Approaches to study the structure and function of nervous systems at the level of gene expression
| Project/Area Number |
61571051
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Biological pharmacy
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| Research Institution | Okayama University, Fuculty of Pharmaceutlcal Sciences |
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Principal Investigator |
TSUDA Masaaki (Associate Professor) Okayama University, Fuculty of Pharmaceutical Sciences, 薬学部, 助教授 (80132736)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMAMOTO Tadashi (Research Associate) Okayama University, Fuculty of Pharmaceutical Sciences, 薬学部, 助手 (90187443)
TSUCHIYA Tomofusa (Professor) Okayama University, Fuculty of Pharmaceutical Sciences, 薬学部, 教授 (80012673)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1987: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1986: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Neurite extension / Synapse formation / Gene expression / Specific mRNA quantitation / Riboprobes / Transfection / レトロウイルスベクター / 脳内への外来遺伝子導入 / マウス脳cDNAバンク / 脳特異的cDNA / 脳内遺伝子発現 / 脳内mRNA定量 / RNA-RNAハイブリダイゼーション / 遺伝子導入 / 遺伝子発現制御 / 神経系ネットワーク |
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| Research Abstract |
For understanding of the structure and function of nervous systems, we have tried some approaches to study the molecular mechanisms controlling the process of synapse formation. Recent advances in the area of molecular biology enable us to study the nervous systems at the level of gene expression.
1. A quatitative procedure by the solution hybridization involving RNA-RNA hybridization kinetics was developed for measurement of specific mRNA accumulated in particular tissues. By adopting the dot-blot hybridization as well, we quantitated the correct amounts of <beta>-tubulin and <beta>-actin mRNAs expressed in mouse brain. Wide application of these methods using RNA probes can be expected to quantitate specific mRNAs expressed in particular regions of brain.
2. We have found that protein kinase inhibitors as well as bibutyryl-cAMP stimulate the neurite extension of mouse neuroblastoma cell N18TG2. In accordance with this stimulation of neurite extension, transient activition of <beta>-tubulin mRNA synthesis was observed. Transfection experiment was applicable to the cells extending neurites, which indicates that it is possible to study the transcription regulation of genes responsible for neurite extension.
3. Using primary cultured cells or tissue slices of mouse brain, we tested the availability of retrovirus vectors to introduce foreign genes into nervus cells of brain. As the result, it was found that foreign genes could be introduced and expressed in the primary cells infected with retroviruses. We are now trying to express foreign genes in mouse brain by transplanting the tissue slices infected with retroviruses or infecting brain directly with retroviruses through microsyringes.
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Report
(2 results)
Research Products
(5 results)
