Llanos M N, Anabalón M C
Unidad de Biología de la Reproducción, INTA, Universidad de Chile, Santiago, Chile.
Mol Reprod Dev. 1996 Nov;45(3):313-9. doi: 10.1002/(SICI)1098-2795(199611)45:3<313::AID-MRD8>3.0.CO;2-V.
Experiments were designed to characterize the effect of progesterone on the hamster sperm acrosome reaction (AR). Progesterone stimulated exocytosis of previously capacitated spermatozoa in a dose-dependent manner. Progesterone-3-(O-carboxymethyl)oxime:BSA conjugate also induced AR when added to capacitated sperm suspensions. EGTA and La3+, added 10 min before progesterone, completely abolished the steroid-stimulatory effect. Benzamidine, a trypsin inhibitor, also inhibited AR when added to sperm cells 10 min before progesterone. This effect was avoided when spermatozoa were treated with the Ca2+ ionophore ionomycin. Conversely, the H+ ionophore FCCP, or the Na+/K+ ionophore nigericin, did not prevent the effect of the inhibitor. Results suggest that progesterone acts on the hamster sperm plasma membrane to stimulate exocytosis, which requires external Ca2+ and presumably Ca2+ influx. In addition, a sperm trypsin-like protease may be part of the mechanism by which progesterone stimulates AR. Since the ionomycin-induced AR does not require this proteolytic activity, the possible involvement of such an enzyme in the progesterone-stimulated Ca2+ influx necessary for the occurrence of AR is discussed.
实验旨在表征孕酮对仓鼠精子顶体反应(AR)的影响。孕酮以剂量依赖的方式刺激先前已获能精子的胞吐作用。孕酮-3-(O-羧甲基)肟:牛血清白蛋白缀合物添加到获能精子悬液中时也可诱导顶体反应。在添加孕酮前10分钟加入乙二醇双四乙酸(EGTA)和La3+,可完全消除类固醇的刺激作用。胰蛋白酶抑制剂苄脒在添加孕酮前10分钟加入精子细胞时也可抑制顶体反应。当精子用Ca2+离子载体离子霉素处理时,这种作用可被避免。相反,H+离子载体羰基氰化物间氯苯腙(FCCP)或Na+/K+离子载体尼日利亚菌素不能阻止抑制剂的作用。结果表明,孕酮作用于仓鼠精子质膜以刺激胞吐作用,这需要细胞外Ca2+,推测还需要Ca2+内流。此外,一种精子胰蛋白酶样蛋白酶可能是孕酮刺激顶体反应机制的一部分。由于离子霉素诱导的顶体反应不需要这种蛋白水解活性,因此讨论了这种酶可能参与孕酮刺激的顶体反应发生所必需的Ca2+内流。