Estradiol coupling to human monocyte nitric oxide release is dependent on intracellular calcium transients: evidence for an estrogen surface receptor.
We tested the hypothesis that estrogen acutely stimulates constitutive NO synthase (cNOS) activity in human peripheral monocytes by acting on an estrogen surface receptor. NO release was measured in real time with an amperometric probe. 17beta-estradiol exposure to monocytes stimulated NO release within seconds in a concentration-dependent manner, whereas 17alpha-estradiol had no effect. 17beta-estradiol conjugated to BSA (E2-BSA) also stimulated NO release, suggesting mediation by a membrane surface receptor. Tamoxifen, an estrogen receptor inhibitor, antagonized the action of both 17beta-estradiol and E2-BSA, whereas ICI 182,780, a selective inhibitor of the nuclear estrogen receptor, had no effect. We further showed, using a dual emission microfluorometry in a calcium-free medium, that the 17beta-estradiol-stimulated release of monocyte NO was dependent on the initial stimulation of intracellular calcium transients in a tamoxifen-sensitive process. Leeching out the intracellular calcium stores abolished the effect of 17beta-estradiol on NO release. RT-PCR analysis of RNA obtained from the cells revealed a strong estrogen receptor-alpha amplification signal and a weak beta signal. Taken together, a physiological dose of estrogen acutely stimulates NO release from human monocytes via the activation of an estrogen surface receptor that is coupled to increases in intracellular calcium.
Stefano
G B
GB
Neuroscience Research Institute, State University of New York, Old Westbury 11568, USA. gstefano@li.nt
Prevot
V
V
Beauvillain
J C
JC
Fimiani
C
C
Welters
I
I
Cadet
P
P
Breton
C
C
Pestel
J
J
Salzet
M
M
Bilfinger
T V
TV
D002118:Calcium / Q000097:blood*; D004958:Estradiol / Q000097:blood* / Q000502:physiology; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D015870:Gene Expression; D006801:Humans; D007424:Intracellular Fluid / Q000378:metabolism*; D009000:Monocytes / Q000378:metabolism*; D009569:Nitric Oxide / Q000097:blood*; D011959:Receptors, Estradiol / Q000502:physiology; D011960:Receptors, Estrogen / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000097:blood* / Q000235:genetics; D013629:Tamoxifen / Q000494:pharmacology
D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D011959:Receptors, Estradiol; D011960:Receptors, Estrogen; D013629:Tamoxifen; D009569:Nitric Oxide; D004958:Estradiol; D002118:Calcium