Name: cystometric and external urethral sphincter measurements in awake rats with implanted catheter and electrodes allowing for repeated measurements
Uploaded: 2018-01-30T13:30:00
Description: Watch this Scientific Journal Video about Cystometric and External Urethral Sphincter Measurements in Awake Rats with Implanted Catheter and Electrodes Allowing for Repeated Measurements at JoVE.com

The authors do not have any acknowledgements.

All procedures described here were approved by the Austrian Governmental Ethics committee for animal research (Bundesministerium f&#252;r Wissenschaft, Forschung und Wirtschaft, WF / V /3b) and were in compliance with the Association for Assessment of Laboratory Animal Care guidelines for animal use. Rats used for this approach were female, 12-week-old Lewis rats. Use sterile instruments throughout the protocol.
1. Material Preparation
<ol>
	<li>Manufacturing of the Catheter...

<ol>
	<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>Schneider, M. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+urodynamic+model+for+lower+urinary+tract+assessment+in+awake+rats.">A novel urodynamic model for lower urinary tract assessment in awake rats.</a> BJU Int. 115, 8-15 (2015).</li><li>Cruz, C. D., Cruz, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+injury+and+bladder+dysfunction:+new+ideas+about+an+old+problem.">Spinal cord injury and bladder dysfunction: new ideas about an old problem.</a> Scientific World Journal. 11, 214-234 (2011).</li><li>de Groat, W. C., Yoshimura, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasticity+in+reflex+pathways+to+the+lower+urinary+tract+following+spinal+cord+injury.">Plasticity in reflex pathways to the lower urinary tract following spinal cord injury.</a> Exp Neurol. 235, 123-132 (2012).</li><li>Matsuura, S., Downie, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+anesthetics+on+reflex+micturition+in+the+chronic+cannula-implanted+rat.">Effect of anesthetics on reflex micturition in the chronic cannula-implanted rat.</a> Neurourol Urodyn. 19, 87-99 (2000).</li><li>Cannon, T. W., Damaser, M. S. <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+anesthesia+on+cystometry+and+leak+point+pressure+of+the+female+rat.">Effects of anesthesia on cystometry and leak point pressure of the female rat.</a> Life Sci. 69, 1193-1202 (2001).</li><li>Patra, P. B., Thorneloe, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhanced+sensitivity+to+afferent+stimulation+and+impact+of+overactive+bladder+therapies+in+the+conscious,+spontaneously+hypertensive+rat.">Enhanced sensitivity to afferent stimulation and impact of overactive bladder therapies in the conscious, spontaneously hypertensive rat.</a> J Pharmacol Exp Ther. 338, 392-399 (2011).</li><li>Herrera, G. M., Meredith, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diurnal+variation+in+urodynamics+of+rat.">Diurnal variation in urodynamics of rat.</a> PloS One. 5, e12298 (2010).</li><li>Chang, H. Y., Havton, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+effects+of+urethane+and+isoflurane+on+external+urethral+sphincter+electromyography+and+cystometry+in+rats.">Differential effects of urethane and isoflurane on external urethral sphincter electromyography and cystometry in rats.</a> Am J Physiol Renal Physiol. 295, F1248-F1253 (2008).</li><li>Yaksh, T. L., Durant, P. A., Brent, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Micturition+in+rats:+a+chronic+model+for+study+of+bladder+function+and+effect+of+anesthetics.">Micturition in rats: a chronic model for study of bladder function and effect of anesthetics.</a> Am J Physiol. 251, R1177-R1185 (1986).</li></ol>

This protocol describes the surgical procedure of a permanent catheter and urethral sphincter electrodes implantation and the cystometric recording technique in awake, lightly restrained rats including both the simultaneous analysis of the urinary bladder and external urethral sphincter.
Critical steps during surgery are the careful implantation of the bladder catheter, avoiding leakage and extensive manipulation. Moreover, a precise implantation of the electrodes bilateral to the urethral ext...

To analyze urinary storage and voiding function and dysfunction, most studies have used rodent models. Through sequential activation of reflexes, micturition is produced. The coordination of these reflexes is essential for efficient voiding1. Cystometric recording techniques provide valuable tools for analyzing the urinary bladder function under its neural control1.
Most conventional cystometries in rats are done as a single, final analysis in anesthesia, mainly urethane2, and focus on the urinary bladder solely. However, in some pathologies like neurogenic lower urinary tract dysfunction (NLUTD), not only the urinary bladder, but also the bladder outlet, the external urethral sphincter, is dysfunctional3,4. This makes NLUTD difficult to follow-up, if only the bladder is examined in a single cystometric measurement. To get reliable results that are comparable to humans, it is essential to accurately measure both the urinary bladder and the external urethral sphincter function and its interactions2. Furthermore, it is crucial to perform functional analyses in awake rats as anesthesia is very likely to alter bladder function2,5,6. A good cystometric recording in awake animals is the basis for the identification of bladder function and malfunction7.
The small animal cystometry station used (e.g., Catamount cystometry station (CCS)) is a unit to perform cystometric analyses in small awake animals8. By means of a permanent bladder catheter and implanted external urethral sphincter electrodes, repetitive measurements can be performed over a longer time periods2. Thus, the CCS provides a valuable tool for non-neurogenic and NLUTD evaluations in the rodent model, in which the pathomechanisms can change during short- or medium-term follow-up. Additionally, this method includes an artefact-reduced cystometric analysis by using a restrainer to conduct bladder measurements in awake rats.
In this paper, we describe the surgical approach to permanently implant a bladder catheter and external urethral sphincter electrodes, along with cystometric measurements in awake rats.

<table>
	<tbody>
		<tr>
			<td>Polyethylene tubing PE 50</td>
			<td>Becton Dickinson</td>
			<td>427411</td>
			<td>Catheter</td>
		</tr>
		<tr>
			<td>Prolene 6-0 (BV-1, 9.3 mm, 3/8c)</td>
			<td>Ethicon</td>
			<td>EH7403H</td>
			<td>Suture</td>
		</tr>
		<tr>
			<td>Teflon coated steel wire</td>
			<td>Cooner wire</td>
			<td>AS631</td>
			<td>Electrode material</td>
		</tr>
		<tr>
			<td>Silver wire 0.250 mm</td>
			<td>World Precision Instruments</td>
			<td>AGW1030</td>
			<td>Electrode material</td>
		</tr>
		<tr>
			<td>Rotilabo - PVC tube</td>
			<td>Carl Roth</td>
			<td>97241</td>
			<td>Harness</td>
		</tr>
		<tr>
			<td>Vicryl rapide 4-0 (P-3, 13 mm, 3/8c)</td>
			<td>Ethicon</td>
			<td>V4940H</td>
			<td>Suture</td>
		</tr>
		<tr>
			<td>Quick Connect Single Harness</td>
			<td>SAI Infusion Technologies</td>
			<td>QCH-23CW</td>
			<td>Harness</td>
		</tr>
		<tr>
			<td>Shrinking tubes</td>
			<td>ChiliTec</td>
			<td>17894</td>
			<td>Electrode soldering</td>
		</tr>
		<tr>
			<td>Soldering wire Pb60 Sn40</td>
			<td>Stannol</td>
			<td>LD0029</td>
			<td>Electrode soldering</td>
		</tr>
		<tr>
			<td>Fluxing agent 157</td>
			<td>Castolin Eutectin</td>
			<td>157 0150</td>
			<td>Electrode soldering</td>
		</tr>
		<tr>
			<td>Conn Unshrouded Header HDR 3 POS, 2.54mm Solder ST Thru-Hole Box</td>
			<td>Preci-dip</td>
			<td>801-87-050-10-001101</td>
			<td>Electrode soldering</td>
		</tr>
		<tr>
			<td>Conn Socket Strip SKT 50 POS 2.54mm, Solder ST Thru-Hole Box</td>
			<td>Preci-dip</td>
			<td>890-18-003-10-001101</td>
			<td>Electrode soldering</td>
		</tr>
		<tr>
			<td>Rat Cystometry Package (contains pump, scale, pressure transducer, hardware for cystometric analysis)</td>
			<td>Catamount Research and Development Inc.</td>
			<td>CAT-CYT-R</td>
			<td></td>
		</tr>
		<tr>
			<td>Differential amplifier with active headstage</td>
			<td>AD instruments</td>
			<td>DP-311</td>
			<td>EMG amplifier</td>
		</tr>
		<tr>
			<td>Restrainer Medium size for rats 200-300 g</td>
			<td>emka Technologies</td>
			<td>HLD-RM</td>
			<td></td>
		</tr>
		<tr>
			<td>Uro Dyn Software</td>
			<td>Z&#252;rich of University</td>
			<td></td>
			<td>MTA-based</td>
		</tr>
		<tr>
			<td>Female rats (Strain Lewis) 12 weeks of age</td>
			<td>Charles River, Sulzfeld, Germany</td>
			<td></td>
			<td>animals</td>
		</tr>
	</tbody>
</table>

cystometric and external urethral sphincter measurements in awake rats with implanted catheter and electrodes allowing for repeated measurements

Lower urinary tract function is mainly assessed by means of cystometric bladder function analysis in rodents. Conventional cystometries are usually performed as terminal analysis under urethane anesthesia. It is well known that anesthetic drugs can influence bladder function. Hence, the aim of this technique is to perform cystometric measurements of the urinary bladder and external urethral sphincter in lightly restrained awake rats. For this purpose, a bladder catheter is implanted into the bladder dome. Subsequently, two electrodes are implanted bilateral to the external urethral sphincter and a ground electrode is sutured to a non-responsive skeletal muscle. The bladder catheter and the three electrodes are finally tunneled subcutaneously to the neck region and affixed to a harness. With this technique, the lower urinary tract can be measured at multiple time points in the same animal to assess lower urinary tract function. The main application of this technique is the follow-up of simultaneous urinary bladder and external urethral sphincter function in awake healthy rats and after induction of a disease or injury. Moreover, subsequent lower urinary tract monitoring can be performed during evaluation of the disease/injury and to monitor treatment efficacy.

A schematic showing the process of awake cystometric measurements is presented in Figure 1 and the internal anatomy for bladder catheter implantation is shown in Figure 2. Surgery takes about 2 h. Postoperative analgesia and antibiotics, as described in the protocol, cover pain and infections over five days after surgery. No signs of any pain were noticed thereafter. Twice daily careful inspection of the abdomen, abdominal suture...

Watch this Scientific Journal Video about Cystometric and External Urethral Sphincter Measurements in Awake Rats with Implanted Catheter and Electrodes Allowing for Repeated Measurements at JoVE.com

Cystometric and External Urethral Sphincter Measurements in Awake Rats with Implanted Catheter and Electrodes Allowing for Repeated Measurements

This protocol first describes the surgical procedure of the permanent implantation of a urinary bladder catheter combined with external urethral sphincter electrodes, and second, the measurement of the function of the urinary bladder and external urethral sphincter in implanted awake animals.

Lower urinary tract function is mainly assessed by means of cystometric bladder function analysis in rodents. Conventional cystometries are usually ...

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