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Functional TRP and ASIC-like channels in cultured urothelial cells from the rat Kullmann FA; Shah MA; Birder LA; de Groat WCAm J Physiol Renal Physiol 2009[Apr]; 296 (4): F892-901Transient receptor potential (TRP) and acid-sensing ion channels (ASIC) are molecular detectors of chemical, mechanical, thermal, and nociceptive stimuli in sensory neurons. They have been identified in the urothelium, a tissue considered part of bladder sensory pathways, where they might play a role in bladder function. This study investigated functional properties of TRP and ASIC channels in cultured urothelial cells from the rat using patch-clamp and fura 2 Ca(2+) imaging techniques. The TRPV4 agonist 4alpha-phorbol-12,13 didecanoate (4alpha-PDD; 1-5 microM) and the TRPA1/TRPM8 agonist icilin (50-100 microM) elicited transient currents in a high percentage of cells (>70%). 4alpha-PDD responses were suppressed by the TRPV4 antagonist HC-010961 (10 microM). The TRPV1 agonist capsaicin (1-100 microM) and the TRPA1/TRPM8 agonist menthol (5-200 microM) elicited transient currents in a moderate percentage of cells ( approximately 25%). All of these agonists increased intracellular calcium concentration ([Ca(2+)](i)). Most cells responded to more than one TRP agonist (e.g., capsaicin and 4alpha-PDD), indicating coexpression of different TRP channels. In the presence of the TRPV1 antagonist capsazepine (10 microM), changes in pH induced by HCl elicited ionic currents (pH 5.5) and increased [Ca(2+)](i) (pH 6.5) in approximately 50% of cells. Changes in pH using acetic acid (pH 5.5) elicited biphasic-like currents. Responses induced by acid were sensitive to amiloride (10 microM). In summary, urothelial cells express multiple TRP and ASIC channels, whose activation elicits ionic currents and Ca(2+) influx. These "neuron-like" properties might be involved in transmitter release, such as ATP, that can act on afferent nerves or smooth muscle to modulate their responses to different stimuli.|Acid Sensing Ion Channels[MESH]|Animals[MESH]|Ankyrins[MESH]|Calcium Channels/metabolism[MESH]|Calcium Signaling[MESH]|Cells, Cultured[MESH]|Female[MESH]|Hydrogen-Ion Concentration[MESH]|Male[MESH]|Membrane Potentials[MESH]|Membrane Transport Modulators/pharmacology[MESH]|Microscopy, Fluorescence[MESH]|Nerve Tissue Proteins/drug effects/*metabolism[MESH]|Patch-Clamp Techniques[MESH]|Rats[MESH]|Rats, Sprague-Dawley[MESH]|Sodium Channels/drug effects/*metabolism[MESH]|TRPA1 Cation Channel[MESH]|TRPC Cation Channels/drug effects/*metabolism[MESH]|TRPM Cation Channels/metabolism[MESH]|TRPV Cation Channels/metabolism[MESH]|Time Factors[MESH]|Urinary Bladder/cytology/drug effects/*metabolism[MESH]|Urothelium/drug effects/*metabolism[MESH] |
