| Project/Area Number |
16500198
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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 |
Neuroscience in general
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| Research Institution | Osaka University |
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Principal Investigator |
AKANEYA Yukio Osaka University, Graduate School of Medicine, Assistant, 医学系研究科, 助手 (40222517)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | mGluR1 / mGluR5 / neuronal activity / synaptic plasticity / raft / dark-rearing / RNA interference / glutamate receptor / 発現制御 / RNAi / siRNA |
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| Research Abstract |
To overcome shortcomings of the conventional "knockout" manipulation, I have established RNAi-iduced Gene Silencing by Local Electroporation (RISLE). The method of RISLE is to introduce siRNA into cells of the targeting brain region by local electroporation. The paradigms of electric pulses are the combination of a preceding poring pulse of high-voltage and short-duration and a following driving pulse of low-voltage and long-duration in tandem. I have made the Homer-knockdown rats using this method. It is known that Homer can bind to GluR1 or GluR5. It is shown that I type metabotropic glutamate receptor-dependent long-term depression of visula cortex is impaired in Homer-knockdown rats, suggesting the involvement of the interaction of Homer and I type metabotropic glutamate receptors. I have obtained the result that the expressions of Homer1-3, GluR1 and GluR5 are regulated by neuronal activity. In the present study, I have employed dark-rearing as a in vivo model of neuronal activity, and have investigated whether and how neuronal activity controls expression of the glutamate receptors and the binding proteins in the membrane microdomains. The membrane contains 'raft' which is cholesterol-rich and is suggested to be involved in the intracellular signal transduction. In the current study, I have carried out the sucrose-gradient method to separate biochemically the component of raft and those of others from the tissues of visual cortex of wild- and knockdown rats which are reared normally or at the completely dark-condition. I have checked localization of glutamate receptors, the binding proteins, the protein kinases and the phosphorylated protein kinases. Finally, I have obtained the conclusion that the localization of these proteins at the raft and at extra-raft is regulated in a activity-dependent manner.
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