| Project/Area Number |
10670004
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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 |
General anatomy (including Histology/Embryology)
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| Research Institution | Gunma University, School of Medicine |
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Principal Investigator |
AOKI Takeo GUNMA UNIVERSITY SCHOOL OF MEDICINE, DEPARTMENT OF ANATOMY, RESEARCH ASSOCIATE, 医学部, 助手 (70150919)
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| Co-Investigator(Kenkyū-buntansha) |
FUJIMOTO Toyoshi NAGOYA UNIVERSITY SCHOOL OF MEDICINE, DEPARTMENT OF ANATOMY, PROFESSOR, 医学部, 教授 (50115929)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1998: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | caveolin / endothelial cells / tyrosine phosphorylation / phosphatase inhibitor / oxidant stress / lipid droplets / vimentin-related antigen / high cholesterol-high lipid diet / 燐酸化カベオリン / チロシンフォスファターゼ / 血管内皮細胞 / IP_3レセプター様蛋白質 / ゲニスタイン / ハービマイシン / 肺胞中隔細胞 / 脂肪細胞 / ゼメンチン / サイトカラシンB / コルセミド / 微細管 / フォドリン |
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| Research Abstract |
TYROSINE PHSPHORYLATION OF CAVEOLIN-1 IN THE ENDOTHELIUM.
1) Tyrosine phosphorylation of caveolin-1 occurred in limited locations, including the endothelium of the continuous capillaries and small venules. 2) Cultured endothelial cells were not labeled by PY14 under a standard culture condition, but became positively labeled when exposed to oxidative stresses and/or tyrosine phosphatase inhibitors. 3) The reaction was prohibited by pretreating with herbimycin A or genistein. Vasoactive reagents or physical stimuli did not cause the phosphorylation. 4) The number of invaginated caveolae decreased drastically, and vesicles labeled intensely for caveolin-1 appeared in the cytoplasm ; the average diameter of the vesicles was larger than that of caveolae. 5) The results implies that tyrosine phosphorylation of caveolin-1 occurred at tyrosine-14 in the normal rat endothelium in vivo and may induce caveolar vesiculation and/or fusion.
VIMEMTIN-RELATED ANTIGEN AROUND LIPID DROPLETE
1) By using a
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monoclonal antibody (mAb A2) raised against the cytoskeleton of goldfish xanthophores, we found that a 190-kDa protein is localized along the rim of lipid droplets in rat pulmonary septal cells. 2) MAb A2 bound to the rim of lipid droplets in the septal cell in vivo. The circular labeling around lipid droplets as well as filamentous labeling in the cytoplasm was observed in cultured septal cells. 3) As the number and size of lipid droplets decreased in cells cultured for several days, the circular labeling became indistinct, whereas the filamentous labeling persisted. 4) In Western blotting, mAb A2 recognized 190-kDa and 58-kDa bands in the cultured septal cell, but the latter turned out to be vimentin. 5) Treatment of septal cells for 4 hr with Colcemid extinguished the labeling around the lipid droplets, and only labeling in a coarse bundle pattern that matched the anti-vimentin labeling pattern remained. 6) After more than 12 hr of Colcemid treatment, the circular labeling reappeared around the lipid droplets. The disappearance of the circular labeling also occurred when septal cells were treated with Taxol or cytochalasin B for more than 4 hr. Less
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