Frequency Of Bladder Contractions Research Articles

Noradrenergic Pathways Involved in Micturition in an Animal Model of Hydrocephalus-Implications for Urinary Dysfunction.

Hydrocephalus is characterized by enlargement of the cerebral ventricles, accompanied by distortion of the periventricular tissue. Patients with hydrocephalus usually experience urinary impairments. Although the underlying etiology is not fully described, the effects of hydrocephalus in the neuronal network responsible for the control of urination, which involves periventricular areas, including the periaqueductal gray (PAG) and the noradrenergic locus coeruleus (LC). In this study, we aimed to investigate the mechanisms behind urinary dysfunction in rats with kaolin-induced hydrocephalus. For that purpose, we used a validated model of hydrocephalus-the rat injected with kaolin in the cisterna magna-also presents urinary impairments in order to investigate the putative involvement of noradrenergic control from the brain to the spinal cord Onuf's nucleus, a key area in the motor control of micturition. We first evaluated bladder contraction capacity using cystometry. Since our previous characterization of the LC in hydrocephalic animals showed increased levels of noradrenaline, we then evaluated the noradrenergic innervation of the spinal cord's Onuf's nucleus by measuring levels of dopamine β-hydroxylase (DBH). We also evaluated the expression of the c-Fos protooncogene, the most widely used marker of neuronal activation, in the ventrolateral PAG (vlPAG), an area that plays a major role in the control of urination by its indirect control of the LC via pontine micturition center. Hydrocephalic rats showed an increased frequency of bladder contractions and lower minimum pressure. These animals also presented increased DBH levels at the Onuf´s nucleus, along with decreased c-Fos expression in the vlPAG. The present findings suggest that impairments in urinary function during hydrocephalus may be due to alterations in descending noradrenergic modulation. We propose that the effects of hydrocephalus in the decrease of vlPAG neuronal activation lead to a decrease in the control over the LC. The increased availability of noradrenaline production at the LC probably causes an exaggerated micturition reflex due to the increased innervation of the Onuf´s nucleus, accounting for the urinary impairments detected in hydrocephalic animals. The results of the study provide new insights into the neuronal underlying mechanisms of urinary dysfunction in hydrocephalus. Further research is needed to fully evaluate the translational perspectives of the current findings.

The aryl hydrocarbon receptor agonist ITE reduces inflammation and urinary dysfunction in a mouse model of autoimmune prostatitis.

Prostate inflammation is linked to lower urinary tract dysfunction and is a key factor in chronic prostatitis/chronic pelvic pain syndrome. Autoimmunity was recently identified as a driver of prostate inflammation. Agonists of the aryl hydrocarbon receptor (AHR), a ligand-activated transcription factor, have been used to suppress autoimmunity in mouse models of colitis, rhinitis, and dermatitis, but whether AHR agonists suppress prostate autoimmunity has not been examined. Here, we test whether ITE (2-(1'H-indole-3'-carbonyl)-thiazole-4-carboxylic acid methyl ester), an AHR agonist, suppresses inflammation, allodynia, and urinary dysfunction in a mouse model of experimental autoimmune prostatitis (EAP). C57BL/6J adult male mice were immunized with rat prostate antigen to induce EAP or TiterMax Gold® adjuvant (uninflamed control). Mice were also treated with ITE (10 mg/kg/day IP) or DMSO (vehicle, 5 mg/kg/day IP) for 6 days. Using the Nanostring nCounter Inflammation Panel, we evaluated the impact of EAP and ITE on prostatic RNA abundance. We validated EAP and ITE-mediated changes in a subset of RNAs by RT-PCR and RNAScope in situ RNA detection. EAP appeared to heighten histological inflammation in the dorsal prostate, induced tactile allodynia, and appeared to increase the frequency of non-voiding bladder contractions. ITE mitigated some actions of EAP. EAP changed abundance of 40 inflammation-related RNAs, while ITE changed abundance of 28 inflammation-related RNAs. We identified a cluster of RNAs for which ITE protected against EAP-induced changes in the abundance of H2-Ab1, S100a8, and S100a9. ITE also increased the abundance of the AHR-responsive Cyp1a1 RNA. These findings support the hypothesis that ITE activates the AHR in the prostate and reduces autoimmune-mediated prostatitis in mice.

Mirabegron and solifenacin are effective for the management of the increased urinary frequency induced by psychological stress in female mice

Evidence to support the effectiveness of β3-adrenoceptor agonist mirabegron and anti-muscarinic solifenacin in the management of bladder dysfunction caused by psychological stress is lacking. This study investigates whether mirabegron or solifenacin reduces the bladder overactivity caused by water avoidance stress (WAS) in mice. Female mice were exposed to WAS for 1 h/day for 10 days and received either placebo, solifenacin or mirabegron in drinking water. Controls were age-matched without stress exposure. Voiding behaviour and functional isolated whole bladder responses during distension and in response to pharmacological agents and electrical field stimulation was investigated. Urinary frequency was significantly increased following stress. Mice treated with mirabegron or solifenacin displayed significantly fewer voiding events compared to the stressed mice, and voiding frequency in drug-treated animals was comparable to unstressed controls. The maximal contractile responses of bladders to carbachol were significantly enhanced by stress and reduced by mirabegron but not solifenacin. The frequency of phasic bladder contractions following stimulation with carbachol was significantly enhanced following stress and remained elevated in the mirabegron treated group. However, treatment with solifenacin significantly reduced the frequency of phasic contractions to unstressed control levels. Solifenacin and mirabegron are beneficial in reducing the overall voiding dysfunction caused by WAS in mice.

Impairment of AMPK-α2 augments detrusor contractions in bladder ischemia.

PurposeIschemia disrupts cellular energy homeostasis. Adenosine monophosphate-activated protein kinase alpha-2 (AMPK-α2) is a subunit of AMPK that senses cellular energy deprivation and signals metabolic stress. Our goal was to examine the expression levels and functional role of AMPK-α2 in bladder ischemia.Materials and MethodsIliac artery atherosclerosis and bladder ischemia were engendered in apolipoprotein E knockout rats by partial arterial endothelial denudation using a balloon catheter. After eight weeks, total and phosphorylated AMPK-α2 expression was analyzed by western blotting. Structural integrity of AMPK-α2 protein was assessed by Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS). Functional role of AMPK-α2 was examined by treating animals with the AMPK activator 5-aminoimidazole-4-carboxamide-1-beta-D ribofuranoside (AICAR). Tissue contractility was measured in the organ bath and bladder nerve density was examined by immunostaining.ResultsTotal AMPK-α2 expression increased in bladder ischemia, while phosphorylated AMPK-α2 was significantly downregulated. LC-MS/MS suggested post-translational modification of AMPK-α2 functional domains including phosphorylation sites, suggesting accumulation of catalytically inactive AMPK-α2 in bladder ischemia. Treatment of rats with AICAR diminished the force of overactive detrusor contractions and increased bladder capacity but did not have a significant effect on the frequency of bladder contractions. AICAR diminished contractile reactivity of ischemic tissues in the organ bath and prevented loss of nerve fibers in bladder ischemia.ConclusionsIschemia induces post-translational modification of AMPK-α2 protein. Impairment of AMPK-α2 may contribute to overactive detrusor contractions and loss of nerve fibers in bladder ischemia. AMPK activators may have therapeutic potential against detrusor overactivity and neurodegeneration in bladder conditions involving ischemia.

An Efficient Noninvasive Neuromodulation Modality for Overactive Bladder Using Time Interfering Current Method.

The present study aimed to evaluate a new tibial nerve stimulation (TNS) modality, which uses interferential currents, in terms of the stimulation electric field penetration efficiency into the body and physiological effectiveness. In silico experiments were performed to analyze the penetration efficiency of proposed interferential current therapy (ICT). Based on this, we performed in vivo experiments to measure excitation threshold of ICT for the tibial nerve, which is related to stimulation field near the nerve. Regarding analysis of the physiological effectiveness, in vivo ICT-TNS was performed, and changes in bladder contraction frequency and voiding volume were measured. The penetration efficiency and physiological effectiveness of ICT were evaluated by comparison with those of conventional TNS using transcutaneous electrical nerve stimulation (TENS). Simulation results showed that ICT has high penetration efficiency, thereby generating stronger field than TENS. These results are consistent with the in vivo results that nerve excitation threshold of ICT is lower than that of TENS. Moreover, ICT-TNS decreased contraction frequency and increased voiding volume, and its performance was profound compared with that of TENS-TNS. The proposed ICT is more efficient in inducing the stimulation field near the tibial nerve placed deep inside the body compared with conventional TENS and shows a good clinical effectiveness for TNS. The high efficiency of ICT increases the safety of noninvasive neurostimulation; therefore, it has clinical potential to become a promising modality for TNS to treat OAB and other peripheral neurostimulations.

Evaluation of a rat model of functional urinary bladder outlet obstruction produced by chronic inhibition of nitric oxide synthase

AimsLigation of the urethra to create partial bladder outlet obstruction has widely been used as an animal model of bladder obstruction, although obstructive bladder dysfunction may be due to both mechanical and functional obstruction. Previous studies in rodents have demonstrated that long-term nitric oxide (NO) deficiency can lead to detrusor overactivity, and lack of NO may thus cause impairment of bladder outlet relaxation. The aim of this study was to define the characteristics of bladder and urethral dysfunction induced by chronic NO deficiency through both in vivo and in vitro investigations. Main methodsRats were divided into two groups, and one group received an NO synthase inhibitor (Nω-nitro-L-arginine methyl ester hydrochloride: L-NAME) in the drinking water for 4 weeks. Bladder and urethral function were evaluated by continuous cystometry and isovolumetric cystometry. In vitro functional studies of detrusor strips and measurement of the mRNA and protein expression of an ischemic marker and a gap junction protein were also performed in separate rats. Key findingsL-NAME administration raised blood pressure and decreased plasma nitrite/nitrate level compared to the control group. L-NAME treatment increased the frequency of bladder contractions and the residual volume, and elevated urethral pressure and bladder contraction pressure. In addition, carbachol-induced contraction was reduced in isolated detrusor strips from the L-NAME group, and bladder expression of HIF-1 and connexin 43 showed upregulation. SignificanceThese findings suggest that chronic administration of L-NAME to rats induces bladder hyperactivity with residual urine, and may provide a useful model of functional bladder obstruction.

Modulatory effects of intravesical P2X2/3 purinergic receptor inhibition on lower urinary tract electromyographic properties and voiding function of female rats with moderate or severe spinal cord injury.

To evaluate the role that intravesical P2X2/3 purinergic receptors (P2X2/3Rs) play in early and advanced neurogenic lower urinary tract (LUT) dysfunction after contusion spinal cord injury (SCI) in female rats. Female Sprague-Dawley rats received a thoracic Th8/Th9 spinal cord contusion with either force of 100 kDy (cN); moderate) or 150 kDy (cN; severe); Sham rats had no injury. Evaluations on urethane-anesthetised rats were conducted at either 2 or 4 weeks after SCI. LUT electrical signals and changes in bladder pressure were simultaneously recorded using cystometry and a set of custom-made flexible microelectrodes, before and after intravesical application of the P2X2/3R antagonist AF-353 (10 μM), to determine the contribution of P2X2/3R-mediated LUT modulation. Severe SCI significantly increased bladder contraction frequency, and reduced both bladder pressure amplitude and intraluminal-pressure high-frequency oscillations (IPHFO). Intravesical P2X2/3R inhibition did not modify bladder pressure or IPHFO in the Sham and moderate-SCI rats, although did increase the intercontractile interval (ICI). At 2 weeks after SCI, the Sham and moderate-SCI rats had significant LUT electromyographic activity during voiding, with a noticeable reduction in LUT electrical signals seen at 4 weeks after SCI. Intravesical inhibition of P2X2/3R increased the ICI in the Sham and moderate-SCI rats at both time-points, but had no effect on rats with severe SCI. The external urethral sphincter (EUS) showed strong and P2X2/3R-independent electrical signals in the Sham and moderate-SCI rats in the early SCI stage. At 4 weeks after SCI, the responsiveness of the EUS was significantly attenuated, independently of SCI intensity. This study shows that electrophysiological properties of the LUT are progressively impaired depending on SCI intensity and that intravesical P2X2/3R inhibition can attenuate electrical activity in the neurogenic LUT at early, but not at semi-chronic SCI. This translational study should be useful for planning clinical evaluations.

Evaluation of Pulse-Width of Spinal Nerve Stimulation in a Rat Model of Bladder Micturition Reflex

The spinal nerve stimulation (SNS) evoked motor threshold (Tmot ) response across different pulse-widths (PWs) was first explored and a subset of selected stimulation PWs were further assessed with respect to bladder reflex contraction (BRC). In anesthetized female rats, wire electrodes were placed under each of the L6 spinal nerves to produce bilateral SNS. The relationship of Tmot response with PW was analyzed using a monoexponential nonlinear regression. A cannula was placed into the bladder via the urethra to ensure an isovolumetric bladder. Saline infusion induced BRC. The chronaxie of the Tmot -PW curve was 0.04 ms. The stimulation charges/energies (current × PW) associated with shorter PWs of 0.02, 0.03, and 0.06 ms were significantly lower than those with longer PW (e.g., >0.15 ms). SNS (Tmot , 10 Hz) at selected PWs from 0.03 to 0.21 ms inhibited the frequency of BRCs. There were no significantly different attenuations among tested PWs. SNS of PWs of 0.03, 0.06, and 0.09 ms decreased bladder contraction frequency from 103 ± 3%, 100 ± 4%, and 103 ± 4% of controls, to 52 ± 16% (n = 8, p = 0.02, paired t-test), 56 ± 15% (n = 11, p = 0.02) and 40 ± 19% (n = 10, p = 0.01), respectively. Effective PWs to produce bladder inhibitory effects in the rat appear much shorter than 0.21 ms typically used with sacral neuromodulation in practice. Potential battery savings manifested by shorter PW while maintaining equivalent efficacy would provide more efficient therapy delivery and increased longevity of the stimulator.

Improvement in Lower Urinary Tract Function Following a Single Bout of Acute Intermittent Hypoxia in Rats with Chronic Spinal Cord Injury

Exposure to acute intermittent hypoxia (AIH) has been reported to improve respiratory and non‐respiratory somatic motor function in rodents and humans and has been proposed as a potential therapeutic intervention for treatment of chronic motor incomplete spinal cord injury (SCI). The goal of the present study is to characterize potential beneficial effects of AIH to improve lower urinary tract (LUT) function following incomplete SCI. To this end, female Sprague Dawley rats (225–250 g; n=11) received moderate midthoracic (T7–T9) spinal cord contusion injuries (200 kilodynes, Infinite Horizon Impactor; n=7) or sham injuries (spinal cord exposed, no injury; n=4) under isoflurane anesthesia. Following SCI, rats were inspected daily for SCI‐related issues (e.g., bladder/urinary tract infections, autophagia, overgrooming), and bladders were manually expressed until spontaneous voiding capability returned. Four weeks following the SCI/sham, bladder intravesical pressure and external urethral sphincter (EUS) EMG activity were recorded under urethane anesthesia (1.4 g/kg) during continuous infusion of saline (0.04 ml/min) into the bladder to elicit reflex micturition events (spontaneously breathing, vagus nerves intact). In addition, EMG data were acquired from the diaphragm and genioglossus muscles to assess effects on inspiratory motor outputs. Following 30–40 minutes of baseline recording, rats were exposed to a single bout of AIH consisting of three five‐minute episodes of hypoxia (10% O2; 90% N2) each separated by five‐minute exposures to room air. Following the AIH exposure, rats continued to spontaneously breathe room air, and data acquisition continued for up to 120 minutes. Prior to AIH, SCI rats exhibited pronounced rhythmic bladder activity consisting of both non‐voiding and voiding contractions. Typically, a series of non‐voiding bladder contractions with increasing amplitude preceded a voiding contraction, and the voiding contractions were incomplete resulting in continuous residual volume and an elevated minimum intravesical pressure. During each hypoxic episode during the AIH, the frequency of rhythmic bladder contraction increased and was associated with increased output volume and decreasing minimum intravesical pressure. In contrast, there was a marked decrease in rhythmic bladder activity during the normoxic period between hypoxic episodes with decreased output volume and increased minimum intravesical pressure. Immediately following the end of AIH, a similar decrease in rhythmic bladder contractions and output volume was observed and persisted for 5–10 minutes after which a more normal pattern of bladder activity (compared to baseline) emerged characterized by fewer non‐voiding contractions, increased output volume per volume per contraction, decreased minimum intravesical pressure, and a threshold‐driven pattern of bladder contraction that persisted up to 120 minutes post AIH. Sham operated rats exhibited a threshold‐driven pattern of bladder activity with efficient voiding and similar hypoxia‐dependent modulation of contraction frequency and increased output volume following AIH. These data suggest that AIH‐induced neuroplasticity results in improved LUT function in rats with chronic incomplete SCI and is a potentially promising therapeutic intervention for humans with SCI.Support or Funding InformationSupport for this project was provided by The Thomas Hartman Center for Parkinson's Research at Stony Brook University and The SUNY Brain Network of Excellence.

Netupitant, a Potent and Highly Selective NK1 Receptor Antagonist, Alleviates Acetic Acid-Induced Bladder Overactivity in Anesthetized Guinea-Pigs.

Introduction. Tachykinins potently contract the isolated urinary bladder from a number of animal species and play an important role in the regulation of the micturition reflex. On the guinea-pig isolated urinary bladder we examined the effects of a new potent and selective NK1 receptor antagonist (netupitant) on the contractions induced by a selective NK1 receptor agonist, SP-methylester (SP-OMe). Moreover, the effects of netupitant and another selective NK1 antagonist (L-733,060) were studied in anesthetized guinea-pigs using two experimental models, the isovolumetric bladder contractions and a model of bladder overactivity induced by intravesical administration of acetic acid (AA).Methods and Results. Detrusor muscle strips were mounted in 5 mL organ baths and isometric contractions to cumulative concentrations of SP-OME were recorded before and after incubation with increasing concentrations of netupitant. In anesthetized female guinea-pigs, reflex bladder activity was examined under isovolumetric conditions with the bladder distended with saline or during cystometry using intravesical infusion of AA. After a 30 min stabilization period, netupitant (0.1–3 mg/kg, i.v.) or L-733,060 (3–10 mg/kg, i.v.) were administered. In the detrusor muscle, netupitant produced a concentration-dependent inhibition (mean pKB = 9.24) of the responses to SP-OMe. Under isovolumetric conditions, netupitant or L-733,060 reduced bladder contraction frequency in a dose-dependent manner, but neither drug changed bladder contraction amplitude. In the AA model, netupitant dose-dependently increased intercontraction interval (ICI) but had no effect on the amplitude of micturition (AM). L-733,060 dose-dependently increased ICI also but this effect was paralleled by a significant reduction of AM.Conclusion. Netupitant decreases the frequency of reflex bladder contractions without altering their amplitude, suggesting that this drug targets the afferent limb of the micturition reflex circuit and therefore may be useful clinically in treating bladder overactivity symptoms.

MP25-07 PROSTATIC INFLAMMATION EVOKES UPREGULATION OF NEUROTROPHINS IN SENSORY GANGLIA: POSSIBLE CONTRIBUTION TO DYSFUNCTIONAL VOIDING

INTRODUCTION AND OBJECTIVES: Chronic prostatitis/ chronic pelvic pain syndrome (CP/CPPS) is a debilitating condition characterized mainly by pelvic pain. In addition, many patients report voiding complaints and lower urinary tract symptoms (LUTS). While the mechanism behind these voiding complaints is not well understood, it is possible that inflammation induces fibrosis within the prostate, as has been hypothesized to occur in benign prostatic hyperplasia (BPH). To better understand this process, we developed two transient infectioninduced prostatitis murine models where LUTS can be studied in the presence or absence of chronic pain. METHODS: C57BL/6 (B6) and NOD/ShiLtJ (NOD) mice (5-7 weeks old) were transurethrally infected with Escherichia coli strain CP1, a bacteria from a CP/CPPS patient that clears from the prostate and bladder of mice by 30 days. At post-infection days 5, 14, and 35, the mice were pain tested for referred pain with von-Frey filaments and the prostates and bladders harvested for analysis (4-8 mice per time point). Additionally, urodynamic evaluation was completed at day 35 under urethane anesthesia. Prostate and bladder specimens were analyzed by flow cytometry and real-time quantitative PCR (QT-PCR) for the presence of cells and markers involved in fibrosis. RESULTS: B6 mice did not develop pain, however, NOD mice developed significant pain at all time points. Cystometry showed increased frequency of bladder contractions in CP-1 infected mice of both strains. By flow cytometry, B6 mice developed an increase in fibrocytes (vimentinþ, CD45þ) in the prostate at day 14 which became statistically significant at day 35. A similar but delayed trend was noted in NOD mice with an increase in fibrocytes at day 35, however, this was not statistically significant. NOD mice did develop a significant increase in vimentinþ cells (mesenchymal cells) at day 14. The bladders of B6 mice demonstrated a significant increase in vimentinþ cells at day 35 and at day 14 in the NOD mice. There was no difference in the presence of fibrocytes in the bladders. B6 mice had a >1 fold increase in expression of collagen 1a1, 1a2, aSMA, and CXCL5 at day 14 by QTPCR. A similar increase in these markers was noted in the NOD mice but at the earlier time point of day 5. CONCLUSIONS: We have demonstrated in infection-induced prostatitis murine models that urinary frequency develops in the presence and absence of pain. This is associated with the concurrent suggestion of fibrosis within the prostate, with altered kinetics of development in each model. Further studies are underway to identify the pathogenesis of LUTS in the context of inflammation and pain.

Mechanisms of Pelvic Organ Cross-Talk: Impact of Urethral Ligation on the Inhibitory Rectovesical Reflex

The existence of an inhibitory rectovesical reflex elicited by noxious colorectal afferent input was previously documented in an isovolumetric cystometry model with a ligated urethra. We compared the effect of noxious colorectal distension on bladder cystometry with an open and a ligated urethra. We used female Sprague Dawley® rats anesthetized with urethane. The effect of noxious (60 mm Hg) colorectal distension on intermittent and continuous cystometry with an open urethra was studied and then compared to inhibiting isovolumetric bladder contractions in the same rat after ligating the urethra. We evaluated volume, the pressure micturition threshold, the intercontraction interval, bladder contraction frequency and amplitude of micturition contractions. Noxious colorectal distension at 60 mm Hg did not significantly influence volume or the pressure micturition threshold during intermittent cystometry. It also did not influence the pressure micturition threshold, the intercontraction interval or bladder contraction frequency during continuous cystometry. After urethral ligation 60 mm Hg colorectal distension inhibited isovolumetric bladder contraction frequency in the same rat (mean ± SEM 0.363± 0.207 vs 0.886 ± 0.106 contractions per minute, p <0.05). This inhibition persisted a mean of 289.08 ± 91.24 seconds after deflating the rectal balloon. The inhibitory rectovesical reflex elicited by noxious colorectal distension clearly occurred in an isovolumetric bladder model with a ligated urethra but only to a negligible extent in filling related voiding contractions. Ourresults suggest that the inhibitory rectovesical reflex, most likely at the levelof the lumbosacral spinal cord, is the result of an additive noxious urethral and colonic afferent stimulus.

Restoring the glycosaminoglycans layer in recurrent cystitis: Experimental and clinical foundations

Restoring the bladder glycosaminoglycans layer has recently been introduced as prophylactic treatment for recurrent urinary tract infections. Herein, we analyze the latest main clinical and experimental studies to support this therapeutic option. An electronic research was carried out in the most common databases in order to identify any published studies. Retrieved studies were categorized as experimental or clinical according to their setting. For the clinical studies, the evidence level was assigned. A total of 13 laboratory studies showed how bladder glycosaminoglycans instillations act: attenuation of the inflammation process, reduction of bladder contraction amplitude and frequency, reduction of epithelium damage, and lower bacterial growth in urine and tissue samples. Likewise, two randomized clinical trials with grade 2 evidence level and two case series with grade 4 evidence level reported glycosaminoglycans as an alternative to reduce episodes and to prolong recurrence time in patients with recurrent urinary tract infections. At least 12 months of follow up was completed. No serious adverse events were reported. Compared with a placebo, in one randomized study a significantly higher maximum cystometric capacity was obtained, whereas in the other study a significant increase in quality of life scores was reported too. An improvement in the urinary symptoms score was reported by the two randomized trials. Although the clinical use of glycosaminoglycans replacement therapy for recurrent urinary tract infections is supported by a small number of clinical studies with different evidence levels, the laboratory studies show that glycosaminoglycans could have a protective role against inflammatory factors, supporting the idea "to restore the glycosaminoglycans bladder layer to prevent chronic disease course".

Can the adrenergic system be implicated in the pathophysiology of bladder pain syndrome/interstitial cystitis? A clinical and experimental study

To evaluate sympathetic system activity in bladder pain syndrome/interstitial cystitis (BPS/IC) patients and to investigate if chronic adrenergic stimulation in intact rats induces BPS/IC-like bladder modifications. Clinical study--In BPS/IC patients and aged and body mass index matched volunteers TILT test was undertaken and catecholamines were measured in plasma and 24 hr urine samples. Experimental study--Phenylephrine was injected subcutaneously (14 days) to female Wistar rats. Pain behavior, spinal Fos expression, urinary spotting, number of fecal pellets expelled, frequency of reflex bladder contractions, and urothelial height were analyzed. Urothelium permeability was investigated by trypan blue staining. Immunoreactivity against caspase 3 and bax were studied in the urothelium and against alpha-1-adrenoreceptor and TRPV1 in suburothelial nerves. Mast cell number was determined in the sub-urothelium. In rats with lipopolysaccharide-induced cystitis, urinary catecholamines, and Vesicular Monoamine Transporter 2 (VMAT2) expression in bladder nerves were analyzed. The TILT test showed an increase of sympathetic activity. Noradrenaline levels in blood at resting conditions and in 24-hr urine samples were higher in BPS/IC patients. Phenylephrine administration increased visceral pain, spinal Fos expression, bladder reflex activity, urinary spotting and the number of expelled fecal pellets. The mucosa showed urothelial thinning and increased immunoreactivity for caspase 3 and bax. Trypan blue staining was only observed in phenylephrine treated animals. Suburothelial nerves co-expressed alpha1 and TRPV1. Mastocytosis was present in the suburothelium. Cystitis increased sympathetic nerve density and urinary noradrenaline levels. Excessive adrenergic stimulation of the bladder may contribute to the pathophysiological mechanisms of BPS/IC.

Stability of the acetic acid-induced bladder irritation model in alpha chloralose-anesthetized female cats.

Time- and vehicle-related variability of bladder and urethral rhabdosphincter (URS) activity as well as cardiorespiratory and blood chemistry values were examined in the acetic acid-induced bladder irritation model in α-chloralose-anesthetized female cats. Additionally, bladder and urethra were evaluated histologically using Mason trichrome and toluidine blue staining. Urodynamic, cardiovascular and respiratory parameters were collected during intravesical saline infusion followed by acetic acid (0.5%) to irritate the bladder. One hour after starting acetic acid infusion, a protocol consisting of a cystometrogram, continuous infusion-induced rhythmic voiding contractions, and a 5 min “quiet period” (bladder emptied without infusion) was precisely repeated every 30 minutes. Administration of vehicle (saline i.v.) occurred 15 minutes after starting each of the first 7 cystometrograms and duloxetine (1mg/kg i.v.) after the 8th. Acetic acid infusion into the bladder increased URS-EMG activity, bladder contraction frequency, and decreased contraction amplitude and capacity, compared to saline. Bladder activity and URS activity stabilized within 1 and 2 hours, respectively. Duloxetine administration significantly decreased bladder contraction frequency and increased URS-EMG activity to levels similar to previous reports. Cardiorespiratory parameters and blood gas levels remained consistent throughout the experiment. The epithelium of the bladder and urethra were greatly damaged and edema and infiltration of neutrophils in the lamina propria of urethra were observed. These data provide an ample evaluation of the health of the animals, stability of voiding function and appropriateness of the model for testing drugs designed to evaluate lower urinary tract as well as cardiovascular and respiratory systems function.

Quantification of effectiveness of bilateral and unilateral neuromodulation in the rat bladder rhythmic contraction model

BackgroundUsing the isovolumetric bladder rhythmic contraction (BRC) model in anesthetized rats, we have quantified the responsiveness to unilateral and bilateral stimulation of the L6 spinal nerve (SN) and characterized the relationship between stimulus intensity and inhibition of the bladder micturition reflex.MethodsA wire electrode was placed under either one or both of the L6 SN roots. A cannula was placed into the bladder via the urethra and the urethra was ligated. Saline infusion induced BRC.ResultsAt motor threshold (Tmot) intensity, SN stimulation of both roots (10 Hz) for 10 min reduced bladder contraction frequency from 0.63 ± 0.04 to 0.17 ± 0.09 contractions per min (26 ± 14% of baseline control; n = 10, p < 0.05). However, the same intensity of unilateral stimulation (n = 15) or sequential stimulation of both SNs (e.g. 5 min per side alternatively for a total of 10 min or 20 min) was less efficacious. The greater sensitivity to bilateral stimulation is not dependent upon precise bilateral timing of the stimulation pulses. Bilateral stimulation also produced both acute and prolonged- inhibition on bladder contractions in a stimulation intensity dependent fashion.ConclusionsUsing the bladder rhythmic contraction model, bilateral stimulation was more effective than unilateral stimulation of the SN. Clinical testing should be conducted to further compare efficacies of unilateral and bilateral stimulation. Bilateral stimulation may allow the use of lower stimulation intensities to achieve higher efficacy for neurostimulation therapies on urinary tract control.

Role of the endogenous opioid system in modulation of urinary bladder activity by spinal nerve stimulation

The role of the endogenous opioid system in modulation of urinary bladder activity by spinal nerve (SN) stimulation was studied in anesthetized female rats, using the rat model of isovolumetric bladder contraction. SN stimulation at a fixed frequency of 10 Hz attenuated bladder contraction frequency; the magnitude of the inhibition was directly proportional to the current intensity. Neither the κ-opioid antagonist nor-binaltorphimine (2 mg/kg iv) nor the δ-opioid antagonist naltrindole (5 mg/kg iv) attenuated the bladder inhibitory response to SN stimulation. In contrast, the μ-opioid receptor antagonist naloxone (NLX; 0.03 mg/kg iv) blocked the inhibitory responses evoked by SN stimulation at therapeutic current intensities at ≤1 × motor threshold current (Tmot). An action at spinal and supraspinal centers was further confirmed by the ability of intrathecal or intracerebroventricular administration of NLX methiodide to attenuate the bladder inhibitory effects of 1 × Tmot SN stimulation. The magnitude of SN-mediated neuromodulation using therapeutically relevant stimulation intensity (Tmot) is equivalent to 0.16 mg/kg of systemically administered morphine, which produces 50% inhibition of bladder contraction frequency. These results suggest that the inhibitory effects of lower intensity SN stimulation may be mediated through the release of endogenous μ-opioid peptides. Additionally, these data suggest that neuromodulation may offer a mode of treating the symptoms of overactive bladder with efficacy equal to the opioid drugs but without their liability for abuse and dependence.

Excitatory effects of neurokinin 2 and bombesin receptor peptide agonists vivo in urinary tract of rats with voiding dysfunctions

Bladder function was examined in anesthetized and awake adult female Sprague Dawley rats. Voiding dysfunction models included STZ‐induced diabetic (streptozotocin; STZ; 65mg/kg i.p., 9–20 weeks post treatment), polyuria (5% sucrose‐fed; &gt;;3 weeks post treatment) and spinal cord injured (SCI; T8‐T9 cut; acute or 3 days – 6 weeks post SCI) rats. Telemetry transmitters were implanted into polyuria group to measure bladder pressure in freely moving rats. [Lys5,MeLeu9,Nle10]‐NKA(4–10) (LMN‐NKA), a neurokinin 2 receptor (NK2R) agonist, gastrin‐releasing peptide (GRP), a bombesin receptor 2 (BBR2) agonist, and neuromedin B (NMB), a BBR1 agonist, were delivered via jugular vein catheters.In urethrane anesthetized STZ and control rats, LMN‐NKA (0.1–100μg/kg), GRP (0.1–300μg/kg) and NMB (0.1–300μg/kg) dose dependently increased bladder contraction (BC, up to 4x) and micturition frequency. In STZ rats, LMN‐NKA reduced residual volume by half. In acute anesthetized SCI rats, LMN‐NKA (&gt;;10μg/kg) induced urine release (voiding efficiency ~40%). In awake polyuria rats, LMN‐NKA and GRP (1–100μg/kg) induced BC that was often associated with voiding. In awake SCI rats, LMN‐NKA (100μg/kg) and GRP (100, 300μg/kg) triggered voiding in 25 – 80% of rats.Activation of NK2R and BBRs enhanced or triggered voiding in naïve rats and in several models of voiding dysfunction including STZ and SCI rats.

Mechanisms of Pelvic Organ Crosstalk: 1. Peripheral Modulation of Bladder Inhibition by Colorectal Distention in Rats

Bladder activity can be inhibited by afferent input from the colorectum (inhibitory rectovesical reflex). We evaluated the functional response of the rat bladder to nonnoxious and noxious colorectal distention, and investigated the mechanical and pharmacological peripheral modulation of this response. In 70 female Sprague-Dawley® rats we evaluated the effect of nonnoxious (20 mm Hg) and noxious (40 and 60 mm Hg) colorectal distention on the micturition volume threshold and on bladder activity in a filled bladder. We also studied the effect of rectal balloon size (1.5 vs 3.5 cm long), and rectal administration of 2% lidocaine jelly or 1 mM allyl isothiocyanate solution on the inhibitory rectovesical reflex. Colorectal distention at 60 mm Hg increased the micturition volume threshold (mean ± SE 0.640 ± 0.056 vs 0.448 ± 0.035 ml in controls, p <0.001). Bladder contraction frequency was significantly decreased by 40 and 60 mm Hg colorectal distention vs controls (mean 0.62 ± 0.06 and 0.33 ± 0.05 per minute, respectively, vs 0.77 ± 0.03, each p <0.001). These effects were reversible and pressure dependent (p <0.001), and more pronounced using a large rectal balloon (mean 40 vs 60 mm Hg colorectal distention 0.35 ± 0.12 vs 0.07 ± 0.04 per minute, p = 0.004). We noted no significant graded inhibition of bladder contraction amplitude or duration. The inhibitory rectovesical reflex was reversibly abolished by intrarectal lidocaine administration. Intrarectal allyl isothiocyanate administration significantly increased the effect of noxious colorectal distention on bladder contraction frequency. Only noxious levels of colorectal distention initiated the inhibitory rectovesical reflex. The effect increased with rectal balloon size and with intrarectal administration of allyl isothiocyanate. It was reversibly abolished by lidocaine. Results suggest that spinal interneurons are the mechanism behind the inhibitory rectovesical reflex.

Effects of Acute Sacral Neuromodulation on Bladder Reflex in Complete Spinal Cord Injury Rats

Neurogenic bladder associated with spinal cord injury (SCI) often results in serious disruption of lower urinary tract function. Compared to conventional therapies, sacral neuromodulation (SNM) may offer an alternative, non-destructive treatment for SCI patients with bladder dysfunction. Understanding bladder reflex changes following SCI and the effects of SNM may yield new insights for innovative use of this promising technique. Using a SCI rat model developed in this study, we investigated: 1) the bladder responses with different grades of bladder filling in intact and SCI rats; and 2) the effects of acute SNM on bladder reflex responses in SCI rats. An SCI rat model with overactive bladder was developed and evaluated in this study to examine the effects of acute SNM on bladder reflex in complete SCI rats. Twelve adult female Sprague-Dawley rats were divided into three groups; group I: spinally intact rats (N = 4), group II: transected (T9-T10) rats (N = 4), i.e., SCI rats, and group III: SCI rats with SNM treatment (N = 4). All rats were anesthetized and set up for continuous saline infusion. Cystometric parameters, including contraction period, contraction duration, bladder peak pressure, and number of uninhibited contractions, were analyzed and compared between groups and between conditions with and without SNM treatment for SCI rats. In the intact rats, the frequency of bladder contraction was dependent upon the rate of bladder filling, while the spinal transected rats exhibited large fluctuation and demonstrated different patterns in response to saline infusion. Moreover, the bladder in SCI rats demonstrated an increased contraction period and a decreased contraction strength compared to the intact rats (all p < 0.05). In SCI rats under acute SNM treatment, bladder contraction period and duration tended to become longer, and the bladder peak pressure was decreased. The accumulating evidence indicated that acute SNM had inhibiting effects for bladder overactivity following SCI. The spinal rat model developed in this study was suitable to investigate the effect of sacral neural stimulation on micturition reflex. The results of present study demonstrated that the micturition reflex can be modulated by sacral neural stimulation.