The origin and distribution of cerebral perivascular nerves containing nitric oxide, a short-acting messenger or neurotransmitter, have been studied in the rat by histochemistry for reduced nicotinamide adenine dinucleotide phosphate-diaphorase activity, a specific marker for neuronal nitric oxide synthase. Positively stained nerve fibers were distributed throughout the major vessels of the cerebral arteries, though the fiber density was higher in the anterior circulation, including the circle of Willis, than in the posterior arteries. Examination using axonal transport methods indicated that nitric oxide-containing neurons in the sphenopalatine ganglion innervate the cerebral arteries bilaterally. Nitric oxide synthase in these ganglionic cells often co-existed with vasoactive intestinal polypeptide. The anatomical information obtained is discussed in terms of non-adrenergic, non-cholinergic neuronal transmission in the cerebral arteries.