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Molecular Endocrinology 16 (5): 938-946
Copyright © 2002 by The Endocrine Society

ERß Has Nongenomic Action in Caveolae

Ken L. Chambliss, Ivan S. Yuhanna, Richard G. W. Anderson, Michael E. Mendelsohn and Philip W. Shaul

Departments of Pediatrics (K.L.C., I.S.Y., P.W.S.) and Cell Biology (R.G.W.A.), University of Texas Southwestern Medical Center, Dallas, Texas 75390; Molecular Cardiology Research Institute (M.E.M.), New England Medical Center, Tufts University School of Medicine, Boston, Massachusetts 02111

Address all correspondence and requests for reprints to: Ken L. Chambliss, Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9063. E-mail: Ken.Chambliss{at}UTSouthwestern.edu.

ER{alpha} and ERß serve classically as transcription factors, and ER{alpha} also mediates nongenomic responses to E2 such as the activation of endothelial nitric oxide synthase (eNOS). In contrast, the nongenomic capacities of endogenous ERß are poorly understood. We evaluated eNOS activation by E2 in cultured endothelial cells that express endogenous ERß to determine whether the ERß isoform has nongenomic action and to reveal the subcellular locale of that function. A subpopulation of ERß was localized to the endothelial cell plasma membrane, overexpression of ERß enhanced rapid eNOS stimulation by E2, and the response to endogenous ER activation was inhibited by the ERß-selective antagonist RR-tetrahydrochrysene (THC). eNOS activation through ERß was reconstituted and shown to occur independent of ER{alpha} in COS-7 cells, and ERß protein in COS-7 was directed to the plasma membrane. THC also blunted E2 activation of eNOS in isolated endothelial cell plasma membranes. Furthermore, ERß protein was detected and THC attenuated E2 stimulation of eNOS in isolated endothelial cell caveolae, and functional ERß-eNOS coupling was recapitulated in caveolae from transfected COS-7 cells. These findings in the ER-eNOS signaling paradigm indicate that endogenous ERß has nongenomic action in caveolae.




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