Estrogen receptor alpha mediates acute potassium channel stimulation in human coronary artery smooth muscle cells.
The pleiotropic effects of estrogen are mediated via stimulation of two estrogen receptor (ER) subtypes, ERalpha and ERbeta. Although a number of studies have identified expression of one or both subtypes in estrogen target tissues, fewer studies have correlated ER expression with a functional role of these proteins in regulating cellular excitability. In the present study, we have combined cellular fluorescence, immunocytochemistry, and molecular expression techniques with single-channel patch-clamp studies to determine which ER mediates estrogen-stimulated potassium channel activity in human coronary artery smooth muscle cells (HCASMC). We had demonstrated previously that estrogen stimulates activity of the large-conductance, calcium- and voltage-activated potassium (BK(Ca)) channel in HCASMC via a nongenomic mechanism. We now demonstrate expression of both ERalpha and ERbeta subtypes in HCASMC. Functionally, however, expression of ERalpha antisense plasmid abolished the acute effect of estrogen on these channels, whereas estrogen retained its ability to stimulate BK(Ca) channels in cells transfected with only green fluorescence protein. In contrast, overexpression of ERalpha enhanced the stimulatory action of estrogen in HCASMC. Transfection with ERalpha antisense/sense plasmid did not alter ERbeta expression. These findings indicate that the ERalpha isoform mediates estrogen-induced stimulation of BK(Ca) channels in HCASMC and thereby provide evidence for a receptor-dependent signaling mechanism that can mediate estrogen-induced inhibition of cellular excitability.
Han
Guichun
G
Dept. Pharmacology and Toxicology, 1120 15th Street, Augusta, GA, USA. ghan@mail.mcg.edu
Yu
Xiuping
X
Lu
Luo
L
Li
Shuyi
S
Ma
Handong
H
Zhu
Shu
S
Cui
Xiuyun
X
White
Richard E
RE
D015153:Blotting, Western; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000378:metabolism*; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology; D047628:Estrogen Receptor alpha / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D047629:Estrogen Receptor beta / Q000032:analysis; D000077267:Fulvestrant; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis; D024681:Potassium Channels, Calcium-Activated / Q000502:physiology*; D012333:RNA, Messenger / Q000032:analysis; D014162:Transfection
D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D024681:Potassium Channels, Calcium-Activated; D012333:RNA, Messenger; D000077267:Fulvestrant; D009569:Nitric Oxide; D004958:Estradiol