| Project/Area Number |
13557203
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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 | 展開研究 |
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| Research Field |
Biological pharmacy
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
AKAIKE Akinori KYOTO UNIVERSITY, Graduate School of Pharmaceutical Sciences Department of Pharmacology, Professor, 薬学研究科, 教授 (80135558)
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| Co-Investigator(Kenkyū-buntansha) |
KUME Toshiaki KYOTO UNIVERSITY, Graduate School of Pharmaceutical Science Department of Pharmacology, Research Associate, 薬学研究科, 助手 (10303843)
KATSUKI Hiroshi KYOTO UNIVERSITY, Graduate School of Pharmaceutical Science Department of Pharmacology, Associate Professor, 薬学研究科, 助教授 (40240733)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 2003: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2002: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2001: ¥8,900,000 (Direct Cost: ¥8,900,000)
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| Keywords | Nitric oxide / Retinal Degeneration / Neuronal death / Apoptosis / Dopamine neuron / Glutamate / 興奮毒性 / 神経栄養因子 / アストロサイト / 網膜裡経節細胞 / ドパミンニューロン / 画像解析 / 中脳切片 / 線条体切片 / ドパミン / カリウムチャネル / 神経線維 |
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| Research Abstract |
We constructed an experimental system for quantitative evaluation of neuronal morphology by on-line image analysis of central neurons in dissociated or organotypic cultures, thereby investigated neuronal degeneration and neuroprotection. (1) We prepared retinal tissue cultures from neonatal rats that received DiI injection into the superior colliculus, and observed morphology of retinal ganglion cells (RGCs). RGCs were identified as bright fluorescent spots aligned in one focal plane within the retina. The number and size of fluorescent spots gradually decreased during the course of cultivation. Cycloheximide or actinomycin D markedly inhibited the decrease in the number of viable RGCs. DEVD, a selective caspase-3 inhibitor, also significantly inhibited the decrease in RGC viability, and DNA fragmentation was found to be restricted in the ganglion cell layer. These results indicate that the gradual decrease in RGC viability results from induction of apoptosis. Moreover, nitric oxide de
… More
rived from neuronal nitric oxide synthase (NOS) was shown to play an important role in induction of apoptosis in RGCs, because an NOS inhibitor L-NAME as well as a relatively selective neuronal NOS inhibitor 7-nitroindazole markedly inhibited the decrease in RGC viability. (2) We found that cultivation of midbrain slice cultures with medium containing high concentrations of Mg^<2+> or that containing NMDA receptor antagonists resulted in a drastic decrease in the number of dopaminergic neurons. There were also prominent atrophic changes in neurites of dopaminergic neurons remaining after these treatments. Co-application of an adenylyl cyclase activator or a cyclic AMP analog abolished the effects of chronic Mg^<2+> and NMDA antagonists. These results suggest that moderate activation of NMDA receptors associated with spontaneous neuronal activity makes significant contribution to early development and maintenance of midbrain dopaminergic neurons, where cyclic AMP-related intracellular signaling plays a key role. Less
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