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The present study explores the possible involvement of a purinergic mechanism in mechanosensory transduction in the bladder using P2X₃ receptor knock-out (P2X₃^−/−) and wild-type control (P2X₃^+/+) mice. Immunohistochemistry revealed abundant nerve fibers in a suburothelial plexus in the mouse bladder that are immunoreactive to anti-P2X₃. P2X₃-positive staining was completely absent in the subepithelial plexus of the P2X₃^−/− mice, whereas staining for calcitonin gene-related peptide and vanilloid receptor 1 receptors remained. Using a novel superfused mouse bladder–pelvic nerve preparation, we detected a release of ATP proportional to the extent of bladder distension in both P2X₃^+/+ and P2X₃^−/− mice, although P2X₃^−/−bladder had an increased capacity compared with that of the P2X₃^+/+ bladder. The activity of multifiber pelvic nerve afferents increased progressively during gradual bladder …