| Project/Area Number |
09670988
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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 |
Psychiatric science
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| Research Institution | Osaka University |
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Principal Investigator |
NAKAMURA Yu Osaka University Medical School, Assistant Professor, 医学部, 助手 (70291440)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Toshihisa Osaka University Medical School, Assistant Professor, 医学部, 助手 (10294068)
KUDO Takashi Osaka University Medical School, Assistant Professor, 医学部, 助手 (10273632)
TAKEDA Masatoshi Osaka University Medical School, Professor, 医学部, 教授 (00179649)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1998: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1997: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | neurofilament / Alzheimer / PKA / PKO / phosphorylation / A Kinase / C Kinase |
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| Research Abstract |
Ser-51 and Ser-55 are the phosphorylation site of PKC and PKA, respectively, in he N-terminal head region of 68 kDa neurofilament subunit protein (NF-L). Site-specific phosphorylation-dependent antibodies (abNFL51 and abNFL55) against those phosphorylation sites were prepared by using affinity-column. The in vitro specificity of those antibodies were appreciated by dot and western blot analyses. Thereafter, The in vivo specificity was studied in rat primary cultured neurons. A little amount of phosphorylated NF-L recognized by abNFL55 was found in them, and the amount was increased by okadaic acid treatment. However, forskolin did not increase the amount, while simultaneous treatment of forskolin and okadaic acid dramatically increased it. These findings suggest that a small part of NF-L is phosphorylated by PKA and that the amount of the phosphorylated NF-L is regulated by okadaic acid-sensitive phosphatase, implying that NF-L dynamics in neurons is regulated by kinases and phosphatases. On the other hand, no signal was found by abNFL51 in cultured neurons, indicating that NF-L is not phosphorylated by PKC in neurons.
15 micrometer thick free-floating sections of Alzheimer and control brains were immunohistochemically studied by abNFL51 and abNFL55. abNFL51 stained neurofibrillary structures, and abNFL55 stained neuronal perikarya and neurites in both brains.
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