Bicarbonate stimulated phospholipid scrambling induces cholesterol redistribution and enables cholesterol depletion in the sperm plasma membrane.
Mammalian sperm cells are activated prior to fertilization by high bicarbonate levels, which facilitate lipoprotein-mediated cholesterol efflux. The role of bicarbonate and cholesterol acceptors on the cholesterol organization in the sperm plasma membrane was tested. Bicarbonate induced an albumin-independent change in lipid architecture that was detectable by an increase in merocyanine staining (due to protein kinase A-mediated phospholipid scrambling). The response was limited to a subpopulation of viable sperm cells that were sorted from the non-responding subpopulation by flow cytometry. The responding cells had reduced cholesterol levels (30% reduction) compared with non-responding cells. The subpopulation differences were caused by variable efficiencies in epididymal maturation as judged by cell morphology. Membrane cholesterol organization was observed with filipin, which labeled the entire sperm surface of non-stimulated and non-responding cells, but labeled only the apical surface area of bicarbonate-responding cells. Addition of albumin caused cholesterol efflux, but only in bicarbonate-responding cells that exhibited virtually no filipin labeling in the sperm head area. Albumin had no effect on other lipid components, and no affinity for cholesterol in the absence of bicarbonate. Therefore, bicarbonate induces first a lateral redistribution in the low cholesterol containing spermatozoa, which in turn facilitates cholesterol extraction by albumin. A model is proposed in which phospholipid scrambling induces the formation of an apical membrane raft in the sperm head surface that enables albumin mediated efflux of cholesterol.
Flesch
F M
FM
Department of Biochemistry and Cell Biology, Utrecht University, Utrecht, 3584 CM, The Netherlands.
Brouwers
J F
JF
Nievelstein
P F
PF
Verkleij
A J
AJ
van Golde
L M
LM
Colenbrander
B
B
Gadella
B M
BM
D020101:Acrosome Reaction / Q000502:physiology; D000418:Albumins / Q000494:pharmacology; D000818:Animals; D001639:Bicarbonates / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D002470:Cell Survival; D002784:Cholesterol / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005372:Filipin; D005434:Flow Cytometry; D005614:Freeze Fracturing; D008051:Lipid Bilayers / Q000378:metabolism; D008297:Male; D021962:Membrane Microdomains / Q000378:metabolism; D010743:Phospholipids / Q000032:analysis / Q000378:metabolism*; D013075:Sperm Capacitation / Q000502:physiology; D013094:Spermatozoa / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D013552:Swine
D000418:Albumins; D001639:Bicarbonates; D008051:Lipid Bilayers; D010743:Phospholipids; D005372:Filipin; D002784:Cholesterol; D002118:Calcium