Comparison of the effects of nitric oxide synthase, guanylate cyclase and potassium channel inhibition on vascular contractions in vitro in the rat.

1. We have investigated the differences between the nitric oxide synthase inhibitor L-NMMA, the guanylate cyclase inhibitor methylene blue and the potassium channel blockers apamin and charybdotoxin or apamin and iberiotoxin, in their abilities to increase vasoconstrictor responses in rat small mesenteric arterial rings. 2. When administered during the maintained contraction to PGF2alpha (10 microM), L-NMMA (100 microM) or the combination of apamin (0.7 microM) and charybdotoxin (0.1 microM) significantly increased the contractile response. Methylene blue (10 microM) increased the contraction, but this did not reach significance. However, apamin (0.7 microM) and iberiotoxin (0.1 microM) also significantly increased the contractile response. 3. The combination of L-NMMA or methylene blue with apamin/charybdotoxin produced significantly greater increases in the contractile response to PGF2alpha than achieved individually. 4. Relaxations to acetylcholine (10 microM) were significantly reduced by L-NMMA or methylene blue but not by apamin in combination with charybdotoxin or iberiotoxin. 5. Since apamin/iberiotoxin had similar effects to apamin/charybdotoxin, it is likely that the actions of these agents involve direct actions on smooth muscle potassium channels rather than inhibition of endothelium-derived hyperpolarising factor (EDHF). These results suggest that endothelium-derived nitric oxide but not EDHF has a major role in modulating vascular tone under these conditions.