Gamete Biology

Cholesterol Efflux Alters Lipid Raft Stability and Distribution During Capacitation of Boar Spermatozoa

Abstract
A reduction in plasma membrane cholesterol is one of the early events that either triggers, or is closely associated with, capacitation of mammalian spermatozoa. In this investigation we have examined the effects of cholesterol efflux on tyrosine phosphorylation, lipid diffusion and raft organisation in boar spermatozoa. Results show that a low level of cholesterol efflux, mediated by 5 mM methyl--cyclodextrin (MBCD), enhances capacitation and induces phosphorylation of two proteins at 26 kDa and 15 kDa without affecting sperm viability. Lipid diffusion rates under these conditions are largely unaffected except when cholesterol efflux is excessive. Low-density Triton X100-insoluble complexes (lipid rafts) were isolated from spermatozoa and found to have a restricted profile of proteins. Capacitation-associated cholesterol efflux has no effect on raft composition but cholesterol depletion destabilises them completely and phosphorylation is suppressed. During MBCD-mediated capacitation the distribution of GM1 gangliosides on spermatozoa changes in a sequential manner from overlying the sperm tail to clustering on the sperm head. It is concluded that there is a safe 'window' for removal of plasma membrane cholesterol from spermatozoa within which protein phosphorylation and polarised migration of lipid rafts take place. A preferential loss of cholesterol from the non-raft pool may be the stimulus that promotes raft clustering over the anterior sperm head.

Key words: Gamete Biology • In vitro fertilization • Sperm capacitation • Sperm maturation

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