Hang H Y, Feng B Y, Zhang Z Y
Department of Endocrinology and Laboratory of Reproductive Biology, Institute of Zoology, Beijing, PRC.
Sci China B. 1990 Nov;33(11):1304-10.
In order to elucidate the biochemical mechanism of sperm capacitation, the relationship between plasmalemma Na,K-ATPase, capacitation and acrosome reaction was investigated. Plasmalemma of guinea pig spermatozoa was isolated by sucrose gradient centrifugation for the determination of Na,K-ATPase activity. As far as the authors are aware, the Na,K-ATPase activity in plasmalemma of the guinea pig has never been detected. By treating sperm plasmalemma with 0.05% DOC (deoxycholate), enzyme activity could be quantitatively measured. After spermatozoa were incubated in capacitation medium for 8 h, Na,K-ATPase activity was found to be decreased to 35.6% as compared with that before incubation. The spermatozoa incubated for 10.5 h in capacitation medium containing 1 and 5 mumol/L ouabain showed 46.5% and 64.4% acrosome reactions respectively, while the acrosome reaction of the control group was only 27.4%. The above results suggest that the decrease in the Na,K-ATPase activity of guinea pig spermatozoa may be a prerequisite for capacitation. Experimental results demonstrated for the first time that Na,K-ATPase activity exists in the sperm plasmalemma of the guinea pig. It was further found that the decrease of Na,K-ATPase activity of plasmalemma enhances sperm capacitation. It is suggested that sperm capacitation in the guinea pig is possibly induced by the decrease in plasmalemma Na,K-ATPase and, as a consequence, the intracellular Na+ is increased, which would benefit the exchange of Na+out/Ca++in and the onset of acrosome reaction.
为阐明精子获能的生化机制,研究了质膜钠钾 - ATP酶、获能与顶体反应之间的关系。通过蔗糖梯度离心法分离豚鼠精子的质膜,以测定钠钾 - ATP酶活性。据作者所知,豚鼠质膜中的钠钾 - ATP酶活性从未被检测到。用0.05%去氧胆酸盐(DOC)处理精子质膜后,可对酶活性进行定量测定。精子在获能培养基中孵育8小时后,发现钠钾 - ATP酶活性相较于孵育前降低至35.6%。在含有1和5 μmol/L哇巴因的获能培养基中孵育10.5小时的精子,其顶体反应分别为46.5%和64.4%,而对照组的顶体反应仅为27.4%。上述结果表明,豚鼠精子钠钾 - ATP酶活性的降低可能是获能的前提条件。实验结果首次证明豚鼠精子质膜中存在钠钾 - ATP酶活性。进一步发现质膜钠钾 - ATP酶活性的降低增强了精子获能。提示豚鼠精子获能可能是由质膜钠钾 - ATP酶活性降低所诱导,结果细胞内钠离子增加,这有利于钠离子外流/钙离子内流的交换及顶体反应的启动。