Meizel S, Turner K O
Department of Cell Biology and Human Anatomy, School of Medicine, University of California - Davis 95616-8643.
Mol Cell Endocrinol. 1991 May;77(1-3):R1-5. doi: 10.1016/0303-7207(91)90080-c.
There has been increasing interest in the relationship between rapid effects of steroids and steroid-plasma membrane interaction. This laboratory has previously reported that progesterone increases human sperm cytosolic free calcium ([Ca2+]i) and thereby initiates the human sperm acrosome reaction (AR) in less than 1 min. Herein, to test whether progesterone acts at the sperm plasma membrane, progesterone 3-(O-carboxymethyl)oxime: bovine serum albumin (BSA) conjugate (free of unconjugated progesterone) was added to capacitated human sperm. Fura-2 assays were used to detect less than 1 min changes in [Ca2+]i, and indirect immunofluorescence was used to assay the AR occurring 1 min after stimulus addition. The conjugate increased [Ca2+]i and the AR (though less than did unconjugated progesterone). Enzyme immunoassays demonstrated that the concentrations of unconjugated progesterone in conjugate-treated sperm suspensions did not increase over those of control suspensions. Since the progesterone: BSA conjugate presumably does not cross the sperm plasma membrane, progesterone must act at that membrane to increase [Ca2+]i and the AR.
人们对类固醇的快速作用与类固醇-质膜相互作用之间的关系越来越感兴趣。本实验室此前曾报道,孕酮可增加人精子胞质游离钙([Ca2+]i),从而在不到1分钟的时间内引发人精子顶体反应(AR)。在此,为了测试孕酮是否作用于精子质膜,将孕酮3-(O-羧甲基)肟:牛血清白蛋白(BSA)缀合物(不含未缀合的孕酮)添加到获能的人精子中。使用Fura-2测定法检测[Ca2+]i在不到1分钟内的变化,并使用间接免疫荧光法检测刺激添加1分钟后发生的AR。该缀合物增加了[Ca2+]i和AR(尽管比未缀合的孕酮增加得少)。酶免疫测定表明,缀合物处理的精子悬液中未缀合孕酮的浓度与对照悬液相比没有增加。由于孕酮:BSA缀合物大概不会穿过精子质膜,因此孕酮必定作用于该膜以增加[Ca2+]i和AR。