Identification of caveolin and caveolin-related proteins in the brain.
Caveolae are 50-100 nm, nonclathrin-coated, flask-shaped plasma membrane microdomains that have been identified in most mammalian cell types, except lymphocytes and neurons. To date, multiple functions have been ascribed to caveolae, including the compartmentalization of lipid and protein components that function in transmembrane signaling events, biosynthetic transport functions, endocytosis, potocytosis, and transcytosis. Caveolin, a 21-24 kDa integral membrane protein, is the principal structural component of caveolae. We have initiated studies to examine the relationship of detergent-insoluble complexes identified in astrocytes to the caveolin-caveolae compartment detected in cells of peripheral tissues. Immunolocalization studies performed in astrocytes reveal caveolin immunoreactivity in regions that correlate well to the distribution of caveolae and caveolin determined in other cell types, and electron microscopic studies reveal multiple clusters of flask-shaped invaginations aligned along the plasma membrane. Immunoblot analyses demonstrate that detergent-insoluble complexes isolated from astrocytes are composed of caveolin-1alpha, an identification verified by Northern blot analyses and by the cloning of a cDNA using reverse transcriptase-PCR amplification from total astrocyte RNA. Using a full-length caveolin-1 probe, Northern blot analyses suggest that the expression of caveolin-1 may be regulated during brain development. Immunoblot analyses of detergent-insoluble complexes isolated from cerebral cortex and cerebellum identify two immunoreactive polypeptides with apparent molecular weight and isoelectric points appropriate for caveolin. The identification of caveolae microdomains and caveolin-1 in astrocytes and brain, as well as the apparent regulation of caveolin-1 expression during brain development, identifies a cell compartment not detected previously in brain.
Cameron
P L
PL
Institute of Molecular Medicine and Genetics, Medical College of Georgia, Augusta, Georgia 30912-3175, USA.
Ruffin
J W
JW
Bollag
R
R
Rasmussen
H
H
Cameron
R S
RS
D000818:Animals; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D001253:Astrocytes / Q000737:chemistry* / Q000648:ultrastructure; D001692:Biological Transport / Q000502:physiology; D015152:Blotting, Northern; D001921:Brain / Q000166:cytology; D051242:Caveolin 1; D022461:Caveolins; D002451:Cell Compartmentation / Q000502:physiology; D002458:Cell Fractionation; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure*; D003902:Detergents; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D007425:Intracellular Membranes / Q000737:chemistry / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D008854:Microscopy, Electron; D008867:Microtomy; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D009836:Oligodendroglia / Q000737:chemistry* / Q000648:ultrastructure; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012995:Solubility
D000911:Antibodies, Monoclonal; C495537:Cav1 protein, rat; D051242:Caveolin 1; D022461:Caveolins; D003902:Detergents; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger