Selective pharmacological blockade of the TRPV1 receptor suppresses sensory reflexes of the rodent bladder.

PURPOSE: We investigated the pharmacological effect of TRPV1 antagonists in anesthetized rodent models of bladder function. MATERIALS AND METHODS: The TRPV1 antagonists JNJ17203212 and JYL1421 were evaluated in the anesthetized rat volume induced micturition reflex model. JNJ17203212 was further evaluated in this model in capsaicin (Sigma) desensitized rats, and in rat capsaicin and mouse citric acid models of irritant induced detrusor overactivity. RESULTS: Systemic JNJ17203212 and JYL1421 administration in the anesthetized rat volume induced micturition reflex model resulted in an increased micturition threshold volume. JNJ17203212 also decreased bladder contraction amplitude but JYL1421 had no effect. Capsaicin desensitization significantly increased baseline micturition threshold volume and decreased bladder contraction amplitude in the volume induced micturition reflex model compared to those in sham treated controls and JNJ17203212 produced no further effect after capsaicin desensitization. JNJ17203212 was also effective in 2 models of irritant induced detrusor overactivity, preventing the decrease in micturition threshold volume and the increase in bladder contraction amplitude observed with intravesical instillation of 10 microM capsaicin, and the decreased voiding interval induced by intravesical citric acid. CONCLUSIONS: The TRPV1 antagonists JNJ17203212 and JYL1421 increased the threshold for activation of the micturition reflex in the anesthetized rat volume induced micturition reflex model. This effect appeared to be mediated by capsaicin sensitive afferents. JNJ17203212 also inhibited detrusor overactivity induced by intravesical capsaicin and intravesical citric acid. These data extend our understanding of the role of TRPV1 in sensory modulation of the micturition reflex under nonirritant and inflammatory conditions.

The role of metabotropic glutamate receptor mGlu5 in control of micturition and bladder nociception.

In micturition control, the roles of ionotropic glutamate (iGlu) receptors NMDA and AMPA are well established, whereas little is known about the function of metabotropic glutamate (mGlu) receptors. Since antagonists for mGlu5 receptors are efficacious in animal models of inflammatory and neuropathic pain, we examined whether mGlu5 receptors play a role in the voiding reflex and bladder nociception and, if so, via centrally or peripherally localized receptors. The mGlu5 receptor antagonist MPEP dose-dependently increased the micturition threshold (MT) volume in the volume-induced micturition reflex (VIMR) model in anesthetized rats. Following doses of 5.2, 15.5 and 51.7micromol/kg of MPEP (intraduodenal), the MT was increased by 24.7+/-5.0%, 97.2+/-12.5% (P<0.01) and 128.0+/-28.3% (P<0.01) from the baseline, respectively (n=4-5; compared with 0.8+/-9.1% in the vehicle group). Infusing MPEP (0.3, 1mM) directly into the bladder also raised MT. However, the efficacious plasma concentrations of MPEP following intravesical dosing were similar to that after intraduodenal dosing (EC(50) of 0.11 and 0.27microM, respectively, P>0.05). MPEP also dose-dependently attenuated the visceromotor responses (VMR, total number of abdominal EMG spikes during phasic bladder distension) in anesthetized rats. The VMR was decreased to 1332.4+/-353.9 from control of 2886.5+/-692.2 spikes/distension (n=6, P<0.01) following MPEP (10micromol/kg, iv). Utilizing the isolated mouse bladder/pelvic nerve preparation, we found that neither MPEP (up to 3microM) nor MTEP (up to 10microM) affected afferent discharge in response to bladder distension (n=4-6). In contrast, MPEP attenuated the responses of the mesenteric nerves to distension of the mouse jejunum in vitro. These data suggest that mGlu5 receptors play facilitatory roles in the processing of afferent input from the urinary bladder, and that central rather than peripheral mGlu5 receptors appear to be responsible.

Pharmacological characterization of canine bradykinin receptors in prostatic culture and in isolated prostate.

The objective of this study was to characterize pharmacologically bradykinin (Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg, BK) receptors in the canine prostate. Primary cultures of canine prostate stromal (PS) and epithelial cells (PE) were established and then characterized using cell-specific antibodies (actin, vimentin and cytokeratin). Cultured cells were assayed for BK receptors using fluorometric imaging plate reader assays. In addition, isolated strips of the canine prostate were studied for BK-induced isometric contraction. PS cells were labeled only with anti-actin and -vimentin antibodies, while the anti-cytokeratin antibodies labeled only the PE cells. In cultured prostate cells, the BK receptor 2 (B2)-preferring agonist BK induced mobilization of intracellular Ca(2+) in a concentration-dependent manner with potencies (log[EC(50)]mid R:PE, pEC(50)) of 8.72+/-0.12 in PS and 8.75+/-0.06 in PE cells. In contrast, the BK receptor 1 (B1)-selective agonist [des-Arg(9)]BK (Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe) did not elicit any significant effect (pEC(50)<5) on Ca(2+) responses. BK agonism (10 nm) was inhibited by HOE-140 (D-arginyl-L-arginyl-L-prolyl-trans-4-hydroxy-L-prolylglycyl-3-(2-thienyl)-L-alanyl-L-seryl-D-1,2,3,4-tetrahhydro-3-isoquinolinecarbonyl-L-(2a,3b,7ab)-octahydro-1H-indole-2-carbonyl-L-arginine), a B2-selective antagonist, with a log[IC(50)] (pIC(50)) of 8.11+/-0.19 and 9.23+/-0.20 in PS and PE cells, respectively. [des-Arg(10)]HOE-140 (d-arginyl-l-arginlyl-l-prolyl-trans-4-hydroxy-l-prolylglycyl-3-(2-thienyl)-L-alanyl-L-seryl-D-1,2,3,4-tetrahydro-3-isoquinolinecarbonyl-L-(2a, 3b,7ab)-octahydro-1H-indole-2-carbonyl), a B1-selective antagonist, displayed weak antagonism with pIC(50) values of 4.87+/-0.23 and 6.38+/-0.16 in PS and PE cells, respectively. Isolated tissue strips of the canine prostate contracted to BK (10 microm) but not to [des-Arg(9)]BK (10 microm). BK-induced contractility was attenuated by HOE-140 (1 microm). In conclusion, canine prostates express functional B2 receptors, with no apparent B1 receptor subtypes.