Membrane and cortical abnormalities in post-ovulatory aged eggs: analysis of fertilizability and establishment of the membrane block to polyspermy

Abstract

Fertilization at increased times after ovulation is associated with poor reproductive outcomes. This study examines the effects of post-ovulatory ageing on egg membrane function through analyses of mouse eggs collected at 13 and 22 h post-HCG ('young' and 'aged' eggs, respectively). Experiments in which fertilized zona pellucida-free young and aged eggs are challenged with additional sperm reveal that aged eggs are less able to establish a membrane block to prevent polyspermy, since sperm penetrate 24% of fertilized aged eggs but are unable to penetrate fertilized young eggs. This is not due to a failure of aged eggs to respond to fertilization, as the extent of sperm-induced cortical granule exocytosis is similar in aged and young eggs. Post-ovulatory ageing also affects egg membrane receptivity to sperm as a subset of zona pellucida-free aged eggs are slow to fertilize or resistant to fertilization. Sperm binding to young and aged eggs is similar, but aged eggs develop cytoskeletal abnormalities that may affect membrane/cortical function, such as the ability of the egg membrane to support sperm-egg fusion. These data demonstrate that the poor reproductive outcomes associated with post-ovulatory ageing could be a result of reduced fertilization, due to reduced egg membrane receptivity to sperm, or a result of increased incidence of polyspermy, due to the reduced ability to establish a membrane block to polyspermy. This analysis of egg membrane function deficiencies provides insights into post-ovulatory ageing and has implications for assisted reproductive technologies.