| Project/Area Number |
63540577
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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 |
動物発生・生理学
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| Research Institution | Kyushu University |
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Principal Investigator |
NOMURA K. Kyushu Univ., Faculty of Science Assistant Professor, 理学部, 助手 (30150395)
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| Co-Investigator(Kenkyū-buntansha) |
YAMANA K. Kyushu Univ., Faculty of Science Professor, 理学部, 教授 (20037162)
小林 英紀 九州大学, 理学部, 助手 (20150394)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1989: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1988: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | E-cadherin / N-cadherin / V-cadherin / Endothelial cell / Calcium / African clawed toad / Amphibian / Cell adhesion / 血管内皮細胞 / カドヘリン |
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| Research Abstract |
In this study, we examined whether cadherin-like molecules are present in frog(Xenopus laevis) developing embryos and in adult tissues or not. By using several different monoclonal and polyclonal antibodies against other vertebrate cadherins, we detected N-cadherin(132kDa) and E-cadherin(120kDa) in frog tissues. The distribution of frog N- and E-cadherin is similar to the distribution of the corresponding cadherin found in other vertebrates and the cDNA clones of these frog cadherins were isolated and analyzed. We also identified a novel cadherin-like molecule on frog endothelial cells. The monoclonal antibody FAD-I which disrupts Ca^<2+> dependent cell-cell adhesion of frog liver and kidney derived cells recognizes epitopes on frog endothelial cells and the molecule is named vascular cadherin(V-cadherin). The cDNA clone of this cadherin was also isolated and analyzed. The DNA sequence of this cadherin is not similar to any previously reported cadherins. The novel cadherin was first de
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tected in gastrula stage embryos and the distribution of the molecule is strictly restricted to the vascular systems or endothelial cells or its precursors. The monoclonal antibody FAD-I against V-cadherin disrupted the cell-cell adhesion of frog preblastula stage cell-cell adhesion in a reversible manner. The antigen recognized by this antibody is not the 140kDa molecule on the endothelial cells but a molecule having less than half the molecular weight of the typical cadherin. The partial amino acid sequence analysis revealed a striking similarity to previously reported intramembrane proteins and we isolated the cDNA clone of this early stage Ca^<2+> dependent cell-cell adhesion molecule by using the cDNA clone of V-cadherin. We are now using the DNA sequences of these frog adhesion molecules to design the antisense oligonucleotides for the injection to oocytes. The injected antisense oligonucleotides will delete the specific maternal mRNAs corresponding to the injected antisense oligos, and the artificially matured and laid eggs are to be inseminated for further development. The analysis of the developing embryos which lacks the specific adhesion molecule mRNAs would enable us to evaluate the roles played by these adhesion molecules in early embryogenesis. Less
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