Name: intracavernosal pressure recording to evaluate erectile function in rodents
Uploaded: 2018-06-06T12:30:00.000+00:00
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这项工作得到了中央大学基础研究基金 (020814380018, 020814380077), 中国奖学金理事会 (CSC, 201606195024 号), 江苏省自然科学基金 (BK20160138) 和重点项目的支持。由南京医科大学科技发展基金会 (2014NJMUZD053) 资助。

本研究采用三月大、18月大的大鼠。所有动物都是按照 NIH 的护理和使用实验动物的指导方针处理的。涉及动物问题的程序得到当地机构动物照料和道德委员会的批准, 努力尽量减少动物的痛苦。这些议定书是由南京理工大学 (中国南京) 机构动物保育和使用委员会 (IACUC) 批准的。
在 ICP 记录前, 根据其年龄和早期表现, 将大鼠分为两组: 青年正常组 (YN 组) 和老年勃起功能障碍组 (AE 组)10。
1. 手术前准备
<ol><li>手动为 ICP 记录制作一对双极性电极 (图 1)。稍微弯曲电极的两端, 并将两个电极之间的距离调整为1-2 毫米宽, 如图 1A所示。</li>
	<li>使用两个鳄鱼钳 (图 1A, B) 将电极连接到刺激器。</li>
	<li>
装配导管系统:首先, 将皮下注射的23G 针连接到带有油管的3路旋塞阀, 然后将旋塞阀连接到压力传感器。接下来, 在旋塞阀的第三端附上一个10毫升的注射器, 以提供肝素生理盐水。</li>
	<li>在用肝素盐 (200 U/毫升) 填充整个系统后, 仔细检查是否有渗漏。然后将3路旋塞阀关闭注射器通道或压力传感器通道(图 1C)。</li>
	<li>提起针20厘米的木垫水平。然后将压力记录系统校准到20厘米 H2O。之后, 移动针的高度, 以验证记录系统的准确性。重复校准, 直到确认准确性。</li>
	<li>将老鼠从动物设施转移到手术室, 让他们习惯于手术室至少30分钟。</li>
	<li>在手术前用70% 乙醇喷洒蒸压设备</li>
</ol>2. 手术程序
<ol><li>麻醉大鼠腹腔注射戊巴比妥钠, 剂量为45毫克/千克体重, 并等待5-10 分钟. 捏脚趾以确认适当的麻醉。</li>
	<li>用电动剃须刀剃去腹部和颈部的皮毛, 然后把老鼠放在暖气垫上。</li>
	<li>用10% 聚维酮碘溶液浸泡棉球, 然后用70% 只乙醇浸泡棉球擦拭手术区。同时, 应用眼科软膏防止眼部干燥。</li>
	<li>Catheterize 左颈动脉。<ol>
<li>用镊子抓住颈部的皮肤, 在颈部中部做水平切口。切开肌肉, 仔细暴露左颈动脉, 并隔离血管的5毫米部分。</li>
		<li>用镊子仔细地将颈动脉与迷走神经分开, 在颈动脉下画一条丝缝线, 并在血管的尾端放一条松散的领带, 然后在血管的颅骨末端再做一个紧的结。</li>
		<li>Caudally 钳夹在缝合线上方的牛头犬钳, 以阻止血液流动。</li>
		<li>用显微外科剪刀在血管上做切口, 在微解剖钩和镊子的协助下, 将动脉导管插入心脏。</li>
		<li>把松动的尾鳍绑在导管上以确保它的安全。取出牛头犬钳以恢复血液流动。</li>
	</ol>
</li>
	<li>
隔离 CN 并放置电极
	<ol>
<li>用钳子抬起腹部的皮肤和肌肉。用解剖剪刀, 切开下腹部到小弟弟做中线切开。</li>
		<li>用拭子轻轻地将肠道推入腹腔上部。</li>
		<li>用一副镊子抓住膀胱, 从腹腔取出膀胱。暴露前列腺的腹叶, 位于尿道的腹部。</li>
		<li>拔出前列腺、精囊和输精管的腹叶, 露出前列腺的背叶。找出输精管和前列腺的粘连点。</li>
		<li>分离前列腺和输精管之间的空间。仔细暴露的纤维胶囊, 这是位于后到连接点的前列腺和输精管。然后找到主要的骨盆神经节 (图 2)。 注: 在前列腺表面可以看到主要的盆腔神经节和海绵状神经。</li>
		<li>小心干燥 CN 区域与无菌拭子。用两极电极仔细隔离并钩住右海绵状神经。</li>
	</ol>
</li>
	<li>
Catheterize 左下肢
	<ol>
<li>用解剖剪刀在小弟弟的皮肤上切开一个小切口, 然后仔细掀的小弟弟的皮肤。</li>
		<li>解剖纹状的小弟弟肌组织。找到耻骨骨的上部分支。</li>
		<li>暴露 bulbospongiosus 的肌肉, 包括 spongious 灯泡。</li>
		<li>用弯钳将 bulbospongiosus 肌从 ischiocavernosus 肌中分离出来。</li>
		<li>小心地将 ischiocavernosus 肌肉与弯曲钳隔离, 然后切开 ischiocavernosus 肌以暴露小腿海绵的白色膜。</li>
		<li>遵循小腿语料库海绵的解剖方向, 小心地将针插入小腿语料库海绵通过白色的膜。 注意: 这是成功导尿的关键步骤。如果针插入正确, 少量的血气盐水可以被注射, 并且应该观察轻微的小弟弟膨胀。</li>
		<li>小心释放针头, 避免任何滑动的针头或中断连接管。检查是否有渗漏。</li>
	</ol>
</li>
</ol>3. 刺激 CN
<ol><li>打开压力信号记录软件程序, 启动压力信号记录。</li>
	<li>设置刺激的参数:15 赫兹, 脉冲宽度为5毫秒, 5 伏, 和六十年代的持续时间。以15赫兹的频率刺激 CN, 脉冲宽度为5毫秒。 注: 在应用电刺激时, 可观察到 ICP 的急剧上升。</li>
	<li>让刺激之间有30分钟的休息时间。每只动物的连续刺激的最大值是三倍。</li>
</ol>4. 结束程序
<ol><li>记录后, 通过注射过量的戊巴比妥钠剂量150毫克/千克体重来管理安乐死。通过检查动脉压力来证实老鼠的死亡。取出老鼠, 清理手术工具。</li>
</ol>5. 数据分析
<ol><li>保存并导出软件中的数据。反应通常表示为 ICP 与全身平均动脉压的比值 (图)。计算了峰值比值/MAP 对勃起功能的评价。</li>
	<li>从至少五只老鼠那里采集数据, 并用统计软件进行分析。当p < 0.05 使用学生的 t 测试时, 差异被认为具有统计学意义。</li>
</ol>

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作为勃起功能的直接措施, ICP 是一种可靠的方法14。它允许获得关于基 icp 备、峰值比较、高原 icp、安装时间和消肿时间、响应时间、等的数据。除这些直接测量参数外, 还有一些其它指标参数: (1) "T80", 达到80% 峰值的时间;(2) "D20", 时间减少到20% 峰值 ICP;(3) "ΔT80", T80 压力增加率 (每秒);(4) "ΔD20", 在 D20 的压力降低率 (每秒)。这些参数允许量化每个阶段和 ICP 反应的质量, 这可...

ED 被定义为无法达到或保持勃起, 并已成为常见的男性性障碍1。实验动物被使用并且提供可再生的模型调查勃起功能2。长期以来, 一些较大的动物模型被用于调查勃起功能3,4,5。虽然啮齿动物比其他动物相对较小, 但它们也被用来研究男性勃起功能障碍, 因为它表现出了几个优点6。首先, 人类的形态和功能性特征在啮齿类动物中概括。第二, 与 ED 研究中使用的大型动物相比, 啮齿动物更经济的购买, 房子和维护。第三, 转基因啮齿动物模型在重现性和后续行为以及神经研究中提供了优势。因此, 啮齿类动物很快就成为了研究男性勃起功能障碍的主要方法。
啮齿动物模型得益于纯遗传背景和一致的文化条件, 提供了一致的可重现数据5,6,7,8。在许多与勃起功能相关的研究中, 阿扑吗啡的勃起反应试验和电刺激诱发的 ICP 反应试验是最广泛使用的方法, 可以可靠地反映勃起函数9,10,11,12。由希顿et 等开发的蛋白诱导的勃起功能测试。13是一种生物检测方法, 它利用阿扑吗啡对大鼠进行勃起和打呵欠的现象。作为一种简便、无创、稳定的生物检测方法, 对勃起功能进行评价, 在许多研究中广泛应用了该蛋白诱导的勃起功能检测。然而, 这项化验并不能充分反映勃起的质量或血液流量的动态变化, 与阳痿的反应14。ICP 测量最初是由昆兰et 等人开发的。15. 在该方法中, 将导管放入颈动脉以测量全身血压, 另一导管插入小腿语料库海绵以记录 ICP。在 ICP 记录之前或期间, 主要骨盆神经节 (加仑) 或 CN 的血管活性剂和/或电场刺激经常被给予老鼠14。这种检测方法是评估 ED 治疗和药物的可靠工具, 在未来的6中很有可能被用作重要的评估手段。
与易于执行的蛋白能诱发的勃起功能测试相比, 成功的 ICP 记录需要注意细节、练习和坚持操作方法。因此, 在这里, 我们提供了如何执行 ICP 记录的详细说明。

<table><tbody><tr><td>&lt;strong&gt;Animal&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Rats</td><td></td><td></td><td>Strain: Sprague-Dawley Age: 2-3 month</td></tr><tr><td>Rats</td><td></td><td></td><td>Strain: Sprague-Dawley Age: 15-18 month</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Reagents&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Saline</td><td>Sigma-Aldrich, S7653</td><td></td><td>dissolve 8.5 gram sodium chloride in distilled water</td></tr><tr><td>Pentobarbital sodium solution</td><td>Sigma-Aldrich, P3761</td><td></td><td>dissolve 1 gram in 100 ml saline</td></tr><tr><td>Povidone-iodine</td><td>BTP Pharmaceutical Co. Limited</td><td></td><td>10% (V/V)</td></tr><tr><td>Ethanol</td><td>China National Pharmaceutical Group Corporation (SINOPHARM)</td><td></td><td>70% (V/V)</td></tr><tr><td>Heparin</td><td>Sigma-Aldrich, H3149</td><td></td><td>dissolve 20000U heparin in 100 ml saline</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Materials&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Hypodermic needle</td><td>Shandong Weigao Group Medical Polymer Co., Ltd.&amp;nbsp;</td><td>23G</td><td></td></tr><tr><td>Syringe</td><td>Shandong Weigao Group Medical Polymer Co., Ltd</td><td>10 ml</td><td></td></tr><tr><td>Three-way stopcock</td><td>Chengdu Instrument factory</td><td>TSK 01</td><td></td></tr><tr><td>Electrode</td><td>Chengdu Instrument factory</td><td>JST-1</td><td></td></tr><tr><td>Catheter tube</td><td>Chengdu Instrument factory</td><td>PE-10, PE-50</td><td></td></tr><tr><td>Operating scissors</td><td>Shanghai operation equipment factory</td><td>J22010, J22020</td><td></td></tr><tr><td>Ophthalmic operating scissors</td><td>Shanghai operation equipment factory</td><td>Y00010, Y00020</td><td></td></tr><tr><td>Ophthalmic forceps</td><td>Shanghai operation equipment factory</td><td>JD1010, JD1020</td><td></td></tr><tr><td>MicroScissors</td><td>World Precision Instruments</td><td>WAA260</td><td></td></tr><tr><td>silk suture</td><td>Shandong Weigao Group Medical Polymer Co., Ltd.&amp;nbsp;</td><td>5-0</td><td></td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Equipment&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Stimulator</td><td>Nanjing medease science and technology co. ltd (model 4C501H)</td><td></td><td>15 Hz, 5 ms pulse, 5 V, 60 s duration and 5 minutes interval</td></tr><tr><td>Multichannel signal collection processing system</td><td>Nanjing medease science and technology co. ltd (model 4C501H)</td><td></td><td>Blood pressure model</td></tr><tr><td>Pressure transducer</td><td>Beijing Xin Hang Xing Ye Technology Trading Company Limited (model YP100)</td><td></td><td>40KPa</td></tr></tbody></table>

intracavernosal pressure recording to evaluate erectile function in rodents

勃起功能障碍 (ED) 被定义为不能达到或保持勃起的小弟弟, 这已经成为一个普遍的男性性障碍。啮齿类动物被许多研究用来研究勃起功能的生理学/病理学。通过测量 intracavernosal 压力 (ICP) 可以评价啮齿动物的勃起功能。在实际应用中, 可以通过电刺激海绵神经 (CNs) 来监测 ICP。应用颈动脉压 (平均动脉压) 作为 ICP 的参考。利用 icp 记录协议, 可以从 icp 反应曲线测量勃起功能的许多关键参数。ICP 测量比阿扑吗啡诱导的勃起试验提供更多的信息, 而且比海绵的人的小弟弟的遥测监测更便宜, 这使得这种方法最受欢迎的一个评价阳痿的功能。然而, 与易于执行的蛋白能诱导勃起功能测试相比, 成功的 ICP 记录需要注意细节, 实践和坚持的操作方法。在这项工作中, 介绍了在大鼠的 ICP 记录, 以有效补充程序。

许多研究表明, 老年男性勃起功能障碍正成为一个普遍的问题。然而, 在管理衰老相关的教育16方面, 医疗治疗受到限制。在衰老相关的啮齿动物模型中, 对老龄大鼠的勃起功能进行了多种治疗。正如我们前面所介绍的那样, ICP 记录试验可用于将 ED 动物与实验动物的总人口区分开来, 这对于量化潜在治疗或药物对勃起功能的影响也很有价值。
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Watch this Scientific Journal Video about Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents at JoVE.com

Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents

Intracavernosal 压力记录对啮齿动物勃起功能的评价

Intracavernosal 压力记录 (ICP) 是评价实验动物勃起功能的重要方法。在这里, 通过置管下肢的小弟弟, 然后电刺激大鼠的海绵神经, 证明了比较 ICP 的记录过程的详细协议。

Erectile dysfunction (ED) is defined as the inability to attain or keep an erection of the penis, and this has become a prevalent male sexual disorder. ...

intracavernosal-pressure-recording-to-evaluate-erectile-function

Research

JoVE Journal

Biology

14.1K 次观看.  Nanjing University. Intracavernosal 压力记录 (ICP) 是评价实验动物勃起功能的重要方法。在这里, 通过置管下肢的小弟弟, 然后电刺激大鼠的海绵神经, 证明了比较 ICP 的记录过程的详细协议。

视频: Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents

Epidural Intracranial Pressure Measurement in Rats Using a Fiber-optic Pressure Transducer

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and cardiac arrest. This elevation may result in further neurological injury, in the form of transtentorial herniation1,2,3,4, midbrain compression, neurological deficit or increased cerebral infarct2,4. Current therapies are often inadequate to control elevated ICP in the clinical setting5,6,7 . Thus there is a need for accurate methods of ICP measurement in animal models to further our understanding of the basic mechanisms and to develop new treatments for elevated ICP.
In both the clinical and experimental setting ICP cannot be estimated without direct measurement. Several methods of ICP catheter insertion currently exist. Of these the intraventricular catheter has become the clinical 'gold standard' of ICP measurement in humans8. This method involves the partial removal of skull and the instrumentation of the catheter through brain tissue. Consequently, intraventricular catheters have an infection rate of 6-11%9. For this reason, subdural and epidural cannulations have become the preferred methods in animal models of ischemic injury. 
Various ICP measurement techniques have been adapted for animal models, and of these, fluid-filled telemetry catheters10 and solid state catheters are the most frequently used11,12,13,14,15. The fluid-filled systems are prone to developing air bubbles in the line, resulting in false ICP readings. Solid state probes avoid this problem (Figure 1). An additional problem is fitting catheters under the skull or into the ventricles without causing any brain injury that might alter the experimental outcomes. Therefore, we have developed a method that places an ICP catheter contiguous with the epidural space, but avoids the need to insert it between skull and brain. 
An optic fibre pressure catheter (420LP, SAMBA Sensors, Sweden) was used to measure ICP at the epidural location because the location of the pressure sensor (at the very tip of the catheter) was found to produce a high fidelity ICP signal in this model. There are other manufacturers of similar optic fibre technologies13 that may be used with our methodology. Alternative solid state catheters, which have the pressure sensor located at the side of the catheter tip, would not be appropriate for this model as the signal would be dampened by the presence of the monitoring screw. 
Here, we present a relatively simple and accurate method to measure ICP. This method can be used across a wide range of ICP related animal models.

A novel technique to record the pressures within the skull is described. The minimally invasive method uses a fibre-optic pressure sensing system to accurately measure intracranial pressure (ICP) in anaesthetized rats without causing significant brain trauma. The technique may be used in a wide range of experimental models.

硬膜外颅内压测量中使用的光纤压力传感器的大鼠

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and ...

科研

医学

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. Despite the discovery of key regulators of uterine contractility, this process is still not fully understood. Transgenic mice provide an ideal model in which to study parturition. Previously, the only method to study uterine contractility in the mouse was ex vivo isometric tension recordings, which are suboptimal for several reasons. The uterus must be removed from its physiological environment, a limited time course of investigation is possible, and the mice must be sacrificed. The recent development of radiometric telemetry has allowed for longitudinal, real-time measurements of in vivo intrauterine pressure in mice. Here, the implantation of an intrauterine telemeter to measure pressure changes in the mouse uterus from mid-pregnancy until delivery is described. By comparing differences in pressures between wild type and transgenic mice, the physiological impact of a gene of interest can be elucidated. This technique should expedite the development of therapeutics used to treat myometrial disorders during pregnancy, including preterm labor.

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

宫内遥测来衡量鼠标收缩压力<em&gt;在体内</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

Novel Approach for Simultaneous Recording of Renal Sympathetic Nerve Activity and Blood Pressure with Intravenous Infusion in Conscious, Unrestrained Mice.

Renal sympathetic nerves contribute significantly to both physiological and pathophysiological phenomena. Evaluating renal sympathetic nerve activity (RSNA) is of great interest in many areas of research such as chronic kidney disease, hypertension, heart failure, diabetes and obesity. Unequivocal assessment of the role of the sympathetic nervous system is thus imperative for proper interpretation of experimental results and understanding of disease processes. RSNA has been traditionally measured in anesthetized rodents, including mice. However, mice usually exhibit very low systemic blood pressure and hemodynamic instability for several hours during anesthesia and surgery. Meaningful interpretation of RSNA is confounded by this non-physiological state, given the intimate relationship between sympathetic nervous tone and cardiovascular status. To address this limitation of traditional approaches, we developed a new method for measuring RSNA in conscious, freely-moving mice. Mice were chronically instrumented with radio-telemeters for continuous monitoring of blood pressure as well as a jugular venous infusion catheter and custom-designed bipolar electrode for direct recording of RSNA. Following a 48-72 hour recovery period, survival rate was 100% and all mice behaved normally. At this time-point, RSNA was successfully recorded in 80% of mice, with viable signals acquired up to 4 and 5 days post-surgery in 70% and 50% of mice, respectively. Physiological blood pressures were recorded in all mice (116&plusmn;2 mmHg; n=10). Recorded RSNA increased with eating and grooming, as well-established in the literature. Furthermore, RSNA was validated by ganglionic blockade and modulation of blood pressure with pharmacological agents. Herein, an effective and manageable method for clear recording of RSNA in conscious, freely-moving mice is described.

Anesthetized mice exhibit non-physiological systemic blood pressure, which precludes meaningful assessment of autonomic tone given the intimate relationship between blood pressure and the autonomic nervous system. Thus, a novel method to simultaneously record renal sympathetic nerve activity and blood pressure with intravenous infusion in conscious mice is outlined.

清醒、无拘无束的小鼠静脉输液同时记录肾交感神经活动和血压的新方法。

Renal sympathetic nerves contribute significantly to both physiological and pathophysiological phenomena. Evaluating renal sympathetic nerve activity ...

Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat

The stimulation of the cavernous nerve (CN) and measurement of intracavernous pressure (ICP) have been used extensively to test and evaluate therapies for erectile dysfunction. However, the methods used vary between laboratories, and pitfalls still exist. The goal of this study was to describe a surgical technique that would provide a reliable and reproducible model. By exposing the ischiocavernosus muscle at its point of insertion on the ischial tuberosity, the penile crus could be cannulated with minimal dissection and injury to the structures involved in erectile function. Repeated stimulation of the CN, without the need for lifting and drying, was achieved by using a 125 &#181;m bipolar silver electrode and biocompatible silicon glue to isolate the electrode-nerve complex. This method prevents neuropraxia by reducing stretching and drying the nerve and provides complete isolation of the nerve, negating electrical leakage and preventing stimulation of alternative pathways.

This study describes a simplified surgical procedure and technique for performing cavernous nerve stimulation with the isolation of the nerve-electrode complex using silicone glue and intracavernous pressure measurement.

大鼠海绵状神经刺激与海绵压力记录

The stimulation of the cavernous nerve (CN) and measurement of intracavernous pressure (ICP) have been used extensively to test and evaluate therapies for ...

Simultaneous Electrocardiography Recording and Invasive Blood Pressure Measurement in Rats

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. Research focusing on cardiovascular disease models, potential cardiovascular therapeutic targets or pharmaceutical agents requires assessment of systemic arterial pressure and heart rhythm changes. In situations where radio telemetry systems are not available or affordable, the technique of femoral artery cannulation is an alternative way to obtain intra-arterial pressure waveform recordings and systemic BP measurements. This technique is economical and can be performed with standard equipment in animal facilities. However, invasive arterial pressure recording requires cannulation of small arteries, which can be a challenging surgical skill. Here, we present step-by-step protocols for femoral artery cannulation procedures. Key procedures include the calibration of the data acquisition system, tissue dissection and femoral artery cannulation, and setup of the arterial cannulation system for pressure recording. Surface electrocardiography recording procedures are also included. We also present examples of BP recordings from normotensive and hypertensive rats. This protocol allows reliable direct recordings of systemic BP with simultaneous electrocardiography.

Here, we describe a setup for simultaneous recording of electrocardiography and intra-arterial blood pressure (BP) in experimental rats, which can be done with standard equipment in animal facilities and can be applied to physiological or pharmacological studies to investigate pathogenic or therapeutic mechanisms in cardiovascular medicine.

大鼠同时心电图记录及侵入性血压测量

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. ...

In Vivo Luminal Measurement of Distension-Evoked Urothelial ATP Release in Rodents

ATP, released from the urothelium in response to bladder distension, is thought to play a significant sensory role in the control of micturition. Therefore, accurate measurement of urothelial ATP release in a physiological setting is an important first step in studying the mechanisms that control purinergic signaling in the urinary bladder. Existing techniques to study mechanically evoked urothelial ATP release utilize cultured cells plated on flexible supports or bladder tissue pinned into Ussing chambers; however, each of these techniques does not fully emulate conditions in the intact bladder. Therefore, an experimental setup was developed to directly measure ATP concentrations in the lumen of the rodent urinary bladder.
In this setup, the bladders of anesthetized rodents are perfused through catheters in both the dome of the bladder and via the external urethral orifice. Pressure in the bladder is increased by capping the urethral catheter while perfusing sterile fluid into the bladder through the dome. Measurement of intravesical pressure is achieved using a pressure transducer attached to the bladder dome catheter, akin to the setup used for cystometry. Once the desired pressure is reached, the urethral catheter's cap is removed, and fluid collected for ATP quantification by luciferin-luciferase assay. Through this experimental setup, the mechanisms controlling both mechanical and chemical stimulation of urothelial ATP release can be interrogated by including various agonists or antagonists into the perfusate or by comparing results between wildtype and genetically modified animals.

In Vivo Luminal Measurement of Distension-Evoked Urothelial ATP Release in Rodents

This protocol describes the procedure for measuring ATP concentrations in the lumen of the bladder in an anesthetized rodent.

体内 啮齿动物膨胀诱发尿路上皮ATP释放的管腔测量

ATP, released from the urothelium in response to bladder distension, is thought to play a significant sensory role in the control of micturition. ...

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.

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.

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

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

生物学

Intracavernous Pressure Recording in a Cavernous Nerve Injury Rat Model

The bilateral cavernous nerve (CN) injury rat model has been extensively used to simulate clinical cavernous nerve injury associated with erectile dysfunction (ED) for evaluating the effect of clinical therapeutic methods. However, the methods of CN injury model construction are flawed and varied in the ED research field. It is CN crush injury that is the most commonly used method in recent years. This study aims to provide a detailed description of the procedure of bilateral CN injury rat model construction and measurement of intracavernous pressure (ICP) recording, providing a reliable and reproducible CN injury rat model. This work successfully developed the CN injury method of hemostat crush injury using a syringe needle as hard support and a hemostat with a rubber sleeve. Also, this method concludes that a voltage of 1.0 V, frequency of 20 Hz, and pulse-width of 5 ms are the optimized stimulation parameters for ICP recording in a bilateral CN injury rat model.

This protocol describes developing a stable bilateral cavernous nerve injury rat model of radical prostatectomy associated with erectile dysfunction and intracavernous pressure measurement.

The bilateral cavernous nerve (CN) injury rat model has been extensively used to simulate clinical cavernous nerve injury associated with erectile ...

Performing Cavernous Nerve Stimulation in Rats to Measure Intracavernous Pressure

Source: Hox, M., et al. <a href="https://app.jove.com/v/56807">Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat.</a> J. Vis. Exp. (2018).
This video demonstrates the isolation and stimulation of the cavernous nerve in an anesthetized rat to study blood pressure changes in the cavernous body.

对大鼠进行海绵状神经刺激以测量海绵体内压力

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

JoVE Encyclopedia of Experiments

Neuroscience

Neurophysiology

Stimulation and Modulation Techniques

Recording Changes in Renal Sympathetic Nerve Activity in Response to Arterial Pressure in Rats

Before arterial cannulation, inspect the pressure-sensing catheter under high magnification to confirm that the catheter is free of bubbles and debris and has an intact meniscus between the fluid and gel-filled components. Prior to each implantation, refill the gel at the distal tip of the catheter, and use a magnet to turn on the transmitter. Place a loose overhand half knot in the proximal suture to briefly occlude the femoral artery, and holding the catheter introducer in the non-dominant hand, use vessel cannulation forceps to grasp the tip of the cannula of the telemetry unit to avoid displacing the gel from the tip.
Use a bent 22-gauge needle tip to puncture a hole in the artery and insert the cannula into the artery as far as possible. Then, secure the pressure catheter with the proximal and distal suture ties, and tuck the body of the telemetry implant inside the flank adjacent to the incision. Gently move the rat into the prone position in a stereotaxic surgery frame with the head between the ear bars.
To isolate the renal sympathetic nerves, connect a wire Renal Sympathetic Nerve Activity or RSNA electrode to a 10X preamplifier in a microelectrode amplifier, and make a scalpel incision extending from 4 to 5 centimeters below the ribs in the caudal direction, slightly lateral to the spine.
Blunt-dissect to visualize the paraspinal muscles and make a very superficial 1 to 2-centimeter rostrocaudal incision where the fat meets the muscle. Using cotton-tipped applicators, spread the fat away from the muscle to visualize the kidney without entering the peritoneal space, and use retractors to gently separate the kidney from the paraspinal muscles to visualize the renal artery and abdominal aorta, and use high magnification to identify the renal nerves in the incision pocket.
The nerve bundles are most easily visible in the right angle formed by the aorta and the renal artery, closely following the renal artery from the aorta to the kidneys. Then, use micro-dissecting tweezers to gently dissect a segment of the nerve bundle to be placed on the recording electrode from the surrounding tissue. Secure the wire RSNA electrode in a holder and lower the electrode to the level of the nerve segment.
Use a nerve hook to gently lift the segment of the renal nerve onto the electrode without stretching the nerve and fill the incision with mineral oil to keep the exposed renal sympathetic nerve hydrated. Attach a grounding clip to the animal, in the other end to the Faraday cage, and use high and low pass filtering to direct the signal to the amplifiers. Then, adjust the gain up to 10 kilohertz, including an audio monitor, to assess the bursting pattern of the RSNA.
To assess the quality of the RSNA recording, evoke the baroreflex with a 100-microliter bolus intravenous injection of 10 micrograms per milliliter of phenylephrine to increase the blood pressure and to inhibit the renal sympathetic nerve activity, adjusting the position of the electrodes to improve the signal as necessary. Once a clear signal has been obtained, aspirate the mineral oil, and secure the electrode in place with silica gel.

Intracavernosal 压力记录对啮齿动物勃起功能的评价

摘要

探索更多视频

此视频中的章节

硬膜外颅内压测量中使用的光纤压力传感器的大鼠

宫内遥测来衡量鼠标收缩压力

清醒、无拘无束的小鼠静脉输液同时记录肾交感神经活动和血压的新方法。

大鼠海绵状神经刺激与海绵压力记录

大鼠同时心电图记录及侵入性血压测量

体内啮齿动物膨胀诱发尿路上皮ATP释放的管腔测量

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

Intracavernous Pressure Recording in a Cavernous Nerve Injury Rat Model

对大鼠进行海绵状神经刺激以测量海绵体内压力

记录大鼠响应动脉压的肾交感神经活动的变化

Intracavernosal 压力记录对啮齿动物勃起功能的评价

摘要

探索更多视频

此视频中的章节

硬膜外颅内压测量中使用的光纤压力传感器的大鼠

宫内遥测来衡量鼠标收缩压力

清醒、无拘无束的小鼠静脉输液同时记录肾交感神经活动和血压的新方法。

大鼠海绵状神经刺激与海绵压力记录

大鼠同时心电图记录及侵入性血压测量

体内 啮齿动物膨胀诱发尿路上皮ATP释放的管腔测量

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

Intracavernous Pressure Recording in a Cavernous Nerve Injury Rat Model

对大鼠进行海绵状神经刺激以测量海绵体内压力

记录大鼠响应动脉压的肾交感神经活动的变化

体内啮齿动物膨胀诱发尿路上皮ATP释放的管腔测量