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Am J Physiol Heart Circ Physiol (December 12, 2002). doi:10.1152/ajpheart.00781.2002
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Articles in PresS, published online ahead of print December 12, 2002
Am J Physiol Heart Circ Physiol, 10.1152/ajpheart.00781.2002
Submitted on September 5, 2002
Accepted on December 4, 2002

Epitope Dependent Localization of Estrogen Receptor {alpha}, but not {beta}, in En Face Arterial Endothelium

Pauline Dan1, Joyce C. Cheung1, David R. Scriven1, and Edwin D. Moore1*

1 Department of Physiology, University of British Columbia, Vancouver, British Columbia, Canada

* To whom correspondence should be addressed. E-mail: edmoore{at}interchange.ubc.ca.

Rapid, non-genomic, effects of 17{beta}-estradiol (E2) in endothelial cells are postulated to arise from membrane-associated estrogen receptors (ER), 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 the cell periphery, where one-third colocalized with caveolin-1. The receptor location was dependent on the antibody's epitope. Human ovarian surface epithelium produced similar results, but in rat myometrium the distribution was epitope-independent and nuclear. ER{beta} distribution was predominately 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 E2. We postulate that the different ER{alpha} conformations at the membrane, in the nucleus and between different cell types allow E2 to trigger cell and location specific signaling cascades.




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