Urothelial cells are reported to express a diverse range of receptors and ion channels similar to those found in sensory nerves, with TRPV1 the most strongly implicated in normal bladder function (1). In addition, these cells are known to release a variety of transmitter/modulator substances, including ATP (2), which can alter bladder afferent nerve activity. Stimulation of sub-urothelial neurones may also release a number of factors that act upon urothelial cells (2). With the potential involvement of these cells in bidirectional signalling with bladder afferent nerves and their possible influence on sub-urothelial myofibroblasts and smooth muscle cells, their complement of channel and receptor expression could have major consequences for normal bladder function. Urothelial cells were isolated from human tissue bank and post-operative samples (3) obtained in accordance with ethical guidelines. Currents were recorded using the whole-cell patch clamp technique at a holding potential of −60mV. Experiments were performed in a physiological bath solution and with an ATP-supplemented pipette solution with a free-calcium concentration of 100nM. All experiments were performed at room temperature (20-22°C). Urothelial cells were exposed to ATP (50μM); forskolin (10μM, adenylate cyclase activator); capsaicin (1μM, TRPV1 activator); 4α-PDD (1μM, TRPV4 activator) and menthol (2mM, TRPM8 activator). Responses were observed in 6/13 cells to ATP (n=5 samples); 5/10 cells to forskolin (n=2 samples); 8/8 cells to menthol (n=3 samples); 0/12 cells to capsaicin (n=3 samples) and 0/15 to 4α-PDD (n=4 samples). Application of ATP elicited a peak increase in normalised current of 3.87±0.95 pA/pF (mean±S.E.M.), that declined to a low plateau. The response to forskolin peaked at 5.57±3.96 pA/pF then desensitised in the maintained presence of agonist. The menthol response peaked at 3.56±1.48 pA/pF and was maintained in the presence of the activator. These data demonstrate urothelial responses to ATP, forskolin and menthol, but not to capsaicin or 4α-PDD. TRPV1 (4) and TRPM8 (5) are expressed on urothelial cells; however, antibody staining of TRPV1 shows it is located mostly in the cytoplasm (4) and not at the plasma membrane. Results presented here suggest that TRPV1, unlike TRPM8, may be non-functional in these cells or that it may play an alternative role in intracellular signalling but not as a plasma membrane localised channel. Further work is required to identify the receptors/channels mediating responses to ATP and forskolin in these cells.