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Research for development and differentiation of nervous system using cultured neurons transfected with drebrin gene

Research Project

Project/Area Number 04670872
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Cerebral neurosurgery
Research InstitutionKeio University

Principal Investigator

TOYA Shigeo  Dept of Neurosurgery, School of Medicine, Keio University, Professor, 医学部, 教授 (40051205)

Co-Investigator(Kenkyū-buntansha) SHIRAO Tomoaki  Dept of Neurobiology ans Behavior, School of Medicine, Gunma University, Profess, 医学部, 教授 (20171043)
IKEDA Keiro  Dept of Neurosurgery, School of Medicine, Keio University, Instructor, 医学部, 助手 (10222879)
TODA Masahiro  Dept of Neurosurgery, School of Medicine, Keio University, Instructor, 医学部, 助手 (20217508)
UYEMURA Keiichi  Dept of Physiology, School of Medicine, Keio University, Professor, 医学部, 教授 (90049792)
矢崎 貴仁  慶應義塾大学, 医学部, 助手 (80200484)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1993: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1992: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordsDrebrin / Cytoskeletal Protein / Cell Adhesion / Neuronal Development / 遺伝子導入 / 神経発症 / SH-SY5Y細胞 / 突起伸展
Research Abstract

Drebrins are developmentally-regulated actin-binding proteins. First, we used neuroblastoma cells as a model for neural differentiation. In undifferentiated cells, drebrin E was scattered as flocculus small dots along the stress fibers. In parallel with the neuronal differentiation by retinoic acid treatmant, drebrin E was accumulated, accompanying filamentous (F) actin, in the submembranous cortical cytoplasm. F-actin with drebrin E was more stable against cytochalasin D than F-actin without it. These results indicate that drebrin E play roles in neuronal differetiation by changing its subcellular localization with stabilized filamentous actin. Second, drebrin expression was induced in nonneuronal cells (L-cells) via transfection with two kinds of drebrin expression vectors containing beta-actin promoter (MiwD-6) and methallothionein-I promoter (MTI-5). In these transfected cells, drebrin expression changed stress fibers into mesh work structure of actin fibers. After treatment with colcemid and cytochalasin D,most of the L-cells shrinked and became round, however three-quarters of the MiwD-6 cells maintained thier processes. After treatment with cytochalasin D,MTI-5 cells with drebrin expression induced by CdSO_4 did'nt make retraction processes in spite of disruption of stress fibers, while most L-cells formed retraction processes. Immunocytochemical analysis using NBD-phallacidin and anti-vinculin antibody showed that cytochalasin D caused disruption of actin fibers in L-cells, MiwD-6 cells, and MTI-5 cells, but that it didn't cause disruption of adhesion plaques in MiwD-6 and MTI-5 cells, in contrast to disappearance of those in L-cells. Because cytochalasin D has no effects on vinculin polymerization, these results suggest that drebrin has effects on actin fibers and adhesion plaques, via an interaction with actin.

Report

(4 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] 池田圭朗: "非神経細胞に発現したドレブリン蛋白が細胞骨格及び細胞接着に及ぼす影響" 神経組織の成長、再生、移植. 5. 33-34 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 池田圭朗: "非神経細胞に発現したドレブリン蛋白が細胞接着に及ぼす影響" 神経化学. 32. 358-359 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 池田圭朗: "非神経細胞に誘導発現されたドレブリン蛋白のF-アクチンに及ぼす影響" 神経組織の成長、再生、移植. 4. 30-31 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 池田圭朗: "非神経細胞に誘導発現されたドレブリン蛋白による突起形成及び細胞骨格の変化" 神経化学. 31. 564-565 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 戸田正博: "ヒトのドレブリンEcDNAクローニング" 神経化学. 31. 342-343 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 浅田英穂: "神経細胞の突起形成にともなうドレブリン蛋白の細胞内局在とF-アクチンの変化" 神経化学. 31. 146-147 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shirao T,Hayashi K,Ishikawa R,Isa K,Asada H,Ikeda K,Uyemura K: "Formation of thick, curving bundles of actin by drebrin A expressed in fibroblasts." Exp Cell Res. 215. 145-153 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ryoki Ishikawa, Kensuke Hayashi, Tomoaki Shirao, Yinhuan Xue, Takashi Takagi, Yo Sasaki, Kazuhiro Kohama: "Drebrin, a Development-associated Brain Protein from Rat Embryo, Causes the Dissociation of Tropomyosin from actin Filaments." J Biol Chem. 269(47). 29928-29933 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Ikeda, T.Shirao, S.Toya, K.Uyemura: "EFFECTS OF DREBRIN EXPRESSION ON CELL-SUBSTRATE ADHESION IN NON-NEURONAL CELLS." Neurochemical Research. 19(7). 886 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] K.Ikeda, T.Shirao, P.Kaub, H.Asada, S.Toya, K.Uyemura: "DREBRIN PROTEINS EXPRESSED IN NON-NEURONAL CELLS PRODUCED MORPHOLOGIKAL CHANGES THROUGH EFFECTS ON CYTOPLASMIC ACTIN NETWORKS." Neurochemical Research. 18(7). 856 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shirao T: "Formation of thick,curving bundles of actin by drebrinA expressed in fibroblasts" Experimental Cell Research. 215. 145-153 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] K.Ikeda: "Effects of Drebrin Expression On Cell-Sobstrate Adhesion In Non-Nevronal Cells" Neuvochemical Research. 19. 886 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 池田圭朗: "非神経細胞に誘導発現されたドレブリン蛋白による突起形成及び細胞骨格の変化" 神経化学. 31. 564-565 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] 池田圭朗: "非神経細胞に誘導発現されたドレブリン蛋白のF‐アクチンに及ぼす影響" 神経組織の成長、再生、移植. 4. 30-31 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] 池田圭朗: "非神経細胞に発現したドレブリン蛋白が細胞骨格及び細胞接着に及ぼす影響" 神経組織の成長、再生、移植. 5. 33-34 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Keiro Ikeda: "DOREBRIN PROTEINS EXPRESSED IN NON‐NEURON.4L CELLS PRODUCED MORPHOLOGICAL CHANGES THROUGH EFFECTS ON CYTOPLASMIC ACTIN NETWORKS" Neurochemical Research. 18. 856-856 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 白尾智明: "神経発生における神経細胞関連蛋白質の役割" 日本薬理学会誌. 100. 465-473 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Tomoaki Shirao: "Cloning of drebrin A and induction of neurite‐like processes in drebrin‐transfected cells" Neuro Report. 3. 109-112 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] 池田 圭朗: "非神経細胞に誘導発現されたドレブリン蛋白による突起" 神経化学. 31. 564-565 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 池田 圭朗: "非神経細胞に誘導発現されたドレブリン蛋白のF-アクチンに及ぼす影響" 神経組織の成長、再生、移植. 4. 30-31 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 浅田 英穂: "神経細胞の突起形成にともなうドレブリン蛋白の細胞内局在とF-アクチンの変化" 神経化学. 31. 146-147 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 白尾 智明: "神経発生における神経細胞関連蛋白質の役割" 日本薬理学会誌. 100. 465-473 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Tomoaki Shirao: "Cloning of drebrinA and induction of neurite-like processes in drebrin-transfected cells" Neuro Report. 3. 109-112 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Peter Kaub: "Identification of rat drebrin embryonic isoform by polymerase chain reaction and its genomic analysis" Bulletin Jap.Neurochem.Soc.31. 148-149 (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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