulus set consisted of colored nine monkey faces, nine human faces and two geometric shapes. Based on the original set, three-stimulus sets were synthesized by image-based material editing methods: high-gloss, low-gloss and style-transferred images. We simultaneously recorded 227 neurons using three 96-channel multi-electrode arrays in the anterior, middle and posterior areas of the inferior temporal gyrus (ITG) of one Japanese monkey. We also recorded 55 neurons using single electrodes in the bank of the superior temporal sulcus (STS) of one rhesus monkey. By analyzing the responses of face-responsive neurons to each facial image in a window from 50 to 450 ms after the stimulus onset, we demonstrated the effects of the material editing on their activities; 47/136 neurons in the ITG and 33/51 neurons in the STS showed significant changes in the strength of the responses to the high-gloss, low-gloss, or style-transferred images as compared to the responses to the original images (paired <I>t</I>-test, p<0.05). These results reveal that the face-responsive neurons in the STS and the ITG code the differences in facial skin textures, suggesting the role of the temporal lobe in information processing about the physical properties of faces.		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-L-559 両眼視野における運動残効の情報処理 Miki Yamashita（山下美季）¹，Naoki Nakajima（中島直樹）¹，Naoki Kogo（向後直樹）²，Takeshi Aihara（相原威）¹，Hiroaki Hayakawa（早川博章）¹ ¹玉川大院工 ²Radboud Univ, The Netherlands In the daily life of animals including humans, the visual system detects two of the wide-field motion associated with one's own exercise and local motion associated with the movement of an object in the outside world is present. The visual system works by integrating information in some stages. Bottom-up visual information from both eyes is thought to be treated subconsciously in the brain. Recently, it was reported that the motion aftereffect was appeared in the reverse direction of each vector addition of the translational motion that constitutes the adaptation stimulus, as a study of bottom-up information processing in the wide-field motion. However, the modification effect of top-down information, attention, for the motion aftereffect is not clear. In this study, to investigate influence of the top-down information relating attention on the bottom-up information processing of two inputs from both eyes in the wide-field motion, random dot-patterns moving upward and rightward were simultaneously and independently applied to the left eye and the right eye using a virtual reality headset, respectively. In addition, to measure the rate of attention paid to right and left eyes, we tagged the right-side and left-side random-dots with dynamic noises, which were oscillated as side-tags at 7.5Hz for the right eye and at that at 5.45Hz for the left eye image. The brain signals were monitored at these frequencies with EEG. The characteristics of the motion aftereffect were measured and the EEG was also analyzed using Fast Fourier Transform(FFT) and Wavelet Transform (WT) during the presentation of random dots superimposed with the dynamic noise. As a result, the responses to the frequencies used for tagging, their harmonics, and the interaction components were confirmed by the FFT and WT analyses. When attention was paid, for example, to the left eye image, the response of the frequency used for tagging the left eye increased and the motion aftereffect sifted reflecting the amount of attention. It showed that the motion aftereffect was influenced by increasing vector of the opposite direction for the visual field motion to which attention was paid. Our results suggested that binocular information-processing was modified by attention (consciousness) through top-down mechanisms.

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-L-560 パルブアルブミン細胞はvertical cellを介した低閾値機械受容性終末から脊髄後角第Ⅰ層投射細胞への入力をシナプス前性・後性に抑制しゲートしている Toshiharu Yasaka（八坂敏一）¹，Kieran A Boyle（Boyle A Kieran）²，Mark A Gradwell（Gradwell A Mark）³，Allen C Dickie（Dickie C Allen）²，Erika Polgar（Polgar Erika）²，Robert P Ganley（Ganley P Robert）²，Desmond Ph Orr（Orr PH Desmond）²，Masahiko Watanabe（渡辺雅彦）⁴，Victoria E Abraira（Abraira E Victoria）⁵，Emily D Kuehn（Kuehn D Emily）⁵，Amanda L Zimmermann（Zimmermann L Amanda）⁵，David D Ginty（Ginty D David）⁵，Robert J Callister（Callister J Robert）³，Brett A Graham（Graham A Brett）³，David I Hughes（Hughes I David）² ¹鹿児島大院医歯免疫 ²Spinal Cord Group, Inst. of Neurosci. and Psychol., Univ. of Glasgow, Glasgow, UK ³Sch. of Biomed. Sci. and Pharmacy, Univ. of Newcastle, Newcastle, Australia ⁴北海道大院医解剖 ⁵Dept. of Neurobiol., Howard Hughes Med Inst., Harvard Med Sch., Boston, USA Dorsal horn neurons are targets for primary afferents that convey sensory information from the skin and other tissues, and that respond to specific types of stimuli (e.g. noxious and non-noxious). Among these afferents, nociceptors terminate superficial area (I-II), while low-threshold mechanoreceptors (LTMRs) terminate relatively deeper laminae (IIi-V). Majority of dorsal horn neurons consist a variety of excitatory and inhibitory interneurons and build complex local circuits that process incoming information within the dorsal horn. Abnormal activity of these circuits is thought to lead pathological pain such as tactile allodynia (touch-evoked pain), a symptom of many chronic pain diseases. We have shown that parvalbumin-expressing inhibitory interneurons (PV interneurons) provide a source of presynaptic (axo-axonic) inputs onto the central terminals of LTMRs. We have also described a putative relay circuit providing LTMR-input to lamina I projection neurons via vertical cells, a type of excitatory interneuron in lamina II. These observations implicate that circuits consisting of these cells are very likely involved in development of tactile allodynia. Here, we show that PV interneurons modulate inputs from LTMRs directly via presynaptic inhibition, and also gate the relay circuit from LTMR input to pain pathway by inhibiting vertical cells. We also show that excitability of PV interneurons is diminished following peripheral nerve injury, while silencing these cells increases network activity in laminae I to IV so that touch stimuli can activate pain pathway. PV interneurons may therefore play an important role in the development of tactile allodynia, and could offer a realistic target for new strategy to manage chronic pain.		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-L-561 自閉スペクトラム症者における刺激の時間的加重と感覚応答性との関係性に関する検討 Ayako Yaguchi（矢口彩子）^1,2,3，Masakazu Ide（井手正和）¹ ¹国立リハ研究所・脳機能部 ²立教大・現代心理学研究科 ³日本学術振興会 Studies reported enhanced perceptual sensitivity (i.e., lower detection threshold) to vibro-tactile stimuli in individuals with autism spectrum disorder (ASD), and this has been speculated to be underlying diagnostic sensory hyper-responsivity. We demonstrated that individuals with ASD with superior sensory processing in discriminating two successive vibro-tactile stimuli tended to have severer sensory hyper-responsivity. This suggests that higher temporal resolution of sensory signals would result in greater amounts of temporal summation of that. If so, individuals with ASD having high temporal resolution would perceive stronger subjective impression of the intensity. In the present study, we addressed that temporal summation by short stimulus presentation is obvious in individuals with ASD. 10 ASD and 13 typically developing (TD) adolescents were participated. We first estimated individuals' detection threshold of vibro-tactile stimuli to the left index fingers. Then, we conducted a detection task with 50-1000 ms duration conditions of vibro-tactile stimuli, using the amplitudes of the estimated threshold in each participant. By fitting the data of response rates in each duration condition to an exponent function, we quantified a `Threshold of stimulus duration' enough to perceive stimulus in supra-threshold level. We also conducted a gap detection task to evaluate temporal resolution and sensitivity of temporal gap of two successive vibro-tactile stimuli. We evaluated the severity of symptomatic hyper/hypo-sensitivity using the adolescent/adult sensory profile (AASP). We found that Threshold of stimulus durations were positively correlated with temporal resolution across all participants (N = 23, r = 0.44, p = 0.04). The sensitivities of temporal gap were positively correlated with scores of sensory seeking in AASP only in ASD group (N = 10, r = 0.71, p = 0.02). These findings indicated that the individuals with high temporal resolution of the stimuli tended to keenly detect it regardless of shorter duration. This effect of temporal summation on perceptual impression of stimulus intensity was shared across ASD and TD individuals. The result that temporal resolutions are associated with sensory seeking only in ASD group was corresponded to our previous finding (Ide et al., 2018). This might suggest temporal resolution which associated strength of perceptual impression of stimuli contributes sensory hyper-responsivity in individuals 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-L-562 恐怖条件づけによる光と体性感覚情報の結合における感覚皮質の応答伝播 Gennosuke Tasaka（田坂源之助）¹，Naoki Nakajima（中島直樹）¹，Muneyoshi Takahashi（高橋宗良）²，Yoshinori Ide（井出吉紀）²，Takeshi Aihara（相原威）¹ ¹玉川大院工 ²玉川大脳研 Fear conditioning by sensory stimulation is often used in the study of emotional memory, and many studies of the plasticity of sensory areas using this paradigm are performed. However, many conventional studies have attempted only a single stimulus and electrical stimulation conditioning. In our previous study, plastic changes induced by a fear conditioning, pairing a pure tone with an electric foot-shock, were measured in the auditory cortex and somatosensory cortex of a guinea pig. It showed that somatosensory and auditory cortices were associatively activated by a single modality of sensory information. However, the association mechanism of sensory cortical signals elicited with the conditioning is still unclear. In this work, in order to investigate the associative binding-way among visual cortical stimuli and an aversive somatosensory-stimulus as a foot-shock, visual and somatosensory cortical activities were simultaneously measured using the optical imaging method with voltage sensitive dye (VSD) after the fear conditioning. As a result, conditioned responses to the light could be observed by monitoring cardiac pulse. Also, we found that neural activities for the foot-shock were propagated to V1 and higher visual cortex after activation of S1 and S2 in conditioned animals, although only S1 and S2 were activated to the foot-shock alone in a naïve animal. Additionally, neural activities to the light were propagated to the S1 and S2 in conditioned animals after or simultaneously activation of V1, although only V1 was activated to the light alone in a naïve animal. These results suggest that two different modalities could be retrieved across each sensory area and higher cortices by one modality of sensory information after the conditioning.		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-L-563 マウス後部頭頂連合野の機能解析 Kohei Yoshitake（吉武講平）¹，Ryuichi Hishida（菱田竜一）¹，Takeshi Yagi（八木健）²，Katsuei Shibuki（澁木克栄）¹ ¹新潟大・脳研・システム脳生理 ²大阪大学大学院・生命機能・心生物学 Sensory integration is important role in higher brain area. Predictive coding is a theoretical concept which helps us to understand higher brain functions. According to this idea, one of essential brain functions is to predict the coming sensory input, and to minimize the prediction error (difference between the prediction and actual sensory inputs). Previously, we have reported that spacial mismatch between whisker and visual inputs were detected in the posterior parietal cortex (PPC) of mice. We directly recorded neuronal activities in PPC using flavoprotein fluorescence imaging. Visual stimulation with grating patters moving forward/backward alone, or forward/backward whisker-shift stimulation alone hardly activated PPC in anesthetized mice. However, anti-phase combination of moving grating patterns and whisker stimulation (grating moving forward plus backward whisker shift, or grating moving backward plus forward whisker shift), which is very unlikely in natural environment for mice, produced prediction error responses in PPC. In contrast, in-phase combination of grating patterns and whisker stimulation failed to produce any clear activity in PPC. Obviously, prediction and prediction errors are expected to be dependent on past experience and learning. To test this possibility, we investigated the prediction error responses in PPC of dark-reared mice from birth to 4 weeks of age. As expected, the dark-reared mice could not show any prediction error responses in PPC. However, this dark reared effect was cancelled after the mice returned to a normal light-dark cycle for 2 additional weeks. These results suggest that prediction error responses in PPC is experience dependent. Recording in awake animal is required to study higher brain functions. We also investigated neuronal activities in awake mice. Flavoprotein fluorescence responses in awake mice were slightly smaller than those in anesthetized mice. Prediction error responses in PPC were similarly observed in awake mice. Clustered protocadherins (cPcdhs) comprising cPcdh-α, -β, and -γ, encode a large family of cadherin-like cell-adhesion molecules specific to brain. Prediction error responses in PPC was impaired in mice with reduced molecular diversity of cPcdh-α. These results suggest that the diversity of cPcdh-α is important for the experience-dependent PPC functions to produce 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-L-564 マウスの時間順序判断におけるGABA-A受容体拮抗薬の効果 Takeshi Atsumi（渥美剛史）¹，Mrinmoy Chakrabarty（Chakrabarty Mrinmoy）²，Shigehiro Miyachi（宮地重弘）³，Yasuo Terao（寺尾安生）¹，Masakazu Ide（井手正和）² ¹杏林大医病態生理 ²国立障害者リハビリテーションセンター脳機能系障害研究部 ³京都大霊長研高次脳 Individuals with autism spectrum disorders (ASD) often show sensory abnormalities (Marco et al., 2011), and an altered γ-aminobutyric acid (GABA)-mediated signaling in brain may explain these symptoms (Cellot & Cherubini, 2014). We previously reported that increased sensory hyper-responsiveness in ASD associates with greater temporal resolution in discriminating two successive sensory stimuli that were temporally spaced by varying intervals (temporal order judgment: TOJ task, Ide et al., 2018). Role of GABAergic neural signaling during TOJ is however, unknown. We hypothesized that lower GABAergic signaling heightens temporal resolution of tactile stimuli. The present animal study examined whether blocking GABA-A receptor improves temporal resolution during TOJ task. We trained three mice to perform the TOJ task. In one trial, the animal was delivered air-puff stimulations successively to its left and right whiskers. If the subject correctly nose-poked the side stimulated latter, a reward (4 μl 10 ~ 11 % sucrose solution) was delivered, otherwise a 5 ~ 14 sec time-out trial was delivered as punishment. We trained the animals with ±300ms of stimulus onset asynchronies (SOA). During testing, we added eight new SOAs (±5 ~ 240 ms) to the conventional sets (baseline) and delivered them randomly. Temporal resolution was calculated by fitting cumulative Gaussian function to the response data in each SOA. Three mice showed 82, 44, and 205ms as the temporal resolution, respectively. We administered a GABA-A antagonist, (+)-Bicuculline (1mg/kg: BIC) and its vehicle (saline with 10 % DMSO) intraperitoneally on different days, prior to the sessions. Either drug/vehicle was injected every second day, and the response data from three days were pooled. All the three mice showed better temporal resolution with BIC (64, 6, and 92ms) than vehicle (234, 11, and 105ms). The preliminary result suggests that administration of GABA-A blocker may improve TOJ performance in mice. There is a possibility that GABAergic neural dysfunction in ASD brain may induce greater neural response to temporal processing of external stimuli, resulting in a heightened perceptual intensity of them. Future studies should aim to reveal the mouse brain areas analogous to humans that relate to temporal resolution of stimuli, and examine whether the GABAergic signaling in that area associates with sensory hyper-responsiveness.		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-L-565 マウス前帯状皮質における歩行運動に関連したニューロン活動の解析 Riga Sachu（サチリガ），Hiroshi Nishimaru（西丸広史），Yusaku Takamura（高村雄策），Jumpei Matsumoto（松本惇平），Taketoshi Ono（小野武年），Hisao Nishijo（西条寿夫）富山大医システム情動（生理１） The prefrontal cortex (PFC) plays a critical role in controlling behavior. Locomotion is one of the fundamental aspects of animal behavior, an important element for exploring the environment for food or escaping from predators. Therefore, controlling the timing and speed of locomotion is crucial for the survival of the animal. However, the role of PFC in controlling locomotion during an emotional behavior still remains unclear. Among the PFC areas, the anterior cingulate cortex (ACC) has been shown to be involved in the preparation and execution of movements. In this study, we investigated how the information for initiation, termination and maintenance of locomotion during goal directed behavior is coded in the ACC by examining the neuronal signals in mice performing a stop-and-go task on a spherical treadmill. Five male mice were implanted with custom-build micro-drive tetrode array in the ACC. After recovery, they were trained to perform a self-initiated locomotor task with their head fixed which they were required to run on the spherical treadmill to obtain reward. In a single trial of this task, the mouse was allowed to start running voluntarily but when it stopped, a visual stimulus was presented, and then it had to immediately start running again within a certain amount of time to obtain sucrose/water solution as a reward. After 1-2 weeks of training, when the success rate of the task reached 60-80%, we performed single unit recording of ACC neurons. Out of 205 sorted single units, the majority of them (135 units, 66 %) showed activity related to the task and the majority of those task-related units (73 units, 36%) were modulated when the mice were running. Half of these locomotion-modulated neurons increased their firing rate when the mice started running while the activity of the others were suppressed while running. These results indicate that the rodent ACC neurons encode information related to initiation and termination of locomotion during goal directed behaviors.

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-L-566 発達期依存的な脊髄の可塑性によって決定される自律的な歩行回路の結合特性 Aya Takeoka（竹岡彩）^1,2,3，Karen Bens（ベンズカレン）^1,3，Hannah Bertels（バーテルズハナ）^1,3 ¹ニューロエレクトロニクス　リサーチ　フランダース ²フランダースバイオテクノロジー研究所 ³ルーベン大学 Spinal cord injury disrupts communication between the brain and spinal cord. Severe injury to a mature nervous system often leads to irreversible paralysis of movements controlled by spinal circuits below the lesion. In contrast, mice that receive a complete transection (cSTX) of spinal cord at low thoracic segments (~T10) at the early post-natal period (~P5) demonstrate proficient weight-supported bipedal treadmill locomotion. Here, we used combinatorial approaches of mouse genetics, viral circuit tracings and kinematic analyses to determine the underlying circuit signatures of P5 cSTX spinal cords that can walk without the brain. We found that expression patterns of intermediate and ventral local excitatory interneuronal subpopulations and their recruitment during locomotion are dependent on age of injury. Much in contrast, expression and recruitment patterns did not differ among intact, adult cSTX or P5 cSTX spinal cords. These results indicate that while a complete isolation of spinal cords from the brain circuits as an adult induces a significant alteration in excitation/inhibition balance, a spinal cord transection as a neonate maintains neuronal excitability and recruitment seemingly similar to that of intact spinal cords. Furthermore, ablation of proprioceptive afferents at the same time of P5 cSTX led to deterioration of spinal locomotion without the brain. Together, our study reveals that proprioceptive afferents serve as a driving force for spinal circuits to learn to walk without 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-L-567 糖尿病モデルラットの横隔神経に生じる軸索変性 Masako Ikutomo（生友聖子）¹，Toru Tamaki（玉木徹）²，Satoshi Shimo（志茂聡）³，Naomi Ooshiro（大城直美）⁴，Masatoshi Niwa（丹羽正利）⁴，Minori Fukuda（福田実乃里）⁴，Ken Muramatsu（村松憲）⁵ ¹東京医療学院大学リハビリテーション学科 ²健康科学大学健康科学部理学療法学科 ³健康科学大学健康科学部作業療法学科 ⁴杏林大学保健学部作業療法学科 ⁵杏林大学保健学部理学療法学科 We previously reported a decrease in the number of retrogradely labeled phrenic motoneurons and appearance of a high-amplitude single motor-unit potential of the diaphragm in diabetic rats. These might have been caused by diabetic motor neuropathy, which is characterized by axonal degeneration. Therefore, in this study, we examined the electrophysiological and morphological alterations of the phrenic nerve in a type 1 diabetic model. In 13-week-old male Wistar rats, we induced diabetes by administering streptozotocin (diabetic group), whereas in age-matched control rats, we injected saline (control group). After 20 weeks of streptozotocin administration, we mounted the phrenic nerves on a bipolar silver hook electrode and inserted bipolar wire electrodes into the diaphragm to record evoked electromyograms. Next, we calculated the motor nerve conduction velocity (MCV) of the phrenic nerve. Subsequently, the proximal portion of the phrenic nerve, which is the part that arises from the brachial plexus, was collected and embedded in EPON after fixation. Transverse thin sections were then stained with toluidine blue, and digital images of each sample were obtained using light microscopy and a digital camera. The cross-sectional areas (CSAs) of the axons were measured on the digital images of the samples using Image J software. Statistical comparisons of the diabetic and control groups were performed using the paired t-test. The data distribution was assessed using the Kolmogorov-Smirnov test. The MCV was significantly lower in the diabetic group than in the control group (p < 0.05). The number of axons was significantly decreased in the diabetic rats (p < 0.01). The ratio of smaller axons was higher and that of larger axons was lower in the diabetic group than in the control group (p < 0.01). The distribution of the CSAs of the axons was significantly different between the diabetic and control groups (p < 0.01). The MCV reflects the severity of axonal degeneration induced by diabetic neuropathy. Additionally, we noted a reduction in the MCV and the number and size of axons in the diabetic group. Our results suggest that phrenic motoneuron disorders in diabetes might be caused by axonal degeneration.

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-L-568 微小デバイスを用いた3D運動ユニットの構築とALS疾患モデル Tatsuya Osaki（大崎達哉）^1,2，Roger D Kamm（Kamm D Roger）^1,3 ¹Dept. Mech. Eng. Massachusetts Institute of Technology, Cambridge, USA ²東京大学生産技術研究所 ³Dept. Biological. Eng. Massachusetts Institute of Technology, Cambridge, USA Recent advances in iPS cell technology, combined with microfluidics, have enabled the development of a wide variety of what are termed "micro-physiological systems", or MPS. These MPS are useful for studying fundamental physiology of organ or tissue systems, but can also be used to create disease models with the potential for use in drug screening. With these goals in mind, we created a microfluidic platform in which a motor neuron-containing neurosphere is placed in close proximity to a skeletal muscle strip, facilitating through the extension of neurites connecting the motor neurons to the myocytes, the formation of a functional motor unit. All cells were derived from pluripotent source cells, the motor neurons from human embryonic stem cell (hESC)-derived neural stem cells and the skeletal muscle from human iPSC-derived skeletal myoblasts. The muscle self-organized around two flexible posts contained in the system so that generated muscle force could be measured. All cells, in the final configuration, were encapsulated in a three-dimensional hydrogel, replicating certain important aspects of real physiology. The motor neurons were engineered to express the channelrhodopsin-2[H134R] gene, and could therefore be activated by light, creating controlled and quantifiable contractile forces in the muscle strip. Substituting for the healthy motor neurons, iPSC-derived neural stem cells from a patient with sporadic Amyotrophic Lateral Sclerosis (ALS), a disease model could also be generated. The two systems, one from a healthy subject and the other from the ALS patient, were used to demonstrate the potential to use this as a functional screen for two drugs currently in clinical trial for 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-L-569 小脳歯状核から視床外側中心核への神経投射の運動機能における役割の解析 Masateru Sugawara（菅原正晃）¹，Nobuyuki Sakayori（酒寄信幸）^1,2，Shigeki Kato（加藤成樹）¹，Susumu Setogawa（瀬戸川将）¹，Hotaka Fukushima（福島穂高）³，Rie Ishikawa（石川理絵）³，Satoshi Kida（喜田聡）³，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東京農大生命科学動物分子生物 The cerebellum regulates complex animal behaviors, such as motor control and spatial recognition, through communication with many other brain regions. The major targets of the cerebellar projections are the thalamic regions including the ventroanterior nucleus (VA) and ventrolateral nucleus (VL). Another thalamic target is the central lateral nucleus (CL), which receives the innervations mainly from the dentate nucleus (DN) in the cerebellum. Although previous electrophysiological studies suggest the role of the CL as the relay of cerebellar functions, the kinds of behavioral functions mediated by cerebellothalamic tracts projecting to the CL remain unknown. Here, we used immunotoxin (IT) targeting technology combined with a neuron-specific retrograde labeling technique, and selectively eliminated the cerebellothalamic tracts of mice. We confirmed that the number of neurons in the DN was selectively decreased by the IT treatment. These IT-treated mice showed normal overground locomotion with no ataxic behavior. However, elimination of these neurons impaired motor coordination in the rotarod test and forelimb movement in the reaching test. These mice showed intact acquisition and flexible change of spatial information processing in the place discrimination, Morris water maze, and T-maze tests. Although the tract labeling indicated the existence of axonal collaterals of the DN-CL pathway to the rostral part of the VA/VL complex, excitatory lesion of the rostral VA/VL did not show any significant alterations in motor coordination or forelimb reaching, suggesting no requirement of axonal branches connecting to the VL/VA complex for motor skill function. Taken together, our data highlight that the cerebellothalamic tracts projecting to the CL play a key role in the control of motor skills, including motor coordination and forelimb reaching, but not spatial recognition and its flexibility.

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-L-570 2光子顕微鏡イメージングで見るパーキンソン病モデルマウスにおける一次運動野同期神経活動 Yoshihisa Tachibana（橘吉寿）¹，Yuki Aoyama（青山友紀）¹，Hiroaki Wake（和氣弘明）^1,2 ¹神戸大院医・システム生理 ²CREST Parkinson's disease is characterized by degeneration of nigral dopaminergic neurons, leading to a wide variety of motor dysfunctions such as the akinesia/bradykinesia, rigidity, tremor, and postural instability. We have previously reported that abnormally synchronized oscillations in the internal globus pallidus (GPi), an output station of the basal ganglia, contribute to the expression of parkinsonian motor signs. We have also demonstrated that, in the dopamine-depleted state, glutamatergic inputs to the subthalamic nucleus (STN) and reciprocal external globus pallidus (GPe)-STN interconnections are both important for the generation and amplification of the oscillatory activity of STN neurons, which is subsequently transmitted to GPi neurons, thus contributing to the symptomatic expression of Parkinson's disease. However, it remains unknown whether the primary motor cortex (M1), which sends the glutamatergic afferents to the STN and also receives the GPi signals through the motor thalamus, shows neuronal synchronization in the parkinsonian state. To answer this question, we performed <I>in vivo</I> two-photon calcium imaging of M1 neurons in parkinsonian mice. To generate the parkinsonian mice, we unilaterally injected dopamine neurotoxin, 6-hydroxydopamine (6-OHDA), into the medial forebrain bundle. Open field tests showed that parkinsonian motor sign (decreased locomotor activity) severely appeared one day after the 6-OHDA injection, and the parkinsonian sign became gradually attenuated within a week. To visualize the neuronal activity in the M1 before and after the 6-OHDA injection, we measured fluorescent signal from GCaMP6f expressing in pyramidal neurons of M1. We found that neuronal synchronization in the parkinsonian state was strongly elevated between neuronal pairs in the superficial layers (layer II/III) of M1, while less evident between M1 deep layer (layer V) neurons. The synchronized activity was positively correlated with the parkinsonian motor sign. We conclude that the neuronal synchronization in the M1 is well associated with the expression of Parkinson's disease, as well as the basal ganglia neurons. We now plan to perform optogenetic manipulation of M1 activity to reveal the causal relationship between synchronized M1 activity and parkinsonian motor signs.		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-L-571 手の運動の制御における一次運動野および内側運動野の神経細胞活動と局所フィールド電位の関与 Yoshihisa Nakayama（中山義久），Osamu Yokoyama（横山修），Eiji Hoshi（星英司）東京都医学総合研 The primary motor cortex (M1) of primates is considered to play a crucial role in executing contralateral hand movements. In addition, medial motor areas, such as the supplementary motor area (SMA) and caudal cingulate motor area (CMAc), are also involved in controlling contralateral hand movements, although they also contribute much to ipsilateral hand movements. However, the specific contributions of the M1 and medial motor areas to contralateral hand movements still remain elusive. In the present study, we recorded neuronal activity and local field potentials (LFPs) from the M1, SMA, and CMAc, while monkeys (<I>Macaca fuscata</I>) performed a button-press movement with either the right or left hand. Three types of movement-related neuronal activity were observed: (1) with only the contralateral hand (contralateral neuron), (2) with only the ipsilateral hand (ipsilateral neuron), and (3) with either hand (bilateral neuron). In the M1, the proportion of contralateral neurons was much larger than that of ipsilateral neurons. In addition, quantitative analyses revealed that neuronal activity in the M1 was biased toward contralateral hand movement. The contralateral bias of M1 neurons was stronger than that of SMA and CMAc neurons. Moreover, we found that the activity of M1 neurons for contralateral movements around the Go signal onset exhibited a stronger correlation with the reaction time than that of SMA and CMAc neurons. We also found a movement-related power increase in the high-gamma (80–120 Hz) and theta (3–8 Hz) bands of the LFPs in the M1. Both high-gamma and theta activity in the M1 most frequently reflected contralateral hand movement. Only a few (≤ 2%) of the changes represented movements of ipsilateral hand movement. The contralateral bias of LFPs in the M1 was stronger than that in the SMA and CMAc for both of the high-gamma and theta bands. Taken together, the results of the present study indicate that the M1 is directly involved in initiating contralateral hand movements, and thus suggest that the M1 is located hierarchically closer to the output stage of motor control than medial motor areas.

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-L-572 内包梗塞後の機能回復により生じる運動皮質の活動変化：機能的近赤外線分光法(fNIRS)によるマカクザルの脳活動計測 Junpei Kato（加藤隼平）^1,2，Toru Yamada（山田亨）¹，Hirishi Kawaguchi（川口拓之）¹，Keiji Matsuda（松田圭司）¹，Noriyuki Higo（肥後範行）¹ ¹国立研究開発法人産業技術総合研究所人間情報研究部門 ²筑波大学大学院人間総合科学研究科 Neuromotor systems have the capacity for functional recovery following damage to the central nervous system, and the recovery can be enhanced by postlesion rehabilitative training. Several lines of evidence have suggested that functional recovery from brain damage stands on compensatory activity changes in related cortical areas. Monitoring of brain activity in such relevant regions will therefore contribute to assess the recovery and optimize the rehabilitative training for maximizing the recovery. Functional near-infrared spectroscopy (fNIRS) has been thought to be a strong candidate to monitor brain activity during rehabilitative training because it allows recording with subjects in non-retrained condition. Using a macaque model of internal capsular infarcts and an fNIRS system for macaque monkeys, both of which we recently established, we here evaluated the effectiveness of fNIRS to detect cortical activity changes after recovery of hand movements. We measured the task-evoked hemodynamic responses in the motor-related cortical areas of macaque monkeys during performing a small-object retrieval task. Before infarction, significant increases in oxygenated hemoglobin (Hb) and decrease in deoxygenated Hb were detected in the primary motor area (M1) contralateral to the hand used. Focal infarcts were thereafter induced in the posterior limb of internal capsule by injecting endothelin-1, a vasoconstrictor peptide. Immediately after the injection, flaccid paralysis was observed in the hand contralateral to the injected hemisphere. Dexterous hand movements were gradually improved to the prelesion level over the subsequent several months. After recovery of hand movements, task-evoked hemodynamic responses increased at the areas within the ventral premotor area (PMv). Interestingly, the response at PMv in the infarcted hemisphere increased in case the damage of the internal capsule was small, whereas PMv in the non-infarcted hemisphere was recruited in case larger damage were induced. The causal role of the newly-recruited areas in recovery of dexterous hand movements was confirmed by means of pharmacological inactivation by muscimol. We here conclude that the fNIRS measurement is applicable to evaluate brain activity changes crucial for functional recovery after brain damage.		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-L-573 自閉スペクトラム症者の到達運動における外部物体中心座標利用の低下 Yumi Umesawa（梅沢侑実）¹，Kanae Matsushima（松島佳苗）²，Takeshi Atsumi（渥美剛史）³，Mrinmoy Chakrabarty（Chakrabarty Mrinmoy）¹，Reiko Fukatsu（深津玲子）¹，Masakazu Ide（井手正和）¹ ¹国立リハ研究所・脳機能部 ²関西医大 ³杏林大医病態生理 Approximately 80 % of individuals with autism spectrum disorder (ASD) are reported to have various motor difficulties (Green et al., 2009), and a part of them relate to bodily movements using visuo-motor coordination. Preferential utilization of allocentric (external landmark-centered) over egocentric (body-centered) coordinates are known to be important for executing visuo-motor functions (Chakrabarty et al., 2016; Uchimura et al., 2013). We studied whether ASD can use allocentric coordinates when performing a reaching task to a memorized target. We then examined whether the task performance would associate with ordinal motor skills. Sixteen ASDs and 19 typically developing individuals (TD) participated in the experiment. We asked them to memorize a target cross presented on a touch screen at a randomized position relative to a square (reference frame) in the background. The frame randomly shifted either right or leftward just before the reaching, as the participants touched either a spatially congruent position with memorized target position in relation to the reference frame (allocentric condition) or a bodily congruent position ignoring the frame (egocentric condition). We calculated the degree of "Touch position deviation"; by subtracting the touched position on the screen from the correct position in accordance with each condition. Motor skills of the participants were evaluated using a clinical assessment tool (Bruininks-Oseretsky Test of motor proficiency, second edition; BOT-2). The degree of the deviation in the allocentric condition tended to be greater in ASD than TD (t = 1.71, p = 0.09, Cohen's d = 0.56), while there were no group-difference of the deviation in egocentric condition (t = -1.0, p = 0.32, Cohen's d = 0.34). By pooling over all participants, individual degrees of the deviation in allocentric condition negatively correlated with respective BOT-2 sub-scores of "fine manual precision" (r = -0.61, p = 0.001, power (1-β) = 0.96) and "upper limb coordination" (r = -0.52, p = 0.008, power (1-β) = 0.87), which assess ordinal motor performances. These results demonstrate that motor skills demanding visual-motor coordination (e.g. drawing lines along a frame, throwing a ball within a frame) are strongly related to the ability of efficiently utilizing allocentric coordinates and further indicate that decrements in this allocentric coding may be a reason for the visuo-motor difficulties in 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-L-574 <I>Lactobacillus</I> species modulates brain gene expression and anxiety-like behavior in <I>Danio rerio</I> Joshua Philippe Olorocisimo（Olorocisimo Joshua Philippe），Harold M. Carag（Carag Harold M.），Michael C. Velarde（Velarde Michael C.） Institute of Biology, University of the Philippines Diliman The gut-brain axis is a bidirectional communication pathway that links the digestive and central nervous systems. Through this axis, an organism's gut microbiome can affect the brain, and induce genetic and behavioral changes. Certain microorganisms and their metabolites have been shown to improve mood, and are termed as psychobiotics which may be used to treat diseases like depression or anxiety disorders. However, given their recent discovery, relatively few psychobiotics have been discovered and fully characterized. To accomplish this, <I>Danio rerio</I> or zebrafish were used because they are easier to maintain and manipulate than mice, have similar genetics and behavior with mammals, and can be treated with various bacteria which make them good model organisms for this research. In this study, we investigated three different species of <I>Lactobacillus</I> that may induce psychobiotic effects in zebrafish. The novel object exploration test and novel tank diving test were used to determine the effects of <I>Lactobacillus</I> on anxiety-like behavior in zebrafish. Then, qPCR was used to analyze changes in brain gene expression related to anxiety. Treatment with <I>Lactobacillus</I> sp. 2 and 3 increased fish exploration rate despite the presence of an unfamiliar object, indicating reduction of anxiety-like behavior in zebrafish. In addition, <I>Lactobacillus</I> sp. 3 treated fish spent more time in the upper half of the novel tank diving test, demonstrating anxiolytic effects in fish exposed to an unfamiliar environment. <I>Lactobacillus</I> sp. 3 also induced an upregulation in <I>gad</I> brain gene expression, suggesting the effects in zebrafish are mediated through the GABAergic pathway. Taken together, <I>Lactobacillus</I> treatment can modulate brain gene expression and behavior of zebrafish. Finally, we have discovered a new psychobiotic species of <I>Lactobacillus</I> that can reduce anxiety.		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-L-575 ストレスによるマウス眼窩前頭皮質―扁桃体シナプス伝達変化とストレス関連行動への寄与 Hiroshi Kuniishi（國石洋）^1,2，Daisuke Yamada（山田大輔）²，Keiji Wada（和田圭司）²，Mitsuhiko Yamada（山田光彦）¹，Masayuki Sekiguchi（関口正幸）² ¹国立精神・神経セ精神保健研精神薬理 ²国立精神・神経セ神経研疾病４ Stress increases the risk for developing various neuropsychiatric disorders such as major depression and post-traumatic stress disorder. However, the mechanisms how stress induces stress-related behaviors were not fully understood. In this study, we focused on a brain network comprising the orbitofrontal cortex (OFC) and the basolateral nucleus of amygdala (BLA) as key regions for stress-related behavior. Frist, we confirmed that repeated tail-shock stress increased immobility in the tail-suspension test and the forced swim test in mice. Next, we isolated excitatory transmission of the OFC-BLA synapse by using optogenetic and whole-cell patch clamp methods, and examined the effects of repeated tail-shock stress on the OFC-BLA synaptic transmission. In this study, greater AMPA/NMDA current ratio and inward-rectification of AMPARs current in the OFC-BLA synapse were observed in mice after the repeated tail-shock stress. Interestingly, these changes in the OFC-BLA synapse and stress-induced behavioral changes were blocked by the PKA inhibition during the stress. Finally, to examine causal relationship between synaptic transmission in the OFC-BLA pathway and stress-related behavior, we performed optogenetic activation of the OFC-BLA pathway during the tail-suspension test in mice. As a result, the activation of the OFC-BLA transmission increased immobility in the tail-suspension test in mice. In conclusion, our data suggest that stress changes the OFC-BLA synaptic transmission, and thereby induces stress-related behaviors 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-L-576 マウス前部帯状回皮質におけるグルココルチコイド受容体発現細胞の特定 Yasushi Hojo（北條泰嗣）¹，Mai Kawata（川田真衣）²，Rina Ito（伊藤吏那）¹，Miki Hashizume（橋爪幹）¹，Takanari Nakano（中野貴成）¹，Takayuki Murakoshi（村越隆之）¹ ¹埼玉医大医　生化学 ²埼玉医大保健医療 Anterior cingulate cortex (ACC), responsible for higher order processing of cognition, attention and emotion, is vulnerable to stress, and its network oscillation is disrupted upon stress or in the brain of patients with mental disorders such as schizophrenia. Upon various forms of stress, glucocorticoid (GC) is secreted from adrenal glands, and chronic exposure to GC induces various deleterious effects on the nervous system, for example, shrinkage of dendrites and neuronal loss in the hippocampus, which may underlie the pathophysiology of disorders including major depression and posttraumatic stress disorder (PTSD). Previously, we revealed that GC enhances the pre-pulse inhibition (PPI) ratio and suppresses dopamine (DA)-induced modulation of oscillation power in the ACC slices, using mice (C57BL/6J,4-5-week-old, male) administrated with corticosterone (25 μg/mL) via drinking water for 7 days. These results suggest that GC affects the functions of ACC via alterations of oscillation power. The site of GC action in the ACC, however, is still unclear. We explored to identify cell types that express glucocorticoid receptors in mouse ACC with immunohistochemical staining. The ACC from 4-5-week-old male C57BL/6J mice were frozen-sliced with a cryostat. After incubation of antibodies against GR and various neuronal markers; namely, anti-NeuN IgG, anti-parvalbumin (PV) IgG and anti-somatostatin (SST) IgG for labeling neurons, PV-positive interneurons and SST-positive interneurons, respectively. The each primary antibody was visualized with secondary antibodies labeled with fluorescent dye, and images were acquired with confocal microscopy. In the Cg1 area of ACC, 91% of neurons (NeuN positive cells), 95% of PV-positive ineterneurons, and 88% of SST-positive interneurons expressed GR. GR may also express in the excitatory neurons because approximate 80% of neurons in the ACC is glutamatergic. In Cg2 area of ACC, 92% of neurons (NeuN positive cells), 96% of PV-positive ineterneurons, and 85% of SST-positive interneurons expressed GR. Neither statistical significance was observed in the area nor cell types, suggesting the ubiquitous expression of GR in the ACC. These results suggest that GC may act GR on these types of neurons (glutamatergic, and PV- or SST-positive inhibitory neurons) in the ACC, resulting in the alteration of network oscillation in the ACC.		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-L-578 JUNK-FOOD DIFFERENTIALLY AFFECTS THE BRAIN REWARD SYSTEM OVER TIME Maria Florencia Andreoli（Andreoli Maria Florencia）^1,2，Gisela P. Lazzarino（Lazzarino Gisela P.）²，Maria F Acutain（Acutain Maria F）³，Maria F Rossetti（Rossetti Maria F）^2,4，Rocio Schumacher（Schumacher Rocio）²，Cora Stoker（Stoker Cora）^2,4，Guillermina Canesini（Canesini Guillermina）²，Jorge G Ramos（Ramos Jorge G）² ¹IDIP Hospital de Ninos de La Plata ²Instituto de Salud y Ambiente del Litoral (ISAL), Facultad de Bioquimica y Ciencias Biologicas, Universidad Nacional del Litoral. CONICET, Santa Fe, Argentina. ³Instituto de Biologia Celular y Neurociencia (IBCN), Facultad de Medicina, Universidad de Buenos Aires. CONICET, Buenos Aires, Argentina. ⁴Departamento de Bioquimica Clinica y Cuantitativa, Facultad de Bioquimica y Ciencias Biologicas, Universidad Nacional del Litoral, Santa Fe, Argentina We analysed the short and long-term effects of the intake of a highly palatable `junk-food' cafeteria diet (CAF) on the expression of key genes of the brain reward system. Female Wistar rats were fed with chow or CAF from weaning for 30 (CAF30) or 77 (CAF77) days. The Ventral Tegmental Area (VTA), two regions of the Accumbens Nucleus (Core, NAcC; and shell, NAcSh), and the Ventral Pallidum (VP) were isolated, and the expression of several genes of the dopaminergic and GABAergic pathway, and the leptin receptor (ObRb) was evaluated by qPCR. Circulating leptin was assessed by radioimmunoanalysis. Data was statistically analysed by two-way ANOVA followed by Tukey post-test. CAF30 increased energy intake and adiposity, without affecting circulating leptin or body weight. In VTA, CAF30 enhanced dopamine active transporter (DAT) expression and decreased both isoforms of glutamate decarboxylase (GAD1 and GAD2), without altering tyrosine hydroxylase (TH) levels. In NAcSh, CAF30 dopamine receptor (DR) 2 mRNA decreased. In NAcC of CAF30, increased levels of DR1 were found. Besides, CAF30 increased GAD2 levels in VP. CAF77 animals further increased energy intake and adiposity, leading to higher body weight, hyperleptinemia, and increased expression of ObRb in VTA, with no changes on the expression of key genes of the reward system. Our results indicate that, in the short-term, CAF deregulates the dopamine pathway, reflecting a reward hyposensitivity state, which promotes the excessive intake of palatable foods to compensate this status. Conversely, this is reverted in the long-term, when the hypercaloric intake could respond to an altered homeostatic control.

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-L-579 神経細胞サブタイプ特異的SIK3欠損マウスの睡眠覚醒 Fuyuki Asano（浅野冬樹）¹，Chika Miyoshi（三好千香）¹，Tomoyuki Fujiyama（藤山知之）¹，Noriko Hotta（堀田範子）¹，Miyo Kakizaki（柿崎美代）¹，Aya Ikkyu（一久綾）¹，Satomi Kanno（管野里美）¹，Shinya Nakata（中田慎也）¹，Seiya Mizuno（水野聖哉）²，Fumihiro Sugiyama（杉山文博）²，Satoru Takahashi（高橋智）^1,2，Hiromasa Funato（船戸弘正）^1,3，Masashi Yanagisawa（柳沢正史）¹ ¹国際統合睡眠医科学研究機構 ²筑波大学医学医療系 ³東邦大学医学部解剖学講座 Sleep is a fundamental behavior conserved from vertebrates to invertebrates. Sleep/wakefulness is regulated by two processes, homeostatic sleep need and circadian rhythm. SIK3 is a member of AMP-activated protein kinase (AMPK) family and is associated with sleep need and circadian rhythm regulation. Gain of function mutation of <I>Sik3<I> gene was identified through the forward genetic screening of randomly mutagenized mice, which results in an increased sleep need and NREM sleep amounts. Conversely, flies and round worms with a hypomorphic mutation in <I>Sik3</I> orthologue showed reduced sleep-like behavior. Most of systemic SIK3 KO mice die neonatally and the few survived mice suffered from several growth retardation and malnutrition, while those survivors demonstrated longer circadian period lengths under constant dark conditions. Despite these previous results, whether endogenous SIK3 is involved in the regulation of sleep/wakefulness in mice and, if so, which neuron types are responsible for the SIK3-dependent sleep/wake regulation remain unknown. Here, we report a sleep/wakefulness of neuron subtype-specific SIK3-deficient mice. Newly developed <I>Sik3-flox</I> mice were crossed with neuron subtype-specific Cre-expressing mouse lines. Neuron subtype-specific SIK3-deficient mice were essentially healthy; we successfully recorded their electroencephalograms/electromyograms and evaluated the circadian variation of sleep/wakefulness. Additionally, we analyzed the spectral power of the electroencephalogram in each sleep/wake state and determined the level of their sleep need.		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-L-580 SIK3キナーゼによる睡眠覚醒制御の分子メカニズム Shinya Nakata（中田慎也）¹，Tomoyuki Fujiyama（藤山知之）¹，Chika Miyoshi（三好千香）¹，Satomi Kanno（管野里美）¹，Aya Ikkyu（一久綾）¹，Fuyuki Asano（浅野冬樹）¹，Haruna Komiya（小宮春奈）¹，Seiya Mizuno（水野聖哉）²，Fumihiro Sugiyama（杉山文博）²，Satoru Takahashi（高橋智）^1,2，Hiromasa Funato（船戸弘正）^1,3，Masashi Yanagisawa（柳沢正史）¹ ¹筑波大学国際統合睡眠医科学研究機構 ²筑波大学医学医療系 ³東邦大学医学部解剖学講座 Sleep is a ubiquitous behavior in animals from vertebrates to invertebrates. Although neural networks switching sleep/wake behaviors are being unveiled, the intracellular signaling for sleep regulation remains unknown. Recently, <I>Sik3</I>, which belongs to the AMP-activated protein kinase (AMPK) family, was identified as a novel gene regulating sleep/wakefulness using forward genetics. A pedigree (<I>Sleepy</I>) which has a causative mutation in <I>Sik3</I> resulting in the skipping of exon 13 shows marked decrease of wake time and prolonged NREM sleep time. The exon 13-encoded region contains a protein kinase A (PKA) phosphorylation site Ser551 (S551). 14-3-3 proteins bind to the SIK3 S551 in a phosphorylation-dependent manner and our previous report showed deletion of exon 13-encoded region results in abolishment of binding of 14-3-3 proteins, suggesting 14-3-3 binding is important for sleep/wake regulation. It has been reported that SIK3 kinase activity is tightly linked with phosphorylation of Thr221 in the kinase domain T-loop of SIK3. Interestingly, the phosphorylation of Thr221 of SIK3 increased after sleep deprivation in wild type mice, suggesting SIK3 activity increases in mice having higher sleep need. However, whether the kinase activity of SIK3 is involved in sleep/wake regulation is still unknown. Here, we investigated the role of SIK3 kinase activity in its phosphorylation status and sleep/wakefulness. First, we examined the phosphorylation status and physical interaction of mutant SIK3 proteins using HEK293T cells. While wild-type SIK3 protein was recognized by anti-PKA phosphorylation motif antibody, the kinase-dead SIK3 (T221A) exhibited a decrease in reactivity to the antibody. Consistently,14-3-3 binding to SIK3 (T221A) was lower than that to wild-type SIK3. On the other hand, constitutive active SIK3 (T221E) exhibited a slightly increased or similar reactivity to anti-PKA phosphorylation motif antibody compared with wild-type SIK3. SIK3 (T221E) exhibited an increased binding to 14-3-3. This result suggests that SIK3 kinase activity is required for the phosphorylation at S551 by PKA and subsequent 14-3-3 binding. To further investigate the role of SIK3 kinase activity in sleep/wakefulness, we have made <I>Sik3</I> gene-modified mice in which the threonine residue (T221) was substituted with glutamate using the CRISPR/Cas9 system. We confirmed the establishment of <I>Sik3 T221E</I> mice using direct sequencing and genotyping.