| Project/Area Number |
16590718
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | Nihon University |
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Principal Investigator |
MUGISHIMA Hideo Nihon University, School of Medicine, Professor, 医学部, 教授 (80183648)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUDA Noboru Nihon University, Advanced Research Institute of the Science and Humanities, Associate Professor, 大学院総合科学研究科, 准教授 (40267050)
MATSUMOTO Taro Nihon University, School of Medicine, Associate Professor, 医学部, 准教授 (50366580)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2006: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2004: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | vascular endothelial cells / angiogenesis / proteomic analysis / 管腔形成 |
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| Research Abstract |
In collagen gel cultures, bovine capillary endothelial (BCE) cells formed tubular morphogenesis in response to FGF-2. The three-dimensional collagen assay has been employed in analyses of gene regulation during the sprouting process, but post transcriptional events critical in capillary formation have not been identified. We have therefore undertaken a proteomic approach to identify key regulatory events at the protein level, in tubular morphogenesis. We found that 21 protein spots were increased during FGF-2-induced BCE cell tubular morphogenesis. By MALDI-TOF MS, some of the proteins were identified such as alpha B crystalline, phosphatidylethanolamine binding protein, Homeobox protein and phosphoglycerate kinase. In addition, we found that approximately 220 and 280 kDa proteins were tyrosine phosphorylated during BCE cell tubular morphogenesis. We identified that these proteins were both ninein, a microtubule-binding protein. In western immunoblot analysis, phosphorylation of ninein was seen in BCE cells cultured in collagen gels from 8 to 48 hours after incubation, corresponding to a period of tubular morphogenesis. Introduction of ninein siRNA into BCE cells significantly inhibited tubular morphogenesis in collagen gel culture. These results suggest that ninein plays an important role in tubular morphogenesis of vascular endothelial cells.
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