Binding of both acrosome-intact and acrosome-reacted guinea pig sperm to the zona pellucida during in vitro fertilization.

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.