Changes in lipid diffusibility in the hamster sperm head plasma membrane during capacitation in vivo and in vitro.
The technique of fluorescence recovery after photobleaching (FRAP) was employed on spermatozoa labeled with the fluorescent lipid analogue C14dil to provide two measures of lateral diffusion in the plane of the sperm plasma membrane during capacitation in vivo and in vitro: the diffusion coefficient (D) for C14dil and the fraction of C14dil that is free to diffuse (%R) within the domain. To evaluate changes in lipid diffusibility during capacitation in vivo, spermatozoa were recovered from the uterus within 30 min after ejaculation or from the oviduct at 2, 4, 6 and 8 hr after mating. To compare the changes which occur in vivo with those which occur during capacitation in vitro, caudal epididymal spermatozoa were incubated under capacitating or non-capacitating (control) conditions for 4 hr. Although transient changes in D occurred during the course of capacitation, there was no net change in D for either anterior (AH) or posterior head (PH) domains following capacitation in vitro or in vivo. Significant differences in the lipid diffusion coefficient between the two head domains were observed during the course of capacitation. A transient decrease in %R was observed for the AH domain during capacitation in vitro and incubation under control conditions, but no significant change in %R was observed in the AH domain during capacitation in vivo. A significant decline in %R of the PH domain was observed for spermatozoa during capacitation in vivo, in vitro and following incubation under non-capacitating conditions. These data indicate that the changes in the lipid diffusibility of the AH and PH domains which occur during capacitation in vivo exhibit both similarities and differences to those which occur during capacitation in vitro.
Smith
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Department of Obstetrics and Gynecology, University of Kentucky, Lexington 40536-0084, USA.
McKinnon-Thompson
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Wolf
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D000818:Animals; D002232:Carbocyanines / Q000378:metabolism*; D002462:Cell Membrane; D006224:Cricetinae; D004058:Diffusion; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism*; D050356:Lipid Metabolism; D008297:Male; D008647:Mesocricetus; D013075:Sperm Capacitation / Q000502:physiology*; D013077:Sperm Head / Q000378:metabolism*
D002232:Carbocyanines; D005456:Fluorescent Dyes; C055316:ditetradecylindocarbocyanine