Epitope-dependent localization of estrogen receptor-alpha, but not -beta, in en face arterial endothelium.
Rapid, nongenomic effects of 17 beta-estradiol (E(2)) in endothelial cells are postulated to arise from membrane-associated estrogen receptors (ERs), which have not been visualized in vascular tissue. To identify membrane ERs, we used multiple site-directed ER alpha or ER beta antibodies to label en face rat cerebral and coronary arterial endothelia. Western blots revealed a novel 55-kDa ER alpha isoform. Three-dimensional images of cells labeled with these antibodies and markers for the nucleus and caveolin-1 were acquired with a wide-field microscope, deconvolved, and numerically analyzed. We found ER alpha in the nucleus and cell periphery, where one-third colocalized with caveolin-1. The receptor location was dependent on the epitope of the antibody. Human ovarian surface epithelium produced similar results; but in rat myometrium, the distribution was epitope independent and nuclear. ER beta distribution was predominantly intranuclear and epitope independent. A small amount of ER alpha colocalized with ER beta within the nucleus. The results were identical in both arterial preparations and insensitive to E(2). We postulate that the different ER alpha conformations at the membrane, in the nucleus, and between different cell types allow E(2) to trigger cell- and location-specific signaling cascades.
Dan
Pauline
P
Department of Physiology, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
Cheung
Joyce C Y
JC
Scriven
David R L
DR
Moore
Edwin D W
ED
D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D001488:Basilar Artery / Q000737:chemistry / Q000648:ultrastructure; D015153:Blotting, Western; D051242:Caveolin 1; D022461:Caveolins / Q000032:analysis; D002460:Cell Line; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000737:chemistry; D002467:Cell Nucleus / Q000737:chemistry; D002536:Cerebral Arteries / Q000737:chemistry / Q000648:ultrastructure; D003331:Coronary Vessels / Q000737:chemistry / Q000648:ultrastructure; D004730:Endothelium, Vascular / Q000737:chemistry*; D004847:Epithelial Cells; D000939:Epitopes / Q000032:analysis*; D004958:Estradiol / Q000097:blood / Q000494:pharmacology; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D005260:Female; D005455:Fluorescent Antibody Technique; D008077:Lipoproteins, LDL / Q000378:metabolism; D010052:Ovariectomy; D010053:Ovary; D011487:Protein Conformation; D051381:Rats; D017207:Rats, Sprague-Dawley; D011960:Receptors, Estrogen / Q000032:analysis* / Q000737:chemistry / Q000276:immunology
D000906:Antibodies; C495537:Cav1 protein, rat; D051242:Caveolin 1; D022461:Caveolins; D000939:Epitopes; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D008077:Lipoproteins, LDL; D011960:Receptors, Estrogen; C032629:acetyl-LDL; D004958:Estradiol