title: Biochemical and imaging methods to study receptor membrane organization and association with lipid rafts.
abstract: Lipid rafts, cell membrane domains with unique composition and properties, modulate the membrane distribution of receptors and signaling molecules facilitating the assembly of active signaling platforms. However, the underlying mechanisms that link signal transduction and lipid rafts are not fully understood, mainly because of the transient nature of these membrane assemblies. Several methods have been used to study the association of membrane receptors with lipid rafts. In the first part of this chapter, a description of how biochemical methods such as raft disruption by cholesterol depletion agents are useful in qualitatively establishing protein association with lipid rafts is presented. The second part of this chapter is dedicated to imaging techniques used to study membrane receptor organization and lipid rafts. We cover conventional approaches such as confocal microscopy to advanced imaging techniques such as homo-FRET microscopy and superresolution methods. For each technique described, their advantages and drawbacks are discussed.
authors.affiliation: ICFO-Institut de Ciencies Fotoniques, Mediterranean Technology Park, Castelldefels (Barcelona), Spain.
authors.forename: Bruno M
authors.initials: BM
authors.lastname: Castro
authors.forename: Juan A
authors.initials: JA
authors.lastname: Torreno-Piña
authors.forename: Thomas S
authors.initials: TS
authors.lastname: van Zanten
authors.forename: Maria F
authors.initials: MF
authors.lastname: Gracia-Parajo
mesh_terms: D002789:Cholesterol Oxidase / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D021962:Membrane Microdomains / Q000737:chemistry*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000295:instrumentation / Q000379:methods*; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000378:metabolism; D015398:Signal Transduction; D013194:Staining and Labeling; D047392:beta-Cyclodextrins / Q000737:chemistry
chemical_list: D043562:Receptors, G-Protein-Coupled; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D002789:Cholesterol Oxidase
keywords: Cholesterol oxidase; Homo-FRET; Methyl-β-cyclodextrin; NSOM; PALM; Protein cluster; Putative raft markers; Receptor oligomerization; STED; STORM