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: 留一些额外的长度隧道和更容易处理。</li>
		<li>耀斑一个结束了一个打火机得到一个圆润的提示。检查导管的最后合适的开口和钝端。</li>
		<li>将一个2毫米长的硅胶管放在导管上方, 直到它位于喇叭的正下方。</li>
	</ol>
</li>
	<li>
电极的制造
	<ol>
<li>准备一个20-25 厘米的聚四氟乙烯涂层钢丝 (长短取决于动物的大小)。</li>
		<li>准备一个银线长度2厘米和扭曲的两端, 直到一个小循环保持。在一端剥去2毫米的聚四氟乙烯绝缘。焊接扭曲的银线结束的剥离钢丝。</li>
		<li>在涂布区应用常规指甲油。在涂敷区域上准备4毫米长的聚乙烯导管, 并通过将热量应用于压缩钳来密封末端。</li>
	</ol>
</li>
</ol>2. 动物制剂
<ol><li>
麻醉
	<ol>
<li>使用适当的麻醉鸡尾酒机构批准的全身麻醉。</li>
		<li>准备麻醉鸡尾酒与 medetomidine (0.15 毫克/千克), 咪唑安定 (0.08 毫克/千克), 和芬太尼 (0.01 毫克/千克), 并注射注1毫升注射器与27测量针。</li>
	</ol>
</li>
	<li>
手术准备
	<ol>
<li>用电动剃须刀刮伤腹部, 包括肩胛骨的生殖器区域和背部区域。</li>
		<li>先用70% 乙醇消毒腹部和颈部区域, 然后用聚维酮碘溶液3交替擦洗。</li>
	</ol>
</li>
</ol>3. 膀胱导管植入
<ol><li>在第三和第四奶嘴 (大约2到2.5 厘米长) 的水平上进行低线剖腹手术, 使用手术刀为皮肤和外科剪刀为腹部肌肉。</li>
	<li>通过在颅尾的方向引导腹壁并将其固定在膀胱的后面, 以避免重新定位, 从而使膀胱显露出来。</li>
	<li>使用6-0 不可吸收的单丝缝线和锥形针尖针在膀胱圆顶周围放置一个包线缝合。</li>
	<li>切开膀胱圆顶在钱包串内, 或者由手术刀尖或18G 针, 插入膀胱导管 (参见协议步骤 1)。</li>
	<li>插入导管预与无菌生理0.9% 氯化钠溶液, 并仔细收回膀胱导管, 直到扩口的导管是位于膀胱圆顶以下。</li>
	<li>在导管周围固定包线缝合, 并在导管体周围进行停止缝合, 以进行进一步的固位。</li>
	<li>通过导管缓慢填充膀胱, 用无菌生理0.9% 氯化钠溶液, 检查膀胱穹窿的渗漏。</li>
</ol>4. 尿道括约肌电极植入术
<ol><li>准备三电极植入 (见协议步骤 1)。</li>
	<li>标记一电极用色的永久毛毡笔为未来空电极的进一步证明。用70% 乙醇消毒电极。 注: 导管不适合于热化学灭菌程序。相反, 冷杀菌的电极24小时。</li>
	<li>用外科剪刀将腹部切口延伸至耻骨, 但不要切开耻骨联合。</li>
	<li>通过尿道两侧的细钳, 识别尿道并创建钝袋, 但要避免血管或神经的损伤。</li>
	<li>确定适当的脂肪囊靠近尿道在这个窗口。</li>
	<li>使用6-0 不可吸收的单丝缝合线, 通过双边方式将电极固定在适当的脂肪袋上。 注意: 电极的最后位置应该是双侧的在尿道的中部区域。</li>
	<li>将两个电极与6-0 不可吸收的单丝缝合在一起。</li>
	<li>将标记的空电极缝合到尿道的距离内的腹壁肌肉。将所有三电极与一条缝合线绑在一起。</li>
</ol>5. 隧道
<ol><li>在肩胛骨之间做一个小的皮肤切口为隧道。</li>
	<li>隧道电极线到颈部, 并做最后检查正确放置, 并验证在隧道后电极的位置。</li>
	<li>隧道膀胱导管到肩胛骨水平-注意膀胱, 而隧道导管, 以避免膀胱扭曲。为了达到这个目的, 在进入膀胱穹窿前按住导管, 以避免膀胱扭曲。</li>
	<li>用 4-0 polyfilament 可吸收缝合线连续或间断缝合, 关闭腹部肌肉。通过中断的床垫缝合关闭皮肤切口。</li>
	<li>将动物伸展至其全长, 使电极线和导管的最大膨胀。</li>
	<li>将导管和电极丝固定在肩部肌肉的缝合线上。</li>
	<li>用4-0 缝线缝合皮肤。</li>
</ol>6. 吊带配件
<ol><li>在动物头上拉上安全带, 将前肢拉到两个硅胶条之间的末端位置, 以适应动物。在与分销商进行手术之前, 检查一下动物的重量。</li>
	<li>隧道的膀胱导管的中心孔的线束和电极通过定制孔的钻头调整, 以电极线的大小。</li>
	<li>通过拉动硅胶条来调节安全带。调整安全带, 使其不太松散, 但确保一些空间仍然保持移动能力的大鼠。</li>
	<li>使用电缆带固定硅条。</li>
	<li>将膀胱导管的长度削减到3厘米以上的线束, 连接到23克塞子, 最后修复它的安全带。</li>
</ol>7. 电极连接器的制造
<ol><li>准备三小的热收缩管 (一个不同的颜色为零电极)。准备两个进一步的热收缩管在一个合适的较大的大小。</li>
	<li>缩短电极导线的长度到最佳长度, 以便能够在以后插入女性插头, 而不太松或太短。剥去三根电线 (约2毫米) 的聚四氟乙烯绝缘, 并将钢丝绳捻成一个字符串。</li>
	<li>将大型热收缩管置于所有电线上, 将小管单独放置在所有三根电线上, 并使用有色管作为空电极。</li>
	<li>焊锡电极到男性3连接插头。将空电极放在中间, 将三小的单个管收缩到焊接区域之上。</li>
	<li>收缩的第一个较大的管在年底的个别管尾和最后大管到男性插头边界。</li>
	<li>将雄性插头连接到固定在吊带上的母接头上, 并用一条胶带固定以进一步安全。</li>
</ol>8. 术后护理
<ol><li>清洁手术区, 用聚维酮碘消毒。</li>
	<li>将动物放在加热垫上直到清醒, 然后根据当地动物护理指南, 在手术期间和术后给0.9% 氯化钠溶液进行水替代。</li>
	<li>管理镇痛药 (昔1毫克/千克) 和抗生素 (sulfadoxinum 200 毫克, trimethoprimum 40 毫克, 15 毫克/千克) 作为联合注射溶液皮下注射。每天两次, 给镇痛药 (早晚), 每天一次给予抗生素, 随后五天。</li>
	<li>继续使用相同剂量的抗生素, 在整个随访期间, 每周注射两到三次。</li>
	<li>每天检查适当的吊带, 并进行外科实地检查, 特别是颈部区域。调整线束, 如果它变得太紧, 仔细地拉在硅胶条。</li>
	<li>每周定期冲洗导管, 避免阻塞。</li>
</ol>9. 压测量的准备
<ol><li>执行第一次压测量后六后手术天。 注: 早期的测量可以受镇痛药和/或尿囊刺激, 由于导管植入。</li>
	<li>打开主开关、计算机和肌电信号放大器。</li>
	<li>用室温加热0.9% 氯化钠填充注射器泵。以下行方式依次打开三路连接器, 从泵开始, 然后冲洗管子。 注: 仔细检查剩余气泡, 因为气泡会改变压的测量。</li>
</ol>10. 校准
<ol><li>启动程序 uroflowmeter 软件, 然后转到校准。</li>
	<li>关闭泵和动物的三路连接器。</li>
	<li>打开阀门到连接的压力表, 并按零在程序中。</li>
	<li>调整压力在压表到100柱, 并按100柱按钮的程序。按 "确认" 以保存校准调整。窗口将自动关闭。</li>
	<li>关闭阀门的压力表和打开三路连接器从泵到动物准备好测量。</li>
</ol>11. 动物数据库 (动物 DB)
<ol><li>要注册动物, 给动物一个简明的 ID。</li>
	<li>输入进一步的数据, 如 SCI 的日期, 开始治疗, 导管植入日期, 实验组, 和动物的出生年月。</li>
	<li>要最终记录数据, 请按 "存储记录" 并保存到文件。</li>
</ol>12. 记录前的测量设置
<ol><li>按 "开始" 按钮启动软件程序。</li>
	<li>选择动物和压重量秤和零压力。 注意: 在将动物放入抑制剂时要注意, 这样就不会有电缆卡住, 因为这可能会导致肌电信号线的移除。</li>
</ol>13. 动物制剂
<ol><li>拔下导管插头, 从吊带上拔下肌电信号插头。</li>
	<li>把老鼠放在抑制剂。关闭抑制剂, 用钳锁安全带。</li>
	<li>把导管和肌电信号插入抑制剂。</li>
	<li>将抑制剂放置在猫单元中。把尾巴穿过管子, 用胶带固定管子, 避免运动。</li>
	<li>将男性肌电信号插头连接到记录女性肌电信号连接器。</li>
	<li>再按零压力, 然后将导管连接到灌装/录音管。检查软件中的压力。 注: 压力应该是正数, 大约5-10 厘米 H2O 在基线。如果压力是负的, 断开导管从套管, 按零压力, 然后重新连接导管。</li>
</ol>14. 记录
<ol><li>如果导管和肌电信号连接, 按下运行泵和录音。</li>
	<li>使灌装速度适应μ l/分钟的实验需要。</li>
	<li>记下记录开始的时间和日志簿中的室温。</li>
	<li>要停止压记录, 请按下运行泵并再次按下记录停止泵。 注: 水泵将关闭, 泵的绿灯熄灭。</li>
	<li>断开导管并使用23G 塞子插头关闭导管末端。断开肌电信号电缆。</li>
	<li>打开抑制剂的前面, 引导老鼠走出抑制剂。 注意: 小心地处理老鼠, 并监视电线, 以避免电缆的任何锁定。</li>
	<li>将导管插回安全带。将肌电插头重新插到吊带上, 并在插头周围放置一条胶带。 注: 如果动物有病理性下尿路状况, 请在末端手动表达膀胱以避免膀胱 overdistention。</li>
	<li>把动物放回笼子里</li>
	<li>按退出键关闭程序。</li>
	<li>清洁抑制剂和烧杯。</li>
	<li>关闭泵和动物出口的三路连接器。</li>
	<li>通过主交换机关闭计算机、肌电信号单元和系统。 注意: 数据保存在单独的文件夹 "记录的数据" 与被测量的鼠 ID 的子文件夹文件。单个记录按日期在单个 rat ID 文件夹中排序。</li>
</ol>

<ol>
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作者没有什么可透露的。

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&amp;uuml;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

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

cystometric-external-urethral-sphincter-measurements-awake-rats-with

Universidad de Cartagena UDEC

Research

JoVE Journal

Biology

15.9K 次观看.  Paracelsus Medical University. 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。

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

Evaluation of Biomaterials for Bladder Augmentation using Cystometric Analyses in Various Rodent Models

Renal function and continence of urine are critically dependent on the proper function of the urinary bladder, which stores urine at low pressure and expels it with a precisely orchestrated contraction. A number of congenital and acquired urological anomalies including posterior urethral valves, benign prostatic hyperplasia, and neurogenic bladder secondary to spina bifida/spinal cord injury can result in pathologic tissue remodeling leading to impaired compliance and reduced capacity1. Functional or anatomical obstruction of the urinary tract is frequently associated with these conditions, and can lead to urinary incontinence and kidney damage from increased storage and voiding pressures2. Surgical implantation of gastrointestinal segments to expand organ capacity and reduce intravesical pressures represents the primary surgical treatment option for these disorders when medical management fails3. However, this approach is hampered by the limitation of available donor tissue, and is associated with significant complications including chronic urinary tract infection, metabolic perturbation, urinary stone formation, and secondary malignancy4,5.
Current research in bladder tissue engineering is heavily focused on identifying biomaterial configurations which can support regeneration of tissues at defect sites. Conventional 3-D scaffolds derived from natural and synthetic polymers such as small intestinal submucosa and poly-glycolic acid have shown some short-term success in supporting urothelial and smooth muscle regeneration as well as facilitating increased organ storage capacity in both animal models and in the clinic6,7. However, deficiencies in scaffold mechanical integrity and biocompatibility often result in deleterious fibrosis8, graft contracture9, and calcification10, thus increasing the risk of implant failure and need for secondary surgical procedures. In addition, restoration of normal voiding characteristics utilizing standard biomaterial constructs for augmentation cystoplasty has yet to be achieved, and therefore research and development of novel matrices which can fulfill this role is needed.
In order to successfully develop and evaluate optimal biomaterials for clinical bladder augmentation, efficacy research must first be performed in standardized animal models using detailed surgical methods and functional outcome assessments. We have previously reported the use of a bladder augmentation model in mice to determine the potential of silk fibroin-based scaffolds to mediate tissue regeneration and functional voiding characteristics.11,12 Cystometric analyses of this model have shown that variations in structural and mechanical implant properties can influence the resulting urodynamic features of the tissue engineered bladders11,12. Positive correlations between the degree of matrix-mediated tissue regeneration determined histologically and functional compliance and capacity evaluated by cystometry were demonstrated in this model11,12. These results therefore suggest that functional evaluations of biomaterial configurations in rodent bladder augmentation systems may be a useful format for assessing scaffold properties and establishing in vivo feasibility prior to large animal studies and clinical deployment. In the current study, we will present various surgical stages of bladder augmentation in both mice and rats using silk scaffolds and demonstrate techniques for awake and anesthetized cystometry.

Surgical stages of bladder augmentation are described using 3-D scaffolds in murine and rat models. To test the efficacy of biomaterial configurations for use in bladder augmentation, techniques for both awake and anesthetized cystometry are presented.

膀胱膀胱分析各种啮齿类动物模型中使用的增广的生物材料评价

Renal function and continence of urine are critically dependent on the proper function of the urinary bladder, which stores urine at low pressure and ...

科研

生物工程

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.

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.

的的膀胱测压小老鼠:A膀胱Chemosensation研究的使用

	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 ...

医学

Making Patch-pipettes and Sharp Electrodes with a Programmable Puller

Glass microelectrodes (also called pipettes) have been a workhorse of electrophysiology for decades. Today, such pipettes are made from glass capillaries using a programmable puller. Such instruments heat the capillary using either a metal filament or a laser and draw out the glass using gravity, a motor or both. Pipettes for patch-clamp recording are formed using only heat and gravity, while sharp electrodes for intracellular recording use a combination of heat, gravity, and a motor. The procedure used to make intracellular recording pipettes is similar to that used to make injection needles for a variety of applications, including cRNA injection into Xenopus oocytes. In general, capillary glass &lt;1.2 mm in diameter is used to make pipettes for patch clamp recording, while narrower glass is used for intracellular recording (outer diameter = 1.0 mm). For each tool, the puller is programmed slightly differently. This video shows how to make both kinds of recording pipettes using pre-established puller programs.

This video shows how to use a programmable puller to make patch pipettes and sharp electrodes for electrophysiology. The same procedure can be used to make a variety of glass tools, including injection needles.

膜片吸液管和夏普电极与一个可编程的拖轮

Glass microelectrodes (also called pipettes) have been a workhorse of electrophysiology for decades.  Today, such pipettes are made from glass capillaries ...

生物学

A Reversible, Non-invasive Method for Airway Resistance Measurements and Bronchoalveolar Lavage Fluid Sampling in Mice

Airway hyperreactivity (AHR) measurements and bronchoalveolar lavage (BAL) fluid sampling are essential to experimental asthma models, but repeated procedures to obtain such measurements in the same animal are generally not feasible.  Here, we demonstrate protocols for obtaining from mice repeated measurements of AHR and bronchoalveolar lavage fluid samples.  Mice were challenged intranasally seven times over 14 days with a potent allergen or sham treated.  Prior to the initial challenge, and within 24 hours following each intranasal challenge, the same animals were anesthetized, orally intubated and mechanically ventilated.  AHR, assessed by comparing dose response curves of respiratory system resistance (RRS) induced by increasing intravenous doses of acetylcholine (Ach) chloride between sham and allergen-challenged animals, were determined.  Afterwards, and via the same intubation, the left lung was lavaged so that differential enumeration of airway cells could be performed.  These studies reveal that repeated measurements of AHR and BAL fluid collection are possible from the same animals and that maximal airway hyperresponsiveness and airway eosinophilia are achieved within 7-10 days of initiating allergen challenge.  This novel technique significantly reduces the number of mice required for longitudinal experimentation and is applicable to diverse rodent species, disease models and airway physiology instruments.  

Repeated measurements of rodent respiratory physiology and sampling of airway inflammatory cells are desirable, but generally not feasible. Here we describe a repeatable method for orally intubating mice that permits repeated measurements of airway hyperreactivity and sampling of airway inflammatory cells.

一个可逆的，非侵入性方法，气道阻力测量和支气管肺泡灌洗小鼠流体取样

Airway hyperreactivity (AHR) measurements and bronchoalveolar lavage (BAL) fluid sampling are essential to experimental asthma models, but repeated ...

Evaluating the Procedure for Performing Awake Cystometry in a Mouse Model

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 laboratories. The goal of this study was to describe the microsurgical procedure used to implant an intravesical tube and the experimental technique for recording urinary bladder pressure in an awake, freely moving mouse. In addition, experimental data is presented to show how surgery, as well as tubing type and size, affect lower urinary tract function and recording sensitivity. The effect of tube diameter on pressure recording was assessed in both polyethylene and polyurethane tubing with different internal diameters. Subsequently, the best performing tube from both materials was surgically implanted into the dome of the urinary bladder of male C57BL/6 mice. Twelve-hour, overnight micturition frequency was recorded in healthy, intact animals and animals 2, 3, 5, and 7 days post-surgery. At harvest, bladders were assessed for signs of swelling using gross observation and were subsequently processed for pathological analysis. The greatest extent of bladder swelling was observed on day 2 and 3, which correlated with behavioral voiding data showing significantly impaired bladder function. By day 5, bladder histology and voiding frequency had normalized. Based on the literature and evidence provided by our studies, we propose the following steps for in vivo recording of intravesical pressure and voided volume in an awake mouse: 1) Perform the surgery using an operating microscope and microsurgical tools, 2) Use polyethylene-10 tubing to minimize movement artifacts, and 3) Perform cystometry on post-operative day 5, when bladder swelling resolves.

This study describes the surgical procedures and experimental techniques for performing awake cystometry in a freely moving mouse. In addition, it provides experimental evidence to support its optimization and standardization.

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 ...

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 ...

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. ...

Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents

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. Rodents are employed by many studies to research the physiology/pathology of erectile function. Erectile function in rodents can be evaluated by measuring the intracavernosal pressure (ICP). In practice, ICP can be monitored following electrical stimulation of the cavernous nerves (CNs). The arterial pressure of the carotid artery (the mean arterial pressure) is used as the reference for ICP. Using ICP recording protocols, many key parameters of erectile function can be measured from the ICP response curve. The ICP measurement provides more information than the apomorphine-induced penile erection test, and is cheaper than telemetric monitoring of the corpus spongiosum penis, making this method the most popular one to evaluate erectile function. However, compared to the easily-performed APO-induced erectile function test, successful ICP recordings require attention to detail, practice, and adherence to the operation method. In this work, an introduction to ICP recording in rats is provided to complement the procedure efficiently.

Intracavernosal pressure recording (ICP) is an important method to evaluate the erectile function of experimental animals. Here, a detailed protocol is demonstrated for the recording procedure of ICP by catheterizing the crura penis and then electrically stimulating the cavernous nerves in rats.

Intracavernosal 压力记录对啮齿动物勃起功能的评价

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. ...

使用氰基丙烯酸酯皮肤屏障对清醒小鼠进行精确尿动力学测量

Optimizing Mouse Urodynamic Techniques for Improved Accuracy

Accurate measurement of urinary parameters in awake mice is crucial for understanding lower urinary tract (LUT) dysfunction, particularly in conditions like neurogenic bladder post-traumatic spinal cord injury (SCI). However, conducting cystometry recordings in mice presents notable challenges. When mice are in a prone and restricted position during recording sessions, urine tends to be absorbed by the fur and skin, leading to an underestimation of voided volume (VV). The goal of this study was to enhance the accuracy of cystometry and external urethral sphincter electromyography (EUS-EMG) recordings in awake mice. We developed a unique method utilizing cyanoacrylate adhesive to create a waterproof skin barrier around the urethral meatus and abdomen, preventing urine absorption and ensuring precise measurements. Results show that after applying the cyanoacrylate, the sum of VV and RV remained consistent with the infused saline volume, and there were no wet areas observed post-experiment, indicating successful prevention of urine absorption. Additionally, the method simultaneously stabilized the electrodes connected with the external urethral sphincter (EUS), ensured stable electromyography (EMG) signals, and minimized artifacts caused by the movement of the awakened mouse and manipulation of the experimenter. Methodological details, results, and implications are discussed, highlighting the importance of improving urodynamic techniques in preclinical research.

作者聚焦:用于神经源性膀胱清醒小鼠精确尿液测量的增强型尿动力学方法

This protocol provides a guide for waterproofing the skin with cyanoacrylate to prevent urine absorption by fur and skin. It includes instructions for applying the glue to the skin, implanting a bladder catheter, and electrodes for cystometry and external urethral sphincter electromyography recordings in awake mice.

优化小鼠尿动力学技术以提高准确性 

Accurate measurement of urinary parameters in awake mice is crucial for understanding lower urinary tract (LUT) dysfunction, particularly in conditions ...

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

摘要

探索更多视频

此视频中的章节

膀胱膀胱分析各种啮齿类动物模型中使用的增广的生物材料评价

的的膀胱测压小老鼠:A膀胱Chemosensation研究的使用

膜片吸液管和夏普电极与一个可编程的拖轮

一个可逆的，非侵入性方法，气道阻力测量和支气管肺泡灌洗小鼠流体取样

评估在鼠标模型中执行唤醒膀胱测压的过程

大鼠海绵状神经刺激与海绵压力记录

体内啮齿动物膨胀诱发尿路上皮ATP释放的管腔测量

Intracavernosal 压力记录对啮齿动物勃起功能的评价

优化小鼠尿动力学技术以提高准确性

优化小鼠尿动力学技术以提高准确性

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

摘要

探索更多视频

此视频中的章节

膀胱膀胱分析各种啮齿类动物模型中使用的增广的生物材料评价

的的膀胱测压小老鼠:A膀胱Chemosensation研究的使用

膜片吸液管和夏普电极与一个可编程的拖轮

一个可逆的，非侵入性方法，气道阻力测量和支气管肺泡灌洗小鼠流体取样

评估在鼠标模型中执行唤醒膀胱测压的过程

大鼠海绵状神经刺激与海绵压力记录

体内 啮齿动物膨胀诱发尿路上皮ATP释放的管腔测量

Intracavernosal 压力记录对啮齿动物勃起功能的评价

优化小鼠尿动力学技术以提高准确性

优化小鼠尿动力学技术以提高准确性

体内啮齿动物膨胀诱发尿路上皮ATP释放的管腔测量