Antibody-specific detection of caveolin-1 in subapical compartments of MDCK cells.
Caveolin-1 is the major structural component of caveolae and is also found in the Golgi complex of many cell types. Occasionally, caveolin-1 has been observed in additional intracellular compartments, including recycling endosomes. Why caveolin-1 expression is detected at these sites only infrequently is not clear. In this study, we test the hypothesis that non-caveolar, non-Golgi pools of caveolin-1 display unique and/or fixation-dependent epitopes. We compared the ability of a panel of antibodies raised against various domains of caveolin-1 to detect distinct subcellular pools of the protein by immunofluorescence microscopy in Madin-Darby canine kidney (MDCK) cells, a cell line where the subcellular localization of caveolin-1 has been extensively characterized. We show that three antibodies directed to the N-terminus of caveolin-1 recognize a previously undetected pool of caveolin-1 in the subapical region of MDCK cells, a localization characteristic of endosomal recycling compartments. The antibodies vary in their ability to label caveolin-1 at the cell surface, and the epitopes detected by each are highly fixation dependent. Our findings suggest that no single caveolin antibody or staining condition is capable of detecting all the caveolin-1 in a cell simultaneously. Consequently, the subcellular distribution of caveolin-1 may be much broader than currently believed.
Bush
William S
WS
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, 718 Light Hall, Nashville, TN 37232, USA.
Ihrke
Gudrun
G
Robinson
John M
JM
Kenworthy
Anne K
AK
D000818:Animals; D051242:Caveolin 1 / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004285:Dogs; D011992:Endosomes / Q000378:metabolism; D000939:Epitopes / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000378:metabolism; D007150:Immunohistochemistry; D007668:Kidney / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D011956:Receptors, Cell Surface / Q000378:metabolism; D016707:Tissue Fixation
D051242:Caveolin 1; D000939:Epitopes; D011956:Receptors, Cell Surface