Role of nNOS in Blood Pressure Regulation in eNOS Null Mutant Mice

Abstract

Abstract —The role of neural nitric oxide synthase (nNOS) in regulating blood pressure (BP) remains uncertain. Recently it was reported that in mice lacking functional endothelial NOS (eNOS) genes (−/−), acute administration of a nonselective NOS inhibitor, N w -nitro- l -arginine, decreased mean BP, suggesting that NO released by non-eNOS isoforms increases BP. Because the inducible NOS isoform is not constitutively expressed and when induced causes hypotension, we hypothesize that it is NO produced by nNOS that increases BP in the absence of eNOS activity. To test this hypothesis, we studied the acute effect of selective and nonselective nNOS inhibitors on BP and cerebellar NOS activity in eNOS (−/−), wild-type (+/+), and heterozygous (+/−) mice as well as in +/+ mice with renovascular hypertension. Because it is not known whether the decrease in BP caused by acute NOS inhibition in −/− mice can occur chronically, we also studied the effect of chronic NOS inhibition on both BP and cerebellar NOS activity. eNOS (−/−) mice had higher BP than +/+ or +/−mice, and acute administration of the selective nNOS inhibitor 7-nitroindazole (7-NI) decreased their mean BP from 137±13 to 124±12 mm Hg ( P <0.01). In +/+, +/−, or renovascular hypertensive +/+ mice, 7-NI caused a small but insignificant rise from 105±5 to 110±6 mm Hg, from 115±9 to 119±13 mm Hg, and from 146±6 to 150±6 mm Hg, respectively. Fifteen minutes after administration of 7-NI, cerebellar NOS activity decreased by 70%; however, this inhibitory effect was brief, since 2 hours after 7-NI administration NOS returned toward control values. Chronic oral or intraperitoneal administration of 7-NI did not inhibit cerebellar NOS activity, whereas the nonselective NOS inhibitor N G -nitro- l -arginine methyl ester (L-NAME) decreased this activity by 50%. Therefore, we studied the effect of chronic L-NAME administration (4 weeks) on BP. In −/− mice, chronic L-NAME administration decreased BP from 135±4 to 120±3 mm Hg ( P <0.05), whereas in +/+ and +/−mice, as expected, it increased BP from 109±2 to 125±3 mm Hg ( P <0.001) and from 107±6 to 119±5 mm Hg ( P <0.02), respectively. After L-NAME administration was stopped, BP returned to baseline. These results suggest that in eNOS −/− mice, NO derived from nNOS increases BP both acutely and chronically.