Evidence suggesting a role for sperm metalloendoprotease activity in penetration of zona-free hamster eggs by human sperm.
Evidence suggesting a role for sperm metalloendoprotease activity in penetration of zona-free hamster eggs by human sperm.
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有证据表明精子金属内蛋白酶活性在人类精子穿透无透明带的仓鼠卵中发挥作用。
DOI：
10.1002/jez.1402480213
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Meizel,S
中科院分区：
文献类型：
--
作者：
Diaz-Perez,E;Thomas,P;Meizel,S
据报道，金属内肽酶（MEP）活性参与体细胞膜融合事件和海胆精子顶体反应（AR）。MEP活性也在人类和其他哺乳动物精子中被证实。本研究关注的是调查是否人精子MEP是重要的精子卵融合所必需的膜事件。将射出的人精子洗涤，体外获能，并与竞争性MEP抑制剂phosphoramidon（50 μM）或CBZ-L-苯丙氨酸（1 mM）、100 μM二亚乙基三胺五乙酸（DTPA）（一种重金属螯合剂）或作为对照，与适当溶剂一起预孵育。用排卵前人卵泡液在体外启动AR，洗涤精子以稀释抑制剂，然后与无精子的金黄地鼠卵共孵育（分别用胰蛋白酶和透明质酸酶去除透明带和卵丘）。0.5 h后清洗卵，计数剩余结合的精子数。进一步孵育2.5小时后，用乙酰鲎素或乙酰地衣红染色卵，并通过计数每个卵的去致密精子头数（穿透指数）和穿透卵的百分比来测定穿透率。抑制剂处理没有降低穿透卵的百分比（范围80-90%），但观察到穿透指数显著降低。Phosphoramidon使渗透指数降低了45%，CBZ-L-苯丙氨酸降低了57%，DTPA降低了56%。当以洗涤抑制剂处理的精子后存在的稀释水平直接添加到顶体反应精子和无精子卵的悬浮液中时，没有抑制剂降低穿透指数或穿透卵的百分比。其他研究表明，MEP抑制剂不抑制精子活力、精子结合、AR的发生、顶体基质和AR囊泡的丢失或精子头部去凝聚。当AR由钙离子载体离子霉素引发时，Phosphoramidon也显著降低了渗透指数。这些结果表明，在AR过程中，人精子MEP参与（直接或间接）修改精子质膜，以增加其与卵质膜融合的潜力。
It has been reported that metalloendopotease (MEP) activity is involved in somatic cell membrane fusion events and in the sea urchin sperm acrosome reaction (AR). MEP activity also has been demionstrated in human and other mammalian sperm. The present study was concerned with investigating whether a human sperm MEP is important in membrane events necessary for sperm egg fusion. Ejaculated human sperm were washed, capacitated in vitro, and preincubated with the competitive MEP inhibitors phosphoramidon (50 μM) or CBZ‐L‐phenylalanine (1 mM), with 100 μM diethylenetriaminepentaacetic acid (DTPA), a heavy metal chelator, or as controls, with the appropriate solvents. The AR was initiated in vitro with preovulatory human follicular fluid and the sperm washed to dilute inhibitors and then coincubated with zona‐free golden hamster eggs (zonae and cumuli removed with trypsin and hyaluronidase, respectively). Eggs were washed after 0.5 h, and the number of sperm remaining bound was counted. After 2.5 h further incubation, the eggs were stained with acetolacmoid or acetoorcein and penetration was assayed by counting the number of decondensed sperm heads per egg (penetration index) and the percent of penetrated eggs. The inhibitor treatments did not decrease the percentage of penetrated eggs (range 80–90%), but a significant reduction in the penetration index was observed. Phosphoramidon reduced the penetration index by 45%, CBZ‐L‐phenylalanine by 57%, and DTPA by 56%. None of the inhibitors decreased the penetration index or the percentage of penetrated eggs when added directly to suspensions of acrosome‐reacted sperm and zona‐free eggs at the diluted levels that would have been present after washing inhibitor‐treated sperm. Other studies demonstrated that the MEP inhibitors did not inhibit sperm motility, sperm binding, the occurrence of the AR, the loss of acrosomal matrix and AR vesicles, or sperm head decondensation. Phosphoramidon also significantly decreased the penetration index when the AR was initiated by the calcium ionophore ionomycin. These results suggest that during the AR, a human sperm MEP is involved (directly or indirectly) in modifying the sperm plasma membrane so as to increase its potential for fusion with the egg plasma membrane.
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影响因子：
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DOI：
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发表时间：
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期刊：
Pharmacology, Biochemistry and Behavior
影响因子：
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作者：
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通讯作者：
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