Use of Bodipy-labeled sphingolipid and cholesterol analogs to examine membrane microdomains in cells.
Much evidence has accumulated to show that cellular membranes such as the plasma membrane, contain multiple "microdomains" of differing lipid and protein composition and function. These domains are sometimes enriched in cholesterol and sphingolipids and are believed to be important structures for the regulation of many biological and pathological processes. This review focuses on the use of fluorescent (Bodipy) labeled analogs of sphingolipids and cholesterol to study such domains. We discuss the similarities between the behavior of Bodipy-cholesterol and natural cholesterol in artificial bilayers and in cultured cells, and the use of Bodipy-sphingolipid analogs to visualize membrane domains in living cells based on the concentration-dependent monomer-excimer fluorescence properties of the Bodipy-fluorophore. The use of Bodipy-D-erythro-lactosylceramide is highlighted for detection of domains on the plasma membrane and endosome membranes, and the importance of the sphingolipid stereochemistry in modulating domain formation is discussed. Finally, we suggest that Bodipy-sphingolipids may be useful in future studies to examine the relationship between membrane domains at the cell surface and domains enriched in other lipids and proteins on the inner leaflet of the plasma membrane.
Marks
David L
DL
Thoracic Diseases Research Unit, Department of Medicine, Mayo Clinic College of Medicine, Stabile 8, 200 First Street, SW, Rochester, MN 55905-0001, USA.
Bittman
Robert
R
Pagano
Richard E
RE
D000818:Animals; D001896:Boron Compounds / Q000737:chemistry*; D002784:Cholesterol / Q000031:analogs & derivatives / Q000032:analysis*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007425:Intracellular Membranes / Q000737:chemistry; D021962:Membrane Microdomains / Q000737:chemistry*; D008567:Membranes, Artificial; D008856:Microscopy, Fluorescence / Q000379:methods*; D013107:Sphingolipids / Q000032:analysis*
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D005456:Fluorescent Dyes; D008567:Membranes, Artificial; D013107:Sphingolipids; D002784:Cholesterol