| Project/Area Number |
15590957
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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 |
Metabolomics
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| Research Institution | Tokyo Women's Medical University |
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Principal Investigator |
OGATA Makiko Tokyo Women's Medical University, Physiology, 医学部, 助手 (10233404)
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| Co-Investigator(Kenkyū-buntansha) |
AWAJI Takeo Tokyo Women's Medical University, Physiology, assistant professor, 医学部, 講師 (60297546)
IWASAKI Naoko Tokyo Women's Medical University, Physiology, assistant professor, 医学部, 講師 (70203370)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2004: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2003: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | insulin secreting cell / green fluorescent protein / Ca^<2+> oscillations / β細胞 / MODY / PH可視化測定 / 細胞内局在 / ミトコンドリア機能 |
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| Research Abstract |
The adjacent pH of the mithochondrial cytochrome IV conjugated with pH sensor green fluorescent protein (Mit-AfpH, Mit-V/FpH) were simultaneously assessed using real-time imaging of living, stably expressed CHO-K1 cells and INS-1 cells.
Mit-AFpH and Mt-VFpH fluorescence was detected in a cytosol and show mithochondrial formation. Stably expressed CHO-K1 cells and INS-1 cells added CCCP showed dropping of pH and surfaced by digitoninn show integrant change according to extracellular pH changing. Even in the Glucose free condition, INS-1 show slightly lower pH than CHO-K1 cells. And gllucose stimulation made no change of mitochondrial pH in CHO-K1 cells but decreasing more from the baseline in INS-1 cells toward Ca oscillation.
These results shows insulin secreting cells were continuously activating mitochondria at base condition and more activating by glucose existence. Normally, insulin shows continuous secreting in base line. Although INS-1 is the insulinoma cell line, the insulin secreting reaction may be more active but simultaneous as normal β cells reaction in glucose stimulation. This stable cell line is the unique and newly tool for mitoghondrial function study using vital cells.
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