| Project/Area Number |
19590260
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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 pharmacology
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| Research Institution | Saitama Medical University |
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Principal Investigator |
AWAJI Takeo Saitama Medical University, 医学部, 講師 (60297546)
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| Co-Investigator(Kenkyū-buntansha) |
HIRASAWA Akira 京都大学, 薬学研究科(研究院), 准教授 (70242633)
OGATA Makiko 東京女子医科大学, 医学部, 講師 (10233404)
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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 |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2009: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2008: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2007: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
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| Keywords | 不飽和脂肪酸受容体 / Green Fluorescent Protein / 生命現象可視化 / 細胞内カルシウムイオン濃度測定 / 水素イオン濃度 / ミトコンドリア機能 / 細胞内シグナル伝達機構 / 三量体共役形受容体 / GPR 40 / GPR120 / 可視化解析 / 細胞内カルシウム濃度 / GPR40 |
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| Research Abstract |
In this study, we evaluate the long chain fatty acid receptor function using visualization of mitochondrial function and intracellular Ca^<2+> concentration. Mitochondrial pH is known to reflect its functional viability. However, there has been no useful system to measure the quantitative and serial mitochondrial pH. Recently, a pH of organum has been successfully calculated by using GFP, but still it is not a direct quantitative calculation. Here, we developed the new system to evaluate mitochondrial pH of living cell with fluorescence ratio between two GFP. This pH probe system could evaluate the wide and preferable pH range. We applied insulin-stimulating substance, such as glucose, tolbutamide, arginine and potassium, and measure mitochondrial pH. In the cultured β-cells, the mitochondrial pH decreased immediately by glucose and potassium, and leisurely decreased by arginine and tolbutamide. A basal level of the mitochondrial pH of β-cells has increased by aeration in a cultured medium. However, GPR40 agonist quickly decreases the mitochondrial pH. This result suggested that the signal transduction of GPR40 is differ from the others. Additionally, we evaluate the agonist of GPR120 using Ca2+ visualization in STC-1 cell.
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