We investigated the possibility of nitric oxide (NO), a powerful vasodilator agent, being synthesized by perivascular nerve fibres. Immunoreactivity and catalytic activity of the NO synthesizing enzyme, NO synthase (NOS), were demonstrated in perivascular nerve fibres of blood vessels receiving autonomic vasodilator innervation, but not of those innervated exclusively by vasoconstrictor nerve fibres. Double-labelling techniques allowed identification of NOS-containing nerve fibres as belonging to the vasoactive intestinal peptide (VIP)/acetylcholine-containing class whereas noradrenergic and substance P-containing perivascular fibres were devoid of NOS. We suggest that, in addition to its endothelial source, NO is a neuronal co-mediator of VIP/cholinergic vasodilation.