Bladder Urothelial Cells From Patients With Interstitial Cystitis Have an Increased Sensitivity to Carbachol

Aims: The presence of muscarinic receptors on bladder urothelial cells (BUC), suggests BUC may be a target for antimuscarinics. This study determined whether human BUC are responsive to a muscarinic agonist and if so, whether responses are altered in interstitial cystitis (IC) BUC. Methods: Primary urothelial cell cultures were established from cystoscopic biopsies. Normal (NB) and IC BUC were studied using calcium imaging techniques as a means to monitor the response to muscarinic receptor activation with the agonist, carbachol (CCh). Changes in intracellular Ca2+ concentration [(Ca2+](i)) were measured with fura-2 ratiometric microfluorimetry. Dose-response curves (CCh dose vs. [Ca2+](i)) were measured in IC and NB BUC. Tolterodine was used to confirm the specificity (muscarinic versus nicotinic) of CCh evoked increases in [Ca2+](i). Results: CCh induced a dose-dependent increase in [Ca2+](i). Potency and efficacy of CCh was significantly greater in IC BUC. The maximal increase in [Ca2+](i) was 136.3 +/- 5.1% over baseline in 78 cells from 4 IC patients versus 92.4 +/- 4.8% over baseline in 67 cells from 4 NB subjects (P < 0.01). The EC50 of the evoked increase was 1.10 +/- 0.14 mu M versus 3.36 +/- 0.72 mu M (P < 0.01) in BUC from IC and NB controls, respectively. Removal of extracellular calcium or application of tolterodine, abolished CCh evoked increase in [Ca2+](i) in IC and NB BUC. Conclusions: The greater sensitivity of IC BUC to CCh suggests that IC patholobiology may also include alterations muscarinic signaling. The physiologic sequelae of muscarinic activation in BUC need to be further investigated. Neurourol. Urodynam. 28:1022-1027, 2009. (C) 2009 Wiley-Liss, Inc.