一般ポスター

一般ポスター神経・グリアの発生と分化

7月8日（土）　12:50-13:50　ポスター会場① 3P②-1 Dynamic observation of the effects of chemicals on neuronal differentiation using the pluripotent Neurosphere 柴田　真希豊橋技術科学大学　応用化学・生命工学系 Maki Shibata Dept Applied Chem & Life Sci, Toyohashi Univ of Technol, Toyohashi, Japan Glyphosate (GLY), an active ingredient in the roundup, is mainly detected in genetically modified food humans are exposed to daily. Its function as a herbicide is to inhibit 5-enolpyruvylsikimate-3-phosphate synthase (EPSPS) in the shikimic acid pathway in plants and bacteria, including intestinal bacteria in animals. Therefore, several neuronal disorders due to GLY exposure have been reported recently. We have confirmed defects and inconsistencies in Purkinje cells and ASD-like behavior by acute exposure 250 mg/kg GLY to pregnant rats. In addition, GLY-administrated offspring showed ASD-like behavior.In this study, we investigated the effect of GLY on Neurosphere differentiation. Neurosphere from the E16 rat cerebral were differentiated into a control group and a GLY group, respectively. We added 1.8×10 ^-5 mol GLY to the differentiation induction medium of the GLY group. The GLY group had less differentiation than the control group. This change in differentiation over time could be observed sequentially using 3D ultrasound microscopy, indicating that GLY may suppress Neurosphere survival.		7月8日（土）　12:50-13:50　ポスター会場① 3P②-2 アルギニンメチル化酵素PRMT1は生後マウスにおいて小脳顆粒細胞の発達を制御する Protein arginine methyltransferase PRMT1 regulates postnatal cerebellar granule cell development in mice 伊藤　彩夏¹, 石川　康平¹, 篠田　健人², 橋本　美涼², 木塚　康彦³, 中川　寅² 1. 岐阜大学大学院自然科学技術研究科, 2. 岐阜大学応用生物科学部, 3. 岐阜大学糖鎖生命コア研究所 Sayaka Ito¹, Kohei Ishikawa¹, Kento Shinoda², Misuzu Hashimoto², Yasuhiko Kizuka³, Tsutomu Nakagawa² 1. Graduate School of Natural Science and Technology, Gifu University, 2. Faculty of Applied Biological Sciences, Gifu University, 3. Institute for Glyco-core Research (iGCORE), Gifu University PurposeCerebellar granule cells are one of the major neurons for the cerebellar function. They develop as cerebellar granule neuron progenitors (CGNPs) at the external granular layer (EGL) proliferate, differentiate and migrate to form the internal granular layer (IGL) during three weeks after birth. However, its molecular control is largely unknown.PRMT1 is a major enzyme catalyzing protein arginine methylation of cellular proteins. We have previously found that Nestin-Cre-mediated PRMT1 knockout mice exhibited abnormal granule cell development, indicating PRMT1 is important for the cerebellum. In this study, to clarify cell-autonomous role of PRMT1 in the CGNPs, we have generated Atoh1-Cre-induced CGNP-specific PRMT1-deficient (cKO) mice.ResultscKO revealed body tremor and its cerebellum was small at postnatal day 8 (P8). Histological analysis of cKO cerebellum showed significant decrease in the number and density of NeuN-positive mature granule cells. NeuN signals were also found out of IGL in cKO, suggesting premature differentiation by loss of PRMT1. Furthermore, the number of Ki67-positive proliferating CGNPs was low at EGL in cKO. We have also found that cell adhesion molecule CHL-1, which regulates CGNP differentiation and migration, was decreased in cKO cerebellum at P21. Our results indicate that PRMT1 in CGNP is essential for cerebellar granule cell development.

7月8日（土）　12:50-13:50　ポスター会場① 3P②-3 SAGによるShh経路活性化を介した顆粒細胞前駆細胞の培養とその応用 Culture of granule cell progenitors and its applications by activating Shh pathway with SAG. 福本　大貴¹, 小西　慶幸^1,2 1. 福井大学　工, 2. 福井大学　ライフセ Hiroki Fukumoto¹, Yoshiyuki Konishi^1,2 1. Fac. Eng., Univ. Fukui, 2. Life Sci. Innov. Cent., Univ. Fukui. In primary neuronal cultures, prepared neural precursor cells rapidly lose their proliferative capacity and differentiate under normal culture conditions. This makes it difficult to follow the early stages of differentiation or to use experimental techniques that exploit the proliferative potential of the cells or homologous recombination. The aim of this study was to develop a culture system in which the proliferative capacity of CGNs is maintained with a Smo agonist (SAG) that activates the Shh pathway, for labeling or drug-selecting genetically manipulated cells. CGN precursors were cultured for several days in the presence of SAG, and proliferative capacity was confirmed by BrdU labeling and from the change in the size of cell aggregates. We next examined whether pCAG-EGxxFP, a reporter for the CRISPR/Cas9 system, functions in these cells. In the presence of gRNA, EGFP signals were higher in cells in aggregations maintained in the presence of SAG compared to differentiated CGNs, indicating that this culture method may be useful as a new research tool.		7月8日（土）　12:50-13:50　ポスター会場① 3P②-4 生後発達期のオリゴデンドロサイト産生におけるDBZの機能解析 Functional analysis of DBZ in oligodendrocyte generation during early postnatal development 清水　尚子, 石野　雄吾, 遠山　正彌, 宮田　信吾近畿大　東洋医学研究所　分子脳科学 Shoko Shimizu, Yugo Ishino, Masaya Tohyama, Shingo Miyata Div. of Mol. Brain Sci., Research Institute of Traditional Asian Medicine, Kindai Univ., Osaka, Japan Oligodendrocytes (OLs) are myelinating cells of the central nervous system (CNS) and arise from oligodendrocyte progenitor cells (OPCs). Developmentally, OPCs are generated during embryonic stages and after birth. OPCs migrate a long distance throughout the CNS, proliferate and then differentiate to mature OLs. Multiple lines of evidence have implicated OLs and myelin abnormalities in neuropsychiatric diseases including schizophrenia, bipolar disorder, and major depression. Previously, we reported that DISC1-binding zinc-finger protein (DBZ) is expressed in OL lineage cells, and positively regulates OL differentiation in vivo and in vitro. DBZ is a binding partner of DISC1. DISC1 is a gene associated with schizophrenia and major mental illness. DBZ KO mice display abnormalities in the myelination of the corpus callosum in early postnatal brain development. However, the underlying molecular mechanisms remain unclear. In this study, we focused on the role of DBZ binding partner, Nuclear Distribution E Homolog 1 (NDE1) in OL generation during early postnatal development. NDE1 is one of the dynein cofactors and expressed in OL lineage cells. In the meeting, we are going to show the results of in vivo experiments.

7月8日（土）　12:50-13:50　ポスター会場① 3P②-5 神経発達障害の外部リスク因子によるアストロサイト発生への影響 Effects of environmental risk factors for neurodevelopmental disorders on the development of astrocytes. 田畑　秀典¹, 宮島　倫生², 久保　健一郎^2,3, 依馬　正次⁴, 永田　浩一¹, 仲嶋　一範² 1. 神経発達障害の外部リスク因子によるアストロサイト発生への影響, 2. 慶應大・医・解剖, 3. 慈恵医大・解剖, 4. 滋賀医大・動物生命科学研究セ Hidenori Tabata¹, Michio Miyajima², Ken-ichiro Kubo^2,3, Masatsugu Ema⁴, Koh-ichi Nagata¹, Kazunori Nakajima² 1. Effects of environmental risk factors for neurodevelopmental disorders on the development of astrocytes. Astrocytes are one of the most abundant cell types in the mammalian brain. During the cortical development, astrocytes play essential roles in synapse formation, maturation, and elimination. Recently, we reported how astrocytes migrate from the ventricular zone to the cortical plate. Astrocyte progenitors adopt two migration modes; erratic migration, in which they move rapidly and irregularly within the developing cortex, and blood vessel-guided migration. We also found CXCR4/7, and integrin β1 regulate the blood vessel-guided migration, and their functional blocking disrupts their final positioning in the cortical gray matter. In this study, we explore the environmental factors that can affect the migration and positioning of astrocytes during the perinatal period. To this end, we use mouse models of hypoxic ischemic encephalopathy and maternal inflammation, in both of which their offspring has been reported to represent abnormal behaviors related to psychiatric disorders. In these model mice, we observed migration defects and abnormal positioning of astrocytes. We will discuss the molecular mechanisms, as well as their relationship with the neuronal phenotypes of these mice. This study will provide new insights into the pathogenesis of neurodevelopmental disorders caused by perturbations of astrocyte development.		7月8日（土）　12:50-13:50　ポスター会場① 3P②-6 高等哺乳動物フェレットアストロサイトの細胞形態、遺伝子発現、機能解析 Isolation of ferret astrocytes reveals their morphological, transcriptional, and functional feature 服部　剛志¹, Jureepon Roboon¹, 石井　宏史¹, 宝田　美佳¹, 観音　隆幸², 前島　隆司³, 新明　洋平⁴, 三枝　理博³, 田島　敦², 河崎　洋志⁴, 堀　修¹ 1. 金沢大学　医　神経解剖, 2. 金沢大学　医　革新ゲノム情報学, 3. 金沢大学　医　統合神経生理, 4. 金沢大・医・脳神経医学 Tsuyoshi Hattori¹, Jureepon Roboon¹, Hiroshi Ishii¹, Mika Takarada¹, Takayuki Kannon², Takashi Maejima³, Yohei Shinmyo⁴, Michihiro Mieda³, Atsushi Tajima², Hiroshi Kawasaki⁴, Osamu Hori¹ 1. Dept. of Neuroanat., Med, Kanazawa Univ, Ishikawa, Japan, 2. Dept. of Bioinfo and Genomics, Med, Kanazawa Univ, Ishikawa, Japan, 3. Dept. of Integrative Neurophysi, Med, Kanazawa Univ, Ishikawa, Japan, 4. Dept. of Medical Neurosci, Med, Kanazawa Univ, Ishikawa, Japan The number of astrocytes in the cerebrum has markedly increased through evolution. However, the manner by which astrocytes change their features during evolution remains unknown. Compared with the rodent brain, the brain of the ferret, a carnivorous animal, has a folded cerebral cortex and higher white to gray matter ratio, which are common features of the human brain. To further clarify the features of ferret astrocytes, we isolated astrocytes from ferret neonatal brains, cultured these cells, and compared their morphology, gene expression, calcium response, and proliferating ability with those of mouse astrocytes. Ferret astrocytes had longer and more branched processes, smaller cell bodies, and different calcium responses to glutamate, as well as had a greater ability to proliferate, compared to mouse astrocytes. RNA sequencing analysis revealed novel ferret astrocyte-specific genes, including several genes that were the same as those in humans. Astrocytes in the ferret brains had larger cell size, longer primary processes in larger numbers, and a higher proliferation rate compared to mouse astrocytes. Our study shows that cultured ferret astrocytes have different features from rodent astrocytes and similar features to human astrocytes, suggesting that they are useful in studying the roles of astrocytes in brain evolution and cognitive functions in higher animals.

7月8日（土）　12:50-13:50　ポスター会場① 3P②-7 脳腫瘍細胞株 U87MG におけるアクチン結合タンパク質 lasp-2 の過剰発現は単離細胞と細胞集団で異なる効果を示す Different effects of lasp-2 overexpression on single cell tracking and wound healing assay of U87MG glioblastoma cells 寺崎　朝子¹, 市毛　鈴乃¹, 米本　秋¹, 佃　聡子¹, 田村　安樹子¹, 吉井　幸恵² 1. 千葉大学　大学院理学研究院, 2. 量子科学技術研究開発機構 Asako G TERASAKI¹, Suzuno Ichige¹, Shu Yonemoto¹, Satoko Tsukuda¹, Akiko Tamura¹, Yukie Yoshii² 1. Graduate School of Science, Chiba University, 2. National Institutes for Quantum and Radiological Science and Technology Lasp-2 is an actin-binding protein highly expressed in nervous tissues and is localized in filopodia, lamellipodia, and focal adhesions (FAs). Lasp-2 also binds to components of FAs, such as paxillin and zyxin protein.We analyzed function of lasp-2 in glioblastoma tissues and a glioblastoma cell line, U87MG. mRNA expression of lasp-2 was significantly lower in tumor tissues in two databases (IVY glioblastoma atlas and TCGA). Semi-quantitative PCR also showed that mRNA expression levels of lasp-2 were lower in U87MG than in human fetal astrocytes; protein expression via immunoblotting was hardly detected.U87MG electroporated with pmCherry-lasp-2 showed that lasp-2 localized in filopodia, lamellipodia, and FAs. In kymograph analysis, lasp-2 was incorporated into FAs just after paxillin. In single cell tracking, the migration speed was promoted, although no changes in area and perimeter were detected. We found that the number of zyxin-positive FAs was increased by lasp-2 expression.We also generated a cell line expressing GFP-lasp-2 using a lentiviral vector and confirmed a similar result in single cell tracking. However, in the wound healing assay, collective cell migration was suppressed by GFP-lasp-2 expression.Our results suggest that lower expression of lasp-2 in glioblastoma tissues and glioblastoma cell lines might promote collective cell migration in invasion.