Myles D G, Hyatt H, Primakoff P
Dev Biol. 1987 Jun;121(2):559-67. doi: 10.1016/0012-1606(87)90191-6.
Mammalian sperm-egg adhesion occurs when sperm bind to the zona pellucida of the egg. In this study with guinea pig gametes, we have asked if sperm can initiate binding to the zona before and after the acrosome reaction and if the sperm surface protein PH-20 is involved in the binding at these two stages. Sperm binding to the zona was examined under a variety of conditions. Sperm were suspended in 0.9% NaCl or capacitated by two different methods. Eggs were immobilized on lectin-coated coverslips, compressed between a coverslip and a glass slide, or free in tissue culture dishes. The sperm-egg interaction was recorded on videotape or assessed after fixation of the eggs with bound sperm. Under all conditions studied, both acrosome-intact and acrosome-reacted sperm initiated binding to the zona. The binding was persistent and not transitory. In particular, acrosome-intact sperm that bound the zona were observed to remain bound for up to 80 min. One acrosome-intact sperm, bound to the zona, was videotaped while it acrosome-reacted. When mixed sperm populations (on the average 24% acrosome-intact and 76% acrosome-reacted) were incubated with eggs for 30 min, an average of 10% of the bound sperm were acrosome-intact. The PH-20 monoclonal antibody has previously been shown to inhibit zona binding by guinea pig sperm of undetermined acrosomal status (P. Primakoff, H. Hyatt, and D. G. Myles (1985), J. Cell Biol. 101, 2239-2244). In this study, when the two populations of sperm were counted separately, PH-20 inhibited the binding of acrosome-reacted but not acrosome-intact sperm. Our results show that both acrosome-intact and acrosome-reacted guinea pig sperm can initiate binding to the zona; however, the binding in the two cases may not occur by the same mechanism.
当精子与卵子的透明带结合时，哺乳动物的精卵黏附就会发生。在这项针对豚鼠配子的研究中，我们探讨了精子在顶体反应前后是否能够启动与透明带的结合，以及精子表面蛋白PH-20是否在这两个阶段的结合过程中发挥作用。我们在多种条件下检测了精子与透明带的结合情况。精子被悬浮于0.9%的氯化钠溶液中，或者通过两种不同的方法进行获能处理。卵子被固定在凝集素包被的盖玻片上，夹在盖玻片和载玻片之间，或者自由放置于组织培养皿中。精卵相互作用过程被录制在录像带上，或者在卵子与结合的精子固定后进行评估。在所有研究的条件下，顶体完整的精子和顶体反应后的精子均能启动与透明带的结合。这种结合是持续的而非短暂的。特别地，观察到顶体完整的精子与透明带结合后能够持续结合长达80分钟。有一个顶体完整且与透明带结合的精子在发生顶体反应时被录像记录下来。当混合的精子群体（平均24%顶体完整，76%顶体反应）与卵子孵育30分钟后，平均10%的结合精子是顶体完整的。之前已经证明，PH-20单克隆抗体能够抑制顶体状态未知的豚鼠精子与透明带的结合（P. Primakoff、H. Hyatt和D. G. Myles（1985年），《细胞生物学杂志》101卷，2239 - 2244页）。在本研究中，当分别对这两种精子群体进行计数时，PH-20抑制了顶体反应后的精子与透明带的结合，但对顶体完整的精子没有影响。我们的结果表明，顶体完整的和顶体反应后的豚鼠精子均能启动与透明带的结合；然而，这两种情况下的结合机制可能并不相同。
