title: Macroscopic domain formation during cooling in the platelet plasma membrane: an issue of low cholesterol content.
abstract: There has been ample debate on whether cell membranes can present macroscopic lipid domains as predicted by three-component phase diagrams obtained by fluorescence microscopy. Several groups have argued that membrane proteins and interactions with the cytoskeleton inhibit the formation of large domains. In contrast, some polarizable cells do show large regions with qualitative differences in lipid fluidity. It is important to ask more precisely, based on the current phase diagrams, under what conditions would large domains be expected to form in cells. In this work we study the thermotropic phase behavior of the platelet plasma membrane by FTIR, and compare it to a POPC/Sphingomyelin/Cholesterol model representing the outer leaflet composition. We find that this model closely reflects the platelet phase behavior. Previous work has shown that the platelet plasma membrane presents inhomogeneous distribution of DiI18:0 at 24 degrees C, but not at 37 degrees C, which suggests the formation of macroscopic lipid domains at low temperatures. We show by fluorescence microscopy, and by comparison with published phase diagrams, that the outer leaflet model system enters the macroscopic domain region only at the lower temperature. In addition, the low cholesterol content in platelets ( approximately 15 mol%), appears to be crucial for the formation of large domains during cooling.
authors.affiliation: Department of Molecular and Cellular Biology, University of California, Davis, CA, USA.
authors.forename: Rachna
authors.initials: R
authors.lastname: Bali
authors.forename: Laura
authors.initials: L
authors.lastname: Savino
authors.forename: Diego A
authors.initials: DA
authors.lastname: Ramirez
authors.forename: Nelly M
authors.initials: NM
authors.lastname: Tsvetkova
authors.forename: Luis
authors.initials: L
authors.lastname: Bagatolli
authors.forename: Fern
authors.initials: F
authors.lastname: Tablin
authors.forename: John H
authors.initials: JH
authors.lastname: Crowe
authors.forename: Chad
authors.initials: C
authors.lastname: Leidy