Isolation and characterization of sea urchin egg lipid rafts and their possible function during fertilization.

TitleIsolation and characterization of sea urchin egg lipid rafts and their possible function during fertilization.
Publication TypeJournal Article
Year of Publication2001
AuthorsBelton RJ, Adams NL, Foltz KR
JournalMol Reprod Dev
Volume59
Issue3
Pagination294-305
Date Published2001 Jul
ISSN1040-452X
KeywordsAnimals, beta-Cyclodextrins, Cell Fractionation, Cyclodextrins, Female, Fertilization, Immunoblotting, Isoenzymes, Male, Membrane Microdomains, Oocytes, Phospholipase C gamma, Sea Urchins, Signal Transduction, Spermatozoa, src-Family Kinases, Type C Phospholipases
Abstract

Specialized membrane microdomains called rafts are thought to play a role in many types of cell-cell interactions and signaling. We have investigated the possibility that sea urchin eggs contain these specialized membrane microdomains and if they play a role in signal transduction at fertilization. A low density, TX-100 insoluble membrane fraction, typical of lipid rafts, was isolated by equilibrium gradient centrifugation. This raft fraction contained proteins distinct from cytoskeletal complexes. The fraction was enriched in tyrosine phosphorylated proteins and contained two proteins known to be involved in signaling during egg activation (an egg Src-type kinase and PLC gamma). This fraction was further characterized as a prototypical raft fraction by the release of proteins in response to in vitro treatment of the rafts with the cholesterol binding drug, methyl-beta-cyclodextrin (M beta CD). Furthermore, treatment of eggs with M beta CD inhibited fertilization, suggesting that egg lipid rafts play a physiological role in fertilization. Mol. Reprod. Dev. 59:294-305, 2001.

DOI10.1002/mrd.1034
Alternate JournalMol. Reprod. Dev.
PubMed ID11424215
Grant ListHD30698 / HD / NICHD NIH HHS / United States