Nongenomic stimulation of nitric oxide release by estrogen is mediated by estrogen receptor alpha localized in caveolae

Biochem Biophys Res Commun. 1999 Sep 16;263(1):257-62. doi: 10.1006/bbrc.1999.1348.


Acute administration of 17beta-estradiol (E(2)) exerts antiatherosclerotic effects in healthy postmenopausal women. The vasoprotective action of E(2) may be partly accounted for by a rapid increase in nitric oxide (NO) levels in endothelial cells (ECs). However, the signaling mechanisms producing this rise are unknown. In an attempt to address the short-term effect of E(2) on endothelial NO production, confluent bovine aortic endothelial cells (BAECs) were incubated in the absence or presence of E(2), and NO production was measured. Significant increases in NO levels were detected after only 5 min of E(2) exposure without a change in the protein levels of endothelial NO synthase (eNOS). This short-term effect of estrogen was significantly blunted by various ligands which decrease intracellular Ca(2+) concentration. Furthermore, plasma membrane-impermeable BSA-conjugated E(2) (E(2)BSA) stimulated endothelial NO release, indicating that in the current system the site of action of E(2) is on the plasma membrane rather than the classical nuclear receptor. The partial antagonist tamoxifen did not block E(2)-induced NO production; however, a pure estrogen receptor alpha (ERalpha) antagonist ICI 182,780 completely inhibited E(2)-stimulated NO release. The binding of E(2) to the membrane was confirmed using FITC-labeled E(2)BSA (E(2)BSA-FITC). Western blot analysis showed that plasmalemmal caveolae possess ERalpha in addition to well-known caveolae-associated proteins eNOS and caveolin. This study demonstrates that the nongenomic and short-term effect of E(2) on endothelial NO release is Ca(2+)-dependent and occurs via ERalpha localized in plasmalemmal caveolae.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism
  • Cattle
  • Cell Membrane / drug effects*
  • Cell Membrane / metabolism*
  • Cells, Cultured
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / metabolism
  • Estradiol / analogs & derivatives
  • Estradiol / metabolism
  • Estradiol / pharmacology*
  • Estrogen Antagonists / pharmacology
  • Estrogen Receptor alpha
  • Female
  • Fluorescein-5-isothiocyanate / analogs & derivatives
  • Fluorescein-5-isothiocyanate / metabolism
  • Fulvestrant
  • Humans
  • Nitric Oxide / biosynthesis*
  • Nitric Oxide Synthase / metabolism
  • Nitric Oxide Synthase Type III
  • Receptors, Estrogen / antagonists & inhibitors
  • Receptors, Estrogen / drug effects*
  • Receptors, Estrogen / metabolism*
  • Serum Albumin, Bovine / metabolism
  • Serum Albumin, Bovine / pharmacology
  • Tamoxifen / pharmacology


  • Estrogen Antagonists
  • Estrogen Receptor alpha
  • Receptors, Estrogen
  • estradiol 6-O-carboxymethyloxime-bovine serum albumin-fluorescein isothiocyanate
  • estradiol-bovine serum albumin
  • Tamoxifen
  • Fulvestrant
  • Serum Albumin, Bovine
  • Nitric Oxide
  • Estradiol
  • NOS3 protein, human
  • Nitric Oxide Synthase
  • Nitric Oxide Synthase Type III
  • Fluorescein-5-isothiocyanate
  • Calcium