Nitric oxide (NO) is a neurotransmitter and neuromodulator in the central nervous system, but this small labile substance also seems to serve as a peripheral neurotransmitter. Abundant evidence is now available that NO, synthesized from L-arginine by NO synthase (NOS), is a nonadrenergic noncholinergic relaxant transmitter of gastrointestinal smooth muscle. Electrically induced nonadrenergic noncholinergic relaxations are antagonized by NOS inhibitors in vitro and in vivo. In a bioassay superfusion system, the release of a substance with the pharmacological characteristics of NO from a gastrointestinal smooth muscle preparation was detected; also, indirect measurements (e.g. of the NO metabolite nitrite or of the co-product of its synthesis L-citrulline) suggest NO release. Immunohistochemistry with antibodies raised against the neuronal NOS showed immunoreactivity in cell bodies of neurones in the myenteric plexus and in nerve fibres in the muscular layer. These data suggest that nerve endings, innervating smooth muscle, are able to release NO that will penetrate the cells to induce relaxation (i.e. nitrergic neurotransmission). It is unlikely that NO as such is stored and it is generally accepted that it is synthesized on demand when the nerve endings are excited, although the possibility of the release of a NO-containing molecule protecting it from degradation in the junction has been proposed. Other sources than neurones (interstitial cells, smooth muscle cells) for the NO involved in nonadrenergic noncholinergic inhibitory transmission have also been proposed. Using NADPH diaphorase as a marker for neuronal NOS, deficiency of the nitrergic innervation has been shown in isolated tissue from patients with infantile hypertrophic pyloric stenosis, achalasia and Hirschsprung's disease, suggesting that a lack of NO release might be involved in these disorders. Evidence in favour of nitrergic neurotransmission to smooth muscle has also been obtained in the respiratory and lower urinary tract, the corpora cavernosa and some blood vessels.