e ポスター

e ポスター 4. 精神疾患の分子基盤 e Poster 4. Molecular basis of pscychiatric disease

2020/9/10　14:00～15:00　オンデマンドB-1 P1-23* 新規SHANK3アイソフォームとトランスクリプトの分子構造の解明およびShank3トランスクリプトのマウス脳における発現解析 Elucidation of molecular structure of novel SHANK3 isoforms and transcripts and expression analysis of multiple Shank3 transcripts in mouse brain 堀江直人¹、横森将輝¹、宇和野水優¹、崎村建司²、内野茂夫¹ 1. 帝京大学大学院理工学研究科、2. 新潟大学脳研究所 Naoto Horie¹, Masaki Yokomori¹, Miyu Uwano¹, Kennji Sakimura², Sigeo Uchino¹ 1. Teikyo University Graduate School of Science and Engineering, 2. Brain Research Institue Niigata University SHANK3 gene is the causative gene of Phelan-McDermid syndrome, which is an autistic disorder characterized by severe language and intellectual disabilities, global development delay, mental retardation, and hypotonia. Since multiple abnormalities, including mutation, deletion, and duplication, in the SHANK3 gene have been found in the genome of patients with autism spectrum disorder (ASD), the SHANK3 gene is considered to be an autism-related gene. Furthermore, Shank3-deficient mice produced by several research groups including us show ASD-like behavior, including social abnormalities, increased anxiety, persistence, indicating the usefulness of Shank3-deficient mice as ASD pathological model mice. The SHANK3 gene consists of 22 exons and encodes a scaffolding protein that interacts with various synaptic molecules, including PSD95 and neuroligin. There are multiple promoters on the 5'-terminal side and within the gene-body, and alternative splicing suggests the presence of several SHANK3 isoforms translated from multiple transcripts, but the molecular structure of these SHANK3 isoforms and transcripts has not been clarified yet. In this study, we investigated the molecular structures of multiple SHANK3 isoforms and their transcripts and analyzed their expression in the mouse brain. As a result, we identified the molecular structure of novel Shank3 transcripts, suggesting encode for the SHANK3d and SHANK3e isoforms, and it was suggested that the expression of each transcript had individual differences among mice.		2020/9/10　14:00～15:00　オンデマンドB-1 P1-24* マウスの脳におけるSHANK3アイソフォームの発現解析 Expression analysis of SHANK3 isoforms in mouse brain 大野創平¹、堀江直人¹、横森将輝¹、井上-上野由紀子²、井上高良²、崎村健司³、内野茂夫¹ 1. 帝京大学大学院理工学研究科、2. 国立精神神経医療研究センター神経研究所、3. 新潟大学脳研究所 Shouhei Ohno¹, Naoto Horie¹, Masaki Yokomori¹, Yukiko Inoue-Ueno², Tkayoshi Inoue², Kenji Sakimura³, Shigeo Uchino¹ 1. Teikyo University Graduate School of Science and Engineering, 2. National Institute of Neuroscience, National Center of Neuroscience and Psyc hiatry, 3. Brain Research Institute, Niigata University SHANK3 is a scaffolding protein that interacts with synaptic functional molecules, including PSD95 and neuroligin. In recent years, the SHANK3 gene has been causative for the Phelan-McDermid syndrome that is characterized by severe language impairment, global development delay, mental retardation, and autistic behavior. In addition, Shank3-deficient mice have shown several autistic-like behaviors, including abnormal social behavior, increased anxiety, and repetitive behavior. Therefore, SHANK3 is thought to be related with the neuropathology of autism spectrum disorder.SHANK3 has multiple isoforms, but the expression profile of each isoform in the brain is largely unknown. The aim of this study is to clarify the expression profile of SHANK3a (full-length SHANK3) and SHANK3c (N-terminal deletion SHANK3) isoforms in the brain. In transgenic mice in which the gfp gene was inserted the downstream of the promoter of each Shank3 isoform gene, cells expressing each SHANK3 isoform were analyzed by immunohistochemical staining with an anti-GFP antibody. As a result, the SHANK3a isoform was confirmed to be expressed in neurons in multiple regions, including layer 2/3 of the cerebral cortex, striatum, hippocampal dentate gyrus, and thalamus. On the other hand, the SHANK3c isoform was confirmed to be expressed in cerebellar granule cells. Furthermore, it was suggested that it was also expressed in parvalbumin-positive basket cells and stellate cells. We currently inject Lucifer Yellow into the neurons under a microscope using a glass pipette to visualize neurons in the layer 2/3 of the cerebral cortex in which the expression of the SHANK3a isoform has been shown, and we are analyzing the differences in synaptic structure between wild-type and Shank3-deficient mice.

2020/9/10　14:00～15:00　オンデマンドB-1 P1-25 自閉スペクトラム症者における末梢血中脂肪酸のVLDL特異的な増大は社会的相互作用と相関する VLDL-specific increases of fatty acids in autism spectrum disorders correlate with social interaction. 松﨑秀夫¹、岩田圭子¹、臼井紀好^2,1 1. 福井大学子どものこころの発達研究センター、2. 大阪大学大学院医学系研究科 Hideo Matsuzaki¹, Keiko Iwata¹, Noriyoshi Usui^2,1 1. Research Center for Child Mental Development, University of Fukui, 2. Graduate School of Medicine, Osaka University Abnormalities of lipid metabolism contribute to ASD pathogenesis has been argued, but the mechanisms are not fully understood. We characterized the lipids metabolism in ASD children by lipidomics, and identified the dramatic alterations of 71 metabolites involved in lipid biosynthesis and metabolism, oxidative stress, and synaptic function. Identified fatty acids (FAs) indicated the correlations with clinical social interaction score of ASD diagnosis as well as lipoproteins-triglyceride concentrations in ASD. We also found the specific reduction of very-low-density lipoprotein (VLDL) in ASD, which showing correlations with a decrease of APOB concentrations in ASD. We further found a significant increase in APOJ known as a sensor for oxidative stress, demonstrating enhancing oxidative stress in ASD is due to VLDL-specific dyslipidemia. These results demonstrate the increases in FAs are due to VLDL-specific degradation, providing novel insights to uncover the mechanisms such as oxidative stress generations via mitochondrial dysfunction underlying ASD pathology.		2020/9/10　14:00～15:00　オンデマンドB-1 P1-26 Lemur kinase 1 (LMTK1) ノックアウトマウスの多動及び衝動性行動 Hyperactive and impulsive behaviors of Lemur kinase 1 (LMTK1) knockout mice. 高橋美由紀^1,2、小林静香³、Wei Ran²、堤弘次^4,2、北一郎⁵、安藤香奈絵²、真鍋俊也³、????????上口裕之⁶、友村美根子⁷、久永眞市² 1. 早稲田大学先進理工学部生命医科学科、2. 東京都立大学生命科学科神経分子機能研究室、3. 東京大学医科学研究所基礎医科学部門神経ネットワーク分野、4. 北里大学理学部生物科学科細胞生物学講座、5. 東京都立大学人間健康科学研究科行動生理学研究室、6. 理化学研究所脳神経科学研究センター神経細胞動態研究チーム、7. 明海大学保健医療学部口腔保健学科　 MIYUKI TAKAHASHI^1,2, Shizuka Kobayashi³, Ran Wei², Koji Tsutsumi^4,2, Ichiro Kita⁵, Kanae Ando², Toshiya Manabe³, Hiroyuki Kamiguchi⁶, Mineko Tomomura⁷, Shin-ichi Hisanaga² 1. Department of Life Science and Medical Bio-Science, Waseda University, 2. Laboratory of Molecular Neuroscience, Department of Biological Sciences, Tokyo Metropolitan University, 3. Division of Neuronal Network, Department of Basic Medical Sciences, Institute of Medical Science, the University of Tokyo, 4. Division of Cell Biology, Department of Biosciences, School of Science, Kitasato University, 5. Laboratory of Behavioral Neuroscience, Department of Human Health Sciences, Tokyo Metropolitan University, 6. Laboratory for Neural Cell Dynamics, RIKEN Center for Brain Science, 7. Department of Oral Health Sciences, Meikai University School of Health Sciences The number and size of spines are carefully regulated depending on synaptic activity. In fact, hyper- and hypo-formation of synapses is reported in patients with psychiatric disorders and neurodegenerative diseases, respectively. Therefore, maintenance of synaptic structure and function is crucial for proper functioning of the brain. Recently, we reported that Lemur kinase 1A (LMTK1A) regulates dendritic spine formation through Rab11 activity. We also found that TBC1D9B, a Rab11A GAP, downstream protein of LMTK1, regulates spine formation. LMTK1 is a novel but uncharacterized Ser/Thr kinase, which is highly expressed in mammalian brain. In neurons, knockdown or knockout (KO) of LMTK1 resulted in longer axons, greater branching of dendrites and spine formation, suggesting that LMTK1 plays a role in neuronal circuit formation. However, the in vivo function of LMTK1 remained to be investigated. Here, we examined the brain structures and behaviors of LMTK1 knockout (KO) mice. LMTK1 was expressed in most neurons throughout the brain. The overall brain structure appeared to be normal in LMTK1 KO mice, but the numbers of synapses were increased. LMTK1 KO mice had a slight impairment in memory formation and exhibited distinct psychiatric behaviors such as hyperactivity, impulsiveness and high motor coordination without social interaction deficits. Some of these abnormal behaviors represent core features of attention deficient hyperactive disorder (ADHD), suggesting the possible involvement of LMTK1 in the pathogenesis of ADHD.

2020/9/10　14:00～15:00　オンデマンドB-1 P1-27 CAPS2を介する有芯小胞放出の欠損はオキシトシン依存的な社会行動とPOMC依存的な摂食行動に影響する A deficiency of the CAPS2-mediated dense-core vesicle release affects Oxytocin-dependent social and POMC-dependent feeding behavior 古市貞一¹、藤間秀平¹、山鹿亮祐¹、南春花¹、雨宮菜月¹、有馬知輝¹、篠田陽²、阿部学³、崎村健司³、佐野良威¹ 1. 東京理科大学、2. 東京薬科大学、3. 新潟大学 Teiichi Furuichi¹, Shuhei Fujima¹, Ryosuke Yamaga¹, Haruka Minami¹, Natsuki Amemiya¹, Tomoki Arima¹, Yo Shinoda², Manabu Abe³, Kenji Sakimura³, Yoshitake Sano¹ 1. Tokyo Univ. of Science, 2. Tokyo Univ. of Pharm. & Life Sci., 3. Niigata Univ. CAPS2 protein facilitates exocytosis of dense-core vesicles which contain neuropeptides/peptide hormones. Intriguingly, CAPS2 KO mice have small body size and low body weight compared with their control mice and show decreased social behavior. Several alterations in CAPS2 gene have been reported in some patients with autism spectrum disorder (ASD). However, it is unknown which bioactive peptide(s) that are associated with behavioral phenotype are regulated by the CAPS2-mediated release mechanism. In this study, we focused on two peptides oxytocin (OXT) and proopiomelanocortin (POMC) which are social- and feeding-related peptides, respectively. CAPS2 was localized in the hypothalamic paraventricular nucleus and posterior pituitary (PP) which produces and releases, respectively, hypophysial OXT. CAPS2 was also localized in POMC neurons of the hypothalamic arcuate nucleus and intermediate pituitary (IP). CAPS2 KO mice showed accumulated levels of OXT in the PP and, conversely, decreased levels of plasma OXT compared to WT mice. Similarly, CAPS2 KO mice showed higher POMC protein levels in the IP than WT mice. These results suggest a deficit of OXT and POMC release in CAPS2 KO mice. Eventually, OXT neuron-specific CAPS2 cKO mice showed impaired social behavior, which could be ameliorated by intranasal treatment with exogenous OXT. Moreover, administration of melanotan-2, an α-MSH receptor agonist, decreased the food intake per body weight in CAPS2 KO but not wild-type mice. Collectively, we suggest that CPAS2 is a critical machinery for both OXT and POMC release, thereby being associated with social and food-intake behavior. We also suggest a possible association between the CAPS2 gene with ASD that is often associated with eating disorder.		2020/9/10　14:00～15:00　オンデマンドB-1 P1-28* 中枢性摂食を制御する一次繊毛からのNPYシグナル経路の解析 Regulation of primary cilia shortening via feeding-related ciliary NPY receptors 西村宣哉¹、小林勇喜¹、斎藤祐見子¹ 1. 広島大学大学院統合生命科学研究科 Nobuya Nishimura¹, Yuki Kobayashi¹, Yumiko Saito¹ 1. Graduate School of Integrated Sciences for Life, Hiroshima Univ. The primary cilium is a sole organelle protruding from plasma membrane. The physiological importance of primary cilia is demonstrated by the gamut of ciliary diseases associated with many syndromes including obesity. Furthermore, stunted cilia within brain subregions have been reported in obese animal models. Recent studies have addressed that a small set of neuronal primary cilia contain a specific inventory of GPCRs, including feeding-related melanin-concentrating hormone (MCH) receptor 1 (MCHR1). We previously revealed that MCH induces cilium length shortening via ciliary MCHR1-Gi/o-Akt pathway in human retinal pigmented epithelial (hRPE1) cells, neurons in rat hippocampal slice culture and human iPS-derived cortical neurons. A recent localization screening using ciliated cells identified a new set of ciliary GPCRs such as neuropeptide Y (NPY) receptor 2 and 5 (Y2 and Y5), which are involved in feeding behavior as well as the MCHR1 system. However, it remains unknown whether NPY affects ciliary status in cells expressing ciliary Y2 and Y5, respectively. Here, we show that NPY caused significant cilia length shortening in a dose-dependent manner both in Y2 and Y5 expressing stable hRPE1 clone. A clear response caused by NPY was observed after 3h in Y2 and 6h in Y5, and this tendency remained constant up to 12h. We next found that an important component in the initial stage of GPCR-mediated cilia shortening was similar between Y2 and MCHR1 but not Y5. Central activation of NPY5R increases caloric intake, whereas activation of the NPY2R decreases caloric intake. Thus, our data suggest that a particular signaling pathway via ciliary Y2 and Y5 might be related to their discrete physiological effects.

2020/9/10　14:00～15:00　オンデマンドB-1 P1-29 【誌上発表】 2種類の一次繊毛保有細胞における繊毛局在型GPCRの特徴付け Characterization of GPCRs selectively localized in primary cilia in two ciliated cellular models 小林勇喜¹、武田茉朋²、徳永優希¹、斎藤祐見子¹ 1. 広島大学大学院統合生命科学研究科、2. 広島大学総合科学部 Yuki Kobayashi¹, Maho Takeda², Yuuki Tokunaga¹, Yumiko Saito¹ 1. Graduate School of Integrated Sciences for Life, Hiroshima University, 2. School of Integrated Arts and Sciences, Hiroshima University G-protein-coupled receptors (GPCRs) comprise the largest and most diverse cell surface receptor family. Many GPCRs show plasma membrane trafficking properties and are primarily localized in the plasma membrane without an agonist. Unexpectedly, in 2008, a limited number of GPCRs such as somatostatin receptor 3 (SSTR3) and melanin-concentrating hormone receptor 1 (MCHR1) were observed to be selectively targeted to an immotile sensory organelle named the primary cilium on several mammalian cell types including neuronal cells. The ciliary membrane is highly enriched in specific signaling molecules, allowing the primary cilium to efficiently convey signaling cascades in a highly ordered microenvironment. Recently, GPCR distribution screening identified that more than 10 GPCRs are localized in primary cilia. At present, both of SSTR3 and MCHR1 are widely recognized as representative ciliary GPCRs, because their localization in the cilium are reproduced in multiple laboratories. In order to establish a definitive list of ciliary GPCRs, we performed follow-up study by using ciliated cellular models. We cloned 8 GPCRs with N-terminus Flag tag, transfected them into hRPE1 cells and NIH3T3 cells, induced ciliogenesis by serum starvation, and observed subcellular localization of GPCRs by immunofluorescent staining. We found that neuropeptide Y (NPY) receptor 2, NPY receptor 5, prolactin-releasing hormone receptor and several dopamine receptors (DRs) are efficiently localized to the primary cilia in both the ciliated cell models. At present, study is undertaken to identify ligand-induced ciliary signaling in ciliary DRs-EGFP-tagged stable clones.		2020/9/10　14:00～15:00　オンデマンドB-1 P1-30 脳や腸菅を介した潰瘍性大腸炎の新たな治療戦略 A new approach for therapeutic strategy against ulcerative colitis via intestine and brain 小山佳久¹、小林悠輝¹、大津巌生²、河野祐介²、鈴木健吾³、臼井紀好¹、近藤誠¹、小林光¹、島田昌一¹ 1. 大阪大学、2. 筑波大学、3. ユーグレナ株式会社 Yoshihisa Koyama¹, Yuki Kobayashi¹, Iwao Ohtsu², Yusuke Kawano², Kengo Suzuki³, Noriyoshi Usui¹, Makoto Kondo¹, Hikaru Kobayashi¹, Shoichi Shimada¹ 1. Osaka University, 2. Tsukuba University, 3. euglena Co., Ltd. Ulcerative colitis (UC) is a non-specific inflammatory bowel disease (IBD) that causes ulcers and erosions in the colonic mucosa and becomes chronic with cycles of amelioration and exacerbation. Apart from gastrointestinal symptoms, it is known that IBD patients have a high risk for mental diseases such as depression and anxiety disorder especially during the exacerbation period, indicating that gastrointestinal inflammation affects brain function via gut-brain axis signals. Therefore, the development of radical medicine effective both intestine and brain is urgent. Because one of the disease pathogenesis is involvement of oxidative stress, it is likely that an appropriate antioxidant drug will be an effective therapeutic drug for UC. Our silicon-based agent made it possible to stably generate a large amount of hydrogen in the intestinal tract continuously by ingestion. Hydrogen is an antioxidant that can selectively extinguish only harmful hydroxyl radicals. In this study, we examined whether the symptoms of UC were alleviated by treatment with the silicone-based agents using dextran sulfate sodium (DSS)-induced UC model mice. As a result, it was revealed that the symptom of UC (such as atrophy of the large intestine, inflammation and loss of the large intestine mucosa) were significantly alleviated in the silicone-based agent-administered group compared to the non-administered group. Surprisingly, it was found that silicon-based agent neutralized the oxidative state in the brain as well as in the large intestine of DSS-induced UC model mice. It is thought that this new silicone-based agent will lead to the development of prophylactic and therapeutic agents for inflammatory bowel disease such as UC.

2020/9/10　14:00～15:00　オンデマンドB-1 P1-31 単回および繰り返し社会的敗北ストレスによる神経活動変化の全脳イメージング解析 Whole-brain mapping analysis of neuronal activation elicited by single and repeated defeat stress relevant to stress-intensity dependent behavioral alteration 勢力薫^1,2、前田駿介¹、平戸祐充¹、笠井淳司¹、橋本均^1,3,4,5,6 1. 大阪大学大学院薬学研究科神経薬理学、2. 大阪大学国際共創大学院学位プログラム推進機構、3. 大阪大学大学院連合小児発達学研究科、4. 大阪大学データビリティフロンティア機構、5. 大阪大学先導的学際研究機構、6. 大阪大学大学院医学研究科分子医薬 Kaoru Seiriki^1,2, Shunsuke Maeda¹, Yumi Hirato¹, Atsushi Kasai¹, Hitoshi Hashimoto^1,3,4,5,6 1. Laboratory of Molecular Neuropharmacology, Graduate School of Pharmaceutical Sciences, Osaka University, 2. Institute for Transdisciplinary Graduate Degree Programs, Osaka University, 3. Molecular Research Center for Children's Mental Development, United Graduate School of Child Development, Osaka University, 4. Institute for Datability Science, Osaka University, 5. Open and Transdisciplinary Research Initiatives, Osaka University, 6. Department of Molecular Pharmaceutical Science, Graduate School of Medicine, Osaka University Even single severe stress event can be a trauma and cause of the post-traumatic stress disorder. Although variety of studies addressed neuronal regulatory mechanisms underlying brain homeostasis and stress-induced brain dysfunctions, differences in neuronal mechanisms of physiological and pathological stress responses remain to be elucidated. To identify the brain regions and circuits contributing to differentiating physiological and pathological stress responses according to the stress intensity, we performed whole-brain mapping of stress-induced c-fos reporter gene expression as an indicator of neuronal activity using c-fos-EGFP mice and our developed high-speed whole-brain imaging system, FAST (block-FAce Serial microscopy Tomography). We found that acutely repeated social defeat stress, which induces social avoidance behavior, caused stress intensity-dependent increase of EGFP-positive cells in subregions in the pons and hypothalamus compared to single social defeat stress. These results suggest that hyperactivation of the multiple subregions in the brain stem can contribute to the stress-induced behavioral alterations. The causal relationship between neuronal activity and stress-related behavioral alterations observed in this study will lead to the better understanding of stress intensity-dependent regulatory mechanisms of the stress responses.