Endothelial vasodilator production by uterine and systemic arteries .2. Pregnancy effects on NO synthase expression

Pregnancy is characterized by elevations in uterine but not omental artery nitric oxide synthase (NOS)-specific activity. We hypothesized that increases in NO production during pregnancy are associated with elevations in protein expression of the constitutive isoform, endothelial cell NOS (ecNOS), in uterine but not systemic arteries. Arterial NOS-specific activity and guanosine 3',5'-cyclic monophosphate (cGMP) production were tested in pregnant sheep in the presence or absence [+5 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N' ,N'-tetraacetic acid] of Ca2+. With the use of Western analysis, ecNOS and neuronal NOS (nNOS) constitutive isoform expressions were evaluated in intact and denuded [vascular smooth muscle (VSM)] uterine and systemic (omental and renal) arteries as well as in isolated endothelium-derived proteins from nonpregnant and pregnant sheep. Uterine and omental artery NOS activity and cGMP production were inhibited 75-85% by Ca2+ removal. ecNOS was localized only in uterine and systemic artery endothelium (not VSM) by immunohistochemistry and Western analysis; nNOS was not detected. Compared with nonpregnant ewes, pregnancy increased expression of ecNOS in uterine [2.1- to 4.2-fold (P < 0.0001)] and omental [1.3- to 2.2-fold (P = 0.032)] but not renal (P = 0.1367) artery endothelium. Increases in uterine were greater than in omental artery endothelium. Levels of plasma and urinary cGMP were elevated (P < 0.01) proportionally (1.8- to 8.0-fold) in pregnant versus nonpregnant ewes. During pregnancy, expression of uterine artery endothelium-derived (not VSM) ecNOS constitutive isoform is increased, whereas expression in systemic vessels shows little or no change.