Polyspermy inhibition in the oyster, Crassostrea virginica

Abstract

Inhibition of polyspermy is a critical response during fertilization which ensures that only one sperm nucleus will fuse with the female pronucleus to restore the diploid state. Oyster (Crassostrea virginica) eggs prevent polyspermy by a process occurring at the cell surface. However, 5 min after fertilization, there are still functional sperm receptor sites available for penetration by supernumerary sperm. Reinsemination experiments indicate that there is no decrease in the number of penetration sites during this interval. Yet, the number of sperm entering eggs is restricted to one per fertilized egg at a sperm:egg ratio of 1000:1. At a sperm:egg ratio of 10(5):1, an average of only six sperm were able to penetrate each egg. Gamete binding experiments indicate that there is a gradual decrease in the number of sperm bound to eggs starting at approximately 75 sec and continuing until all sperm are detached. Since eggs are fertilized within seconds of insemination and this process takes at least 12 min, it is considered an unlikely mechanism for the polyspermy block. There are no ultrastructural correlates to the polyspermy block nor to the unbinding of sperm, such as secretion of cortical granules or fertilization envelope formation. Based on reinsemination experiments, kinetic data, and ultrastructural observations, we suggest a physiological block to polyspermy which prevents fusion of gamete plasma membranes, and is mediated by an inhibitory effect directly upon the sperm.