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P2X 3 receptor localizes into lipid rafts in neuronal cells
Author(s) -
Vacca Fabrizio,
Amadio Susanna,
Sancesario Giuseppe,
Bernardi Giorgio,
Volonté Cinzia
Publication year - 2004
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.20069
Subject(s) - lipid raft , microbiology and biotechnology , receptor , transfection , biology , ganglioside , biochemistry , cell fractionation , membrane , gene
P2X receptors are a family of seven (P2X 1–7 ) cation channels gated by extracellular ATP, widely expressed in neurons and nonneuronal cells. Lipid rafts are cholesterol/sphingolipid‐rich membrane domains, involved in many cellular processes, including transmembrane receptor signaling, vesicle traffic, and protein sorting. We provide direct biochemical evidence that P2X 3 receptor localizes into lipid rafts, in primary cultures of cerebellar granule neurons as well as in brain and dorsal root ganglia extracts. We show that P2X 3 exhibits all the characteristics distinctive of a protein associated with lipid rafts. These characteristics include resistance to detergent extraction at 4°C, solubility after extraction of cholesterol from membranes with either saponin or methyl‐β‐cyclodextrin, and partitioning to low buoyant density fractions after sucrose gradient centrifugation in both detergent‐containing and detergent‐free conditions. Furthermore, P2X 3 localizes in raft‐containing fractions in transiently transfected SH‐SY5Y neuroblastoma cells. The present finding contributes to the characterization of the functional localization of P2X 3 in neurons and provides a novel potential mechanism for correct targeting and dynamic activation of this receptor. © 2004 Wiley‐Liss, Inc.