Mrsny R J, Meizel S
J Cell Biol. 1981 Oct;91(1):77-82.
The role of a K+ ion influx and Na+,K+-ATPase activity in the hamster sperm acrosome reaction (AR) was examined, using a range of concentrations of K+,K+ ionophores and a Na+,K+-ATPase inhibitor. Washed epididymal hamster sperm, capacitated in vitro in an artificial medium containing 2 mM Ca2+, 147 mM Na+, and 3, 6, 12, 18, or 24 mM K+, began undergoing the AR after 3 h of incubation. Sperm incubated in low K+ (0.9 mM) failed to undergo the AR even after 5 h of incubation. Sperm in 0.9 mM K+ could be induced to undergo the AR when either K+ (12 mM) alone or K+ (12 mM) with 0.1 microM nigericin was added after 3.5 h of incubation. The addition of K+ alone stimulated the AR in 30 min, whereas nigericin plus K+ stimulated the AR 15 min after addition. Neither nigericin added alone (0.9 mM K+) nor nigericin plus 12 mM K+ added to a low Ca2+ (0.35 mM) system resulted in acrosome reactions. Valinomycin (1 nM) did not stimulate the AR when added together with K+ (3-24 mM) to sperm incubated in 0.9 mM K+ for 3.5 h but markedly decreased sperm motility. Micromolar levels of ouabain blocked the AR when added between t = 0--3 h to sperm incubated with 3-24 mM K+. Inhibition of AR by the addition of 1 microM ouabain to sperm incubated with 3 mM K+ was completely reversed by the addition of 0.1 microM nigericin at t = 3.5 h. These results suggest that Na+,K+-ATPase activity and the resulting K+ influx are important for the mammalian sperm AR. Some similarities between requirements for the hamster sperm AR and secretory granule exocytosis are discussed.
利用一系列不同浓度的钾离子、钾离子载体以及一种钠钾ATP酶抑制剂,研究了钾离子内流和钠钾ATP酶活性在仓鼠精子顶体反应(AR)中的作用。将附睾洗涤后的仓鼠精子在含有2 mM钙离子、147 mM钠离子以及3、6、12、18或24 mM钾离子的人工培养基中进行体外获能,孵育3小时后开始发生顶体反应。在低钾(0.9 mM)环境中孵育的精子,即使孵育5小时也未能发生顶体反应。在孵育3.5小时后,单独添加钾离子(12 mM)或添加钾离子(12 mM)与0.1微摩尔尼日利亚菌素,可诱导处于0.9 mM钾离子环境中的精子发生顶体反应。单独添加钾离子在30分钟内刺激顶体反应,而尼日利亚菌素加钾离子在添加后15分钟刺激顶体反应。单独添加尼日利亚菌素(0.9 mM钾离子)或在低钙(0.35 mM)系统中添加尼日利亚菌素加12 mM钾离子均未导致顶体反应。将缬氨霉素(1 nM)与钾离子(3 - 24 mM)一起添加到在0.9 mM钾离子中孵育3.5小时的精子中时,并未刺激顶体反应,但显著降低了精子活力。在t = 0 - 3小时之间,向与3 - 24 mM钾离子一起孵育的精子中添加微摩尔水平的哇巴因可阻断顶体反应。在t = 3.5小时时,向与3 mM钾离子一起孵育的精子中添加1微摩尔哇巴因对顶体反应的抑制作用可通过添加0.1微摩尔尼日利亚菌素完全逆转。这些结果表明,钠钾ATP酶活性以及由此产生的钾离子内流对哺乳动物精子顶体反应很重要。文中还讨论了仓鼠精子顶体反应与分泌颗粒胞吐作用需求之间的一些相似之处。