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PMID:19144954

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Citation

Miranda, PV, Allaire, A, Sosnik, J and Visconti, PE (2009) Localization of low-density detergent-resistant membrane proteins in intact and acrosome-reacted mouse sperm. Biol. Reprod. 80:897-904

Abstract

Mammalian sperm become fertile after completing capacitation, a process associated with cholesterol loss and changes in the biophysical properties of the sperm membranes that prepares the sperm to undergo the acrosome reaction. Different laboratories have hypothesized that cholesterol efflux can influence the extent and/or movement of lipid raft microdomains. In a previous study, our laboratory investigated the identity of sperm proteins putatively associated with rafts. After extraction with Triton X-100 and ultracentrifugation in sucrose gradients, proteins distributing to the light buoyant-density fractions were cored from polyacrylamide gels and microsequenced. In this study, a subset of these proteins (TEX101, basigin, hexokinase 1, facilitated glucose transporter 3, IZUMO, and SPAM1) and other molecules known to be enriched in membrane rafts (caveolin 2, flotillin 1, flotillin 2, and the ganglioside GM3) were selected to investigate their localization in the sperm and their behavior during capacitation and the acrosome reaction. These molecules localize to multiple sperm domains, including the acrosomal cap (IZUMO, caveolin 2, and flotillin 2), equatorial segment (GM3), cytoplasmic droplet (TEX101), midpiece (basigin, facilitated glucose transporter 3, and flotillin 2), and principal piece (facilitated glucose transporter 3). Some of these markers modified their immunofluorescence pattern after sperm incubation under capacitating conditions, and these changes correlated with the occurrence of the acrosome reaction. While GM3 and caveolin 2 were not detected after the acrosome reaction, flotillin 2 was found in the equatorial segment of acrosome-reacted sperm, and IZUMO distributed along the sperm head, reaching the post- and para-acrosomal areas. Taking into consideration the requirement of the acrosome reaction for sperm to become fusogenic, these results suggest that membrane raft dynamics may have a role in sperm-egg membrane interaction.

Links

PubMed PMC2804839 Online version:10.1095/biolreprod.108.075242

Keywords

Acrosome Reaction/physiology; Animals; Blotting, Western; Caveolin 2/metabolism; Cell Compartmentation; Detergents; G(M3) Ganglioside/metabolism; Immunoglobulins/metabolism; Immunohistochemistry; Male; Membrane Microdomains/metabolism; Membrane Proteins/isolation & purification; Membrane Proteins/metabolism; Mice; Octoxynol; Spermatozoa/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MOUSE:FLOT1

located_in

GO:0045121: membrane raft

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:HXK1

located_in

GO:0045121: membrane raft

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:BASI

located_in

GO:0002080: acrosomal membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:GTR3

located_in

GO:0002080: acrosomal membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:HYALP

located_in

GO:0045121: membrane raft

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:FLOT2

located_in

GO:0002080: acrosomal membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:ADCYA

located_in

GO:0016020: membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:IZUM1

located_in

GO:0002080: acrosomal membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:TX101

located_in

GO:0002080: acrosomal membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:CAV2

located_in

GO:0002080: acrosomal membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:HXK1

part_of

GO:0045121: membrane raft

ECO:0000314: direct assay evidence used in manual assertion

C

part_of:(CL:0000019)

Seeded From UniProt

complete

MOUSE:IZUM1

part_of

GO:0002080: acrosomal membrane

ECO:0000314: direct assay evidence used in manual assertion

C

part_of:(CL:0000019)

Seeded From UniProt

complete

See also

References

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