The decompensated detrusor V: molecular correlates of bladder function after reversal of experimental outlet obstruction.

PURPOSE: Calcium ion homeostasis has a significant role in smooth muscle function. Its regulation requires complex storage and release mechanisms via ion pumps and channels located within intracellular storage sites (sarcoplasmic reticulum) and at the plasma membrane. We have previously reported a dramatic loss of the 2 major sarcoplasmic reticulum proteins sarcoplasmic endoplasmic reticulum calcium magnesium adenosine triphosphatase (SERCA2) and the ryanodine sensitive ion channel, also called the ryanodine receptor, after outlet obstruction. In our current study we investigated the correlation of the expression of these 2 major sarcoplasmic reticulum components with bladder function recovery after the reversal of outlet obstruction. METHODS AND METHODS: Standard partial bladder outlet obstruction was created in adult New Zealand White rabbits. Voiding patterns were monitored 2 and 4 weeks postoperatively, and rabbits were selected for outlet obstruction reversal based on a voiding pattern consistent with a decompensated state, as indicated by a frequency of greater than 30 voids daily and an average voided volume of less than 4 cc. Bladder biopsy was done when outlet obstruction was reversed. Voiding performance was monitored postoperatively and the animals were sacrificed 2 weeks later. Voiding patterns and muscle strip studies enabled us to define 2 functional outcome categories after reversal, namely normal versus minimally improved. Microsomal membrane protein fractions were prepared from the same bladder tissues before and after reversal, and probed by Western blot analysis for SERCA2 and ryanodine receptor expression. RESULTS: Western blot analysis revealed a major loss of SERCA2 and ryanodine receptor expression at the time of reversal and biopsy. In 65% of bladders obstruction reversal resulted in a normalized voiding pattern with a recovery of ryanodine receptor expression that was 15% to 65% of control values. In contrast, in the 35% of bladders with persistent voiding symptoms there was minimal recovery of ryanodine receptor expression. SERCA2 expression increased slightly in each group after reversal but did not differ in bladders with normalized versus improved function. CONCLUSIONS: Bladder decompensation is highly associated with a loss of sarcoplasmic reticulum function. Furthermore, the decompensated detrusor recovers function after obstruction reversal, which is associated with the recovery of these sarcoplasmic reticulum components.

Effects of tadenan pretreatment on bladder physiology and biochemistry following partial outlet obstruction.

PURPOSE: Tadenan is a pharmaceutical agent used in the treatment of BPH. Prior studies demonstrated that pretreatment of rabbits with Tadenan significantly reduced the contractile dysfunction following two weeks of partial outlet obstruction. The specific aim of the present study was to determine the effect of Tadenan pretreatment on the time course of the response to partial outlet obstruction and correlate the effect of Tadenan on the contractile responses to field stimulation, bethanechol, and KCl with both mitochondrial enzyme activity (citrate synthase) and sarcoplasmic reticular function (calcium-ATP'ase activity). MATERIALS AND METHODS: Sixty male New Zealands white rabbit (3 to 5 kg.) were separated into 12 groups of 5 rabbits each. Each rabbit in groups 1-6 received Tadenan orally at 100 mg./kg./day for three weeks; each rabbit in groups 7-12 received vehicle (peanut oil). Each rabbit in groups 2-6 and 8-12 received a partial outlet obstruction as described below. One group of Tadenan treated and one group of vehicle-treated rabbits were euthanized at 1, 3, 5, 7, and 14 days following partial outlet obstruction. The non-obstructed groups were studied after 4 weeks of drug or vehicle treatment. Each bladder was rapidly removed and weighed, and 3 longitudinal strips prepared and mounted in individual baths for contractile studies. The remainder of the bladder was frozen for biochemical analysis. The contractile responses to field stimulation, bethanechol, and KCl were determined; and the enzyme activities of citrate synthase (marker for mitochondrial function) and calcium-ATP'ase (marker for sarcoplasmic reticulum) were determined. RESULTS: 1) Tadenan did not reduce the effect of partial outlet obstruction on bladder mass. 2) Although the contractile responses to forms of stimulation were reduced at 1 day following partial outlet obstruction, Tadenan pretreatment resulted in a significant protective effect on the contractile responses to field stimulation, bethanechol, and KCl at 3, 5, 7, and 14 days of obstruction. 3) The activities of both citrate synthase and calcium ATP'ase were reduced significantly at 1 day following obstruction for both Tadenan treated and vehicle treated groups. The activities of both enzymes returned to near normal levels at 7 and 14 days for the Tadenan groups whereas the activities of both enzymes remained significantly reduced in the vehicle treated groups. CONCLUSIONS: These results clearly demonstrate that Tadenan pretreatment protected the bladder from both the contractile and metabolic dysfunctions induced by partial outlet obstruction.

Protective effect of Tadenan on bladder function secondary to partial outlet obstruction.

PURPOSE: Tadenan (DEBAT, Paris, France) is a pharmaceutical agent used in the treatment of benign prostatic hyperplasia (BPH). The specific aim of this study was to determine if pretreatment of rabbits with Tadenan reduced either the hypertrophic response of the bladder to partial outlet obstruction or the accompanying contractile dysfunction. MATERIALS AND METHODS: Twenty-five male New Zealand rabbits (3 to 5 kg.) were separated into 5 groups of 5 rabbits each. Each rabbit in groups 1,2, and 3 received Tadenan orally at 1, 10 and 100 mg./kg./day for 3 weeks. Group 4 received vehicle only (peanut oil); Group 5 were controls. The bladders were evaluated (in vitro studies) after 2 weeks of obstruction. RESULTS: 1) Tadenan did not reduce the effect of partial outlet obstruction on bladder mass. 2) Tadenan pretreatment resulted in a significant protective effect on the contractile responses to field stimulation, bethanechol and KCl. CONCLUSIONS: These results clearly demonstrate that Tadenan pretreatment protected the bladder from the contractile dysfunctions induced by partial outlet obstruction.

Comparative studies on urethral function.

Urethral pressure profiles (in vivo), opening pressures, and flow rates at opening pressure (in vitro) were determined for female, male, pregnant, ovariectomized (OVX), OVX and then estrogen-treated, and OVX and then progesterone-treated rabbits. Using the isolated whole-urethra preparation, we determined the opening pressures and flow rates as well as the effects of 250 microM phenylephrine, 250 microM bethanechol, and 120 mM KCl on the urethral opening pressure and flow rate. The results demonstrated that (1) the urethral pressure profiles were similar for male and female rabbits, (2) ovariectomy and pregnancy decreased the urethral pressure profiles, (3) estrogen therapy partially reversed the effect of ovariectomy on the urethra, and (4) progesterone therapy had little effect on the urethral pressure profile. With regard to opening pressure and flow, (1) flow at opening pressure was lower in the male than in the female; (2) the opening pressure was increased by ovariectomy; (3) phenylephrine and KCl stimulated a greater response in the male than in the female, whereas the response to bethanechol was significantly lower in the male than in the female; (4) the male had the greatest resistance to flow among all the groups; and (5) ovariectomy increased the resistance to flow and estrogen treatment reversed the effect of ovariectomy. In conclusion, ovariectomy had significant effects on urethral function that were reversed by estrogen therapy but not by progesterone therapy.

Effects of the peptide leukotriene receptor antagonist ICI 198,615 on the in vivo and in vitro changes in guinea pig bladder function which occur after sensitization with ovalbumin.

PURPOSE: The present studies were designed to determine the effects of in vivo and in vitro administration of ICI 198,615 (ICI), a leukotriene receptor antagonist, on the inflammatory changes that occur in the bladder after sensitization with ovalbumin. MATERIALS AND METHODS: The effect of intravenous administration of ICI on urodynamic changes after instillation of ovalbumin to sensitized guinea pigs was evaluated by in vivo cystometry. Responses of in vitro bladder muscle strips to contractile stimuli and ovalbumin were also evaluated in the presence of ICI. RESULTS: In sensitized guinea pigs, in vivo cystometry with ovalbumin induced a marked decrease in bladder capacity and increase in intravesical pressure. Pretreatment with ICI prior to cystometry prevented the ovalbumin-induced changes in capacity and intravesical pressure. There were no significant differences between control and sensitized animals in the responses of in vitro bladder muscle strips to field stimulation or bethanechol. However, maximal contractile responses to ovalbumin were significantly greater in the strips from sensitized animals than in controls. Preincubation with ICI, indomethacin, or pyrilamine alone was unable to inhibit the contractile responses to ovalbumin. However, combined administration of ICI, indomethacin and pyrilamine completely blocked the responses. CONCLUSIONS: In vivo administration of the leukotriene receptor antagonist ICI 198,615 reversed the urodynamic changes induced by ovalbumin challenge in sensitized guinea pigs. These results indicate that leukotrienes are primarily responsible for the changes in in vivo bladder function associated with sensitization.

Effect of mucosal removal on the response of the feline bladder to pharmacological stimulation.

The urothelium plays an important role in the maintenance of normal bladder function. It provides a nonpermeable barrier to the contents of urine. The urothelium is directly involved in the transduction of both intravesical pressure and intravesical volume information to the afferent nerve fibers located within the lamina propria area. A third function may be to modulate bladder contractile function through local secretion of bioactive substances into the muscularis layers adjacent to the urothelium. To test this last hypothesis, the following experiments were performed: Strips of female cat bladders were isolated from the bladder body, base and urethra. The mucosa of alternate adjacent strips was removed, and the contractile response to field stimulation (FS), bethanechol (body), phenylephrine (base, urethra) and KCI was determined. For the bladder body, the strips without mucosa responded to FS, bethanechol, adenosine triphosphate (ATP) and KCI significantly greater than the strips with mucosa intact. For the bladder base and urethra, the contractile responses to FS, KCI and phenylephrine were significantly greater for the strips with mucosa removed as compared with the strips with mucosa intact. For the urethra and bladder base, FS in the presence of phenylephrine produced a relaxation. For the bladder base, the degree of FS relaxation of the isolated strips with mucosa removed was significantly greater than the strips with mucosa intact. For the urethra, FS relaxation was similar for the two groups. In conclusion, removal of the urothelium significantly and substantially increased the contractile response to FS, KCI, bethanechol and phenylephrine. Field stimulation relaxation in the bladder base was also enhanced. Thus in the cat, the mucosa has a significant inhibitory effect on the contractile response of the bladder to stimulation. The mechanism of this activity is not clear at the present time but will be the subject of further study.

The role of the mucosa in the in vitro changes in guinea pig bladder function which occur after sensitization with ovalbumin.

Bladder strips from sensitized guinea pigs respond to ovalbumin challenge with a contraction accompanied by release of histamine, prostaglandin, and leukotriene. Histamine and prostaglandin release occurs preferentially from the bladder mucosa, while leukotrienes are released by the smooth muscle. This study investigated the effects of removal of the bladder mucosa on contractile responses of strips from control and sensitized guinea pigs to electrical stimulation and to contractile agonists. Removal of the mucosa potentiated the contractile response to electrical stimulation at low voltages and pulse durations. The presence of ovalbumin had no effects on the response to electrical stimulation. Removal of the mucosa had no effect on the contractile responses of bladder strips to bethanechol, histamine, KCl, or ovalbumin, although after correction for the decreased strip mass after mucosa removal there was a tendency for the responses to increase. We conclude that the responsiveness of bladder strips from sensitized guinea pigs to ovalbumin challenge results primarily from effects which occur within the smooth muscle.