title: Cholesterol depletion mimics the effect of cytoskeletal destabilization on membrane dynamics of the serotonin1A receptor: A zFCS study.
abstract: Single-point fluorescence correlation spectroscopy (FCS) of membrane-bound molecules suffers from a number of limitations leading to inaccurate estimation of diffusion parameters. To overcome such problems and with the overall goal of addressing membrane heterogeneities, we performed z-scan FCS (zFCS) of the serotonin(1A) receptor. We analyzed the results according to FCS diffusion laws that provide information on the organization of the diffusing species. Analysis of our results shows that the diffusion coefficients of the receptor and a fluorescently labeled phospholipid are similar when probed at length scales approximately 210 nm. We discuss the significance of the spatiotemporal evolution of dynamics of membrane-bound molecules in the overall context of membrane domains and heterogeneity. Importantly, our results show that the serotonin(1A) receptor exhibits confinement in cell membranes, possibly due to interaction with the actin cytoskeleton. Surprisingly, depletion of membrane cholesterol appears to reduce receptor confinement in a manner similar to that observed in the case of cytoskeletal destabilization, implying possible changes in the actin cytoskeleton induced upon cholesterol depletion. These results constitute the first report on G-protein-coupled receptor dynamics utilizing a combination of zFCS and the FCS diffusion laws, and present a convenient approach to explore cell membrane heterogeneity at the submicron level.
authors.affiliation: Centre for Cellular and Molecular Biology, Council of Scientific and Industrial Research, Hyderabad, India.
authors.forename: Sourav
authors.initials: S
authors.lastname: Ganguly
authors.forename: Amitabha
authors.initials: A
authors.lastname: Chattopadhyay
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D001896:Boron Compounds / Q000378:metabolism; D016466:CHO Cells; D002138:Calibration; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002784:Cholesterol / Q000172:deficiency* / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D004058:Diffusion; D044282:Receptor, Serotonin, 5-HT1A / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012701:Serotonin / Q000494:pharmacology; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D000199:Actins; D001896:Boron Compounds; D011993:Recombinant Fusion Proteins; D044282:Receptor, Serotonin, 5-HT1A; D012701:Serotonin; D002784:Cholesterol
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