1994 Fiscal Year Final Research Report Summary
Research for development and differentiation of nervous system using cultured neurons transfected with drebrin gene
| Project/Area Number |
04670872
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Cerebral neurosurgery
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| Research Institution | Keio University |
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Principal Investigator |
TOYA Shigeo Dept of Neurosurgery, School of Medicine, Keio University, Professor, 医学部, 教授 (40051205)
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| 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)
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| Project Period (FY) |
1992 – 1994
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| Keywords | Drebrin / Cytoskeletal Protein / Cell Adhesion / Neuronal Development / 遺伝子導入 |
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| 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.
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