The L-arginine analogues NG-monomethyl-L-arginine (L-NMMA, 10(-4) M) and NG-nitro-L-arginine methyl ester (L-NAME, 10(-4) M), which specifically inhibit the synthesis of nitric oxide from L-arginine, significantly reduced acetylcholine-induced endothelium-dependent relaxations in rings of human omental arteries. The inhibitory potency of L-NMMA and L-NAME was similar. Addition of L-NMMA or L-NAME to the organ bath did not induce any significant changes in the resting tension of the tissues. The effects of L-NMMA were reversed by L-arginine (3 x 10(-4) M). The L-NMMA enantiomer, D-NMMA (10(-4) M), did not influence either the basal tone of the preparation or the relaxing effects of acetylcholine. Arterial relaxations induced by sodium nitroprusside (10(-6) M) were not influenced by incubation with L-NMMA or L-NAME. These results suggest that endothelium-dependent relaxations in human omental arteries are mediated by the endogenous and substrate-specific generation of nitric oxide from L-arginine.