Furuya S, Endo Y, Osumi K, Oba M, Suzuki S
Department of Obstetrics and Gynecology, Keio University School of Medicine, Tokyo, Japan.
Fertil Steril. 1993 Jun;59(6):1285-90.
To examine the effects of stimulators or inhibitors of protein kinase C on capacitation and protein phosphorylation in human sperm.
Capacitated sperm treated with or without modulators of protein kinase C were monitored by the chlortetracycline fluorescence assay. Capacitation was confirmed by the ability of sperm to undergo the acrosomal reaction in response to mouse zonae pellucidae. 32P-labeled sperm phosphoproteins were analyzed by one-dimensional gel electrophoresis to detect the effect of protein kinase C stimulator, 12-O-tetradecanoyl-phorbol-13-acetate, on protein phosphorylation.
The treatment of sperm with protein kinase C stimulators resulted in the following: [1] the rapid appearance of the clear perimeter pattern, featuring distribution of fluorescence over the entire head exhibiting a bright perimeter and bright midpiece; [2] an accelerated ability to undergo the acrosomal reaction; and [3] an enhanced phosphorylation of 57.5-kd sperm phosphoprotein. Furthermore, these stimulatory effects were inhibited by protein kinase C inhibitors.
Protein phosphorylation mediated by protein kinase C may be involved in the regulation of human sperm capacitation.
研究蛋白激酶C的激活剂或抑制剂对人精子获能及蛋白磷酸化的影响。
采用金霉素荧光测定法监测经蛋白激酶C调节剂处理或未处理的获能精子。通过精子对小鼠透明带发生顶体反应的能力来确认获能。用一维凝胶电泳分析32P标记的精子磷蛋白,以检测蛋白激酶C激活剂12 - O - 十四酰佛波醇 - 13 - 乙酸酯对蛋白磷酸化的影响。
用蛋白激酶C激活剂处理精子会导致以下结果:[1]快速出现清晰的周边模式,其特征是荧光分布在整个头部,呈现明亮的周边和明亮的中段;[2]顶体反应能力加快;[3]57.5-kd精子磷蛋白的磷酸化增强。此外,这些刺激作用被蛋白激酶C抑制剂抑制。
蛋白激酶C介导的蛋白磷酸化可能参与人精子获能的调节。