title: Cholesterol depletion modulates detergent resistant fraction of human serotonin(1A) receptors.
abstract: Insolubility of membrane components in non-ionic detergents such as Triton X-100 at low temperature is a widely used biochemical criterion to identify, isolate and characterize membrane domains. In this work, we monitored the detergent insolubility of the serotonin(1A) receptor in CHO cell membranes and its modulation by membrane cholesterol. The serotonin(1A) receptor is an important member of the G-protein coupled receptor family. It is implicated in the generation and modulation of various cognitive, behavioral and developmental functions and serves as a drug target. Our results show that a significant fraction (∼28%) of the serotonin(1A) receptor resides in detergent-resistant membranes (DRMs). Interestingly, the fraction of the serotonin(1A) receptor in DRMs exhibits a reduction upon membrane cholesterol depletion. In addition, we show that contents of DRM markers such as flotillin-1, caveolin-1 and GM₁ are altered in DRMs upon cholesterol depletion. These results assume significance since the function of the serotonin(1A) receptor has previously been shown to be affected by membrane lipids, specifically cholesterol. Our results are relevant in the context of membrane organization of the serotonin(1A) receptor in particular, and G-protein coupled receptors in general.
authors.affiliation: Centre for Cellular and Molecular Biology, Council of Scientific and Industrial Research, Hyderabad, India.
authors.forename: Santosh Kumar
authors.initials: SK
authors.lastname: Sahu
authors.forename: Roopali
authors.initials: R
authors.lastname: Saxena
authors.forename: Amitabha
authors.initials: A
authors.lastname: Chattopadhyay
mesh_terms: D000818:Animals; D016466:CHO Cells; D051242:Caveolin 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002784:Cholesterol / Q000737:chemistry* / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D003902:Detergents / Q000737:chemistry*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017830:Octoxynol / Q000737:chemistry*; D044282:Receptor, Serotonin, 5-HT1A / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D051242:Caveolin 1; D003902:Detergents; D008565:Membrane Proteins; C106354:flotillins; D044282:Receptor, Serotonin, 5-HT1A; D017830:Octoxynol; D002784:Cholesterol