| Project/Area Number |
12670589
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurology
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| Research Institution | Akita University |
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Principal Investigator |
TOYOSHIMA Itaru Akita Univ School of Medicine Lecturer ., 医学部, 講師 (80108951)
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| Co-Investigator(Kenkyū-buntansha) |
WADA Chizu Akita Uaiversity School of Medicine Researcher (Clinical), 医学部, 医員(臨床)
SUGAWARA Masashiro Akita Uaiversity School of Medicine Researcher (Clinical), 医学部, 医員(臨床)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2000: ¥2,800,000 (Direct Cost: ¥2,800,000)
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| Keywords | motor neuron disease / SOD1 / axonal tranport / DTC microscopy / video-enhanced fluorescent micrpscopy / green fluorescent protein / red fluorescent protein / adeno virus transtection / 緑蛍光タンパク / ALS / SOD1 / GFP / RFP / 培養神経細胞 / 蛍光ビデオ顕微鏡 / 脊髄神経節 |
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| Research Abstract |
Co-aggregation of mutant and wild-type SOD1: Background: One of the causes of familial amyotrophic lateral sclerosis (FALS) is mutation of superoxide dismutase-1 (SOD-1). Mutant SOD-1 forms aggregate in neurons, co-aggregating with several species of molecules other than SOD-1.
Objective: Here we report that the co-aggregation of wild-type SOD-1 with mutant SOD-1 in COS 7 cells, using video-enhanced fluorescent microscope and double labeling of wild-type and mutant SOD-1 with different colors. Methods: The wild-type SOD-1 was labeled with green fluorescent protein (GFP) and 4 mutant SOD-ls; A4T, G85R, G93A, and d126p, were labeled with red fluorescent protein (RFP). Simultaneous expression of both wild-type SOD-1 and mutant SOD-1 in COS 7 cells were observed by a fluorescent microscope.
Results: All of RFP labeled mutant SOD-1s formed aggregates around nuclei and adsorbed wild-type SOD-1 in various extents. Cytoplasmic background of mutant SOD-1 in COS 7 cells was reduced in the presence of aggregates, whereas the background of wild-type SOD-1 was remained even in late stages.
Conclusions: These indicate that adsorption of wild-type SOD-1 by mutant SOD-1s does not cause depletion of healthy SOD-1 in COS 7 cells.
Axonal transport in transgenic mouse of mutant SOD1: Aged spinal ganglion cells of Tg mice were successfully cultured to extend axons. Visualized fast axonal transport using a DIC microscope indicated unaltered axonal transport in mutant Tg axons compared with control age matched mice.
Adeno virus transfection: Transfected adeno virus harboring mutant SOD1 coupled witn GFP did not resulted in alteration of axonal transport in chick spinal ganglion cells.
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