| Project/Area Number |
10680627
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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 |
Biophysics
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| Research Institution | Tokyo University of Agriculture and Technology |
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Principal Investigator |
OHTA Yoshihiro Tokyo University of Agriculture and Technology, Faculty of Engineering Associate Proffessor, 工学部, 講師 (10223843)
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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 |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1998: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | mitochondria / ATP / fluorescence microscopy / imaging / permeability transition / membrane potential / NADH / flavin / 蛍光画像 / ATP合成 / フラビン蛋白質 |
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| Research Abstract |
The mitochondrial activities related to ATP production were measured in single mitochondria with fluorescence microscopy. So far, studies on mitochondria have been performed with mitochondrial suspension. Therefore, it has been impossible to detect asynchronous phenomena like the opening and the closing of a channel. In the present study, we developed the technique to measure the activity of single mitochondria to detect the phenomena that has not been detected with mitochondria suspension.
To observe single mitochondria with fluorescence microscopy, mitochondria were adsorbed on the cover slip coated with polyphenolic protein. Since the adsorption did not decrease the ATP production rate in mitochondria, this method for adsorption was elucidated to be useful for the measurements of mitochondrial activity. Upon addition of malate as a respiratory substrate, the time-courses of NADH formation and Flavin reduction in single mitochondria was similar to the results obtained for mitochondrial suspension. On the other hand, mitochondria showed the generation and disruption of the membrane potential due to the opening and the closing of the permeability transition pore. The opening and the closing depends on the redox state of the electron transport chain.
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