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대한생식의학회지 제27권 제1호 2010년
T-형 $Ca^{2+}$ 채널 길항제인 Mibefradil을 첨가한 인간 정자의 첨체반응 관찰
미래와희망 산부인과;서강대학교 이과대학 생명과학과;서강대학교 이과대학 생명과학과;미래와희망 산부인과;비뇨기과 분당제생병원 대진의료재단;서강대학교 이과대학 생명과학과;
이재호;손원영;이정하;이인선;김영찬;한징택;,
Observation of the Incidence of Acrosome Reaction in Human Spermatozoa Treated with Mibefradil as a T-type $Ca^{2+}i$ Channels Inhibitor
Lee, Jae-Ho;Son, Weon-Young;Lee, Jung-Ha;Lee, In-Sun;Kim, Young-Chan;Han, Ching-Tack;
Mirae & Heemang Obstetrics & Gynecology Clinic;Department of Life Science, Sogang University;Department of Life Science, Sogang University;Mirae & Heemang Obstetrics & Gynecology Clinic;Department of Urology, Pundang Je-Saeng General Hospital, Dae-Jin Medical Center;Department of Life Science, Sogang University;
Objective: The sperm acrosome reaction is a $Ca^{2+}$-dependent exocytotic event that is triggered by adhesion to the mammalian egg's zona pellucida. Previous studies suggested a role of $Ca^{2+}$ 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 ${\mu}M$ $Ca^{2+}$ A23187 $(Ca^{2+}i)$; Group 3 where spermatozoa were exposed 5 ${\mu}M$ $Ca^{2+}i$ and mibefradil; Group 4 where spermatozoa were exposed 5 ${\mu}M$ $Ca^{2+}i$ and nifedipine, and Group 5 where spermatozoa were treated with 5 ${\mu}M$ $Ca^{2+}i$ and both of mibefradil and nifedipine. Spermatozoa in all groups were retrieved after incubation for 15 and 30 minutes at $37^{\circ}C$. After staining with PSA-FITC, fluorescence was observed under a fluorescence microscope, and AR was evaluated on a total>100 spermatozoa/side. 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 ${\mu}M$ mibefradil and the decrease of acrosome reaction inhibition rate 45%. Nifedipine in acrosome reaction inhibition rate was only about 25%. The $Ca^{2+}i$-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.
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