title: The neck of caveolae is a distinct plasma membrane subdomain that concentrates insulin receptors in 3T3-L1 adipocytes.
abstract: Insulin receptors (IRs) segregate on plasma membrane microvilli, but in cells devoid of microvilli, such as adipocytes, the localization of IRs is a matter of controversy. In the present study, we examined the distribution of IRs in the plasma membrane of 3T3-L1 adipocytes. Quantitative electron microscopy indicates that IRs are predominantly associated with the neck, but not the bulb, of caveolae. Caveola necks represent distinct microdomains of the plasma membrane. Indeed, as shown by freeze-fracture analysis, intramembrane particles are concentrated as necklaces around the craters of caveolae. In addition, subcellular fractionation suggests that the neck and the bulb of caveolae present a different resistance to detergent solubility. Finally, cytoskeletal components, including actin, are highly enriched in the membrane area underlying the neck part of caveolae. IRs coimmunoprecipitate with cytoskeletal components, and disruption of the actin cytoskeleton alters IRs expression, localization, and signaling, thus supporting the notion that caveola necks are involved in intracellular signaling by IRs. Together, these results suggest that cytoskeletal proteins anchor IRs to microdomains in the caveola necks of 3T3-L1 adipocytes. By homology with IR localization in other cell types, we suggest that the necks of caveolae may represent the counterpart of microvillar domains in cells poor in microvilli such as adipocytes and that they play an important role as signaling platforms.
authors.affiliation: Department of Cellular Physiology and Metabolism, Faculty of Medicine, University of Geneva, CH-1211 Geneva, Switzerland. Michelangelo.Foti@medecine.unige.ch
authors.forename: Michelangelo
authors.initials: M
authors.lastname: Foti
authors.forename: Geneviève
authors.initials: G
authors.lastname: Porcheron
authors.forename: Margot
authors.initials: M
authors.lastname: Fournier
authors.forename: Christine
authors.initials: C
authors.lastname: Maeder
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Carpentier
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000378:metabolism; D047468:Immunoprecipitation; D007328:Insulin / Q000378:metabolism; D051379:Mice; D046529:Microscopy, Electron, Transmission; D011485:Protein Binding; D011972:Receptor, Insulin / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D003598:Cytoskeletal Proteins; D007328:Insulin; D011972:Receptor, Insulin