Effects of chronic hypoxia on soluble guanylate cyclase activity in fetal and adult ovine cerebral arteries

Abstract

A broad variety of evidence obtained largely in pulmonary vasculature suggests that chronic hypoxia modulates vasoreactivity to nitric oxide (NO). The present study explores the general hypothesis that chronic hypoxia also modulates cerebrovascular reactivity to NO, and does so by modulating the activity of soluble guanylate cyclase (sGC), the primary target for NO in vascular smooth muscle. Pregnant and nonpregnant ewes were maintained at either sea level or at 3,820 m for the final 110 days of gestation, at which time middle cerebral arteries from term fetal lambs and nonpregnant adults were harvested. In both fetal and adult arteries, NO-induced vasodilatation was attenuated by chronic hypoxia and completely inhibited by 10 μM 1 H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), a selective inhibitor of sGC. sGC abundance (in ng sGC/mg protein) measured via Western immunoblots was ∼10-fold greater in fetal (17.6 ± 1.6) than adult (1.7 ± 0.3) arteries but was not affected by chronic hypoxia. The specific activity of sGC (in pmol cGMP·μg sGC−1·min−1) was similar in fetal (255 ± 64) and adult (280 ± 75) arteries and was inhibited by chronic hypoxia in both fetal (120 ± 10) and adult (132 ± 26) arteries. Rates of cGMP degradation (in pmol cGMP·mg protein−1·min−1) were similar in fetal (159 ± 59) and adult (134 ± 36) arteries but were not significantly depressed by chronic hypoxia in either fetal (115 ± 25) or adult (108 ± 25) arteries. The cGMP analog 8-( p-chlorophenylthio)-cGMP was a more potent vasorelaxant in fetal (pD2= 4.7 ± 0.1) than adult (pD2= 4.3 ± 0.1) arteries, but its ability to promote vasodilatation was not affected by chronic hypoxia in either age group. Together, these results reveal that hypoxic inhibition of NO-induced vasodilatation is attributable largely to attenuation of the specific activity of sGC and does not involve significant changes in sGC abundance, cGMP-phosphodiesterase activity, or the vasorelaxant activity of protein kinase G.