Shear-stress-induced molecular dynamics of endothelial cell membrane
| Project/Area Number |
19300155
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | The University of Tokyo |
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Principal Investigator |
YAMAMOTO Kimiko The University of Tokyo, 大学院・医学系研究科, 講師 (00323618)
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| Co-Investigator(Kenkyū-buntansha) |
ANDO Joji 獨協医科大学, 医学部, 特任教授 (20159528)
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| Project Period (FY) |
2007 – 2009
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| Project Status |
Completed (Fiscal Year 2009)
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| Budget Amount *help |
¥18,980,000 (Direct Cost: ¥14,600,000、Indirect Cost: ¥4,380,000)
Fiscal Year 2009: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2008: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2007: ¥9,880,000 (Direct Cost: ¥7,600,000、Indirect Cost: ¥2,280,000)
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| Keywords | 血管 / 剪断応力 / 内皮細胞 / カルシウム・シグナリング / メカノトランスダクション / ATP合成酵素 / カベオラ / 細胞・組織 / 生理学 / バイオメカニクス / 機械刺激 / ナノイメージング |
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| Research Abstract |
Endothelial cells (ECs) release ATP in response to shear stress, a mechanical force generated by blood flow, and the ATP released modulates EC functions through activation of purinoceptors. In this study, we have demonstrated that cell-surface ATP synthase is involved in shear-stress-induced ATP release. Immunofluorescence staining of human pulmonary artery ECs (HPAECs) showed that cell-surface ATP synthase is distributed in lipid rafts and co-localized with caveolin-1, a marker protein of caveolae. Immunoprecipitation indicated that the cell-surface ATP synthase and caveolin-1 are physically associated. Measurement of the extracellular metabolism of ^3H-labeled ADP confirmed that cell surface ATP synthase is active in ATP generation. When exposed to shear stress, HPAECs released ATP in a dose-dependent manner, and the ATP release was markedly suppressed by membrane-impermeable ATP synthase inhibitors, angiostatin and piceatannol, and by an anti-ATP synthase antibody. Depletion of plasma membrane cholesterol with methyl-β cyclodextrin (MβCD) disrupted lipid rafts and abolished co-localization of ATP synthase with caveolin-1, which resulted in a marked reduction in shear-stress-induced ATP release. Down-regulation of caveolin-1 expression by transfection of caveolin-1 siRNA also markedly suppressed ATP-releasing responses to shear stress. These results suggest that the localization and targeting of ATP synthase to caveolae/lipid rafts, is critical for shear stress-induced ATP release by HPAECs.
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Report
(4 results)
Research Products
(104 results)
