1988 Fiscal Year Final Research Report Summary
Development of fluorescence microscopy for 2-D monitering of membrane potential in cultured neuronal networks
| Project/Area Number |
62880026
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
分子遺伝学・分子生理学
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| Research Institution | Osaka University |
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Principal Investigator |
KASAI Michiki Osaka University, Professor, 基礎工学部, 教授 (40022595)
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| Co-Investigator(Kenkyū-buntansha) |
TAGUCHI Takahisa Osaka University, Technical Staff, 基礎工学部, 教務職員 (10197246)
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| Project Period (FY) |
1987 – 1988
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| Keywords | Cultured cells / Brain neuron / Neuronal network / 膜電位感受性色素 |
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| Research Abstract |
Many neurons in the brain from a network to function as a super computor, and it is essentially important to elucidate a principle of information processing in multiple cellular system. In the present, however, it is very difficult to study on a principle of neuronal function using native brain because of its high complexity. Hence, it is worth an effort to establish a brain neuronal network on culture dish which is a much simpler and much more accessible object of study than that in brain, and to develop a system to measure two-dimensional membrane potential of cultured neuron.
In the first step, we established the method of primary culture of chick embryonic (E5) telencephalic neuron in serum free defined medium. Using the method, it is possible to continue the culture at least for 3 weeks. After 5 days culture in CO_2 incubator, each neuron elongates long neurite (more than 100 um), touches each other and increases the activity of cholineacetyltransferase which synthesises one of the dominant transmitter, acetylchline.
In parallel, we developed the fluorescene microscopy system for detectins fluorescence change in cultured neuron stained with voltage sensitive dye. This system is sensitive to about 1% fractional fluorescence change. Since several papers has already shown that membrane potential change causes more than 10% of fluorescese of some dyes, the system would be available to detect neuronal activity in a optical manner. Therefore, we have continued to screen fluorescent dyes as well as to improve the system to be more sensitive to small fluorescent change using photodiode for detector.
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