| Project/Area Number |
05454670
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurochemistry/Neuropharmacology
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| Research Institution | Osaka University (1994-1995)
Tokyo Metropolitan Institute for Neuroscience (1993) |
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Principal Investigator |
YOSHIKAWA Kazuaki Osaka University, Institute for Protein Research, Professor, たんぱく質研修所, 教授 (30094452)
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| Co-Investigator(Kenkyū-buntansha) |
UETSUKI Taichi Osaka University, Institute for Protein Research, Instructor, たんぱく質研究所, 助手 (20260309)
TANIURA Hideo Osaka University, Institute for Protein Research, Instructor, たんぱく質研究所, 助手 (80263325)
高城 啓一 (財)東京都神経科学総合研究所, 分子神経生物学研究部門, 主事研究員 (30250202)
杉浦 弘子 (財)東京都神経科学総合研究所, 分子神経生物学研究部門, 主事研究員 (40162870)
矢尾板 芳郎 (財)東京都神経科学総合研究所, 分子神経生物学研究部門, 副参事研究員 (00166472)
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| Project Period (FY) |
1993 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥7,200,000 (Direct Cost: ¥7,200,000)
Fiscal Year 1995: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1994: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1993: ¥5,600,000 (Direct Cost: ¥5,600,000)
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| Keywords | Gene transfer / Neurons / Differentiation / Necdin / Cell division / Gene expression / Promoter / Transcription factors / 神経分化 / 分裂終了 / 特異的cis配列 / 転写制御因子 / trans因子 / P19細胞 / 核 / 個体発生 / 系統発生 / 遺伝子 |
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| Research Abstract |
Neurons in the brain withdraw from the cell cycle immediately after differentiation from their proliferative precursors (reuroepithelial stem cells) , and enter the postmitotic state, in which the cells remain undivided throughout the lifetime. We have investigated the mechanisms underlying the permanent arrest of cell growth displayd by differentiated neurons using gene transfer techniques. Necdin is a 325 amino acid residue protein localized to the muclei of postmitotic neurons, which withdraw permanently form the cell cycle. To examine whether necdin confers the postmitotic phenotype, necdin cDNA was stably transfected into NIH3T3 fibroblasts using a eukaryotic lac repressor-operator expression system. When the transfectants were induced to express ectopic necdin, cell growth was arrested without appreciable reduction in cell viability. In order to elucidate the mechanisms underlying necdin gene expression, we have isolated and characterized the mouse necdin gene. The necdin gene contains no intron, and its upstream region lacks canonical TATA and CAAT boxes. Deletion analysis of the promoter region using neurally differentiated P19 cells transfected with luciferase reporter genes demonstrated that a neuron-restrictive core promoter is localized to positions -80 to -35, in which a G+C-rich domain and a putative binding site for transcription factors with PAS (per, arnt, and single-minded) dimerization domain are comprised. These results suggest that necdin induces growth arrest during neuronal differntiation of the neuroepithelial stem cells in which expression of this gene is mediated by the specific cis-acting elements located at the 5' upstream region.
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