| Project/Area Number |
09440274
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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 |
動物生理・代謝
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
TAKAHATA Masakazu Hokkaido Univ., Grad. School of Sci., Pro., 大学院・理学研究科, 教授 (10111147)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥10,400,000 (Direct Cost: ¥10,400,000)
Fiscal Year 1999: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1998: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1997: ¥6,100,000 (Direct Cost: ¥6,100,000)
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| Keywords | crayfish / unanesthetized whole animal preparation / synaptic activity / synaptic current / whole-cell recording / voltage clamp experiment / central neurons / in situ recording / 歩行運動 / 姿勢制御 / トレッドミル / 細胞内記録法 / ブラインド・パッチ電極法 / ノンスパイキング介在神経 / 樹状突起 / 微小電極法 |
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| Research Abstract |
In order to consolidate the experimental background for applying the whole-cell recording technique to nonspiking interneurons in the central nervous system of crayfish, Procambarus clarkii Girard, performing behavioral acts, I have focused this study on 1) new development of an unanesthetized whole animal preparation to which micro- and patch electrodes could be applicable, 2) electrophysiological analyses of membrane current of nonspiking interneurons using the single electrode voltage clamp (SEVC) technique, and 3) application of patch electrodes to central neurons in situ under a dissecting microscope. Using the newly developed treadmill system, the synaptic activity of nonspiking interneurons could be recorded intracellular by a glass microelectrode from an animal walking on it. The SEVC experiments have revealed that the dendritic membrane of nonspiking interneurons possesses three types of depolarization-dependent potassium conductances. Under a dissecting microscope, spontaneous activities of single potassium channels could be recorded by a patch electrode applied to the cell bodies of central neurons in situ exposed by mechanical desheathing of the abdominal ganglion. In invertebrate animals, only a few studies have been carried out so far using the in situ whole-cell recording technique. The results of this study indicate that it is technically possible to apply patch electrodes for whole cell recording to the central neurons in situ in the animal performing behavioral acts so that the synaptic currents associated with specific behavior can be analyzed electrophysiologically.
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