一般ポスター

一般ポスターグリア-3

7月8日（土）　12:50-13:50　ポスター会場① 3P③-1 PRMT1欠損マウスにおけるオリゴデンドロサイト前駆細胞の増殖能評価 Evaluation of oligodendrocyte progenitor cell proliferation in PRMT1-deficient mice 八木　敦¹, 芝辻　奈那保¹, 橋本　美涼², 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 Atsushi Yagi¹, Nanaho Shibatsuji¹, Misuzu Hashimoto², Tsutomu Nakagawa² 1. Grad Sch of Natural Science and Technology, Univ of Gifu, 2. Faculty of Applied Biological Science, Univ of Gifu Oligodendrocytes (OL) develop from the fetal period and mature to form myelin by second week of birth in mice. Protein arginine methyltransferase (PRMT1) methylates histone and non-histone proteins and regulates various cellular functions. We have previously generated central nervous system-specific PRMT1 knock-out mice (nKO) and found dramatic hypomyelination, indicating that PRMT1 is essential for OL. However, it remains largely unknown when and how PRMT1 regulates OL lineage progression. Here, to elucidate the effects of PRMT1 deficiency in OL development, we examined the proliferation of oligodendrocyte progenitor cells (OPCs) in vivo and in vitro. First, we found by Western blotting that myelin basic protein (MBP) was dramatically suppressed in the spinal cord of nKO at postnatal day 8 suggesting that PRMT1 is essential for postnatal OL development in the spinal cord as well as in the brain. Next, to investigate the timing of abnormal development of OL lineage cells, we performed immunolabeling of OLIG2 in the fetal spinal cord and found that OLIG2-positive cells tended to decrease in nKO before birth. Furthermore, primary cortical OPCs of nKO revealed dramatic decrease of cell proliferation. Our data suggest that PRMT1 is essential for development of OL lineage cells through its regulation of proliferation of OPCs from the embryonic period.		7月8日（土）　12:50-13:50　ポスター会場① 3P③-2 Microtubule affinity-regulating kinase 4はオリゴデンドロサイトの分化とミエリン形成を制御している Microtubule affinity-regulating kinase 4 regulates oligodendrocyte differentiation and myelination 斎藤　太郎, 安藤　香奈絵東京都立大学　理学部　生命科学科 Taro Saito, Kanae Ando Dept. of Biol. Sci., Tokyo Metro. Univ., Tokyo, Japan Oligodendrocytes (OLs) are glial cells that form the myelin sheaths wrapping around neuronal axons in the central nervous system. OLs are generated by the differentiation of oligodendrocyte precursor cells (OPCs) in both developmental stages and adulthood. Microtubule affinity-regulating kinase 4 (MARK4) is a protein kinase essential in the phosphorylation of microtubule-associated protein tau in neurons. MARK4 is also expressed in glial cells. However, its function has not yet been well studied. Here, we investigated the role of MARK4 in OPCs and OLs.The expression of MARK4 was found to be upregulated during the differentiation of primary mouse OPCs. Knockdown of MARK4 during differentiation unexpectedly enhanced the expression of myelin oligodendrocyte glycoprotein (MOG), expressed in mature OLs relating to myelination, indicating that MARK4 may regulate OL myelination. We also found that alternative splicing of MARK4 harboring different C-terminal domains occurs during differentiation. Now the role of the splicing variants of MARK4 in OL differentiation and myelination is currently under investigation.

7月8日（土）　12:50-13:50　ポスター会場① 3P③-3 The molecular mechanism of CNS myelination through the cell adhesion of teneurin-4 and its effect on the diseases 福原　美月, 林　千香子, 棟方　勇貴, 大野　玲奈, 山田　渚湖, 山田　桃奈, 鈴木　喜晴東京医科歯科大学　臨床分析・分子生物学分野 Mizuki Fukuhara, Chikako Hayashi, Yuki Munakata, Reina Ohno, Nanako Yamada, Momona Yamada, Nobuharu Suzuki Department of Clinical bioanalysis and molecular biology The transmembrane protein teneurin-4 (Ten-4) is expressed in neurons and oligodendrocytes (OLs) and is required for CNS myelination. Ten-4 is predicted as a cell adhesion and its gene ( TENM4 ) mutations/SNPs have been reported in neurological and psychological disorders, but these remain unsolved. Therefore, we aimed to elucidate the cell adhesion function of Ten-4 in myelination and the effects of disease-derived mutations. First, we found that CHO cells overexpressing mouse Ten-4 formed larger cell aggregates than control cells. In the axon-mimicking nanofibers assay, the extracellular domain (ECD) of Ten-4 increased OL adhesion spots with MBP expression along nanofibers, which was inhibited by a Src kinase inhibitor. These indicate that Ten-4 promotes MBP expression via Src kinase, such as Fyn, in OL-axon adhesion. We next evaluated cell adhesion activity of human mutant Ten-4. Wild-type of Ten-4 was localized in the cell-cell contact region with F-actin and showed adhesion activity in K562 cells. Some of mutant Ten-4 affected the activity, particularly the mutation R1952H reduced the adhesion activity. Taken together, Ten-4 plays a critical role in cell adhesion between OLs and axons through the Fyn-MBP pathway and the reduction of cell adhesion by the gene mutations may be associated with deficits in the diseases.		7月8日（土）　12:50-13:50　ポスター会場① 3P③-4 PKCによりリン酸化されたL-MPZの末梢ミエリン形成期における局在変化 Distribution changes of PKC-phosphorylated L-MPZ during peripheral myelin formation 高村　敬太, 鈴木　海斗, 大滝　博和, 山口　宜秀東京薬科大学　薬学部　機能形態学教室 Keita Takamura, Kaito Suzuki, Hirokazu Ohtaki, Yoshihide Yamaguchi Dept. of Funct. Neurobiol, Tokyo Univ of Pharm and Life Sci Large myelin protein zero (L-MPZ) is a translational readthrough molecule in the peripheral nervous system. L-MPZ is synthesized from the peripheral myelin protein zero (MPZ/P0) mRNA by physiological readthrough of the canonical stop codon. Therefore, L-MPZ has additional 63 amino acid residues to P0 protein at the cytoplasmic C-terminus. The L-MPZ-specific extra domain contains an additional protein kinase C (PKC)-dependent phosphorylation site with an original PKC phosphorylation site in P0 protein, which is thought to be related to the cell adhesion property. Previously, we reported that phosphorylated L-MPZ (pL-MPZ) was significantly increased during a period of active myelin formation in mouse sciatic nerve (peak at postnatal day 21 to 28). However, the function of pL-MPZ during myelin formation is still unclear. To estimate the function of L-MPZ, the analysis of the distribution of pL-MPZ is required. To investigate the changes of the distribution of pL-MPZ during myelin formation, immunohistological analyses of developmental mouse sciatic nerves using anti-pL-MPZ specific antibody were performed. Strong positive signals of anti-pL-MPZ were detected along nearby formed Schmidt-Lanterman incisures during myelin formation. These results suggest that pL-MPZ may be significant for formation of Schmidt-Lanterman incisures in peripheral myelin.

7月8日（土）　12:50-13:50　ポスター会場① 3P③-5 Systemic inflammation leads to changes in the intracellular localization of KLK6 in oligodendrocytes in spinal cord 古部　瑛莉子, 扇谷　昌宏, 吉田　成孝旭川医科大学　解剖学講座　機能形態学分野 Eriko Furube, Masahiro Ohgidani, Shigetaka Yoshida Asahikawa Medical University, Department of Functional Anatomy and Neuroscience Axonal injury and demyelination occur in demyelinating diseases, such as multiple sclerosis, and the detachment of myelin from axons precedes its degradation. Paranodes are the areas at which each layer of the myelin sheath adheres tightly to axons. The destruction of nodal and paranodal structures during inflammation is an important pathophysiology of various neurological disorders. However, the underlying pathological changes in these structures remain unclear. Kallikrein 6 (KLK6), a serine protease produced by oligodendrocytes, is involved in demyelinating diseases. In the present study, we intraperitoneally injected mice with LPS for several days and examined changes in the localization of KLK6. Transient changes in the intracellular localization of KLK6 to paranodes in the spinal cord were observed during LPS-induced systemic inflammation. However, these changes were not detected in the upper part of brain white matter. LPS-induced changes were suppressed by minocycline, suggesting the involvement of microglia. Moreover, nodal lengths were elongated in LPS-treated wild-type mice, but not in LPS-treated KLK6-KO mice. These results demonstrate the potential involvement of KLK6 in the process of demyelination.		7月8日（土）　12:50-13:50　ポスター会場① 3P③-6 The molecular mechanism of axonal survival in the murine dysmyelination 齋藤　日菜子, 高橋　りこ, 藤里　澪, 細田　ゆき奈, 林　千香子, 鈴木　喜晴東京医科歯科大学　医歯学総合研究科　臨床分析・分子生物学分野 Hinako Saito, Riko Takahashi, Mio Fujisato, Yukina Hosoda, Chikako Hayashi, Nobuharu Suzuki Dept. of Clinical bioanalysis and molecular biology, Univ. of TMDU, Tokyo, Japan In the CNS, myelin supports axonal maintenance due to supplying lactate and pyruvate via monocarboxylate transporter (MCT)1 and MCT2 in myelin and axon, respectively. Therefore, demyelination eventually leads to axonal degeneration and loss in multiple sclerosis. Teneurin-4 (Ten-4)-deficient mice exhibited myelination defect in the CNS, but the characteristics of axons remained unclear. Thus, in this study, we first performed an electron microscopic analysis in the spinal cord of Ten-4-deficient mice and unexpectedly found that unmyelinated axons were maintained and tightly adhered each other. We observed increased expression of MCT1 and MCT2 in the area of marked demyelination. A comprehensive proteomics analysis identified the Ten-4-deficient tissue-specific proteins: HSPA12A, PDHA1, and LSAMP. HSPA12A and PDHA1 are characterized as lactate/pyruvate metabolizing proteins. LSAMP is a cell adhesion protein, and in our primary neuron culture, a decrease in neurite outgrowth and fasciculation was observed in the presence of recombinant LSAMP. To demonstrate the more detailed mechanism, we attempted functional analysis by overexpression of the proteins using a lentiviral vector and an AAV vector in primary culture systems. Elucidation of the mechanism of unmyelinated axon maintenance in the present study is expected to be applied for a novel treatment in demyelinating diseases.

7月8日（土）　12:50-13:50　ポスター会場① 3P③-7 Hydrogen Sulfide Pathway in Schwann Cells During Peripheral Nerve Degeneration : Neuropharmaco-informatics approaches Yoo Lim Chun Dept. of Anatomy and Neurobiology, Kyung Hee Univ., Seoul, Korea N-ethylmaleimide (NEM) inhibits peripheral nerve degeneration (PND) by targeting Schwann cells in a hydrogen sulfide H₂S-pathway-dependent manner, but the underlying molecular and pharmacological mechanisms are unclear. We investigated the effect of NEM, a α, β-unsaturated carboxyl compound, on H₂S signaling in in vitro- and ex vivo-dedifferentiated Schwann cells using global proteomics and transcriptomics (whole-genome and small RNA-sequencing) methods. The multi-omics analyses identified several genes and proteins related to oxidative stress, such as Sod1, Gnao1, Stx4, Hmox2, Srxn1, and Edn1. The responses to oxidative stress were transcriptionally regulated by several transcription factors, such as Atf3, Fos, Rela, and Smad2. In a functional enrichment analysis, cell cycle, oxidative stress and lipid/cholesterol metabolism were enriched, implicating H₂S signaling in Schwann cell dedifferentiation, proliferation, and myelination. NEM-induced changes in the H₂S signaling pathway affect oxidative stress, lipid metabolism, and the cell cycle in Schwann cells. Therefore, regulation of the H₂S signaling pathway by NEM during PND could prevent Schwann cell demyelination, dedifferentiation, and proliferation.