Parrish J J, Susko-Parrish J, Winer M A, First N L
Department of Meat and Animal Science, University of Wisconsin, Madison 53706.
Biol Reprod. 1988 Jun;38(5):1171-80. doi: 10.1095/biolreprod38.5.1171.
Capacitation of bovine sperm was evaluated by determining the ability of sperm to fertilize bovine oocytes in vitro and to undergo an acrosome reaction upon exposure to lysophosphatidylcholine (LC). Incubation of sperm with heparin (10 micrograms/ml) increased the percentage of oocytes fertilized, but this required exposing sperm to heparin for at least 4 h before adding them to oocytes. There was no effect on the percentage of motile or acrosome-reacted sperm after exposure of noncapacitated sperm to 100 micrograms/ml LC for 15 min. When sperm were incubated for 4 h with heparin, exposure to 100 micrograms/ml LC for 15 min had no effect on the percentage of sperm that were motile, but the percentage of acrosome-reacted sperm increased from less than 10% to over 70%. The acrosome reactions (ARs) induced by LC were synchronous, reached maximal levels within 15 min, and differed (p less than 0.001) between sperm incubated under capacitating (with heparin) and noncapacitating conditions (without heparin). The time course required for heparin to capacitate sperm as judged by in vitro fertilization and to render sperm sensitive to LC induction of the AR were found to be similar. The percentage of ARs induced by LC and percentage of oocytes fertilized by sperm were found to be heparin-dose-dependent, with the maximum responses occurring at 5-10 micrograms/ml heparin. The correlation between the mean fertilization and LC-induced AR percentages was 0.997 (p less than 0.01). These studies demonstrate capacitation of bovine sperm by heparin requires at least a 4-h exposure of sperm to heparin and suggest that plasma membrane changes prior to an AR can be detected by exposure of bovine sperm to LC.
通过测定精子在体外使牛卵母细胞受精的能力以及在暴露于溶血磷脂酰胆碱(LC)时发生顶体反应的能力,对牛精子的获能情况进行了评估。用肝素(10微克/毫升)孵育精子可提高受精的卵母细胞百分比,但这需要在将精子添加到卵母细胞之前,先将精子暴露于肝素中至少4小时。将未获能的精子暴露于100微克/毫升的LC中15分钟后,对活动精子或发生顶体反应的精子百分比没有影响。当精子与肝素孵育4小时后,暴露于100微克/毫升的LC中15分钟,对活动精子的百分比没有影响,但发生顶体反应的精子百分比从不到10%增加到超过70%。LC诱导的顶体反应(ARs)是同步的,在15分钟内达到最高水平,并且在获能(用肝素)和未获能条件(不用肝素)下孵育的精子之间存在差异(p<0.001)。通过体外受精判断肝素使精子获能以及使精子对LC诱导的AR敏感所需的时间进程相似。发现LC诱导的AR百分比和精子使卵母细胞受精的百分比呈肝素剂量依赖性,最大反应出现在5-10微克/毫升的肝素浓度下。平均受精率与LC诱导的AR百分比之间的相关性为0.997(p<0.01)。这些研究表明,肝素使牛精子获能需要精子至少暴露于肝素4小时,并表明在AR之前的质膜变化可以通过将牛精子暴露于LC来检测。