| Project/Area Number |
18580280
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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 |
Applied animal science
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| Research Institution | Shizuoka University |
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Principal Investigator |
KOHSAKA Tetsuya Shizuoka University, Faculty of Agriculture, Professor (10186611)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥4,010,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥510,000)
Fiscal Year 2007: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2006: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | Relaxin / Testis / Sperm / Capacitation / Phosphorvlation / Boar |
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| Research Abstract |
This research project was planed to elucidate protein tyrosine phosphorylation during boar sperm capacitation and acrosome reaction mediated by relaxin-like protein (seminal relaxin). The outcome of this study as follows.
The present study demonstrated that seminal relaxin actually induced boar sperm capacitation and acrosome reaction with high sperm viability. In relaxin-treated boar sperm, the activity of protein tyrosine kinase was increased with progress of the incubation time, which reached a plateau (10U/10^8 cells) at 4 h of incubation. Furthermore, Western blot analysis revealed a specific band of phosphorylated protein corresponding to the molecular size of 30 kDa in the capacitated/acrosome reacted boar sperm induced by relaxin, suggesting that relaxin induces tyrosine phosphorylation in boar sperm. Also proteome analysis characterized it as interesting and unusual. When polyclonal antibody was generated to the recombinant phosphorylated protein-MBP fusion protein expressed in E. coli, immunohistochemical analysis provided clear-cut evidence of the localization of phosphorylated protein in the sperm head and midpiece.
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