Micturition Pressure Research Articles

Induction of detrusor underactivity by extensive vascular endothelial damages of iliac arteries in a rat model and its pathophysiology in the genetic levels

We tried to establish a reliable detrusor underactivity (DUA) rat model and to investigate pathophysiology of chronic bladder ischemia (CBI) on voiding behavior and bladder function. Adult male rats were divided into five groups. The arterial injury (AI) groups (AI-10, AI-20, AI-30) underwent vascular endothelial damage (VED) of bilateral iliac arteries (with 10, 20, and 30 bilateral repetitions of injury, respectively) and received a 1.25% cholesterol diet. The sham group underwent sham operation and received the same diet. Controls received a regular diet. After 8 weeks, all rats underwent unanesthetized cystometrogram. Bladder tissues were processed for organ bath investigation, immunohistochemistry staining, and genome-wide gene expression analysis. Awake cystometry analysis showed that frequency of voiding contractions and micturition pressure were lower in the AI-30 group than in sham group (p < 0.01). Contractile responses to various stimuli were lower in AI-20 and AI-30 groups (both p < 0.001). In the AI-20 and AI-30 groups, atherosclerotic occlusion in the iliac arteries, tissue inflammation, fibrosis, denervation, and apoptosis of bladder muscle were prominent compared to the sham. Mechanistically, the expression of purinergic receptor P2X-1 was reduced in the AI-30 group, and the genome-wide gene expression analysis revealed that genes related to IL-17 and HIF-1 signaling pathways including INF-γ receptor-1 and C-X-C motif chemokine ligand-2 were upregulated in the CBI-induced DUA rat model. A rat model of progressive VED successfully induced DUA. Abnormal tissue inflammation, fibrosis, denervation, and bladder muscle tissue apoptosis may be involved in CBI-induced DUA pathophysiology.

Effects of the combination of vibegron and imidafenacin on bladder function in urethane-anesthetized rats

The combination of a β3-adrenoceptor agonist and an antimuscarinic agent was revealed to be more effective than monotherapy for patients with overactive bladder and in animal models. However, its influence on voiding functions has not been well documented. Therefore, during intermittent-cystometry, we studied the effects of vibegron (a novel β3-adrenoceptor agonist) and imidafenacin (an antimuscarinic agent) alone to determine their dose levels for the combination study. Then, the effects of the combination on voiding functions were investigated in urethane-anesthetized rats (1.0 g/kg s.c.). Independently, vibegron (0.3–3 mg/kg, i.v.) and imidafenacin (0.001 and 0.003 mg/kg, i.v.) dose-dependently increased bladder capacity and voided volume, without affecting voiding functions such as residual volume, voiding efficiency, and micturition pressure. However, vibegron also increased bladder compliance. The combination of vibegron (3 mg/kg) and imidafenacin (0.003 mg/kg) significantly increased bladder capacity and voided volume when compared to those with monotherapy using each individually. The combination did not change residual volume, voiding efficiency, and micturition pressure, compared to those in the vehicle group. We identified no responses in resiniferatoxin (RTX)-treated rats, as opposed to those identified after administering vibegron (3 mg/kg), imidafenacin (0.003 mg/kg), or both to non-RTX-treated rats. These outcomes might have resulted from the combination of the increased effect of vibegron on bladder compliance and the inhibitory effect of both vibegron and imidafenacin on the activation of bladder afferent nerves.

Role of p38 MAP kinase signaling pathways in storage and voiding dysfunction in mice with spinal cord injury.

To investigate the role of p38 MAP kinase in lower urinary tract dysfunction in mice with spinal cord injury (SCI). Cystometry and external urethral sphincter-electromyography were performed under an awake condition in 4-week SCI female mice. Two weeks after SCI, a catheter connected to an osmotic pump filled with a p38 mitogen-activated protein kinase (MAPK) inhibitor or artificial cerebrospinal fluid (CSF) was implanted into the intrathecal space of L6-S1 spinal cord for continuous intrathecal instillation at infusion rate of 0.51 μL/h for 2 weeks before the urodynamic study. L6 dorsal root ganglia were then removed from CSF and p38 MAPK inhibitor-treated SCI mice as well as from CSF-treated normal (spinal intact) mice to evaluate the levels of transient receptor potential cation channel subfamily V member 1 (TRPV1), tumor necrosis factor-α (TNF-α), and inducible nitric oxide synthase (iNOS) transcripts by real-time polymerase chain reaction. In p38 MAPK inhibitor-treated SCI mice, nonvoiding contractions during bladder filling, bladder capacity, and post-void residual volume were significantly reduced while micturition pressure and voiding efficiency were significantly increased in comparison to these measurements in CSF-treated SCI mice. The expression of TRPV1, TNF-α, and iNOS messenger RNA was increased in SCI mice compared with expression in spinal intact mice and significantly decreased after p38 MAPK inhibitor treatment. The p38 MAPK signaling pathway in bladder sensory neurons or in the spinal cord plays an important role in storage and voiding problems such as detrusor overactivity and inefficient voiding after SCI.

AMPK Alters Detrusor Contractility During Emptying in Normal Bladder and Hypertrophied Bladder with Partial Bladder Outlet Obstruction via CaMKKβ.

AMP-activated protein kinase (AMPK) has been implicated in contractility changes in bladders with partial bladder outlet obstruction (PBOO), but the role of AMPK in the contractile response of normal bladder remains unclear. We investigated the phosphorylation of AMPKα and expression of the involved upstream AMPK kinases (AMPKKs) in a model of bladders with PBOO and sought to determine whether the pharmacological inhibition of these two factors affected detrusor contractility in normal bladders, using female Sprague–Dawley rats. Cystometry and Western blot analysis were performed in rats that were subjected to PBOO induction or a sham operation. Cystometry was performed in normal rats that received selective inhibitors of AMPKα and Ca2+/calmodulin-dependent protein kinase kinase (CaMKKβ) (compound C and STO-609, respectively) at doses determined in the experiments. In the PBOO bladders, bladder weight and micturition pressure (MP) were higher and AMPKα phosphorylation (T172) and CaMKKβ expression was significantly reduced. Compound C and STO-609 increased MP. The increased contractile response in bladders with PBOO-induced hypertrophy was related to decreased CaMKKβ/AMPK signaling activity, and the pharmacological inhibition of this pathway in normal bladders increased detrusor contractility, implying a role of CaMKKβ/AMPK signaling in the bladder in the regulation of detrusor contractility.

The effects of neuromodulation in a novel obese-prone rat model of detrusor underactivity.

Obesity is a global epidemic associated with an increased risk for lower urinary tract dysfunction. Inefficient voiding and urinary retention may arise in late-stage obesity when the expulsive force of the detrusor smooth muscle cannot overcome outlet resistance. Detrusor underactivity (DUA) and impaired contractility may contribute to the pathogenesis of nonobstructive urinary retention. We used cystometry and electrical stimulation of peripheral nerves (pudendal and pelvic nerves) to characterize and improve bladder function in urethane-anesthetized obese-prone (OP) and obese-resistant (OR) rats following diet-induced obesity (DIO). OP rats exhibited urinary retention and impaired detrusor contractility following DIO, reflected as increased volume threshold, decreased peak micturition pressure, and decreased voiding efficiency (VE) compared with OR rats. Electrical stimulation of the sensory branch of the pudendal nerve did not increase VE, whereas patterned bursting stimulation of the motor branch of the pudendal nerve increased VE twofold in OP rats. OP rats required increased amplitude of electrical stimulation of the pelvic nerve to elicit bladder contractions, and maximum evoked bladder contraction amplitudes were decreased relative to OR rats. Collectively, these studies characterize a novel animal model of DUA that can be used to determine pathophysiology and suggest that neuromodulation is a potential management option for DUA.

MP26-04 IMPACT OF DAI-KEN-CHU-TOU ON URINARY FREQUENCY INDUCED BY COLD STRESS IN RATS

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Basic Research & Pathophysiology I1 Apr 2017MP26-04 IMPACT OF DAI-KEN-CHU-TOU ON URINARY FREQUENCY INDUCED BY COLD STRESS IN RATS Tetsuichi Saito, Tetsuya Imamura, Tomonori Minagawa, Takashi Nagai, Teruyuki Ogawa, and Osamu Ishizuka Tetsuichi SaitoTetsuichi Saito More articles by this author , Tetsuya ImamuraTetsuya Imamura More articles by this author , Tomonori MinagawaTomonori Minagawa More articles by this author , Takashi NagaiTakashi Nagai More articles by this author , Teruyuki OgawaTeruyuki Ogawa More articles by this author , and Osamu IshizukaOsamu Ishizuka More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.775AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Cold stress produced by sudden change or continuous exposure to low temperature exacerbates lower urinary tract symptoms (LUTS), such as urinary urgency, frequency, and nocturia. Stimulation of C-fibers and TRPM8 channels have been reported as mechanisms of cold stress related LUTS. Dai-ken-chu-tou, a chinese herbal medicine has been traditionally used for improvement of bowel conditions. In basic research, dai-ken-chu-tou has been reported to influence intestinal tracts by decreasing adrenomeduline and decreasing the stimulation of the pathway of TRPV1 and TRPA1, which are also important pathways with bladder function. We examined whether dai-ken-chu-tou improves cold stress related LUTS in rats. METHODS A total of 22 Sprague-Dawley rats at postnatal week 10 were used in the experiments. The animals sere randomly divided into 2 gtoups, which were kept with Dai-ken-chu-tou-including food (2700mg/kg) or normal food for 4 weeks. After 4 weeks, cystometography (CMG) was performed under awake condition. CMG was first performed in room temperature (RT) for 20 minuets. Rats were then put into low temperature (LT) for 40 minutes. After LT, rats were put into RT for 20 minutes. After CMG, the whole bladder was removed and real time PCR was performed. RESULTS Results of the CMG are shown in Fig 1. Basal pressure and micturition pressure did not show a difference between control rats and Dai-ken-chu-tou rats, but change rate with cold stress in voiding interval and micturition volume did show a significant difference. Results of real time PCR are shown in Fig 2. Significant decrease of P2X3, TRPV1, and TRPM8 in the bladder was seen in Dai-ken-chu-tou rats. CONCLUSIONS Dai-ken-chu-tou improved cold stress related frequency in rats. Down regulation of P2X3, TRPV1 and TRPM8 may have a relation with the improvement in cold stress related frequency in rats. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e320 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Tetsuichi Saito More articles by this author Tetsuya Imamura More articles by this author Tomonori Minagawa More articles by this author Takashi Nagai More articles by this author Teruyuki Ogawa More articles by this author Osamu Ishizuka More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

PD70-12 IMPAIRMENT OF MICRO BLOOD FLOW IN THE BLADDER MUCOSA AND DERMIS UP-REGULATES TRPM8 CHANNELS AND INDUCE COLD STRESS RELATED FREQUENCY IN OVARIECTOMIZED RATS.

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Basic Research & Pathophysiology III1 Apr 2017PD70-12 IMPAIRMENT OF MICRO BLOOD FLOW IN THE BLADDER MUCOSA AND DERMIS UP-REGULATES TRPM8 CHANNELS AND INDUCE COLD STRESS RELATED FREQUENCY IN OVARIECTOMIZED RATS. Tetsuichi Saito, Tetsuya Imamura, Takashi Nagai, Toshiro Suzuki, Tomonori Minagawa, Teruyuki Ogawa, and Osamu Ishizuka Tetsuichi SaitoTetsuichi Saito More articles by this author , Tetsuya ImamuraTetsuya Imamura More articles by this author , Takashi NagaiTakashi Nagai More articles by this author , Toshiro SuzukiToshiro Suzuki More articles by this author , Tomonori MinagawaTomonori Minagawa More articles by this author , Teruyuki OgawaTeruyuki Ogawa More articles by this author , and Osamu IshizukaOsamu Ishizuka More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.3166AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Cold stress produced by sudden change or continuous exposure to low temperature exacerbates lower urinary tract symptoms (LUTS), such as urinary urgency, frequency, and nocturia. Stimulation of C-fibers and TRPM8 channels have been reported as mechanisms of cold stress related LUTS. Menopause women are known to be sensitive to cold stress, but the mechanism is not clearly known. The aim of this study is to show that menopause causes impairment of micro blood flow in the bladder mucosa and dermis under the skin and up-regulates TRPM8 channels which induces cold stress related frequency in rats. METHODS A total of 18 Spontaneously Hypertensive rats at postnatal week 10 were used in the experiments. The rats were randomly divided into 2 groups, including 9 with sham operation and 9 with bilateral ovariectomy. At 4 weeks after surgery, cystometography (CMG) was performed. CMG was first performed in room temperature (RT) for 20 minuets. Rats were then put into low temperature (LT) for 40 minutes. After LT, rats were put into RT for 20 minutes. After CMG, the whole bladder and dermis under the lumbar skin was harvested and real-time RT-PCR was performed. Immunohistochemistry was also performed and impairment of micro blood flow was evaluated by hypoxia-inducible factor-1 (HIF-1) staining. RESULTS Results of the CMG are shown in Fig 1. Basal pressure and micturition pressure did not show a difference between control rats and ovariectomy rats, but change rate with cold stress in voiding interval and micturition volume showed a significant exacerbate. Results of real-time RT-PCR are shown in Fig 2. Up-regulation of TRPM8 in the dermis and TRPV1 in the bladder mucosa was seen in ovariectomy rats. Immunohistochemistry showed increase of HIF-1 in the bladder mucosa and dermis. CONCLUSIONS Ovariectomy causes impairment of micro blood flow in the bladder mucosa and dermis which induces cold stress frequency by up-regulating TRPM8 channels. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e1354-e1355 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Tetsuichi Saito More articles by this author Tetsuya Imamura More articles by this author Takashi Nagai More articles by this author Toshiro Suzuki More articles by this author Tomonori Minagawa More articles by this author Teruyuki Ogawa More articles by this author Osamu Ishizuka More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

MP82-06 DETRUSOR UNDERACTIVITY IN AN OBESE-PRONE RAT MODEL

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Basic Research & Pathophysiology II1 Apr 2017MP82-06 DETRUSOR UNDERACTIVITY IN AN OBESE-PRONE RAT MODEL Eric Gonzalez and Warren Grill Eric GonzalezEric Gonzalez More articles by this author and Warren GrillWarren Grill More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.2553AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Detrusor underactivity (DUA) is an understudied health concern that affects up to 45% of men and women in secondary care. The clinical management of DUA is inadequate and fails to improve the quality of life of these patients. The limited availability of animal models that exhibit the integrated pathophysiology of DUA impedes the development of new therapeutic approaches. The current studies characterized the bladder function of an obesity model of DUA to increase our understanding of the initiation and progression of urinary retention. METHODS Animals: Eight-week old female obese-prone (OP) and obese-resistant (OR) rats purchased from Charles River (Boston, MA) were housed two per cage and maintained in standard laboratory conditions with food and water available ad libitum. Experimental procedures were approved by the Duke University IACUC and experimentation was conducted in accordance with the NIH Guide for the Care and Use of Laboratory Animals (8th ed.). High-fat feeding and glucose measurements: OP and OR rats were fed a 45% fat diet (D12451, Research Diets, Inc.) from 9-21 weeks and a 60% fat diet (D12492, Research Diets, Inc.) from 21-24 weeks. Whole blood was collected from the tail vein for glucose analysis at 8 and 24 weeks. Surgical preparation and instrumentation: At 24 weeks, OP and OR rats were anesthetized with urethane (1.2 g/kg s.c. and supplemented as needed). The bladder was exposed through a midline abdominal incision and a flared PE-60 catheter was inserted into the bladder dome. The catheter was secured and connected via a 3-way stopcock to a pressure transducer and infusion pump. A paddle with platinum iridium contacts was placed between the pubic symphysis and the EUS to record EMG signals. Pressure and EMG signals were amplified, filtered, and sampled on a PowerLab acquisition unit with LabChart 7 Pro. RESULTS OP rats weighed significantly more than OR rats (450 ± 11.1 vs. 270 ± 6.7 g, respectively; p ≤ 0.001); however, blood glucose was not altered between OP (169.1 ± 7.11 mg/dl) or OR (176.5 ± 6.09 mg/dl; p ≥ 0.05) rats. Compared to OR rats, OP rats have significantly increased volume threshold (0.7574 ± 0.07 vs. 0.529 ± 0.04 ml; p ≤ 0.05), decreased maximum micturition pressure (16.62 ± 1.2 vs. 23.94 ± 1.1 cmH2O; p ≤ 0.001), and decreased voiding efficiency (8.743 ± 2.58 vs. 42.19 ± 6.5 %; p ≤ 0.001). CONCLUSIONS OP rats exhibit DUA and decreased voiding efficiency. This animal model may be used to understand the pathophysiology underlying DUA and enable the development of novel therapeutic approaches to recover efficient voiding. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e1099 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Eric Gonzalez More articles by this author Warren Grill More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

MP82-01 A GENETIC FEMALE MOUSE MODEL WITH CONGENITAL GENITOURINARY ANOMALIES AND URINARY INCONTINENCE

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Basic Research & Pathophysiology II1 Apr 2017MP82-01 A GENETIC FEMALE MOUSE MODEL WITH CONGENITAL GENITOURINARY ANOMALIES AND URINARY INCONTINENCE Akbari Pedram, Ali Fathollahi, Rong Mo, Michael Chua, Michael Kavran, Nicole Episalla, Chi-Chung Hui, Walid Farhat, and Adonis Hijaz Akbari PedramAkbari Pedram More articles by this author , Ali FathollahiAli Fathollahi More articles by this author , Rong MoRong Mo More articles by this author , Michael ChuaMichael Chua More articles by this author , Michael KavranMichael Kavran More articles by this author , Nicole EpisallaNicole Episalla More articles by this author , Chi-Chung HuiChi-Chung Hui More articles by this author , Walid FarhatWalid Farhat More articles by this author , and Adonis HijazAdonis Hijaz More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.2548AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Hedgehog signaling pathway is known to have important role in the urogenital development. Transcription mediators of the pathway, Gli2 and Gli3, have been shown to be heavily involved in proper urogenital sinus formation. Both Gli2 and Gli3 null mice are non-viable, and display severe urogenital ad hindgut malformations. Here, we have generated a compound genetic mutant, Gli2+/-;Gli3Δ699/+, that is viable well into adulthood, and displaying variable urogenital malformations including urinary incontinence in its females. We aim to characterize the urinary incontinence observed in Gli2+/-; Gli3Δ699/+ female mice and assess its functional, anatomical, and histological characteristics. METHODS Gli2+/- and Gli3Δ699/+ mice were crossed to generate the double mutant (Gli2+/-; Gli3Δ699/+) female mice and wild type female mice were used as comparison controls; which all were verified via Polymerase Chain Reactions. Void measurements, Cystometrogram (CMG) and leak point pressure (LPP) were performed in all genotypes to assess bladder functions. The mice were then sacrificed to harvest the bladders and urethras for gross characterization via ink injection and histological assays. Differences were reported as mean and standard errors of mean (SEM) and analyzed using univariate analysis. Statistical significance set at 0.05. RESULTS No significant differences between the mutant and wild type mice were detected for 24 hour urinary output [(n= 13) mean 26.5cc±5 vs ( n=7) mean 22.15cc±6, p=0.13]. CMG studies revealed a decrease in peak micturition pressure values and significantly reduced LPP in Gli2+/-; Gli3Δ699/+ mice compared to wild type mice [(n=5) 4.28 cmH2O±2.4 vs (n=4) 20.24 cmH2O±6.45, p<0.0001; (n=5) 6.66 cmH2O±1.6 vs (n=5) 26.5cmH2O±5, p<0.05; respectively]. Gross characterization revealed that the ano-genital distance was severely reduced in double mutant mice; however, the urethra, vagina, and anus all remain separate and distinctly identifiable in these mice. Histological analyses revealed Gli2+/-; Gli3Δ699/+ mice exhibited a widened urethra and a decrease in smooth muscle layer thickness in the bladder outlet and urethra, with increased mucosal folding. CONCLUSIONS Gli2+/-; Gli3Δ699/+ female mice display persistent urinary incontinence with evident malformation of the bladder outlet and urethra. This presents a genetic mouse model for female urinary incontinence and alludes to potential genetic factors involved in the human condition. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e1097 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Akbari Pedram More articles by this author Ali Fathollahi More articles by this author Rong Mo More articles by this author Michael Chua More articles by this author Michael Kavran More articles by this author Nicole Episalla More articles by this author Chi-Chung Hui More articles by this author Walid Farhat More articles by this author Adonis Hijaz More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

A genetic female mouse model with congenital genitourinary anomalies and adult stages of urinary incontinence.

To characterize the urinary incontinence observed in adult Gli2+/- ; Gli3Δ699/+ female mice and identify the defects underlying the condition. Gli2+/- and Gli3Δ699/+ mice were crossed to generate: wild-type, mutant Gli2 (Gli2+/- ), mutant Gli3 (Gli3Δ699/+ ), and double mutant (Gli2+/- ; Gli3Δ699/+ ) female mice, verified via Polymerase Chain Reactions. Bladder functional studies including cystometrogram (CMG), leak point pressure (LPP), and voiding testing were performed on adult female mice. Female bladders and urethras were also analyzed via ink injection and histological assays. CMG tracing showed no signal corresponding to the filling of the Gli2+/- ; Gli3Δ699/+ bladders. LPP were significantly reduced in Gli2+/- ; Gli3Δ699/+ mice compared to wild-type mice. CMG studies revealed a decrease in peak micturition pressure values in Gli2+/- ; Gli3Δ699/+ mice compared with all other groups. No significant differences between mutant and wild-type mice were detected in urinary output. Histological analyses revealed Gli2+/- ; Gli3Δ699/+ mice exhibited a widened urethra and a decrease in smooth muscle layer thickness in the bladder outlet and urethra, with increased mucosal folding. Gli2+/- ; Gli3Δ699/+ adult female mice display persistent urinary incontinence due to the malformation of the bladder outlet and urethra. This presents a consistent and reliable genetic mouse model for female urinary incontinence and alludes to the key role of genetic factors involved in the condition.

An optimized transurethral catheterization cystometry in mice and comparison with classic suprapubic catheterization cystometry.

The aim of this study is to establish an optimized, minimally invasive transurethral catheterization cystometry (TUCC) and a novel urethral pressure profile (UPP) measurement for mice. The optimized TUCC and the UPP measurement were first established. This optimized TUCC was then performed in 16 anesthetized female mice and compared with the suprapubic catheterization cystometry (SCC) in parallel after suprapubic catheters implantation (SCI; on zero, third, and seventh day, respectively). Finally, the optimized TUCC and novel UPP measurement were applied to investigate in another eight mice of partial bladder outlet obstruction (pBOO) model. The urodynamic parameters including micturition pressure (MP), basal pressure (BP), threshold pressure (TP), bladder capacity (BC), micturition volume (MV), residual urine (RV), bladder compliance (COM), maximum urethral pressure (MUP), bladder pressure curve and UPP were recorded. Statistical cross-comparisons of parameters for two kinds of cystometries and pBOO model were performed. Compared with the optimized TUCC before SCI, the MV, RV, BC, and MP decreased significantly on the seventh day after SCI (270.4-132.5 µL, 46.13-20.09 µL, 316.4-152.5 µL, 30.01-24.34 cmH2 0, respectively). After SCI, the BP, MP, TP, MV, RV, BC, and COM showed no significant difference between the TUCC and SCC at the same time point. The MUP increased significantly after pBOO operation (19.1-46.6 cmH2 0, P < 0.05). The minimally invasive TUCC along with UPP measurement could be widely applied to study the bladder function of mice as a feasible, repeatable, and accessible method.

Cystometric analysis of the transplanted bladder

ABSTRACTObjective Cystometric evaluation of the bladder after autotransplant and isogeneic transplant in female rats.Material and Methods Two groups were constituted: (A) bladder autotransplant with two subgroups: R1 – (control) and R2 – (bladder transplant); (B) isogeneic bladder transplant with three subgroups; T1 – (control); T2–T3, two subgroups observed for 30 and 60 days after transplant, respectively. All animals underwent cystometric evaluation. Afterwards, the bladders were removed for histological study.Results The transplanted bladders did not show significant changes in filling/storage and emptying/micturition functions after 30 and 60 days of evolution. Upon macroscopical evaluation, there was good revascularization and the tissue was well preserved. Cystometry results: Did not show significant differences in the micturition pressure in subgroups T2-T3, but did between subgroups R1−R2, T1−T2, and T1−T3. Significant differences were verified in the micturition interval between T1−T3, T2−T3, but not between R1−R2, T1−T2. There was significant difference in the micturition duration between T1−T3 but not between R1−R2, T1−T2 and T2−T3. No fistula was noted on the suture site nor leakage of urine in the abdominal cavity or signs of necrosis or retraction were observed.Conclusions Transplant of the bladder was shown to be a viable procedure. The results indicate that there was structural and functional regeneration of transplanted bladders, and these results indicate that it is possible that vascular endothelium growth and neurogenesis factors are involved and activated in the process of the preservation or survival of the transplanted organ.

Effect of filling rate on cystometric parameters in young and middle aged mice.

OBJECTIVESTo investigate the effect of changing the bladder filling rate during cystometry in younger (2–3 months) and older (13–14 months) C57BL/6J male mice.METHODSCystometry was performed on mice under anesthesia. Voiding cycles were established in each mouse at a pump delivery rate of 17 μl/min. After 30 min, the rate was increased sequentially to 25, 33, 41 and 49 μl/min. Each rate was maintained for 30 min. The following cystometric parameters were quantified: peak pressure amplitude, intercontractile interval (ICI), compliance, micturition pressure threshold and voiding efficiency.RESULTSBladder weights were significantly greater in older mice (42 mg vs. 27 mg, P < 0.01), but functional capacities were not different. The pressure amplitudes did not change as filling rate increased, nor did they differ between the 4-month and 13-month-old males. ICIs were not significantly different between young and mature mice. However, both groups exhibited a non-linear reduction in ICI with increasing filling rate, best described by a power curve (R2 > 0.93). Compliance was higher in the older mice at low filling rates (17 and 25 μl/min) but this difference diminished at higher rates. Compliance decreased with increasing flow rate in a non-linear manner, again with greater effects at low filling rates. Micturition pressure thresholds increased with increasing flow rate in a linear manner and older mice began voiding at higher pressures than younger. Both young and old mice exhibited voiding efficiencies of ~70%.CONCLUSIONSThe rate of volume delivery has complex effects on the timing of voiding and compliance. These findings argue for greater standardization of cystometry protocols and further investigation into afferent signaling to higher centers at different filling rates.

Pharmacological Characterization of a Novel Beta 3 Adrenergic Agonist, Vibegron: Evaluation of Antimuscarinic Receptor Selectivity for Combination Therapy for Overactive Bladder.

Although the physiologic role of muscarinic receptors in bladder function and the therapeutic efficacy of muscarinic antagonists for the treatment of overactive bladder are well established, the role of β3-adrenergic receptors (β3ARs) and their potential as therapeutics is just emerging. In this manuscript, we characterized the pharmacology of a novel β3AR agonist vibegron (MK-4618, KRP-114V) and explored mechanistic interactions of β3AR agonism and muscarinic antagonism in urinary bladder function. Vibegron is a potent, selective full β3AR agonist across species, and it dose dependently increased bladder capacity, decreased micturition pressure, and increased bladder compliance in rhesus monkeys. The relaxation effect of vibegron was enhanced when combined with muscarinic antagonists, but differentially influenced by muscarinic receptor subtype selectivity. The effect was greater when vibegron was co-administered with tolterodine, a nonselective antagonist, compared with coadministration with darifenacin, a selective M3 antagonist. Furthermore, a synergistic effect for bladder strip relaxation was observed with the combination of a β3AR agonist and tolterodine in contrast to simple additivity with darifenacin. To determine expression in rhesus bladder, we employed a novel β3AR agonist probe, [3H]MRL-037, that selectively labels β3 receptors in both urothelium and detrusor smooth muscle. Vibegron administration caused a dose-dependent increase in circulating glycerol and fatty acid levels in rhesus and rat in vivo, suggesting these circulating lipids can be surrogate biomarkers. The translation of our observation to the clinic has yet to be determined, but the combination of β3AR agonists with M2/M3 antimuscarinics has the potential to redefine the standard of care for the pharmacological treatment of overactive bladder.

Influence of urothelial or suburothelial cholinergic receptors on bladder reflexes in chronic spinal cord injured cats

The effects of intravesical administration of a muscarinic receptor agonist (oxotremorine-M, OXO-M) and antagonist (atropine methyl nitrate, AMN) and of a nicotinic receptor agonist (nicotine) and antagonist (hexamethonium, C6) on reflex bladder activity were investigated in conscious female chronic spinal cord injured (SCI) cats using cystometry. OXO-M (50μM) decreased bladder capacity (BC) for triggering micturition contractions, increased maximal micturition pressure (MMP), increased frequency and area under the curve of pre-micturition contractions (PMC-AUC). Nicotine (250μM) decreased BC, increased MMP, but did not alter PMC-AUC. The effects of OXO-M on BC and PMC-AUC were suppressed by intravesical administration of AMN (50–100μM), and the effects of nicotine were blocked by hexamethonium (1mM). Antagonists infused intravesically alone did not alter reflex bladder activity. However, AMN (0.2mg/kg, subcutaneously) decreased PMC-AUC. 8-OH-DPAT (0.5mg/kg, s.c.), a 5-HT1A receptor agonist, suppressed the OXO-M-induced decrease in BC but not the enhancement of PMC-AUC. These results indicate that activation of cholinergic receptors located near the lumenal surface of the bladder modulates two types of reflex bladder activity (i.e., micturition and pre-micturition contractions). The effects may be mediated by activation of receptors on suburothelial afferent nerves or receptors on urothelial cells which release transmitters that can in turn alter afferent excitability. The selective action of nicotine on BC, while OXO-M affects both BC and PMC-AUC, suggests that micturition reflexes and PMCs are activated by different populations of afferent nerves. The selective suppression of the OXO-M effect on BC by 8-OH-DPAT without altering the effect on PMCs supports this hypothesis. The failure of intravesical administration of either AMN or hexamethonium alone to alter bladder activity indicates that cholinergic receptors located near the lumenal surface do not tonically regulate bladder reflex mechanisms in the SCI cat.

Urethane anesthesia in acute lower urinary tract studies in the male rat.

Urethane is a widely used anesthetic in animal lower urinary tract research. Our objective was to investigate the quality of anesthesia and the correlation between bladder (voiding) contractions, micturition pressure, bladder capacity and urethane dosage and body weight. Urethane was given subcutaneously and/or intraperitoneally (1.0-1.2 g/kg). The bladder was filled with saline and the bladder pressure was recorded continuously. Animals in which the subcutaneous/intraperitoneal ratio was higher needed less urethane. Heavier animals needed less extra urethane. In animals, in which no bladder contractions could be evoked, the total amount of urethane given was similar to that in those that did show contractions. In the animals that did void, the bladder never emptied completely and residual volumes remained. There was no relationship between animal weight or total amount of urethane and mean capacity. Anesthesia lasted up till 14 h, during which bladder (voiding) contractions could be recorded. Considering all results, we conclude that urethane is a well suited anesthetic for acute lower urinary tract physiological research in the intact rat.

MP68-05 COMPARISON OF THE EFFECTS OF BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) AND NERVE GROWTH FACTOR (NGF) INHIBITION ON BLADDER FUNCTION OF MICE WITH SPINAL CORD INJURY (SCI)

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Basic Research & Pathophysiology II1 Apr 2016MP68-05 COMPARISON OF THE EFFECTS OF BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) AND NERVE GROWTH FACTOR (NGF) INHIBITION ON BLADDER FUNCTION OF MICE WITH SPINAL CORD INJURY (SCI) Naoki Wada, Takahiro Shimizu, Shun Takai, Nobutaka Shimizu, Pradeep Tyagi, William de Groat, Anthony Kanai, Hidehiro Kakizaki, and Naoki Yoshimura Naoki WadaNaoki Wada More articles by this author , Takahiro ShimizuTakahiro Shimizu More articles by this author , Shun TakaiShun Takai More articles by this author , Nobutaka ShimizuNobutaka Shimizu More articles by this author , Pradeep TyagiPradeep Tyagi More articles by this author , William de GroatWilliam de Groat More articles by this author , Anthony KanaiAnthony Kanai More articles by this author , Hidehiro KakizakiHidehiro Kakizaki More articles by this author , and Naoki YoshimuraNaoki Yoshimura More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.1342AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES BDNF and NGF are reportedly involved in changes in neural pathways to induce lower urinary tract (LUT) dysfunction such as detrusor overactivity (DO) and inefficient voiding following SCI. However, it has not been well clarified how these two growth factors differentially affect storage and voiding functions after SCI. Therefore, we investigated the effects of anti-NGF or anti-BDNF antibody treatment on cystometric parameters in SCI mice. METHODS SCI was produced by complete transection of the Th8/9 spinal cord in female C57BL/6N mice. Three weeks later, an osmotic pump was placed subcutaneously to administer vehicle (group A), 10µg/kg/hr of anti-BDNF antibody (group B) or 10µg/kg/hr of anti-NGF antibody (group C) for 1 week. Four weeks after transection, SCI mice were evaluated using continuous or single-filling cystometry under an awake condition. RESULTS In continuous cystometry, voiding efficiency was significantly increased in the groups B (21%) and C (26%) compared to the group A (14%), and postvoid residual volume (PVR) was lower in the group C (0.14ml) vs. the group A (0.25ml). There was also a tendency of the reduction in non-voiding contractions (NVC) in the group C without statistical significance. Micturition pressure (MP) or intercontraction intervals were not different among the groups. In single cystometry, voided volume (0.085 vs. 0.038 ml) and voiding efficiency (42% vs. 18%) in the group B were significantly increased compared to the group A, and PVR of the group C was lower than that of the group A (0.11 vs. 0.15 ml). MP, maximum cystometric capacity or NVC was not different among groups. CONCLUSIONS Both anti-BDNF and anti-NGF antibody treatments improved voiding dysfunction as shown by increased voiding efficiency due to increased voided volume or decreased PVR in SCI mice. In addition, the anti-NGF, but not anti-BDNF, treatment marginally improved NVCs. These results suggest that BDNF and NGF are involved in SCI-induced voiding dysfunction resulting in inefficient voiding, but have minor roles in storage dysfunction to induce DO in mice, in contrast to the previous findings in SCI rats showing the greater contribution of bladder NGF or BDNF overexpression to DO. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e890-e891 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Naoki Wada More articles by this author Takahiro Shimizu More articles by this author Shun Takai More articles by this author Nobutaka Shimizu More articles by this author Pradeep Tyagi More articles by this author William de Groat More articles by this author Anthony Kanai More articles by this author Hidehiro Kakizaki More articles by this author Naoki Yoshimura More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

MP17-18 ROLE OF BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) IN LOWER URINARY TRACT DYSFUNCTION OF MICE WITH SPINAL CORD INJURY (SCI)

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Neurogenic Voiding Dysfunction1 Apr 2016MP17-18 ROLE OF BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) IN LOWER URINARY TRACT DYSFUNCTION OF MICE WITH SPINAL CORD INJURY (SCI) Naoki Wada, Takahiro Shimizu, Shun Takai, Nobutaka Shimizu, Pradeep Tyagi, William de Groat, Anthony Kanai, Hidehiro Kakizaki, and Naoki Yoshimura Naoki WadaNaoki Wada More articles by this author , Takahiro ShimizuTakahiro Shimizu More articles by this author , Shun TakaiShun Takai More articles by this author , Nobutaka ShimizuNobutaka Shimizu More articles by this author , Pradeep TyagiPradeep Tyagi More articles by this author , William de GroatWilliam de Groat More articles by this author , Anthony KanaiAnthony Kanai More articles by this author , Hidehiro KakizakiHidehiro Kakizaki More articles by this author , and Naoki YoshimuraNaoki Yoshimura More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2686AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Brain-derived neurotrophic factor (BDNF) is reportedly involved in the plasticity of spinal neuronal pathways that induces lower urinary tract dysfunction in SCI. Although pervious BDNF studies have mainly been performed in rats, we recently reported that bladder and urethral activity is urodynamically quite different between rats and mice with SCI. Therefore, this study investigated the effects of anti-BDNF antibody treatment on storage and voiding dysfunction in mice with SCI. METHODS The Th8/9 spinal cord was transected to produce SCI in female C57BL/6N mice. Three weeks later, an osmotic pump was placed subcutaneously to administer vehicle (group A) or 10µg/kg/hr of anti-BDNF antibody (group B) for 1 week. Four weeks after transection, mice were evaluated using continuous or single-filling cystometry under a conscious condition. Then, the bladder was removed to measure BDNF levels by the ELISA method and to compare with those of spinal intact mice. RESULTS In continuous cystometry, voiding efficiency was significantly increased in the group B vs. the group A (21.4 vs. 14.1%) whereas there were no significant differences in micturition pressure (MP), intercontraction intervals or non-voiding contractions (NVC) between two groups. In single cystometry, voided volume (0.085 vs. 0.038 ml) and voiding efficiency (42.3 vs. 18.4%) in the group B were significantly increased compared to the group A. MP, maximum cystometric capacity or NVC was not different between two groups. The BDNF level of the group A bladder was significantly higher than that of normal mice (5.29±0.45 vs. 1.38±0.18 pg/mg tissue); however, it was decreased in the group B bladder (1.69±0.26 pg/mg tissue) compared to the group A. CONCLUSIONS The anti-BDNF antibody treatment, which reduced bladder BDNF expression, improved voiding dysfunction as shown by increases in voided volume and voiding efficiency, but did not affect storage dysfunction as evidenced by unaltered NVCs in SCI mice. These results suggest that increased BDNF after SCI contributes to inefficient voiding, possibly due to impaired coordinating activity of bladder and urethra during voiding. Thus, BDNF-targeting therapies could be effective for treating voiding problems in SCI patients. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e191 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Naoki Wada More articles by this author Takahiro Shimizu More articles by this author Shun Takai More articles by this author Nobutaka Shimizu More articles by this author Pradeep Tyagi More articles by this author William de Groat More articles by this author Anthony Kanai More articles by this author Hidehiro Kakizaki More articles by this author Naoki Yoshimura More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Functional and morphological alterations of the urinary bladder in type 2 diabetic FVBdb/db mice

AimsDiabetic bladder dysfunction (DBD) has been extensively studied in animal models of type 1 diabetes. We aimed to examine the functional and morphological alterations of the urinary bladder in a type 2 diabetes model, FVBdb/db mice. MethodsFVBdb/db mice and age-matched FVB/NJ control mice were tested at either 12, 24 or 52weeks of age. Body weight, blood glucose and glycated hemoglobin (HbA1c) levels were measured. Bladder function was assessed by measurement of 24-h urination behavior and conscious cystometry. Bladder was harvested for Masson's Trichrome staining and morphometric analysis. ResultsThe body weights of FVBdb/db mice were twice as those of FVB/NJ control mice. The blood glucose and HbA1c levels were higher in FVBdb/db mice at 12 and 24weeks, but not at 52weeks. A significant increase in the mean volume per void, but decrease in the voiding frequency, in FVBdb/db mice was observed. Cystometry evaluation showed increased bladder capacity, voided volume, and peak micturition pressure in FVBdb/db mice compared with FVB/NJ mice. Morphometric analysis revealed a significant increase in the areas of detrusor muscle and urothelium in FVBdb/db mice. In addition, some FVBdb/db mice, especially males at 12 and 24weeks, showed small-volume voiding during 24-h urination behavior measurement, and detrusor overactivity in the cystometry measurement. ConclusionsThe FVBdb/db mouse, displaying DBD characterized by not only increased bladder capacity, void volume, and micturition pressure, but also bladder overactivity, is a useful model to further investigate the mechanisms of type 2 diabetes-related bladder dysfunction.

Epigallocatechin Gallate Attenuates Partial Bladder Outlet Obstruction-induced Bladder Injury via Suppression of Endoplasmic Reticulum Stress-related Apoptosis—In Vivo Study

To investigate the protective effect of epigallocatechin gallate (EGCG), a green tea extract, on partial bladder outlet obstruction (pBOO)-induced bladder injury in a rat model. The female Sprague-Dawley rats underwent sham or BOO procedures, and were divided into several groups (sham with saline injection, sham with EGCG treatment, BOO with saline injection, and BOO with EGCG treatment). The rats in each group were randomized into 2 groups (48 hours and 30 days after the BOO procedure) for when their bladders were harvested. EGCG (4.5 mg/kg/day) and saline were administered via intraperitoneal injection after the BOO procedure during the study period. Bladder tissue was examined for inflammation, endoplasmic reticulum (ER) stress-related apoptotic markers by Western blot, and histological staining. BOO induced acute bladder injury (hemorrhage, edema, and neutrophil infiltration) after 48 hours. In addition, cystometry showed a decrease in micturition pressure and intercontractile interval. We also observed increased expressions of cyclooxygenase-2, poly(ADP-ribose) polymerase at 48 hours, as well as ER stress markers such as caspase-12 and CCAAT/-enhancer-binding protein homologous protein (CHOP). Treatment with EGCG significantly improved pBOO-induced histologic changes, bladder dysfunction, and the overexpression of cyclooxygenase-2, CHOP, and caspase-12 at 48 hours. Similarly, EGCG treatment for 30 days effectively recovered compliance and intercontractile interval, submucosal ER stress-related apoptosis (CHOP and caspase-12) at 30 days after pBOO. EGCG alleviate pBOO-induced bladder injury and dysfunction via suppression of inflammation and ER stress-related apoptosis.