The P2X₃ receptor subunit, a member of the P2X family of ATP‐gated ion channels, is almost exclusively localized in sensory neurons. In the present study, we sought to gain insight into the role of P2X₃ and P2X₃‐containing neurons in sensory transmission, using immunohistochemical approaches. In rat dorsal root ganglia (DRG), P2X₃‐immunoreactivity (‐ir) was observed in small‐ and medium‐sized neurons. Approximately 40% of DRG neuronal profiles in normal rats contained P2X₃‐ir. In rats that had received neonatal capsaicin treatment, the number of P2X₃‐positive neurons was decreased by approximately 70%. Analysis of the colocalization of P2X₃‐ir with cytochemical markers of DRG neurons indicated that approximately 94% of the P2X₃‐positive neuronal profiles were labelled by isolectin B₄ from Bandeiraea simplicifolia, while only 3% contained substance P‐ir, and 7% contained somatostatin‐ir. In dorsal horn of rat spinal cord, P2X₃‐ir was observed in the inner portion of lamina II and was reduced subsequent to dorsal rhizotomy, as well as subsequent to neonatal capsaicin treatment. Finally, P2X₃‐ir accumulated proximal to the site of sciatic nerve ligation, and was seen in nerve fibres in skin and corneal epithelium. In summary, our results suggest that P2X₃ is expressed by a functionally heterogeneous population of BSI‐B₄‐binding sensory neurons, and is transported into both central and peripheral processes of these neurons.