Project/Area Number |
09470002
|
Research Category |
Grant-in-Aid for Scientific Research (B)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
General anatomy (including Histology/Embryology)
|
Research Institution | Gunma University |
Principal Investigator |
ISHIKAWA Harunori Gunma University School of Medicine, Professor, 医学部, 教授 (90010058)
|
Co-Investigator(Kenkyū-buntansha) |
MURAKAMI Tohru Gunma University School of Medicine, Lecturer, 医学部, 講師 (10239494)
HIJIKATA Takao Gunma University School of Medicine, Assistant, 医学部, 助手 (70189786)
FUJIMAKI Noboru Gunma University School of Medicine, Lecturer, 医学部, 講師 (10008261)
|
Project Period (FY) |
1997 – 1999
|
Project Status |
Completed (Fiscal Year 1999)
|
Budget Amount *help |
¥10,900,000 (Direct Cost: ¥10,900,000)
Fiscal Year 1999: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1998: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1997: ¥8,300,000 (Direct Cost: ¥8,300,000)
|
Keywords | Plasmalemma / cytoskeleton / plasmalemmal undercoat / electron microscopy / skeletal muscle / actin / intermediate filament / plectin / 平滑筋細胞 / 神経周膜 / 歯根膜 / アストロサイト / 骨格筋 |
Research Abstract |
The present research project was to elucidate the molecular relationships between the plasmalemma and cytoskeleton by immuno-fluorescence, immunoelectron microscopy and biochemical analysis. The results obtained are as follows. 1) The relationship between the plasmalemmal undercoat and extracellular matrix : The crude surface membranes prepared from rat skeletal muscles were ultrastructurally examined to elucidate the dystrophin-glycoprotein complex and its structural relationships to laminin in muscle cells. The results obtained have ultrastructurally proved the molecular model that dystrophin is connected with laminin through dystroglycan complex, dystrophin-associated membrane proteins. 2) The relationship between the plasmalemmal undercoat and intermediate filaments : Desmin and plectin were colocalized with dystrophin along the sarcolemma in rat skeletal muscle cells. Immunoelectron microscopy of skined muscle cells showed that immunogold labels for dystrophin were localized on the
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membrane side of the undercoat whereas those for plectin were found on the cytoplasmic side, where bundles of desmin intermediate filaments were seen to run. Thus, it was concluded that desmin filaments were anchored to the undercoat through fine filamentous strucutures of plectin molecules. 3) The relationship between the intermediate filaments and Z-discs : In immunoelectron microscopy of rat skined skeletal muscle cells, gold labels for desmin labeled intermediate filaments whereas those for plectin were localized on fine filamentous structures which linked intermediate filaments to Z-discs. Thus, plectin molecules were identified as fine filamentous structures linking between intermediate filaments and Zdiscs. From these series of analyses, it can be concluded that the molecular axis of dystrophin-system comprises laminin, α-dystroglycan, β-dystroglycan and dystrophin, and further extends to plectin and intermediate filaments in rat skeletal muscle cells. In addition, the molecular relationships between the plasmlemma and cytoskeleton were also analyzed for rat smooth muscle cells and astrocytes in the central nervous system. Less
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