| Project/Area Number |
10044227
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| Research Category |
Grant-in-Aid for Scientific Research (A).
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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 | Asahikawa Medical College |
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Principal Investigator |
KIYAMA Hiroshi Asahikawa Medical College, Professor, 医学部, 教授 (00192021)
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| Co-Investigator(Kenkyū-buntansha) |
SEO-KIRYU Sumiko Asahikawa Medical College, Research Assistant, 医学部, 助手 (70311529)
KATO Hidemasa Asahikawa Medical College, Research Assistant, 医学部, 助手 (50292123)
EMSON Piers C. ベイブラハム研究所, 外国人特別研究員
SKYNNER Michael J. ファイザー製薬株式会社, 外国人特別研究員
ALLEN Nicholas D. ベイブラハム研究所, 外国人特別研究員
EMSON Piers ベイブラハム研究所, 外国人特別研究員
SKYNNER Mich ベイブラハム研究所, 外国人特別研究員
ALLEN Nichol ベイブラハム研究所, 外国人特別研究員
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥13,000,000 (Direct Cost: ¥13,000,000)
Fiscal Year 2000: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1999: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1998: ¥5,900,000 (Direct Cost: ¥5,900,000)
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| Keywords | injured neuron / adenovirus / neuron-specific / SCGIO / Cre recombinase / GAP-43 / nestin / c-jun / LacZ / トランスジェニックマウス / atf-3 / サイレントノックアウト / c-Jun / 神経再生 / Cre / LoxP / 神経細胞損傷 / 遺伝子相同組換え |
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| Research Abstract |
The current experiment can be dissected down into two principles. First, it is essential for the recombinational events to occur only in the cells in question (ie, injured neurons in this case). Second, it is of large benefit to restrict the number of gene manipulation before observing the outcome. When a non-restrictive promoter is used to drive the expression from the adenoviruses, we were unable to target the Cre expression solely to the axonally-damaged neurons but would widely rearrange the neighboring cells. If the gene in question is closely related to the survival and/or maintenance of a cell, it is crucial to restrict the expression in a strict temporal and spatial manner. We then tried to gain some neuronal specificity within the adenovirus' promoter. As the size matters when recombining into the expression module of an adenovirus, we initially chose the relatively small SCG10/REST system. This has led to a relatively neuronal restrictive expression of a reporter gene. Howeve
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r, when tested with a Cre expressing system, the switch of the expressivity wasn't sharp enough to restrict the recombination to the damaged neurons. We next rested transgenic mice with a pan-neuronal specific promoter such as GAP-43 or nestin enhancer. These in turn showed some problems of their own. Nevertheless, it is of utmost importance to strictly regulate the gene expression and we hope to realize this through some refinements of these. We additionally obtained some interesting insights of the role of the target genes during nerve injury. c-Jun (especially its phosphorylation) is believed to play a part in the decision of cell fate during neuronal cell death. We do agree on this principle and therefore chose the molecule for the target of our silent-knockout (=the background mice for the conditional gene targeting). In an independent experiment, we showed that an additional member of the AP-1 family, Atf-3 might be as much crucial and cooperate with c-jun during death/survival decision. We are now Flaming to apply the current experiment paradigm for this molecule as well. Less
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