Nitric oxide is a free radical, which is produced in several tissues of the body and is thought to be the first of a new class of neural messenger molecules and a retrograde modulator of synaptic transmission in the brain. Nitric oxide synthase (NOS) is the enzyme that produces nitric oxide from the substrate l-arginine. The pattern of the distribution of neuronal isoform of NOS was investigated in neurones and fibres in the brain stem using standard immunocytochemistry. In our results, NOS positive neurones and processes were seen in the spinal trigeminal nucleus, gracile nucleus, nucleus of the solitary tract, nucleus ambiguus, reticular nuclei and lateral to the pyramidal tract of the medulla. In the pons, heavily labelled NOS containing neurones were seen in the pedunuclopontine tegmental nucleus, ventral tegmental nucleus and in the laterodorsal tegmental nucleus. The localization of neuronal NOS expressing neurones suggests a widespread neuromodulatory role for the nitric oxide in the central nervous system of rat.