Name: evaluating the procedure for performing awake cystometry in a mouse model
Uploaded: 2017-05-20T13:00:00
Description: Watch this Scientific Journal Video about Evaluating the Procedure for Performing Awake Cystometry in a Mouse Model at JoVE.com

This study was funded by the Department of Surgery University of Vermont, Danish Council for Independent Research, and by the Odense University Hospital.

根据机构指导，动物被安置在佛蒙特州动物保护基金会大学。所有的动物实验都是按照国家卫生研究院的指南进行的，用于护理和使用实验动物。 膀胱内管植入<ol><li> 准备外科手术的管道和器械 <ol><li>切割一个7厘米的PE10管，使导管植入。 </li><li>通过缓慢地将端部朝着明亮的火焰推进，在PE10管的一端形成一个耀斑。 注意：一旦发生耀斑，就可以快速取出?...

<ol>
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最佳材料和大小的膀胱内管为了确定管径对压力记录的影响，我们测试了不同的微流体管; PE50（0.58mm ID），聚氨酯PU027（0.4mm ID），PE25（0.46mm ID）和PE10（0.28mm ID）。对于每个管，记录输注泵的压力为1mL / h，同时将管垂直地从0延伸至30cm。初始体内实验尝试使用PE50管，但是由于与小鼠膀胱相比，管的尺寸不成功（ 图4A ）。虽然这支持了Smith和Kuchel的发现，他们建议在...

填充性膀胱测量术（FC）是一种诊断方法，其涉及将膀胱放置在膀胱中以在缓慢的膀胱充盈期间记录压力。首先在1927年作为评估下尿路功能的临床诊断方法，它一直被广泛使用。 1在研究应用中，FC可用于测试健康和患病动物模型中的膀胱功能，并研究药理作用的影响。啮齿类动物模型通常用于调查下尿路功能。 2在这组哺乳动物中，FC首先开发用于大鼠。 3在这里，将管子植入膀胱并执行FC的方法已被许多研究人员充分描述和使用，具有可接受的重现性水平。转基因和敲除菌株的可用性使得小鼠成为许多研究领域的宝贵物种，包括下尿路功能障碍的领域。用于执行小鼠膀胱测压术的方法在实验室之间变化明显，使得难以比较结果。 五 与离体模型相比，FC保留了下尿路解剖学，允许评估排尿周期的储存和排尿期间膀胱及其出口之间的协调作用。以前的研究表明，许多常用的麻醉药抑制排尿收缩。维持膀胱平滑肌收缩的药物（氨基甲酸酯，α-氯醛酸，氯胺酮和赛拉嗪），允许动物排尿，仍然显着降低功能性膀胱容量并抑制神经传递。 6，7，8，9虽然在技术上更具挑战性，FC在aw中表现出色ake行动动物保留排尿反射的功能完整性。 下尿路功能受多种因素的影响，包括手术后膀胱壁肿胀，由于疼痛和不适引起的压力，以及环境影响。使用在管植入期间最小化组织损伤的手术技术和减少管运动的记录方法，同时允许动物自由行走，对于获得准确和可再现的记录是必需的。 如果充分进行，在体内 FC在自由移动的动物中可以提供可靠地反映生理性膀胱功能的数据。自由移动动物中的10个 FC可以提供以下参数的数据;基础或基线压力：两次排尿之间的最小压力。间歇压力：两次排尿之间的平均压力。阈值压力：膀胱内压排尿之前。最大压力：排尿期间膀胱压力最大。自发活动（或平均间歇性振荡压力）：排气压力减去基础压力。无空洞的收缩：在充盈阶段增加膀胱内压，与流体释放无关。膀胱依从性：膀胱容量除阈值压力减去基础压力。排尿频率：每单位时间的排尿次数。间隔间隔：两个最大排尿压力之间的周期。膀胱容量：输注量除以排尿次数。以前已经公布了这些参数和标准化术语的详细描述。 11 FC可以使用连续或单循环膀胱输液法进行。连续膀胱测量允许记录多个排尿周期并选择代表性的数据重现性。由于未知剩余体积，其测量膀胱容量的准确性受到限制。此外，在自由行走的小鼠中收集小的空隙体积（其基于应变和性别在30和184μL之间变化）是具有挑战性的。使用这种方法记录空洞体积与麻醉制剂相比不太准确，但是优于其避免麻醉剂对膀胱功能的抑制作用。单循环膀胱测量应用于评估膀胱容量。在这种方法中，膀胱在输注之前通过抽吸排空，并且根据输注速率乘以时间到最大压力的函数计算容量。 虽然已经出版了在小型啮齿动物中进行膀胱测压的技术，但它描述了在大鼠中进行的手术，并建议在氨基甲酸乙酯麻醉下进行小鼠膀胱测压。这个沟通的目标是to描述了用于将膀胱管植入膀胱圆顶的显微手术技术和用于在清醒的自由移动的小鼠中连续膀胱充盈和排尿期间在体内记录下尿路功能的实验装置。此外，进行实验以解决管长度，直径和材料以及在体内 FC中执行的方法是否影响记录。该实验方案总结了以前发表的技术，并提出了一些基于实验结果的修改。

<table border="0" cellpadding="0" cellspacing="0" width="946">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Polyethylene (PE) 10 tubing</td>
			<td style="width:259px;">Instech</td>
			<td style="width:119px;">BTPE-10</td>
			<td style="width:191px;">Fits 30G connectors/plugs</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Polyethylene (PE) 50 tubing</td>
			<td style="width:259px;">Instech</td>
			<td style="width:119px;">BTPE-50</td>
			<td>Fits 22G connectors/plugs</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">22ga single channel stainless steel swivel</td>
			<td style="width:259px;">Instech</td>
			<td style="width:119px;">375/22</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">High Carbon Steel Utility Extension Spring (9/64&#34; OD)</td>
			<td style="width:259px;">Grainger</td>
			<td style="width:119px;">1NAH1</td>
			<td>Protects PE50 tubing - Cut to length</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">22G connector</td>
			<td style="width:259px;">Instech</td>
			<td style="width:119px;">SP22/12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Yutaoz Professional Hot Melt Adhesive Glue Gun</td>
			<td style="width:259px;">Yutaoz</td>
			<td style="width:119px;"></td>
			<td>Use low temperature setting (100&#176;C) - Any hot melt glue gun with an adjustable temperature range will work</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Surebonder DT-2010 all purpose glue stick</td>
			<td style="width:259px;">Surebonder</td>
			<td style="width:119px;"></td>
			<td>Any all purpose hot glue will work</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Dumont #5 curved microforceps</td>
			<td style="width:259px;">World Precision Instruments</td>
			<td align="right">500232</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Dumont #7 curved microforceps</td>
			<td style="width:259px;">World Precision Instruments</td>
			<td align="right">14188</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mini dissecting scissors - straight</td>
			<td style="width:259px;">World Precision Instruments</td>
			<td align="right">503240</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro mosquito forceps (12.5cm)</td>
			<td style="width:259px;">World Precision Instruments</td>
			<td align="right">500451</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dissecting scissors - straight</td>
			<td style="width:259px;">World Precision Instruments</td>
			<td align="right">14393</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Castroviejo Needle Holder</td>
			<td style="width:259px;">World Precision Instruments</td>
			<td align="right">503258</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Isoflurane, USP</td>
			<td style="width:259px;">Phoenix</td>
			<td style="width:119px;"></td>
			<td>2%, 1 L/min flow rate</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Buprenorphine</td>
			<td style="width:259px;"></td>
			<td style="width:119px;"></td>
			<td>0.05mg/kg</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">0.9% Sodium Chloride Irrigation, USP</td>
			<td style="width:259px;">Baxter</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">6-0 Ethilon black monofilament, non-absorbable suture</td>
			<td style="width:259px;">Ethicon</td>
			<td style="width:119px;"></td>
			<td>Bladder tie</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">6-0 Vicryl violet braided, absorbable suture</td>
			<td style="width:259px;">Ethicon</td>
			<td style="width:119px;"></td>
			<td>Muscle suture, running</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">6-0 Prolene blue monofilament, non-absorbable suture</td>
			<td style="width:259px;">Ethicon</td>
			<td style="width:119px;"></td>
			<td>Skin suture, vertical mattress, buried interrupted</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">KD Legato 210 infuse/withdraw pump</td>
			<td style="width:259px;">KD Scientific</td>
			<td style="width:119px;"></td>
			<td>1.5ml/hr</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Disposable pressure transducer</td>
			<td style="width:259px;">Digitimer</td>
			<td style="width:119px;">NL108T2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Pressure Amplifier</td>
			<td style="width:259px;">Digitimer</td>
			<td style="width:119px;">NL108A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Power1401-3 data acquisition interface</td>
			<td style="width:259px;">Digitimer</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Spike2&#160;</td>
			<td style="width:259px;">Cambridge Electronic Design Limited</td>
			<td style="width:119px;"></td>
			<td>PC pressure recording software</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:379px;">Leica MZ6 surgical operating microscope (3.2-20X)</td>
			<td style="width:259px;">Leica Microsystems</td>
			<td style="width:119px;"></td>
			<td>Magnification</td>
		</tr>
	</tbody>
</table>

evaluating the procedure for performing awake cystometry in a mouse model

唤醒灌注膀胱测量法已经被用于评估自由移动小鼠的膀胱功能，但是实际使用的具体方法有所不同。这项研究的目的是描述用于植入膀胱内管的显微外科手术和用于在清醒，自由移动的小鼠中记录膀胱压力的实验技术。此外，还提供了实验数据，以显示手术以及管道类型和尺寸如何影响下尿路功能和记录敏感性。在具有不同内径的聚乙烯和聚氨酯管中评估管直径对压力记录的影响。随后，将来自两种材料的最佳试管手术植入雄性C57BL / 6小鼠膀胱的圆顶。手术后2,3,5和7天在健康，完整的动物和动物中记录了12小时，隔夜排尿次数。在收获时，气囊w用大量观察评估肿胀征象，随后进行病理分析。在第2天和第3天观察到膀胱肿胀最大程度，与行为排尿数据相关，显示膀胱功能受损明显。到第5天，膀胱组织学和排尿频率正常化。根据我们研究提供的文献和证据，我们提出了在醒来的小鼠体内记录膀胱内压和空洞体积的以下步骤：1）使用手术显微镜和显微外科手术进行手术，2）使用聚乙烯-10管道以最小化运动伪影，以及3）在膀胱肿胀消退后，在术后第5天进行膀胱测压。

在管堵塞期间，管材料与直径之间的压力上升和系统内的压力一致性没有显着差异。膀胱管内植入的膀胱壁肿胀对于聚乙烯（PE）和聚氨酯（PU）材料都是显着的。在第2天，严重粘膜下肿胀。它占据膀胱横截面的一半，导致管腔阻塞。在第5天，水肿完全解决，使黏膜下层区域渗入部分侵入肌层的炎症细胞。在第7天，炎症浸润显着减少，膀胱壁组织学恢复正常（ 图8</...

Watch this Scientific Journal Video about Evaluating the Procedure for Performing Awake Cystometry in a Mouse Model at JoVE.com

Evaluating the Procedure for Performing Awake Cystometry in a Mouse Model

本研究描述了在自由移动的小鼠中进行清醒膀胱测压术的外科手术和实验技术。此外，它还提供实验证据来支持其优化和标准化。

评估在鼠标模型中执行唤醒膀胱测压的过程

Awake filling cystometry has been used for a long time to evaluate bladder function in freely moving mice, however, the specific methods used, vary among ...

Research

JoVE Journal

Medicine

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在接受深部脑刺激手术术中唤醒人类行为任务

Deep brain stimulation (DBS) is a surgical procedure that directs chronic, high frequency electrical stimulation to specific targets in the brain through ...

科研

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

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	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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Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder resulting in progressive degeneration of motoneurons. Peak of onset is around 60 ...

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Intranasal administration is a method of delivering  therapeutic agents to the central nervous system (CNS). It is  non-invasive and allows large ...

Performing Subretinal Injections in Rodents to Deliver Retinal Pigment Epithelium Cells in Suspension

视网膜下进行注射的鼠害提供视网膜色素上皮细胞悬浮

The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment ...

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The SOD1-G93A transgenic mouse is the most widely used animal model of amyotrophic lateral sclerosis (ALS). At ALS TDI we developed a phenotypic screening ...

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The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the ...

The Monoiodoacetate Model of Osteoarthritis Pain in the Mouse

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A major symptom of patients with osteoarthritis (OA) is pain that&#160;is triggered by peripheral as well as central changes within the pain pathways. The ...

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气道局部麻醉清醒用于内镜插管使用喷雾为你去技术在高氧流量

A patient&#39;s willingness to cooperate is an absolute precondition for successful awake intubation of the trachea. Whilst drug-sedation of patients can ...