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

作者没有任何致谢。

这里描述的所有规程由奥地利政府道德委员会批准动物研究 (Bundesministerium Wissenschaft, Forschung 和 Wirtschaft, WF/伏/3b) 和符合协会为实验室评估动物使用动物保育指南。用于这种方法的老鼠是雌性, 12 周大的路易斯鼠。在整个协议中使用无菌仪器。
1. 材料制备
<ol><li>
导管的制造
	<ol>
<li>将导管 (聚乙烯导管 PE-50) 在适当长度 (20 到25厘米) 上, 以适合动物的大小。 注: ...

<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>

本协议描述了永久性导管和尿道括约肌电极植入的手术过程和压记录技术在清醒, 轻度抑制大鼠包括同时分析膀胱和尿道外括约肌。
手术中的关键步骤是仔细植入膀胱导管, 避免渗漏和广泛操作。此外, 对尿道外括约肌的双侧电极进行精确的植入, 对于外括约肌的声音测量至关重要。在植入后对手术场进行严密检查对维持动物的健康也是必不可少的。后续的抗生素覆盖有助于预...

为了分析尿液的储存和排尿功能和功能障碍, 大多数研究都使用了啮齿动物模型。通过连续的反应激活, 排尿产生。这些反射的协调对于有效地排尿1至关重要。压记录技术为分析其神经控制下的膀胱功能提供了有价值的工具1。
大多数传统的 cystometries 在大鼠是作为一个单一的, 最终分析麻醉, 主要是聚氨酯2, 并专注于膀胱完全。然而, 在某些病症中如神经源性下尿路功能障碍 (NLUTD), 不仅膀胱, 而且排尿口, 外部尿道括约肌, 是功能失调的3,4。这使 NLUTD 困难跟随-, 如果仅膀胱在一个单一压测量被审查。为了获得与人类相媲美的可靠结果, 必须准确测量膀胱和尿道外括约肌功能及其相互作用2。此外, 对清醒大鼠进行功能分析非常重要, 因为麻醉很可能会改变膀胱功能2,5,6。一个好的压记录在醒的动物是依据为识别膀胱作用和故障7。
使用的小动物 cystometry 驻地 (例如, 猫 cystometry 驻地 (CCS)) 是一个单位执行压分析在小醒的动物8。通过永久性膀胱导管和植入的外部尿道括约肌电极, 可在较长时间内进行重复性测量2。因此, CCS 为啮齿动物模型中的非神经源和 NLUTD 评价提供了一个有价值的工具, 在这种模式中, pathomechanisms 可以在短期或中期随访中改变。此外, 该方法包括通过使用抑制剂在清醒大鼠中进行膀胱测量来减少压分析。
在本文中, 我们描述了永久性植入膀胱导管和外尿道括约肌电极的手术方法, 以及在清醒大鼠的压测量。

<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

下尿路功能主要通过对啮齿动物的压膀胱功能分析来评估。常规 cystometries 通常在聚氨酯麻醉下进行终端分析。众所周知, 麻醉药能影响膀胱功能。因此, 该技术的目的是进行压测量的膀胱和外尿道括约肌轻度克制清醒大鼠。为此, 膀胱导管被植入膀胱圆顶。随后, 两个电极被植入双侧的外部尿道括约肌和一个接地电极缝合的非反应性骨骼肌。膀胱导管和三电极最终通过隧道皮下注射到颈部区域, 并贴在安全带上。采用这种技术, 可以在同一动物的多个时间点测量下尿路, 以评估下尿路功能。该技术的主要应用是在清醒健康的大鼠和诱导的疾病或损伤后同时膀胱和外括约肌功能的随访。此外, 随后的下尿路监测可以进行评估的疾病/伤害和监测治疗效果。

在图 1中显示了唤醒压测量过程的示意图, 并在图 2中显示了膀胱导管植入的内部解剖。手术需要2小时, 术后镇痛和抗生素, 在手术后五天内包括疼痛和感染。此后没有发现任何疼痛的迹象。每天两次仔细检查腹部、腹部缝合和颈部缝合是维持动物健康所必需的。线束控制 (位置和松紧度) 应在头五天和以后定期进行一次。腹部缝?...

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

本协议首先介绍了膀胱导管与外尿道括约肌电极联合永久性植入的手术方法, 其次, 对膀胱及外尿道功能的测量在植入的清醒动物的括约肌。

压和外尿道括约肌测量在清醒大鼠植入导管和电极允许重复测量

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