PT - JOURNAL ARTICLE AU - Radhika R Tunstall AU - Praveen Shukla AU - Anna Grazul-Bilska AU - Chengwen Sun AU - Stephen T O'Rourke TI - MT2-Receptors Mediate the Inhibitory Effects of Melatonin on Nitric Oxide-Induced Relaxation of Porcine Isolated Coronary Arteries AID - 10.1124/jpet.110.174482 DP - 2010 Jan 01 TA - Journal of Pharmacology and Experimental Therapeutics PG - jpet.110.174482 4099 - http://jpet.aspetjournals.org/content/early/2010/10/19/jpet.110.174482.short 4100 - http://jpet.aspetjournals.org/content/early/2010/10/19/jpet.110.174482.full AB - Previous studies from our laboratory demonstrated that melatonin inhibits nitric oxide (NO)-induced relaxation in porcine coronary arteries. The present study was designed to further characterize the mechanisms underlying this inhibitory effect of melatonin. Western immunoblot studies identified the presence of melatonin type-2 (MT2)-receptors, but not MT1- or MT3-receptors, in porcine coronary arteries. Immunohistochemical analysis revealed that MT2-receptors co-localized with alpha-actin in the smooth muscle cell layer. In coronary arterial rings suspended in organ chambers for isometric tension recording, melatonin (10-7 M) inhibited relaxations induced by the exogenous NO-donor, sodium nitroprusside (SNP; 10-9 - 10-5 M), and by the alpha2-adrenoceptor agonist, UK14,304 (10-9 - 10-5 M), an endothelium-dependent vasodilator. The inhibitory effect of melatonin on SNP- and UK14,304-induced relaxations was abolished in the presence of the selective MT2-receptor antagonists, 4P-PDOT (10-7 M) or luzindole (10-7 M). In contrast to melatonin, the selective MT3-receptor agonist, 5-MCA-NAT (10-7 M), had no effect on the concentration-response curves to either SNP or UK14,304. Melatonin (10-7 M) had no effect on coronary artery relaxation induced by 8-Br-cGMP, but it significantly attenuated the increase in intracellular cyclic GMP levels in response to SNP (10-5 M). This effect of melatonin was abolished in the presence of 4P-PDOT (10-7 M). Taken together, these data support the view that melatonin acts on MT2-receptors in coronary vascular smooth muscle cells to inhibit NO-induced increases in cyclic GMP and coronary arterial relaxation, thus demonstrating a novel function for MT2-receptors in the vasculature.