1996 Fiscal Year Final Research Report Summary
Development of analyzing softwere of neuronal death model in primary cultured cells
| Project/Area Number |
07557328
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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 | 試験 |
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| Research Field |
General pharmacology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
AKAIKE Akinori Kyoto University, Faculty of Phamaceutical Sciences, Professor, 薬学部, 教授 (80135558)
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| Co-Investigator(Kenkyū-buntansha) |
MAEDA Takehiko Kyoto University, Faculty of Phamaceutical Sciences, Assistant, 薬学部, 助手 (50271010)
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| Project Period (FY) |
1995 – 1996
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| Keywords | Zinc / Nitric Oxide / N-methyl-D-asparate / Glutamate / Neuronal Death / Cortex / Methylcobalamine / Retina |
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| Research Abstract |
In this study, we developed the software for taking the microscopic pictures of cultured cells in a computer to automatically analyze cell viability by counting cultured cells. The automated analysis of cell viability accelerated the speed of cell-counting experiments. Using this softwere, we performed experiments to elucidate the mechanism underlying glutamate neurotoxicity in cultured rat cortical neurons and in cultured rat retinal neurons. Moreover, we searched neuroprotective factors against glutamate neurotoxicity. 1.The effect of methylcobalamin, a vitamin B^<12> analog, on glutamate neurotoxicity was examined using cortical cultures. The results suggests that methylcobalamin promotes intracellular methylation with S-adenosylmethionine, which is formed in the metabolic pathway of methylcobalamin. It is also suggested that methylcobalamin protects cortical cultures against glutamate neurotoxicity by reducing neurotoxic action of nitric oxide (NO). 2.The effect of Zn^<2+> on glutamate neurotoxicity was examined using retinal cultures. Zn^<2+> protected the cultures against glutamae neurotoxicity mediated by N-methyl-D-aspartate (NMDA) receptor. 3.The role of NO in glutamate neurotoxicity was analyzed using retinal cultures. A low concentration of NO induced a protective action against glutamate neurotoxicity by reducing the NMDA receptor-mediated currents and that elevated concentrations of NO,interacting with oxygen radicals, became toxic and enhanced glutamate neurotoxicity in the cultures. These results suggest that high concentration of glutamate released in retinal ischemia causes formation of large amount of NO,which is toxic to neurons when it is present in excess. The results in this study will offer basic reference materials to drive forward developmental research for neuroprotective drugs against many neurodegeneration diseases in central nervous system.
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