Name: preparation of peripheral nerve stimulation electrodes for chronic implantation in rats
Uploaded: 2020-07-14T13:00:00
Description: Watch this Scientific Journal Video about Preparation of Peripheral Nerve Stimulation Electrodes for Chronic Implantation in Rats at JoVE.com

这项工作由德克萨斯大学达拉斯分校和UT摄政委员会资助。我们感谢所罗门·戈尔丁、比拉尔·哈桑、玛吉·贾尼和清慈宗的技术援助。

本议定书中描述的所有程序都按照《美国国家卫生和植物保护与使用指南》执行，并经达拉斯德克萨斯大学机构动物护理和使用委员会批准。
1. 刺激袖口电极制造
<ol><li>准备袖口管。<ol>
<li>使用剃须刀刀片，切割一块 2.5 毫米长的聚合物管。插入钳子尖或回形针通过管子，并使用刀片使一个狭缝通过管壁的一侧袖口。</li>
		<li>从管子上取下钳子，在袖口的中线插入一根大缝?...

<ol>
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R., 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=Vagus+nerve+stimulation+reverses+the+extinction+impairments+in+a+model+of+PTSD+with+prolonged+and+repeated+trauma.">Vagus nerve stimulation reverses the extinction impairments in a model of PTSD with prolonged and repeated trauma.</a> Stress. , (2019).</li><li>Noble, L. J., Souza, R. R., McIntyre, C. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vagus+nerve+stimulation+as+a+tool+for+enhancing+extinction+in+exposure-based+therapies.">Vagus nerve stimulation as a tool for enhancing extinction in exposure-based therapies.</a> Psychopharmacology. , (2019).</li><li>Childs, J. E., Kim, S., Driskill, C. 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在这里，我们描述了一种简单、低成本的方法，用于组装用于啮齿动物的慢性植入刺激袖口电极，从而促进对这种新兴疗法的临床前研究。这种简化的方法不需要专门的培训或设备，并且使用少量的工具和用品，大多数研究实验室都,很容易获得，与其他方法16、26、27、28相比26,27，降低了设备制造的货币?...

最近，对慢性植入袖口电极刺激周围神经的需求越来越大，因为研究越来越证明这种技术对治疗许多炎症,性疾病,,,,,,1、2、3,2和神经系统疾病44、5、6、7、8、9、10、11、12、13、14、15,14,,11,12的临床7,8105前有用。3 6913例如，慢性VNS已被证明可在各种学习环境中增强新皮质可塑性，改善运动康复,54，5，6，7，8，灭绝学习,6,7,810，11，12，13，14，,11,12,13,14和感官歧视15。10市售的外设神经袖口电极通常与订单履行时间延长和成本相对较高有关，从而限制其可访问性。另外，长期植入袖口电极的"内部"制造协议仍然有限，啮齿动物解剖由于其体积小而面临特殊挑战。目前为慢性啮齿动物实验构建袖口电极的协议通常需要使用复杂的设备和技术，以及经过广泛培训的人员。在该协议中，我们演示了一种基于以前发布和广泛使用的方法16、17,的袖口电极制造的简化方法。我们验证了我们长期植入大鼠电极的功能，证明在左颈椎迷走神经周围植入袖口时，对袖口电极的刺激成功地使SpO2的呼吸和下降停止。刺激的肺受体血管纤维是众所周知的从事赫林-布劳尔反射，其中抑制几个呼吸核在脑干的结果抑制灵感18。因此，停止呼吸与赫林-布劳尔反射一致，以及由此导致的 SpO2 下降，为麻醉大鼠正确植入电极和袖口功能提供了直接的测试。为了验证慢性植入袖口电极的长期功能，在植入时测量反射反应，并与植入后六周内在同一动物获得的反应进行比较。第二组大鼠在操纵杠杆任务的行为训练后植入了VNS袖口电极。在这些大鼠中，VNS与正确的任务性能配对，产生了皮质运动图的重组，与之前发表的,研究19、20、21、22,2021,一致。在麻醉下发生5~10周麻醉运动皮质映射时，通过确认VNS成功诱导呼吸停止和SpO2下降5%以上，进一步验证了VNS治疗动物的袖口功能。
最近出版的儿童类17和里奥斯等人的协议为简化袖口电极制造方法提供了一个经过验证的起点，因为这种流行的方法已被多个实验室使用,，在啮齿动物,,,,,,11、2、3、4、5、6、7、8、9、10、11中进行慢性,27,8610,VNS研究。349原始方法涉及几个高精度步骤来操纵细微线，使袖口电极制造需要一个多小时才能完成，并进行了广泛的培训才能可靠地执行。此处描述的简化方法所需的材料和工具要少得多，只需经过最少培训的人员，只需在一小时内完成。

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			<td height="21" style="height:21px;width:441px;">Biocompatible polyurethane-based polymer tubing, 0.080&#34; OD x 0.040&#34; ID</td>
			<td style="width:178px;">Braintree Scientific</td>
			<td style="width:174px;">MRE080 36 FT</td>
			<td style="width:542px;"></td>
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			<td height="21" style="height:21px;">Dissecting microscope</td>
			<td style="width:178px;">AM Scopes</td>
			<td style="width:174px;">#SM-6T-FRL</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fine Serrated Scissors, straight, 22mm cutting edge</td>
			<td style="width:178px;">Fine Science Tools</td>
			<td style="width:174px;">#14058-09</td>
			<td>for cutting Pt/Ir wire and suture thread</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Forceps, #5 Dumont forceps, straight, 11 cm, 0.1 x 0.06 mm tip</td>
			<td style="width:178px;">Fine Science Tools</td>
			<td style="width:174px;">#11626-11</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Forceps, ceramic tipped forceps, 0.3 mm x 30 mm tips</td>
			<td style="width:178px;">Electron Microscopy Sciences</td>
			<td style="width:174px;">#78127-71</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Gold Pins, PCB Press Fit Socket</td>
			<td style="width:178px;">Mill-Max</td>
			<td style="width:174px;">#1001-0-15-15-30-27-04-0</td>
			<td>or similar small pins for connecting cuff leads to headcap</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Isobutane lighter</td>
			<td style="width:178px;">BIC</td>
			<td style="width:174px;">#LCP21-AST</td>
			<td>for de-insulating Pt/Ir wire</td>
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		<tr height="42">
			<td height="42" style="height:42px;">Micro strip connector with latch, 4-pin</td>
			<td style="width:178px;">Omnetics</td>
			<td style="width:174px;">A24002-004 / PS1-04-SS-LT</td>
			<td></td>
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			<td height="21" style="height:21px;">Pipette tip, 10 uL</td>
			<td style="width:178px;">VWR</td>
			<td>89079-464</td>
			<td></td>
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			<td height="21" style="height:21px;">Platinum-Iridium (90/10%) Wire, 0.001&#34; (diameter) x 9 strands, PTFE insulated</td>
			<td style="width:178px;">Sigmund Cohn</td>
			<td style="width:174px;">10IR9/49T</td>
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			<td height="21" style="height:21px;">Razor Blade, Single Edge, Surgical Carbon Steel No.9</td>
			<td style="width:178px;">VWR</td>
			<td style="width:174px;">#55411-050</td>
			<td>for cutting MicroRenathane tubing</td>
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			<td height="21" style="height:21px;">Sewing needle, ca. 4.0 cm length x 0.7 mm diameter (size 6-7)</td>
			<td style="width:178px;">Singer</td>
			<td style="width:174px;">00276</td>
			<td>Smaller needle for threading Pt/Ir wire</td>
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			<td height="21" style="height:21px;">Sewing needle, ca. 4.5 cm length x 0.8 mm diameter (size 2-3)</td>
			<td style="width:178px;">Singer</td>
			<td style="width:174px;">00276</td>
			<td>Larger needle for pinning cuff during assembly and for threading suture</td>
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			<td height="21" style="height:21px;">Small foam board</td>
			<td style="width:178px;">Juvo+/Amazon</td>
			<td>B07C9637SJ</td>
			<td>for fabrication platform; our dimensions are ca. 2.5&#34; x 3.5&#34; x 1&#34; (L x W x H)</td>
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			<td height="21" style="height:21px;">Solder, multicore lead-free, 0.38mm diameter</td>
			<td style="width:178px;">Loctite/Multicore</td>
			<td style="width:174px;">#796037</td>
			<td></td>
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			<td height="21" style="height:21px;">Soldering station</td>
			<td style="width:178px;">Weller</td>
			<td style="width:174px;">WES51</td>
			<td>or similar soldering iron compatible with long conical tips (this part has been discontinued)</td>
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			<td height="21" style="height:21px;">Soldering tip, long conical, 0.01&#34; / 0.4 mm</td>
			<td style="width:178px;">Weller</td>
			<td style="width:174px;">1UNF8</td>
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			<td height="21" style="height:21px;">Suture, nonabsorbable braided silk ,size 6/0</td>
			<td style="width:178px;">Fine Science tools</td>
			<td style="width:174px;">#18020-60</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">UV (405 nm) spot light</td>
			<td style="width:178px;">Henkel/Loctite</td>
			<td style="width:174px;">#2182207</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">UV Light Cure Adhesive 25 ml</td>
			<td style="width:178px;">Henkel/Loctite</td>
			<td style="width:174px;">AA 3106</td>
			<td>or similar biocompatible UV cure adhesive</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Wire wrapping wire, 30 AWG</td>
			<td style="width:178px;">Digikey</td>
			<td style="width:174px;">K396-ND</td>
			<td></td>
		</tr>
	</tbody>
</table>

preparation of peripheral nerve stimulation electrodes for chronic implantation in rats

外周神经袖口电极早已用于神经科学和相关领域，以刺激，例如，迷走神经或坐骨神经。最近的几项研究表明，慢性VNS在提高中枢神经系统可塑性以改善运动康复、灭绝学习和感官歧视方面的有效性。由于大鼠体型小，长期植入装置的构建具有挑战性，典型的方案需要人员的广泛培训和耗时的微制造方法。或者，可以以更高的成本购买市售的植入式袖口电极。在该协议中，我们提出了一种简单、低成本的方法，用于构建用于大鼠的小型、慢性植入的外周神经袖口电极。我们验证了袖口电极的短期和长期可靠性，证明氯胺酮/xylazine麻醉大鼠的 VNS 在植入时和装置植入后 10 周内都会产生与激活 Hering-Breer 反射一致的呼吸速率降低。通过将 VNS 与熟练的杠杆压机性能配对以诱导电机皮质贴图可塑性，我们进一步演示了袖口电极在慢性刺激研究中的适用性。

根据先前公布的外科手术17、19、20、21、22，迷走神经,19袖口,电极和21头盖被长期植入大鼠体内。在植入之前，在袖口引线上测量 1 kHz 的阻抗，袖口管浸入盐水中（阻抗 = 1.2 = 0.17 kΩ [均值 = std];N = 9）。在盐水中只植入阻抗小于2 kΩ的袖口;所有袖口都符合此标准（不包括 0/9 ?...

Watch this Scientific Journal Video about Preparation of Peripheral Nerve Stimulation Electrodes for Chronic Implantation in Rats at JoVE.com

Preparation of Peripheral Nerve Stimulation Electrodes for Chronic Implantation in Rats

现有的用于建造用于小型啮齿动物的可长期植入的外周神经袖口电极的方法通常需要专门的设备和/或训练有素的人员。在该协议中，我们演示了一种简单、低成本的制造慢性植入袖口电极的方法，并演示了它们对大鼠迷走神经刺激（VNS）的有效性。

大鼠慢性植入的外周神经刺激电极的制备

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. ...

外周神经刺激作为治疗各种疾病（包括炎症性疾病和神经紊乱）的研究日益受到关注。为小型实验室动物建造慢性植入外周神经袖口电极既耗时又昂贵。该协议描述了一种简单、低成本的方法，用于构建用于大鼠的慢性植入外周神经刺激电极。与类似的技术相比，该方法需要很少且易于获取的材料和用品，而且需要减少危险或技术要求的步骤数量，既降低了设备成本，又降低了培训人员和设备装配所需的时间。这项技术旨在使临床前研究，测试慢性拉斯维加斯神经刺激在中风恢复或其他类型的地铁功能障碍的有效性。首先准备袖口管。使用剃刀刀片切割 2.5 毫米的聚合物管。然后插入回形针或力插头穿过管子，然后通过袖口一侧的管子壁纵向滑动。从管子上取下钳子，在袖口的中线插入一根大缝纫针。将针头放入泡沫板中，在其余装配步骤中将袖口固定到位。在植入过程中，要放置用于固定袖口的缝合线，请插入一根小缝纫针穿过袖口的墙，从一侧顶部狭缝处约五毫米处穿过袖口壁。然后通过针头的眼睛插入两厘米的六零缝合线，将针头拉过管子的墙壁，将缝合线插入袖口。沿着袖口中线穿过管壁，大约在第一个孔下方的五毫米处刺穿第二个孔。缝合线穿过针头的眼睛，将针头拉过管壁。将少量紫外线固化胶粘剂涂抹到从下孔延伸的缝合线短端，并拉长缝合线端，直到下尾与管材的外墙几乎齐平。然后用紫外线棒固化胶粘剂。重复此过程，将另一个缝合线放在袖口的另一侧。下一个位置铂金虹线引线。使用小缝纫针在袖口壁上打四个洞。然后再次插入缝纫针，这一次通过第一个铅孔从外部到内部工作。将大约五厘米的 7.5 厘米铂金线插入针头的眼睛，并将针头拉过管子，将导线穿过袖口墙。调整导线，使袖口外侧延伸约 4.5 厘米。再次插入针头穿过第一个引线孔，然后通过第二个引线孔。将导线短端的五厘米点插入针头，将针头拉过管子，将导线穿过袖口壁。重复此过程，将导线穿过第三个和第四个引线孔。使用丁烷打火机，小心地从五至六毫米的导线中去除绝缘层，从第二个和第四个引线孔延伸，然后轻轻地拉扯从第一个孔延伸的导线的端端，直到导线的未绝缘部分与孔齐平。与其他引线重复此操作，以将螺纹线未绝缘端通过第三和第四个引线孔对齐。将少量紫外线固化胶在袖口外侧的第一和第三个引线孔的导线环上涂抹。然后使用紫外线棒固化粘合剂，并固定引线到位。使用小移液器尖端将未绝缘的导线导线推到袖口的内壁上。将电线的一毫米尾巴平放在袖口的外表面上，注意不要把它们短路在一起。然后应用少量的紫外线粘合剂，只覆盖两条尾巴并固化它。从泡沫板上取下带袖口组件的大针。三厘米长的六零缝合线穿过针头的眼睛，将针头拉过管子，将缝合线穿过中点袖口的底部。插入小缝纫针通过同一中线孔工作从内部到外部，以避免变形，管道和电线引线。然后通过针头的眼睛插入缝合线的外表尾，将针头拉过袖口壁，在袖口边缘形成缝合环。通过将缝合线末端和袖口外侧的半结绑在一起，在袖口的另一端周围创建第二个环。将结平放在管子上时，将少量紫外线粘合剂涂抹在半结上并固化。小心地切割缝合线的末端，尽可能靠近结。然后，使用丁烷打火机从每个铂金钛线引线端的大约三毫米中去除绝缘层。将金针的杯面焊接到每个引线未内化的端。在进行植入测试之前，先使用组装装置的阻抗，以验证组装的袖口在一千赫下是否阻抗小于两千兆欧姆。组装头盖和根据手稿方向植入袖口电极后，在大鼠清醒时刺激迷走神经。通过植入的头盖将大鼠连接到刺激发生器并调整刺激设置。对于电机皮质图的 VNS 诱导重组，提供一列 15 个双相脉冲，每个脉冲的宽度为 100 微秒，振幅为 800 微安培，频率为 30 赫兹。在 10 到 30 分钟的培训课程中，每次成功的操纵压机检测成功后，将立即提供刺激列车。使用示波器监测电流刺激的成功传递。该协议，用于长期植入迷走神经袖口电极和头盖在大鼠。在植入过程中，通过测试VNS诱导的血氧饱和度下降，对所有袖口进行功能验证。为了唤起这种反应，10秒的30赫兹脉冲，每个脉冲的宽度为100微秒，振幅为800微安培，通过袖口引线。对于所有袖口，我们观察到一个VNS诱导停止呼吸的10秒刺激，伴随着血氧饱和度下降至少5%确认袖口功能和适当的植入。为了测试这些装置的长期功能，大鼠在植入后六周被麻醉，VNS再次应用。对于九分之七的设备来说，刺激的幅度引起血氧饱和度的改变，与最初植入时观察到的没有区别，这表明持续出色的性能。其余两个装置，将刺激幅度提高至1.6毫安，足以使血氧饱和度可靠降低至少5%，表明这些装置继续发挥作用。但是阻抗、神经损伤或袖口方向的变化可能导致性能降低。为了进一步测试慢性植入装置的长期功能，第二组大鼠接受了技术熟练的伸手操纵杆按压任务的简化版本培训。老鼠被要求在训练室外达到两厘米，以完全按下操纵杆，然后在两秒钟内释放它。与先前的研究表明，迷走神经刺激驱动任务相关运动图表示的扩展一致，VNS治疗的大鼠表现出比Sham治疗大鼠大得多的近位前肢表示。该协议可以很容易地适应，以适应慢性刺激实验在其他实验室物种或其他周围神经位点，包括坐骨神经或囊神经。例如。

Research

JoVE Journal

Neuroscience

Surgical Implantation of Chronic Neural Electrodes for Recording Single Unit Activity and Electrocorticographic Signals

用于记录单机活动和Electrocorticographic信号的慢性神经电极植入手术

The success of long-term electrophysiological recordings often depends on the quality of the implantation surgery.  Here we provide useful information for ...

科研

神经科学

Fiber-optic Implantation for Chronic Optogenetic Stimulation of Brain Tissue

光纤植入慢性Optogenetic刺激脑组织

Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard ...

Transplantation of Olfactory Ensheathing Cells to Evaluate Functional Recovery after Peripheral Nerve Injury

嗅鞘细胞移植周围神经损伤后，以评估功能恢复

Olfactory ensheathing cells (OECs) are neural crest cells which allow growth and regrowth of the primary olfactory neurons. Indeed, the primary olfactory ...

Microelectrode Guided Implantation of Electrodes into the Subthalamic Nucleus of Rats for Long-term Deep Brain Stimulation

微电极引导电极植入到大鼠丘脑底核的长期深部脑刺激

Deep brain stimulation (DBS) is a widely used and effective therapy for several neurologic disorders, such as idiopathic Parkinson&#8217;s disease, ...

Fabrication of High Contact-Density, Flat-Interface Nerve Electrodes for Recording and Stimulation Applications

高接触密度的制作，平面接口神经电极用于记录和刺激的应用

Many attempts have been made to manufacture multi-contact nerve cuff electrodes that are safe, robust and reliable for long term neuroprosthetic ...

Chronic Transcranial Electrical Stimulation and Intracortical Recording in Rats

大鼠慢性经颅电刺激及 Intracortical 记录

Transcranial electrical stimulation (TES) is a powerful and relatively simple approach to diffusely influence brain activity either randomly or in a ...

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

常见马莫塞的全皮质电皮质阵列的慢性植入

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...

Chronic Implantation of Multiple Flexible Polymer Electrode Arrays

多柔性聚合物电极阵列的慢性植入

Simultaneous recordings from large populations of individual neurons across distributed brain regions over months to years will enable new avenues of ...

Implantation and Control of Wireless, Battery-free Systems for Peripheral Nerve Interfacing

用于周围神经接口的无线、无电池系统的植入和控制

Peripheral nerve interfaces are frequently used in experimental neuroscience and regenerative medicine for a wide variety of applications. Such interfaces ...

联合周围神经刺激和可控脉搏参数 TMS

Combined Peripheral Nerve Stimulation and Controllable Pulse Parameter Transcranial Magnetic Stimulation to Probe Sensorimotor Control and Learning

作者聚焦： 结合周围神经刺激和可控脉搏参数经颅磁刺激以探测感觉运动控制和学习  

结合周围神经刺激和可控脉搏参数经颅磁刺激探测感觉运动控制和学习

Skilled motor ability depends on efficiently integrating sensory afference into the appropriate motor commands. Afferent inhibition provides a valuable ...