Sperm motility hyperactivation facilitates penetration of the hamster zona pellucida.
Sperm motility hyperactivation facilitates penetration of the hamster zona pellucida.
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DOI：
10.1095/biolreprod53.6.1280
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发表时间：
1995-12
影响因子：
3.6
通讯作者：
C. Stauss;Timothy J Votta;S. Suarez
中科院分区：
文献类型：
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作者：
C. Stauss;Timothy J Votta;S. Suarez
进行这些实验以确定精子活力超活化是否促进卵母细胞透明质膜的穿透。采用了两种办法。首先，仓鼠精子在含有2.9或25.0 mM碳酸氢钠的获能培养基中孵育4.0-4.25 h。在这些培养基中，精子变得同样具有获能性，这一点可以通过它们在暴露于溶血磷脂酰胆碱或完整的透明带时经历顶体反应的能力来证明;然而，精子仅在含有25.0 mM碳酸氢盐的培养基中变得过度活化。当这些精子被添加到卵丘的卵母细胞在体外，只有2 88卵母细胞被穿透的精子预孵育在2.9 mM碳酸氢盐，而31 86卵母细胞被穿透的精子在25.0 mM碳酸氢盐。结果发现，在两种培养液中，相同数量的精子与卵母细胞结合，相同数量的精子顶体反应于卵透明带表面。对于第二种方法，将精子在含有25 mM碳酸氢盐的获能培养基中孵育。当> 70%被过度激活时，将等份试样添加到三组卵母细胞中。在10分钟后，允许精子附着和顶体反应，加入超活化抑制剂，并在固定和检查精子穿透前将精子和卵母细胞再孵育20分钟。在用抑制剂维拉帕米或Cd 2+处理的培养皿中，42个卵母细胞中的1个和42个卵母细胞中的0个分别被穿透，而对照组中40个卵母细胞中的25个被穿透。因此，似乎超活化促进仓鼠透明细胞的渗透。
These experiments were conducted to determine whether or not sperm motility hyperactivation facilities penetration of the zona pellucida of the oocyte. Two approaches were used. For the first, hamster sperm were incubated for 4.0-4.25 h in a capacitating medium that contained either 2.9 or 25.0 mM sodium bicarbonate. In these media, sperm became equally capacitated as evidenced by their ability to undergo the acrosome reaction when exposed to lysophosphatidyl choline or intact zonae pellucidae; however, sperm became hyperactivated only in the medium containing 25.0 mM bicarbonate. When these sperm were added to cumulus-free oocytes in vitro, only 2 of 88 oocytes were penetrated by sperm preincubated in 2.9 mM bicarbonate, while 31 of 86 oocytes were penetrated by sperm in 25.0 mM bicarbonate. It was found that equal numbers of sperm were bound to the oocytes and that equal numbers were acrosome-reacted on the surface of the zonae in the two media. For the second approach, sperm were incubated in a capacitating medium containing 25 mM bicarbonate. When > 70% were hyperactivated, aliquots were added to three sets of oocytes. After 10 min had been allowed for sperm to attach and acrosome-react, inhibitors of hyperactivation were added and the sperm and oocytes were incubated for an additional 20 min before fixation and examination for zona penetration. In the dishes treated with the inhibitors verapamil or Cd2+, 1 of 42 and 0 of 42 oocytes were penetrated, respectively, compared with 25 of 40 in controls. Therefore, it appears that hyperactivation facilitates penetration of the hamster zona pellucida.