This work was supported by grants from the Belgian Federal Government (IUAP P6/28), the Research Foundation-Flanders (F.W.O.) (G.0565.07 and G.0686.09), the Astellas European Foundation Award 2009 and the Research Council of the KU Leuven (GOA 2009/07, EF/95/010 and PFV/10/006). P.U. and W.E. are doctoral fellows of the Research Foundation-Flanders (FWO). M.B. is a Marie Curie fellow. D.D.R. a fundamental-clinical fellow of the FWO.

1. Laboratory Animals
<ol>
 <li>Animals (mice, rats) are housed in a specialized animal facility with a 12-hr light-dark cycle and ad libitum access to water and standard food pellets. Both age and sex of the animals are important parameters that should be standardized according to the needs. We typically perform cystometry in 10 - 12 week old female animals5,6.</li>
 <li>All animal experiments are carried out in accordance with the European Union Community Council guidelines and approved by the loc...

<ol>
	<li>Everaerts, W., Gevaert, T., Nilius, B., De Ridder, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+origin+of+bladder+sensing:+Tr(i)ps+in+urology.">On the origin of bladder sensing: Tr(i)ps in urology.</a> Neurourol. Urodyn. 27, 264-2673 (2008).</li><li>Fry, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+and+their+use+in+understanding+lower+urinary+tract+dysfunction.">Animal models and their use in understanding lower urinary tract dysfunction.</a> Neurourol. Urodyn. 29, 603-608 (2010).</li><li>Gevaert, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deletion+of+the+transient+receptor+potential+cation+channel+TRPV4+impairs+murine+bladder+voiding.">Deletion of the transient receptor potential cation channel TRPV4 impairs murine bladder voiding.</a> J. Clin. Invest. 117, 3453-3462 (2007).</li><li>Andersson, K. E., Soler, R., Fullhase, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+for+urodynamic+investigation.">Rodent models for urodynamic investigation.</a> Neurourol. Urodyn. 30, 636-646 (2011).</li><li>Everaerts, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+the+cation+channel+TRPV4+improves+bladder+function+in+mice+and+rats+with+cyclophosphamide-induced+cystitis.">Inhibition of the cation channel TRPV4 improves bladder function in mice and rats with cyclophosphamide-induced cystitis.</a> Proc. Natl. Acad. Sci. U.S.A. 107, 19084-19089 (2010).</li><li>Everaerts, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+capsaicin+receptor+TRPV1+is+a+crucial+mediator+of+the+noxious+effects+of+mustard+oil.">The capsaicin receptor TRPV1 is a crucial mediator of the noxious effects of mustard oil.</a> Curr. Biol. 21, 316-321 (2011).</li><li>Yoshiyama, M., Roppolo, J. R., Thor, K. B., de Groat, W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+LY274614,+a+competitive+NMDA+receptor+antagonist,+on+the+micturition+reflex+in+the+urethane-anaesthetized+rat.">Effects of LY274614, a competitive NMDA receptor antagonist, on the micturition reflex in the urethane-anaesthetized rat.</a> Br. J. Pharmacol. 110, 77-86 (1993).</li><li>Yoshiyama, M., Roppolo, J. R., de Groat, W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+MK-801+on+the+micturition+reflex+in+the+rat--possible+sites+of+action.">Effects of MK-801 on the micturition reflex in the rat--possible sites of action.</a> J. Pharmacol. Exp. Ther. 265, 844-850 (1993).</li><li>Boudes, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+Characterization+of+a+Chronic+Cyclophosphamide-Induced+Overactive+Bladder+Model+in+mice.">Functional Characterization of a Chronic Cyclophosphamide-Induced Overactive Bladder Model in mice.</a> Neurourol. Urodyn. , (2011).</li><li>Yoshiyama, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sex-related+differences+in+activity+of+lower+urinary+tract+in+response+to+intravesical+acid+irritation+in+decerebrate+unanesthetized+mice.">Sex-related differences in activity of lower urinary tract in response to intravesical acid irritation in decerebrate unanesthetized mice.</a> Am. J. Physiol. Regul. Integr. Comp. Physiol. 295, R954-R960 (2008).</li><li>McMahon, S. B., Abel, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+model+for+the+study+of+visceral+pain+states:+chronic+inflammation+of+the+chronic+decerebrate+rat+urinary+bladder+by+irritant+chemicals.">A model for the study of visceral pain states: chronic inflammation of the chronic decerebrate rat urinary bladder by irritant chemicals.</a> Pain. 28, 109-127 (1987).</li><li>Du, S., Araki, I., Yoshiyama, M., Nomura, T., Takeda, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transient+receptor+potential+channel+A1+involved+in+sensory+transduction+of+rat+urinary+bladder+through+C-fiber+pathway.">Transient receptor potential channel A1 involved in sensory transduction of rat urinary bladder through C-fiber pathway.</a> Urology. 70, 826-831 (2007).</li><li>Streng, T., Santti, R., Talo, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Similarities+and+differences+in+female+and+male+rat+voiding.">Similarities and differences in female and male rat voiding.</a> Neurourol. Urodyn. 21, 136-141 (2002).</li><li>Igawa, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cystometric+findings+in+mice+lacking+muscarinic+M2+or+M3+receptors.">Cystometric findings in mice lacking muscarinic M2 or M3 receptors.</a> J. Urol. 172, 2460-2464 (2004).</li><li>Schroder, A., Newgreen, D., Andersson, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detrusor+responses+to+prostaglandin+E2+and+bladder+outlet+obstruction+in+wild-type+and+Ep1+receptor+knockout+mice.">Detrusor responses to prostaglandin E2 and bladder outlet obstruction in wild-type and Ep1 receptor knockout mice.</a> J. Urol. 172, 1166-1170 (2004).</li><li>Chen, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Function+of+the+lower+urinary+tract+in+mice+lacking+alpha1d-adrenoceptor.">Function of the lower urinary tract in mice lacking alpha1d-adrenoceptor.</a> J. Urol. 174, 370-374 (2005).</li><li>May, V., Vizzard, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bladder+dysfunction+and+altered+somatic+sensitivity+in+PACAP-/-+mice.">Bladder dysfunction and altered somatic sensitivity in PACAP-/- mice.</a> J. Urol. 183, 772-779 (2010).</li><li>Thorneloe, K. S., Meredith, A. L., Knorn, A. M., Aldrich, R. W., Nelson, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Urodynamic+properties+and+neurotransmitter+dependence+of+urinary+bladder+contractility+in+the+BK+channel+deletion+model+of+overactive+bladder.">Urodynamic properties and neurotransmitter dependence of urinary bladder contractility in the BK channel deletion model of overactive bladder.</a> Am. J. Physiol. Renal. Physiol. 289, 604-610 (2005).</li></ol>

No conflicts of interest declared.

The cystometry technique presented here allows performing in vivo measurements of bladder function in animal models. Rats are probably the most used animal model. Mice are more difficult to handle, but offer the advantage of using genetically manipulated animals. Because of the technical difficulty of using conscious mice, which tend to be very active resulting in loosening of the implanted catheter and changes in the intra-abdominal pressures that may influence the intravesical pressure, we advise to kee...

<table border="1">
	<tbody>
 <tr>
 <td>Name of the reagent</td>
 <td>Commercial name, company,</td>
 <td>Catalogue number</td>
 <td>Comments </td>
 </tr>
 <tr>
 <td>urethane</td>
 <td>Urethane, Sigma-Aldrich</td>
 <td>315419</td>
 <td>group 2B carcinogen</td>
 </tr>
 <tr>
 <td>isoflurane</td>
 <td>Isoba, Schering-Plough Animal Health</td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>polyethylene catheter</td>
 <td>Intramedic Polyethylene tubing PE50, Becton Dickinson </td>
 <td>427411</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>surgical microscope</td>
 <td>Op-Mi 6, Carl Zeiss</td>
 <td>Op-Mi 6</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>purse-string suture</td>
 <td>Prolene 6/0, Ethicon</td>
 <td>8610H</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>fascia and skin suture</td>
 <td>Ethilon 4/0 or 5/0, Ethicon</td>
 <td>662G or 661G</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>postoperative analgesics</td>
 <td>Temgesic, Schering-Plough Animal Health</td>
 <td>&nbsp;</td>
 <td>dosage for rats: 0.05 mg/kg</td>
 </tr>
 <tr>
 <td>amplifier</td>
 <td>78534c monitor, Hewlett Packard</td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>analytical balances and balance data acquisition software</td>
 <td>FZ 300i, A&amp;D</td>
 <td>FZ-300i</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>infusion pumps</td>
 <td>pump 33, Harvard apparatus</td>
 <td>HA33</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>cystometry recording system</td>
 <td>Dataq instruments, DI-730 series and Windaq/Lite</td>
 <td>DI-730-USB Windaq/Lite</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>temperature registration</td>
 <td>Fluke 52 KJ thermometer</td>
 <td>52 KJ</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>pressure transducers</td>
 <td>Edwards Lifesciences, pressure monitoring set</td>
 <td>T322247A</td>
 <td>&nbsp;</td>
 </tr>
	 </tbody>
 </table>

the use of cystometry in small rodents: a study of bladder chemosensation

The lower urinary tract (LUT) functions as a dynamic reservoir that is able to store urine and to efficiently expel it at a convenient time. While storing urine, however, the bladder is exposed for prolonged periods to waste products. By acting as a tight barrier, the epithelial lining of the LUT, the urothelium, avoids re-absorption of harmful substances. Moreover, noxious chemicals stimulate the bladder's nociceptive innervation and initiate voiding contractions that expel the bladder's contents. Interestingly, the bladder's sensitivity to noxious chemicals has been used successfully in clinical practice, by intravesically infusing the TRPV1 agonist capsaicin to treat neurogenic bladder overactivity1. This underscores the advantage of viewing the bladder as a chemosensory organ and prompts for further clinical research. However, ethical issues severely limit the possibilities to perform, in human subjects, the invasive measurements that are necessary to unravel the molecular bases of LUT clinical pharmacology. A way to overcome this limitation is the use of several animal models2. Here we describe the implementation of cystometry in mice and rats, a technique that allows measuring the intravesical pressure in conditions of controlled bladder perfusion.
	After laparotomy, a catheter is implanted in the bladder dome and tunneled subcutaneously to the interscapular region. Then the bladder can be filled at a controlled rate, while the urethra is left free for micturition. During the repetitive cycles of filling and voiding, intravesical pressure can be measured via the implanted catheter. As such, the pressure changes can be quantified and analyzed. Moreover, simultaneous measurement of the voided volume allows distinguishing voiding contractions from non-voiding contractions3.
	Importantly, due to the differences in micturition control between rodents and humans, cystometric measurements in these animals have only limited translational value4. Nevertheless, they are quite instrumental in the study of bladder pathophysiology and pharmacology in experimental pre-clinical settings. Recent research using this technique has revealed the key role of novel molecular players in the mechano- and chemo-sensory properties of the bladder.

Watch this Scientific Journal Video about The Use of Cystometry in Small Rodents: A Study of Bladder Chemosensation at JoVE.com

The Use of Cystometry in Small Rodents: A Study of Bladder Chemosensation

Cystometry is an efficient technique to measure bladder function of small animals in vivo. The bladder is continuously infused at rates controlled through an intravesical catheter, whereas the urethra is left free for micturition. This allows for repetitive filling and emptying of the bladder, while intravesical pressure and voided volume are recorded.

	The lower urinary tract (LUT) functions as a  dynamic reservoir that is able to store urine and to efficiently expel it at a  convenient time. While ...

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22.2K vues.  KU Leuven, Belgium. Cystomanométrie est une technique efficace pour mesurer la fonction de la vessie de petits animaux In vivo. La vessie est perfusé en continu à des vitesses contrôlées par un cathéter intra-vésicale, tandis que l'urètre est laissée libre pour la miction. Ceci permet de répétition de remplissage et de vidange de la vessie, tandis que la pression intravésicale volume évacué et sont enregistrées.