1999 Fiscal Year Final Research Report Summary
The molecular biological analysis for the release mechanism of endothelial-derived vasoactre substances.
| Project/Area Number |
10670631
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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 | The University of Tokyo |
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Principal Investigator |
SHIN Wee Soo Instructor, Health Service Center, 保健管理センター, 助手 (10211971)
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| Co-Investigator(Kenkyū-buntansha) |
TOYO-OKA Teruhiko Professor of Medicine and Director of Health Service Center, 医学部, 教授 (00146151)
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| Project Period (FY) |
1998 – 1999
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| Keywords | Vascular endothelium / nitre oxide / IPィイD23ィエD2 / intracellular calcium ions / antisense aligouucleotides |
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| Research Abstract |
1. Anti, IP3 receptor antibody : made from C-terminal of type I IP3 receptor, showing single band on the Western Blotting and immunologically positive for bovine endothelial cells and rat cerebellum (membrane fraction)
2. Effects of anti-IP3 receptor Ab on [CaィイD12+ィエD1]ィイD2iィエD2 ATP-induced CaィイD12+ィエD1 elevation was inhibited by anti IP3 receptor antibody dose-dependently. It inhibited both initial peak and sustained phase of CaィイD12+ィエD1 elevation.
3. Effects of NOS inhibitor l-NMMA : Cotreatment of l-NMMA with anti IP3 receptor antibody is not enough for complete block of IPィイD23ィエD2 induced CaィイD12+ィエD1 release, revealing the autocrine action of NO. These results have shown that this method can analyze each signal transduction step in endothelial cells using intracellular calcium ions as a biological marker and also can apply more precise mechanism of vasoactive substances such as NO or endothelin.
4. Transfection experiments : As a result of transfection of a marker gene (LacZ) into smooth muscle cell line, Lipofection is superior to HVJ-liposome method or to plasmid alone. Currently, antisense ODNs to NOS are transfected into VSMC or EC and [CaィイD12+ィエD1]ィイD2iィエD2 are measured.
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