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

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....

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Bibliographic Details
Published in:Molecular reproduction and development 2001-07, Vol.59 (3), p.294-305
Main Authors: Belton Jr, Robert J., Adams, Nikki L., Foltz, Kathy R.
Format: Article
Language:English
Subjects:
Animals
beta-Cyclodextrins
Biological and medical sciences
Cell Fractionation
Cell physiology
Cyclodextrins - pharmacology
Echinoidea
Female
Fertilization - drug effects
Fertilization - physiology
Fundamental and applied biological sciences. Psychology
Immunoblotting
Isoenzymes - metabolism
Male
membrane microdomains
Membrane Microdomains - chemistry
Membrane Microdomains - enzymology
Membrane Microdomains - metabolism
Molecular and cellular biology
Oocytes - chemistry
Oocytes - metabolism
Oocytes - ultrastructure
Phospholipase C gamma
PLCγ
Sea Urchins - physiology
Signal transduction
Signal Transduction - physiology
Spermatozoa - metabolism
Src
src-Family Kinases - metabolism
Type C Phospholipases - metabolism
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Summary: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 PLCγ). 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‐β‐cyclodextrin (MβCD). Furthermore, treatment of eggs with MβCD inhibited fertilization, suggesting that egg lipid rafts play a physiological role in fertilization. Mol. Reprod. Dev. 59:294–305, 2001. © 2001 Wiley‐Liss, Inc.
ISSN:1040-452X
1098-2795
DOI:10.1002/mrd.1034