Membrane fluidity changes in goat sperm induced by cholesterol depletion using beta-cyclodextrin.
Cholesterol efflux from membranes promotes acrosome reaction in goat spermatozoa. In 1 h of incubation of sperm in the presence of beta-cyclodextrin (beta CD), all the interchangeable cholesterol is desorbed from sperm membranes, although acrosome reaction is fully accomplished only after 3-4 h of incubation, as previously published. In the present paper we investigate the effect of cholesterol removal from mature goat spermatozoa on the overall membrane "fluidity" of live cell membranes and of liposomes from sperm lipid extracts. Using steady state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene (DPH), we studied the average thermotropic behaviour of membrane lipids, after incubation of live sperm for 1 h in BSA-free medium with the presence/absence of 8 mM beta-cyclodextrin, as a cholesterol acceptor. Unimodal and bimodal theoretical sigmoids fitted best to the experimental thermotropic profiles of liposomes and whole cells, respectively. In the case of whole sperm, two phase transitions, attributable to different lipid domains, were clearly separated by using the fitting parameters. After cholesterol removal, important changes in the relative anisotropy range of the two transitions were found, indicating an increase in the "fluidity" of some of the lipid microdomains of sperm membranes. These changes in sperm lipid dynamics are produced before the onset of sperm acrosome reaction.
Unitat de Biofísica, Departament de Bioquímica i de Biologia Molecular, Facultat de Medicina, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.
Mònica
M
Companyó
Antoni
A
Iborra
Joaquim
J
Villaverde
Paz
P
Martínez
Antoni
A
Morros
D020101:Acrosome Reaction / Q000187:drug effects / Q000502:physiology*; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D006041:Goats; D008297:Male; D008560:Membrane Fluidity / Q000187:drug effects / Q000502:physiology*; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*; D047392:beta-Cyclodextrins / Q000008:administration & dosage*
D047392:beta-Cyclodextrins; D002784:Cholesterol; C031215:betadex