Development of the topographic map of the somatosensory cortex of rodents appears to depend on fine-tuned patterns of neuronal activity. Nitric oxide (NO) has been described as a potent messenger in the modulation of neural activity associated with synaptic plasticity. To evaluate the role of NO in the murine somatosensory pathway, we investigated NO synthase activity by NADPH diaphorase histochemistry at crucial developmental stages. At birth, NADPH diaphorase activity was detected in the cortical plate of the developing somatosensory cortex. At day 3, diffuse NADPH diaphorase activity increased within the emerging layer 4 in the future barrel field hollows. This staining was most intense at day 6 in the barrel field hollows and became undetectable by the end of the second postnatal week. The appearance of the diffuse NADPH diaphorase staining pattern was also observed in a similar time course and topography in the ascending relays of the somatosensory cortex, specifically in the barreloids within the ventrobasal nucleus of the thalamus and the barrelettes of the trigeminal nucleus of the brainstem. Lesioning the C row of whiskers at day 1 (i.e. during the critical period of barrel formation) led to fused C barrels of diffuse NADPH diaphorase activity in the barrel fields. In addition, highly NADPH diaphorase activity-positive individual cells present in the deeper layers of the somatosensory cortex at days 0 and 3 became visible in the upper layers at day 6 and remained until day 15. In layer 4, these cells were predominantly localized in the septa at day 6 and 9. No positive individual cells were detected in barrelettes or barreloids at any age. We conclude that NADPH diaphorase activity is present during experience-dependent consolidation of synaptic contacts in the somatosensory pathway.