Abstract
Cholesterol is an essential component of mammalian cells. It is the major lipid constituent of the plasma membrane and is also abundant in most other organelle membranes. In the plasma membrane cholesterol plays critical physical roles in the maintenance of membrane fluidity and membrane permeability. It is also important for membrane trafficking, cell signalling, and lipid as well as protein sorting. Cholesterol is essential for the formation of liquid ordered domains in model membranes, which in cells are known as lipid nanodomains or lipid rafts. Cholesterol depletion is widely used to study the role of cholesterol in cellular processes and can be performed over days using inhibitors of its synthesis or acutely over minutes using chemical reagents. Acute cholesterol depletion by methyl-β-cyclodextrin (MBCD) is the most widely used method and here we describe how it should be performed to avoid the common side-effect cell death.
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References
Maxfield FR, van Meer G (2010) Cholesterol, the central lipid of mammalian cells. Curr Opin Cell Biol 22:422–429
Huang J, Feigenson GW (1999) A microscopic interaction model of maximum solubility of cholesterol in lipid bilayers. Biophys J 76:2142–2157
Mahammad S, Parmryd I (2008) Cholesterol homeostasis in T cells. Methyl-beta-cyclodextrin treatment results in equal loss of cholesterol from Triton X-100 soluble and insoluble fractions. Biochim Biophys Acta 1778:1251–1258
Taraboulos A, Scott M, Semenov A, Avrahami D, Laszlo L et al (1995) Cholesterol depletion and modification of COOH-terminal targeting sequence of the prion protein inhibit formation of the scrapie isoform. J Cell Biol 129:121–132
Esfahani M, Bigler RD, Alfieri JL, Lund-Katz S, Baum JD et al (1993) Cholesterol regulates the cell surface expression of glycophospholipid-anchored CD14 antigen on human monocytes. Biochim Biophys Acta 1149:217–223
Nazih-Sanderson F, Pinchon G, Nion S, Fruchart JC, Delbart C (1997) HDL3-signalling in HepG2 cells involves glycosyl-phosphatidylinositol-anchored proteins. Biochim Biophys Acta 1346:45–60
Cerneus DP, Ueffing E, Posthuma G, Strous GJ, van der Ende A (1993) Detergent insolubility of alkaline phosphatase during biosynthetic transport and endocytosis. Role of cholesterol. J Biol Chem 268:3150–3155
Nguyen DH, Taub DD (2003) Inhibition of chemokine receptor function by membrane cholesterol oxidation. Exp Cell Res 291:36–45
Nishijo J, Moriyama S, Shiota S (2003) Interactions of cholesterol with cyclodextrins in aqueous solution. Chem Pharm Bull (Tokyo) 51:1253–1257
Loftsson T, Magnusdottir A, Masson M, Sigurjonsdottir JF (2002) Self-association and cyclodextrin solubilization of drugs. J Pharm Sci 91:2307–2316
Behnke O, Tranum-Jensen J, van Deurs B (1984) Filipin as a cholesterol probe. II. Filipin-cholesterol interaction in red blood cell membranes. Eur J Cell Biol 35:200–215
Lopez CA, de Vries AH, Marrink SJ (2011) Molecular mechanism of cyclodextrin mediated cholesterol extraction. PLoS Comput Biol 7:e1002020
Leventis R, Silvius JR (2001) Use of cyclodextrins to monitor transbilayer movement and differential lipid affinities of cholesterol. Biophys J 81:2257–2267
Davis ME, Brewster ME (2004) Cyclodextrin-based pharmaceutics: past, present and future. Nat Rev Drug Discov 3:1023–1035
van de Manakker F, Vermonden T, van Nostrum CF, Hennink WE (2009) Cyclodextrin-based polymeric materials: synthesis, properties, and pharmaceutical/biomedical applications. Biomacromolecules 10:3157–3175
Li Z, Wang M, Wang F, Gu Z, Du G et al (2007) Gamma-cyclodextrin: a review on enzymatic production and applications. Appl Microbiol Biotechnol 77:245–255
Ohtani Y, Irie T, Uekama K, Fukunaga K, Pitha J (1989) Differential effects of alpha-, beta- and gamma-cyclodextrins on human erythrocytes. Eur J Biochem 186:17–22
Puglisi G, Ventura CA, Spadaro A, Campana G, Spampinato S (1995) Differential effects of modified beta-cyclodextrins on pharmacological activity and bioavailability of 4-biphenylacetic acid in rats after oral administration. J Pharm Pharmacol 47:120–123
Ohvo H, Slotte JP (1996) Cyclodextrin-mediated removal of sterols from monolayers: effects of sterol structure and phospholipids on desorption rate. Biochemistry 35:8018–8024
Levitan I, Christian AE, Tulenko TN, Rothblat GH (2000) Membrane cholesterol content modulates activation of volume-regulated anion current in bovine endothelial cells. J Gen Physiol 115:405–416
Christian AE, Haynes MP, Phillips MC, Rothblat GH (1997) Use of cyclodextrins for manipulating cellular cholesterol content. J Lipid Res 38:2264–2272
Mahammad S, Dinic J, Adler J, Parmryd I (2010) Limited cholesterol depletion causes aggregation of plasma membrane lipid rafts inducing T cell activation. Biochim Biophys Acta 1801:625–634
Cox BE, Griffin EE, Ullery JC, Jerome WG (2007) Effects of cellular cholesterol loading on macrophage foam cell lysosome acidification. J Lipid Res 48:1012–1021
Nguyen DH, Espinoza JC, Taub DD (2004) Cellular cholesterol enrichment impairs T cell activation and chemotaxis. Mech Ageing Dev 125:641–650
Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST et al (1994) Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84:1415–1420
Acknowledgements
This work was supported by grants from Magnus Bergvall’s Foundation, Signhild Engkvist’s Foundation, and O.E. and Edla Johansson’s Foundation.
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Mahammad, S., Parmryd, I. (2015). Cholesterol Depletion Using Methyl-β-cyclodextrin. In: Owen, D. (eds) Methods in Membrane Lipids. Methods in Molecular Biology, vol 1232. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1752-5_8
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DOI: https://doi.org/10.1007/978-1-4939-1752-5_8
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