2004 Fiscal Year Final Research Report Summary
Role of μ-Calpain in Human Sperm for Fertilization
| Project/Area Number |
14571577
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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 |
Obstetrics and gynecology
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| Research Institution | Nagoya City University |
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Principal Investigator |
OZAKI Yasuhiko Nagoya City University, Graduate School of Medical Sciences, Research Associate, 大学院・医学研究科, 助手 (50254280)
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| Co-Investigator(Kenkyū-buntansha) |
SUGIURA Mayumi (OGASAWARA Mayumi) Nagoya City University, Graduate School of Medical Sciences, Associate Professor, 大学院・医学研究科, 助教授 (30264740)
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| Project Period (FY) |
2002 – 2004
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| Keywords | calpain / human sparm / acrosome reaction / calcium ion / proteolysis |
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| Research Abstract |
We evaluated the role of μ-calpain in human sperm for fertilization.
Semen collected manually from healthy donors with informed consent was liquefied and following percoll gradient centrifugation. After exposure to different concentration of progesterone (P) or under hypoxic condition (H), the samples were used for immunostaining, SDS-PAGE and western blot analysis. The increase of calcium ion concentration in the sperm was observed by fluorescent microscope (ARGUS-50CA) system using Fura 2-AM. The role of calpain for fertilization was speculated from the results of Hamster penetration test, Hamilton Motility Analyzer, Triple satin method and Acrobeads test using calpain inhibitors.
Immunostaining in human sperm was observed using antibodies against the μ-calpain in the types of whole acrosome, equatorial segment, head segment, neck segment or neck and tail segment. Most of these demonstrated acrosome type staining with anti-pro μ-calpain antibody. Western blot analysis revealed P or H treatment to cause a concentration or time-dependent activation of μ-calpain.
In addition, calpain inhibitors significantly reduced the acrosome reaction and penetration.
The results suggest that μ-calpain in human sperm plays an important role for the fertilization.
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Research Products
(3 results)
