With the immunofluorescence technique, nitric oxide synthase (NOS)‐like immunoreactivity (LI) was found in a few medium‐sized and small sensory neurons in lumbar (L) 4 and L5 dorsal root ganglia (DRG) of normal rat, and in most of these neurons, NOS‐LI coexisted with calcitonin gene‐related peptide and sometimes with substance P and galanin. NOS‐immunoreactive nerve fibers, terminals and small neurons were also located in the dorsal horn of the segments 4 and 5 of the rat lumbar spinal cord with the highest density in inner lamina II. Many NOS‐positive neurons and fibers were seen in the area around the central canal. A sparse network of NOS‐immunoreactive nerve fibers was found in the ventral horn. After unilateral sciatic nerve cut in the rat, the number of NOS‐positive neurons increased in the ipsilateral L4 and L5 DRGs, mainly in medium and small neurons, but also in some large neurons and very small neurons. NOS‐LI could now also be seen in the ipsilateral dorsal roots, and in an increased number of fibers and terminals in both outer and inner lamina II of the ipsilateral dorsal horn. The number of NOS‐immunoreactive neurons in lamina II of the ipsilateral dorsal horn was reduced. In the monkey L4 and L5 DRGs, many small neurons were NOS‐immunoreactive, but only a few weakly stained nerve fibers and terminals were found in laminae I‐IV of the dorsal horn at L4 and L5 lumbar levels. A few NOS‐positive neurons were present in lamina X. The number of NOS‐immunoreactive neurons was somewhat reduced in DRGs 14 days after peripheral axotomy, but no certain effect was seen in the dorsal horn. These results, together with earlier in situ hybridization studies, demonstrate that axotomy in rat induces a marked upregulation of NOS synthesis in primary sensory neurons, thus suggesting a role for NO in lesioned sensory neurons. In contrast, no such effect was recorded in monkey, perhaps indicating distinct species differences. © 1993 Wiley‐Liss, Inc.