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PMID: 19012288 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Mechanisms underlying the micron-scale segregation of sterols and GM1 in live mammalian sperm.

Journal of cellular physiology ·Vol. 218 ·No. 3 ·2009-03-00 ·Pages 522-36

Selvaraj V, Asano A, Buttke DE, Sengupta P, Weiss RS, Travis AJ

Abstract

We demonstrate for the first time that a stable, micron-scale segregation of focal enrichments of sterols exists at physiological temperature in the plasma membrane of live murine and human sperm. These enrichments of sterols represent microheterogeneities within this membrane domain overlying the acrosome. Previously, we showed that cholera toxin subunit B (CTB), which binds the glycosphingolipid, G(M1), localizes to this same domain in live sperm. Interestingly, the G(M1) undergoes an unexplained redistribution upon cell death. We now demonstrate that G(M1) is also enriched in the acrosome, an exocytotic vesicle. Transfer of lipids between this and the plasma membrane occurs at cell death, increasing G(M1) in the plasma membrane without apparent release of acrosomal contents. This finding provides corroborative support for an emerging model of regulated exocytosis in which membrane communications might occur without triggering the "acrosome reaction." Comparison of the dynamics of CTB-bound endogenous G(M1) and exogenous BODIPY-G(M1) in live murine sperm demonstrate that the sub-acrosomal ring (SAR) functions as a specialized diffusion barrier segregating specific lipids within the sperm head plasma membrane. Our data show significant differences between endogenous lipids and exogenous lipid probes in terms of lateral diffusion. Based on these studies, we propose a hierarchical model to explain the segregation of this sterol- and G(M1)-enriched domain in live sperm, which is positioned to regulate sperm fertilization competence and mediate interactions with the oocyte. Moreover, our data suggest potential origins of subtypes of membrane raft microdomains enriched in sterols and/or G(M1) that can be separated biochemically.

MeSH Terms
Acrosome/metabolism,ultrastructure Animals Annexin A5/metabolism Boron Compounds Cell Death Cell Membrane/ultrastructure Cell Survival Cholera Toxin/metabolism Cytoskeleton/metabolism Diffusion G(M1) Ganglioside/metabolism Humans Male Mammals/metabolism Mice Protein Transport Spermatids/cytology,metabolism Spermatozoa/cytology,metabolism,ultrastructure Staining and Labeling Sterols/metabolism
Chemicals
4,4-difluoro-4-bora-3a,4a-diaza-s-indacene Annexin A5 Boron Compounds Sterols G(M1) Ganglioside Cholera Toxin
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Selvaraj Vimal
Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853, USA.
Asano Atsushi
Buttke Danielle E
Sengupta Prabuddha
Weiss Robert S
Travis Alexander J
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Article Info
Journal
Journal of cellular physiology
Abbr.
J Cell Physiol
ISSN
1097-4652
DOI
10.1002/jcp.21624
Published
2009-03-00
Pages
522-36
Language
English
Region
United States
NLM ID
0050222
PMCID
PMC2692964
Subset
IM
Grants
NICHD NIH HHS · R01 HD045664 · United States
NICHD NIH HHS · R01 HD045664-05 · United States
NICHD NIH HHS · R01-HD-045664 · United States
External Links
PubMed source DOI: 10.1002/jcp.21624 PMC Full Text View journal details
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