1. Isolated transverse and longitudinally oriented preparations of sheep urethra precontracted with noradrenaline responded to electrical field stimulation (EFS) with stimulus-dependent non-adrenergic, non-cholinergic (NANC) relaxations. 2. Exogenous nitric oxide (NO) (acidified NaNO2), S-nitroso-L-cysteine (NC), sodium nitroprusside (SNP), 8-Br-cGMP, dibutyryl-cAMP, forskolin and isoprenaline each relaxed precontracted transverse urethral preparations in a concentration-dependent manner in order of potency: NC > forskolin > isoprenaline = SNP > NO > 8-Br-cGMP = dibutyryl-cAMP. Longitudinally oriented preparations responded to NO and NC with concentration-dependent relaxation, no different from that observed in transverse strips. 3. Methylene blue (MB) and oxyhaemoglobin (HbO2) mechanism independent of NO generation. 9. A dense network of NADPH diaphorase-positive fibres associated with both the circular and longitudinal smooth muscle layers of sheep urethra was found. 10. The possibility that a nitrosyl compound which releases NO in the target tissue is the NANC transmitter in sheep urethra is discussed. The data also suggest a role for increased cGMP levels in the relaxation mechanisms, whereas changes in cAMP levels, phosphoinositide hydrolysis or K+ channel activation do not seem to be involved.