title: Molecular diffusion in sperm plasma membranes during epididymal maturation.
abstract: Fluorescence recovery after photobleaching (FRAP) analysis has been used to measure lipid diffusion in different regions of the sperm plasma membrane. Our goal has been to understand how some membrane components are confined to specific surface domains, whilst others are freely diffusing and in some cases are able to migrate against large concentration gradients. Results with a variety of fluorescent lipid reporter probes (ODAF, NBD-PC, NBD-cholesterol) show that diffusion coefficients (D) are generally three to four times higher on the sperm acrosome than on the principal piece of the tail and increase significantly during epididymal maturation (ram, mouse, goat, dog and monkey sperm). Cholesterol diffusion is approximately 10 times faster on the sperm head than the tail and has a heterogenous distribution when detected with filipin. Lipid diffusion is very temperature sensitive but remarkably insensitive to changes in external pH and osmotic pressure. There was no evidence that the posterior ring or annulus functioned as diffusion barriers to lipids. On this basis it was possible to construct models of increasing complexity to describe the behaviour of a lipid molecule on the sperm surface, beginning with simple linear diffusion progressing to random diffusion and eventually to constrained diffusion.
authors.affiliation: Gamete Signalling Laboratory, The Babraham Institute, Cambridge CB2 4AT, UK.
authors.forename: Yonka
authors.initials: Y
authors.lastname: Christova
authors.forename: Peter
authors.initials: P
authors.lastname: James
authors.forename: Alan
authors.initials: A
authors.lastname: Mackie
authors.forename: Trevor G
authors.initials: TG
authors.lastname: Cooper
authors.forename: Roy
authors.initials: R
authors.lastname: Jones
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism; D004058:Diffusion; D004822:Epididymis / Q000166:cytology / Q000254:growth & development*; D036681:Fluorescence Recovery After Photobleaching; D005456:Fluorescent Dyes / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008297:Male; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism*; D008954:Models, Biological; D009997:Osmotic Pressure; D013094:Spermatozoa / Q000166:cytology* / Q000378:metabolism; D013696:Temperature
chemical_list: D005456:Fluorescent Dyes; D008563:Membrane Lipids; D002784:Cholesterol