Detrusor overactivity induced by intravesical application of adenosine 5′-triphosphate under different delivery conditions in rats

doi:10.1016/j.urology.2005.06.099

Urology

Volume 66, Issue 6, December 2005, Pages 1332-1337

https://doi.org/10.1016/j.urology.2005.06.099 Get rights and content

Abstract

Objectives

To investigate the effects of intravesical application of adenosine 5′-triphosphate (ATP) on bladder activity to elucidate the role of urothelial barrier function and ecto-ATPase activity in the ATP-mediated mechanism inducing detrusor overactivity.

Methods

Continuous cystometry by an intravesical catheter inserted from the bladder dome was performed in conscious female rats.

Results

ATP solutions adjusted to pH 6.0 did not elicit significant detrusor overactivity at a concentration of 60 mM. However, in bladders pretreated with protamine sulfate (10 mg/mL) to increase urothelial permeability, ATP solution (pH 6.0) induced detrusor overactivity by decreasing the intercontraction intervals. These irritant effects of ATP after protamine treatment were antagonized by P2X receptor antagonists, such as pyridoxal-5-phosphate-6-azophenyl-2′,4′-disulfonic acid (70 μmol/kg) and 2′,3′-O-(2,4,6, trinitrophenyl) ATP (30 μmol/kg). These were also suppressed in rats pretreated with systemic capsaicin (125 mg/kg subcutaneously). Alpha,beta-methylene ATP (5 mM, pH 6.0) or ATP (60 mM, pH6) after intravesical infusion of 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (5 mM, pH 6.0), an ecto-ATPase inhibitor, induced detrusor overactivity without protamine pretreatment, but the reduction in intercontraction intervals was smaller compared with that with ATP after protamine treatment.

Conclusions

Low permeability of bladder epithelium and ecto-ATPase activity can prevent ATP activation of subepithelial P2X receptors to induce bladder overactivity. Thus, enhanced penetration of endogenous ATP owing to urothelial damage may contribute to urinary frequency and bladder pain in hypersensitive bladder disorders such as interstitial cystitis.

Section snippets

Animals

Female Sprague-Dawley rats (weighing 170 to 240 g) were used. The protocol for this study complied with the Guide for the Care and Use of Laboratory Animals, published by the National Institute of Laboratory Animal Resources, and was approved by the University of Pittsburgh Institutional Animal Care and Use Committee.

Cystometry

Under halothane anesthesia, a PE-10 polyethylene catheter (Clay Adams, Parsippany, NJ) was inserted into the right jugular vein, and the free end of the catheter was tunneled

Effect of Intravesical ATP Adjusted to pH 6.0

PBS (pH 6.0) alone when infused into the bladder did not alter any cystometric parameters. Similarly, application of ATP solutions (20 to 60 mM), adjusted to pH 6.0, did not elicit significant detrusor overactivity (Table I).

Effect of ATP (pH 6.0) After PS Infusion

When PS was infused intravesically, bladder overactivity was observed, as evidenced by decreased ICIs (Fig. 1A). After PS infusion, PBS (pH 6.0) was infused intravesically, which partially normalized the cystometric parameters. Intravesical instillation of ATP (20, 40, and

Comment

The bladder permeability barrier is located in the apical membrane of the superficial layer of epithelial cells, so-called umbrella cells.¹⁰ Intravesical application of PS has been used to induce bladder epithelial injury,⁹ which permeabilizes the apical membranes of the urothelium.⁹ Acetate, propionate, butyrate, or succinate salts at pH 4.4, but not at pH 5.0, alters the transepithelial permeability of the rabbit urothelium.¹¹ In our study, ATP (pH 6.0) alone did not induce significant

Conclusions

Intravesically applied ATP in pH 6.0 solution does not penetrate the intact bladder epithelial layer in the rat because of permeability barrier and ecto-ATPase of the urothelium. Therefore, increased permeability of the bladder epithelium is needed for exogenously applied ATP to activate subepithelial P2X receptors to sensitize C-fiber afferents and induce detrusor overactivity.

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Adenosine triphosphate (ATP) contributes to generating of unstable detrusor muscle contractions. The Nishiguchi et al. [25] study revealed that botulin toxin inhibits ATP and acetylocholine release from urinary bladder strips in animals. The inhibition of ATP release by botulin toxin is complex within urothelium, afferent nerves and efferent nerves.
The aim of the study was to estimated the clinical efficacy of intravesical injections of botulinum toxin type A (BTX-A) in refractory idiopathic overactive bladder (OAB) in the aspect of the quality of life. The potential mechanism of BTX-A actions were described.
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In patients with refractory idiopathic OAB a decrease in the general impact of LUTS/OAB symptoms domain of KHQ, and also the negative impact of LUTS/OAB on daily indoor/outdoor, physical and social activity were observed within 9 months follow-up after intravesical injections of BTX-A. Personal relationships also improved. Moreover, BTX-A has a positive effect on the patients emotions. Sleep difficulties and fatigue also decreased. The severity of LUTS/OAB symptoms were alleviated up to 9^th month after BTX-A injections, however we observed a statistically insignificant increase of LUTS/OAB severity in 9^th month after treatment.
Intravesical injections of BTX-A is an effective therapy in refractory idiopathic OAB over the period of 9 months leading to significant improvement of quality of life in many aspects.
Intravesical ATP instillation induces urinary frequency because of activation of bladder afferent nerves without inflammatory changes in mice: A promising model for overactive bladder
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For analysis of cystometry and ultrasonography before treatment, parameters were evaluated 10 h after anesthesia [9,10]. For accurate analysis of ATP (Tokyo Chemical Industry, Tokyo, Japan), we prepared pH-adjusted 50 mM ATP solution (approximately pH 6.4) with NaOH to eliminate the strong acidic characteristics [11]. Parameters were evaluated in the stable state approximately 40 min after intravesical treatment.
ATP in the suburothelial layer is released from the bladder urothelium by mechanical stimuli. ATP directly activates purinergic receptors that are expressed on primary bladder afferent neurons and induces the micturition reflex. Although ATP is also released to the bladder lumen from the bladder urothelium, the role of ATP in the bladder lumen is unknown. Recently, clinical studies have reported that urinary ATP levels are much higher in patients with an overactive bladder than healthy controls. These results suggest that ATP in the bladder lumen is also involved in the micturition reflex. In this study, we performed intravesical ATP instillation in the mouse bladder. We evaluated urinary function with novel reliable methods using improved cystometry and ultrasonography, which we previously established. We found that intravesical ATP instillation induced urinary frequency because of activation of bladder afferent nerves without inflammatory changes in the bladder or an increase in post-void residual urine. These results suggest that not only ATP in the suburothelial layer, but also ATP in the bladder lumen, are involved in enhancement of the micturition reflex.
Ba-Wei-Die-Huang-Wan (Hachimi-jio-gan) can ameliorate cyclophosphamide-induced ongoing bladder overactivity and acidic adenosine triphosphate solution-induced hyperactivity on rats prestimulated bladder
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Rats were infused with ATP solution for 1.5 h, starting after baseline cystometric data were collected using normal saline. In order to ensure stable voiding patterns, data was not analyzed from the initial 20 min of ATP solution instillation (Lee et al., 2012; Nishiguchi et al., 2005). Following the stabilization period, data from 1-hr of reproducible micturition cycles were then collected and used for evaluation.
Ba-Wei-Die-Huang-Wan (BWDHW) is the traditional Chinese medicine formula containing eight ingredients, namely Rehmannia glutinosa (Gaetn.) DC., root, steamed & dried; Cornus officinalis Siebold & Zucc., fructus, dried; Dioscorea oppositifolia L., root, dried; Alisma plantago-aquatica, subsp. orientale (Sam.) Sam., tuber, dried; Poria cocos (Fr.) Wolf., sclerotium, dried; Paeonia×suffruticosa Andrews, bark, dried; Cinnamomum cassia (Nees & T.Nees) J. Presl, bark, dried; Aconitum carmichaeli Debeaux, lateral root, dried & processed. It has been used for diabetes and urinary frequency treatments.
We investigate effects of BWDHW on cyclophosphamide (CYP)-induced ongoing bladder overactivity and acidic adenosine triphosphate (ATP) solution-induced hyperactivity on rat’s prestimulated bladder.
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As compared to the controls, the CYP group showed significantly decreased mean cystometric intercontractile interval and increased micturition frequency, whereas the CYP/BWDWH group did not. The CYP group had significant protein overexpression in mucosal M₂, M₃, P2X₂, and P2X₃ receptors as well as detrusor M₂ and M₃ receptors. However, the CYP/BWDWH group had insignificant changes from controls. In the provoking test, the control/BWDHW and CYP/BWDHW groups were less affected by acidic ATP stimulation of intercontractile interval changes than the control group. Compared to the control group, the control/BWDHW group showed significantly lower mucosal P2X₃ protein expression and the CYP group showed significant mucosal TRPV1 protein upregulation after the provoking test.
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Functions of the lower urinary tract to store and periodically eliminate urine are regulated by a complex neural control system in the brain, spinal cord, and peripheral autonomic ganglia that coordinates the activity of smooth and striated muscles of the bladder and urethral outlet. Neural control of micturition is organized as a hierarchic system in which spinal storage mechanisms are in turn regulated by circuitry in the rostral brainstem that initiates reflex voiding. Input from the forebrain triggers voluntary voiding by modulating the brainstem circuitry. Many neural circuits controlling the lower urinary tract exhibit switch-like patterns of activity that turn on and off in an all-or-none manner. The major component of the micturition switching circuit is a spinobulbospinal parasympathetic reflex pathway that has essential connections in the periaqueductal gray and pontine micturition center. A computer model of this circuit that mimics the switching functions of the bladder and urethra at the onset of micturition is described. Micturition occurs involuntarily during the early postnatal period, after which it is regulated voluntarily. Diseases or injuries of the nervous system in adults cause re-emergence of involuntary micturition, leading to urinary incontinence. The mechanisms underlying these pathologic changes are discussed.
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: This work was supported by NIH DK68557, DK55045, and DK66138, and the Fishbein Family CURE-IC.

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Basic scienceDetrusor overactivity induced by intravesical application of adenosine 5′-triphosphate under different delivery conditions in rats

Abstract

Objectives

Methods

Results

Conclusions

Section snippets

Animals

Cystometry

Effect of Intravesical ATP Adjusted to pH 6.0

Effect of ATP (pH 6.0) After PS Infusion

Comment

Conclusions

Trends Pharmacol Sci

J Urol

J Urol

J Urol

Biochem Biophys Res Commun

Neuroscience

J Urol

Prog Neurobiol

J Urol

J Urol

ATP is released from rabbit urinary bladder epithelial cells by hydrostatic pressure changes—a possible sensory mechanism?

J Physiol

Feline interstitial cystitis results in mechanical hypersensitivity and altered ATP release from bladder urothelium

Am J Physiol Renal Physiol

P2X3 knock-out mice reveal a major sensory role for urothelially released ATP

J Neurosci

Basic science
Detrusor overactivity induced by intravesical application of adenosine 5′-triphosphate under different delivery conditions in rats