Non-adrenergic non-cholinergic nerves (NANC) are thought to be important in the relaxation of corpus cavernosum for erectile function. On the other hand, it has been shown that nitric oxide (NO) is synthesized from L-arginine and released by the endothelium of blood vessels and accounts for the activity of endothelium-derived relaxing factor (EDRF). We studied whether electrical field stimulation (EFS) of isolated strips of human corpus cavernosum released NO and whether it could act as a NANC neurotransmitter. Human penile tissue specimens were mounted in an organ bath and the isometric tension recorded. EFS was applied to precontracted strips between two parallel platinum electrodes. Relaxation induced by EFS was not influenced by guanethidine or atropine, but was markedly inhibited by NG-nitro-L-arginine which is an analog of L-arginine that inhibits the conversion of L-arginine to NO, and hemoglobin which is an agent that inhibits the biological actions of NO. Methylene blue which is an inhibitor of cytosolic guanylate cyclase also reduced the relaxation of EFS, but not completely. The inhibitory effect of NG-nitro-L-arginine was reversed by addition of excess L-arginine. Exogenous NO relaxed human corpus cavernosum. The relaxation was only a transient response and concentration-dependent. The profile of responses is similar to that evoked by EFS. Our data clearly demonstrate the release of NO during EFS and the function of NO as a NANC neurotransmitter. Human penile erection may be mediated by NO generated in response to NANC neurotransmission.