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대한생식의학회지 제27권 제1호 2010년
T-형 Ca2+ 채널 길항제인 Mibefradil을 첨가한 인간 정자의 첨체반응 관찰
1미래와희망 산부인과, 2비뇨기과 분당제생병원 대진의료재단, 3서강대학교 이과대학 생명과학과
이재호1, 손원형3, 이정하3, 이인선1, 김영찬2, 한징택3,
Observation of the Incidence of Acrosome Reaction in Human Spermatozoa Treated with Mibefradil as a T-type Ca2+I Channels Inhibitor
Jae-Ho Lee1, Weon-Young Son3, Jung-Ha Lee3, In Sun Lee1, Young Chan Kim2, Ching-Tack Han3
1Mirae & Heemang Obsterics & Gynecology Clinic, 2Dpartment of Urology, Pundang Je-Saeng General Hospital, Dae-Jin Medical Center, 3Department of Life Science, Sogang University
Objective: The sperm acrosome reaction is a Ca2+-dependent exocytotic event that is triggered by adhesion to the mammalian egg’s zona pellucida. Previous studies suggested a role of Ca2+ channels in acrosome reactions. This study was conducted to investigate the T-type calcium channel is operated in acrosome reaction of human spermatozoa. Method: Human semen samples were obtained from healthy donors with normal criteria. The spermatozoa were divided into five groups: Group 1 were non-treated as a control; Group 2 where spermatozoa were exposed to 5 ㎛ Ca2+ A23187 (Ca2+i); Groups 3 where spermatozoa were exposed 5 ㎛ Ca2+i and mibefradil; Group 4 where spermatozoa were exposed 5 ㎛ Ca2+i mibefradil and nifedipine. Spermatozoa in all groups were retrieved after incubation for 15 and 30 minutes at 37°C. After staining with PSA-FITC, fluorescence was observed under a fluorescence microscope, and AR was evaluated on a total > 100 spermatozoa/slide. Result and Conclusion: We observed on acrosome reaction inhibition rate in human spermatozoa the various of concentration of mibefradil, nifedipine. Maximum response was noted with 1.0 ㎛ mibefradil and te decrease of acrosome reaction inhibition rate 45%. Nifedipine in acrosome reaction inhibition rate was only about 25%. The Ca2+i-induced AR of spermatozoa was significantly suppressed by mibefradil. Incidence of the suppression was depending on concentration of mibefradil. Results from the present study suggest that the human spermatozoa possess T-type channel. The observation that reversible inhibitor of T channels in male germ cells provides a new mechanism of contraceptive action.
키워드 : Human spermatozoa, Acrosome reaction, T-type Ca2+ Channel inhibitor, Contraceptive action
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