1995 Fiscal Year Final Research Report Summary
Evaluation of human sperm acrosome reastion by the activation of acroshin, and its clinical application
| Project/Area Number |
06671690
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | TOKYO DENTAL COLLEGE |
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Principal Investigator |
KANEKO Satoru Tokyo Dental College, Lecturer, 歯学部, 講師 (40214457)
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| Project Period (FY) |
1994 – 1995
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| Keywords | Acrosin / acrosome reaction / Concanavalin A / Cell affinity chromatography / Equilibrium sedimentation |
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| Research Abstract |
1. The acrosome reacted human sperm with progressive motility was purified by using cell affinity chromatography on Concanavalin A Sepharose.
2. The un-acrosome reasted sperm was separated by means of the continuous-step density gradient with an inner capiral, to avoid induction of the acrosome reaction, Percoll was made isotonic with Ca, NaHCO_3 free Hanks solution.
3. Hystochemical staining with FITC-Con A achieved to distinguish the acrosome reacted and un reacted sperm, the inner acrosome membrane exposed in consequence of the acrosome reaction had affinity against the agent.
4. Equilibrium sedimentation in Percoll revealed that the acrosome reacted sperm had lighter apparent density than those un-reacted.
5. Human sperm contained at least two arginine amidases. The above results, however, suggested it is difficult to monitor the acrosome reaction by activation of acrosin, which bound to the inner acrosome membrane.
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