这项工作是由该中心法国国家科学研究（合同UPR3212），斯特拉斯堡大学和由NARSAD青年研究者格兰特从脑与行为研究基金会（以IY）的支持。出版费用由Neurex网络（计划区域间四莱茵河上游）的支持。

协议已被批准的“科米特德ETHIQUE恩MATIERE德实验ANIMALE斯特拉斯堡”（CREMEAS）。 1。基线缩足阈值的测定<ol><li>允许小鼠启动测试程序之前习惯于给动物设施至少10天至2周。 </li><li>习惯于小鼠到的von Frey试验的设置和于在第4部分所述的冯弗雷过程。 </li><li>手术前，评估如4.3节所述的机械缩足阈值与冯弗雷细丝。注：直到至少三个稳定连续的值都为缩足阈值获得的重复在不同的日子的过程。 </li><li>分配小鼠到不同的实验组，使这些基团不最初有所不同缩爪阈值。 </li></ol> 2，手术过程的袖口Implantati上<ol><li>称量动物。注意：小鼠体重应超过20克以下说明的压脉袋的插入过程。 </li><li>麻醉动物通过腹膜内注射4毫升/千克氯胺酮（17毫克/毫升）和甲苯噻嗪在0.9％NaCl中，它提供了大约45分钟的麻醉（2.5毫克/毫升）的混合物。 </li><li>检查没有爪子反射的按捏一个后掌用的镊子，并检查没有眼睛反射的，以确保动物是完全麻醉。 </li><li>从膝盖刮右腿使用电动剃须刀的臀部。 </li><li>涂上保护眼部凝胶液对眼睛有棉签。 </li><li>将动物在它的左侧，然后将右后肢上一个小枕头，并保持右后肢用胶带贴在枕头上。 </li><li>用纱布或棉签用洗必泰和70％乙醇消毒外科领域。 </li><li>使用食指发现股骨并进行大约0.5厘米的切口，平行于股骨和大约1.5mm前到股骨。 </li><li>分开接近股骨两个蒸压棒的肌肉。注意事项：切勿切断肌肉。通常，肌肉层容易分离，没有任何出血和坐骨神经然后可见。如有出血，用无菌棉签吸收血液。 </li><li>将下面的坐骨神经2支高压灭菌揭露其主要分支，滋润神经，用无菌的生理溶液（0.9％氯化钠）。 </li><li>持有分裂PE-20聚乙烯管（袖）0.38 mm内径/1.09毫米外径，用尖棒钢的帮助和斗牛犬钳事先准备好的无菌2毫米部分。 <ol><li>将尖钢粘成的袖口，这将略微打开它。 </li><li>用袖带侧开口，插入犬在袖带的一端，并且平行于所述压脉袋。旋转斗牛犬（180°），使其保持袖口的的相反侧的侧面开口。关闭斗牛犬，取出钢尖棒。注：旋转做是为了让持有，在袖口中的插入的最佳位置，牛头犬钳也有助于保持袖口部分开放。该模型和犬夹具的大小是该过程的这一步骤的关键。 </li></ol></li><li>具有第二实验者将两个支神经下，轻轻分开枝，以方便访问到坐骨神经是大约4毫米长的一段。 </li><li>插入周围的坐骨神经的主要分支的2毫米袖带，开始通过插入袖带即远端周围的神经，即最接近于髋关节的一部分的犬的一部分。 </li><li>轻轻合上袖带通过施加其上，用钳子2前端侧的压力，不挤或改变管的形式。把袖口，以确保其被正确关闭。 </li><li>缝合剃光皮肤层的智慧ħ外科结。 </li><li>把鼠标在它的左侧一个干净的家笼。保持下的加热灯，直到鼠标是清醒的。 </li><li>添加额外的水，直接将一些州城在家里笼。 </li></ol> 3，手术过程深水控制<ol><li>应用上述步骤2.1步骤2.9，然后按照与步骤2.15〜2.17相同的手术过程。对于假手术对照，省略步骤2.10至2.14，只有关注的袖口插入。 </li></ol> 4。冯弗雷测试<ol><li>有孔将小鼠清除个别箱（7厘米×9厘米×7厘米），在光滑的不锈钢（1米×50 cm时，5毫米圆孔穿孔边框之间2.5毫米）升高的穿孔板。注：最多12只小鼠可以在此设置了可以相伴测试。手术动物可以在手术后的第二天进行测试。然而，恢复3天，建议削弱手术后超敏反应观察假手术对照。 </li><li>让动物习惯于15分钟测试之前。 </li><li>应用的von Frey细丝在一系列递增的势力每个后爪的足底表面。注：冯弗雷细丝塑料毛标定直径。它们是5厘米长，并固定于手持式涂胶器。的长丝应用的速度，弯曲和应用程序的持续时间的程度可以影响与此试验3中获得的阈值。与小鼠本程序中，被最经常使用的长丝的0.16，0.4，0.6，1，1.4，2，4，6，8，和10克。 <ol><li>应用所选择的长丝向左爪的足底面，直至灯丝只是弯曲。重复该步骤四点五十七连续次，然后做同样的右爪。一旦灯丝已经过测试，在两个爪子，测试下的动物。注意事项：避免它可以是更敏感的爪子横向边界。预期的响应是一个爪子停药，突然退缩或舔爪子。考虑到反应为阳性，如果至少有三个期望的响应被观察出五审判。一个给定的爪子总是测试三次，但第四和第五的试验完成后只有1或2的返回结果是（是）在头三个试验观察。在C57BL/6J小鼠，开始手术前的测试与1.4克长丝。手术后，开始与0.4克灯丝的测试。如果一个肯定的响应与所述第一测试灯丝观察到，在步骤4.3.2测试下的力（而不是更大）的长丝。 </li><li>按照4.3.1程序应用相同的灯丝到下一个动物。一旦所有的动物进行测试，在用更大的力量下一个细丝的第一个动物重新开始。重复上述步骤，直到所有小鼠给予积极的回应。注：测试每个动物，直到连续两个灯丝给予积极的回应。考虑到较低的灯丝，给了一个肯定的答复为爪子机智克价值hdrawal门槛，这种动物。 </li></ol></li></ol>

作者宣称，他们有没有竞争的财务权益。

在“袖口”模式最初是在大鼠获得标准化和可再现性损伤与周围的坐骨神经6多个袖口的植入。它然后修改，以植入单袖7,8，即使有些研究小组仍然使用多个袖口插入19-22。这在当时适应小鼠9,23，其中开通使用转基因动物的可能性。袖带通常为2毫米长，但其它长度也已在大鼠22使用。聚乙烯管材取决于物种：PE-20在小鼠9，和在大鼠的PE-60 24,25或PE-90 7,8,26,27。 机械异常性疼痛是测量的von Frey毛。在该试验中，为缩爪阈值的绝对值可以取决于该动物代表28或后丝弯曲3的持续时间的表面上，但是这些因素不影响神经性异常性疼痛的检测。 在“袖口”模型是对神经病理性疼痛机制的研究兴趣。它被用来研究髓鞘和无髓鞘纤维6,29，和感觉神经元，初级传入和脊髓神经元19,21,22,30-35功能变化的形态变化。它使示范胶质激活和神经阴离子梯度转移中心参与变化的活动，并在脊髓痛觉神经元的反应和神经性异常性疼痛24,36-38。谷氨酸受体烟碱受体46和阿片受体16,42-45的7,39-41的影响下，也研究了这个模型。 该模型的另一利益是其对神经性疼痛， 即得 。，gabapentinoids和抗抑郁药目前的治疗。类似的临床观察：gabapentinoids同时显示一个Ñ​​急性短暂的镇痛作用在高剂量和被处理后，三环类抗抑郁药和选择性血清素和去甲肾上腺素再摄取的几天抑制剂具有在相关的剂量没有急性止痛效果观察到，但它们显示延迟的持续缓解延迟的持续缓解动作动作，需要1〜2周的治疗，并选择性血清素再摄取抑制剂氟西汀是无效16。该模型是如此适合研究的分子机制这些治疗相关16-18,44,45,47，这可能揭示新的治疗靶点，以测试患者48-51。 最后，该模型还允许的神经性疼痛的anxiodepressive后果的研究。在临床上，这些后果周围神经病理性疼痛患者的三分之一影响但临床前研究较少比疼痛的感觉方面。在这个模型中，焦虑样和抑郁的时间依赖性发展西伯样表型存在52和相关机制，因此可以解决。 标准化的袖口和程序，在神经性疼痛该小鼠模型导致低的个体差异为机械性异常疼痛。以使用转基因动物17,18,44-47,52中，长效性异常性疼痛的可能性，则响应于用于临床治疗和anxiodepressive症状随时间变化的发展使这一模型适用于各个方面的研究，并神经病理性疼痛及其治疗后果，这已经带来了宝贵的资料，这一领域的研究。

神经性疼痛通常是慢性的，并产生作为病变或疾病影响的体感系统的结果。中枢神经系统内的受伤的感觉神经元和沿整个痛觉通路适应不良的变化参与这一复杂的综合征。各种模型已经开发了啮齿动物用于研究神经性疼痛及其治疗1-3。 基于已知的病因，神经性疼痛的目标模型在模仿在糖尿病中观察到的多神经病，在受伤外周神经，中央受伤，三叉神经痛，连续的化疗中，带状疱疹后神经痛等的神经病变的不同型号外周神经损伤的啮齿动物集中在坐骨神经。这些模型依赖于压缩和/或该神经的截面。的确，坐骨神经能提供相对容易的外科手术和允许基于缩爪反射的测试。莫慢性神经压迫德尔斯包括例如：慢性缩窄性损伤（CCI）4,5，坐骨神经掴，如图6-9所示，坐骨神经部分结扎（PSL）10，脊神经结扎（SNL）11，或共同腓总神经结扎12。被称为“幸免神经损伤”（SNI）模型也被广泛使用。它们由一个紧结扎和两个出坐骨神经的三个末端分支的轴突切断的，而第三分支保持不变13-15。各种型号的神经病理性疼痛，其中针对坐骨神经，导致慢性机械性异常性疼痛（一个正常的非伤害性刺激伤害性反应）在受伤后爪。 在这里，详细的手术过程为“袖口模式”小鼠神经病理性疼痛的描述。它由在围绕坐 ​​骨神经6-9的主分支的聚乙烯箍的植入。钍E使用的的von Frey细丝也被描述。这些细丝允许评估机械痛这是一个长期持久的疼痛症状出现在这个模型。

<table border="0" cellpadding="0" cellspacing="0" style="width:633px;" width="633">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Name of Reagent/ Equipment</td>
			<td style="width:111px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:188px;">Comments/Description</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">PE-20 polyethylene tubing</td>
			<td>Harvard apparatus</td>
			<td>PY2-59-8323</td>
			<td>Splitted before surgery</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ketamine</td>
			<td>Centravet</td>
			<td>IMA004</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Xylazine HCl</td>
			<td>Sigma</td>
			<td>X1251</td>
			<td>Freshly prepared before surgery</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ocry-gel</td>
			<td>Centravet</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Plier</td>
			<td>FST</td>
			<td>11003-12</td>
			<td>52.5 mm straight</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bulldog clamp</td>
			<td>FST</td>
			<td>p130 18038-45</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Perforated plate</td>
			<td>CTTM</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">von Frey filaments</td>
			<td>Bioseb</td>
			<td>NC-12775</td>
			<td></td>
		</tr>
	</tbody>
</table>

the sciatic nerve cuffing model of neuropathic pain in mice

神经性疼痛的产生作为病变或疾病影响的体感系统的结果。这种综合征的结果从在受伤的感觉神经元和沿中枢神经系统内的整个痛觉通路适应不良的变化。它通常是慢性的和具有挑战性的治疗。为了研究神经性疼痛及其治疗方法，不同的模型已经发展在啮齿类动物。这些模型从已知的病因推导，从而再生周围神经损伤，中枢损伤，代谢，感染性或化疗相关的神经病变。周围神经损伤的小鼠模型往往针对坐骨神经这是很容易访问，并允许伤害性测试的后爪。这些模型依赖于压缩和/或部分。在这里，详细的手术过程为“袖口模式”小鼠神经病理性疼痛的描述。在这个模型中，标准化的长度（2毫米）的PE-20聚乙烯管材的袖口是我单方面mplanted周围的坐骨神经的主要分支。它诱导长期持久的机械性异常性疼痛， 即 ，，以一个通常的非伤害性刺激，可以通过使用von Frey细丝来评价一个伤害性反应。除了详细的手术和测试程序，这款型号为神经病理性疼痛的机制，对于神经病理性疼痛的感觉和anxiodepressive方面的研究，研究和神经病理性疼痛治疗的研究兴趣进行了讨论。

数据为平均值±SEM表示。统计学分析用方差多因素分析（ANOVA）或不成对t-检验按照实验设计进行。对于这些分析，在深水和袖口手术组以及盐水与药物治疗被认为是组间因素。当适当时，重复测量分析是用于时间过程的数据。在事后比较采用邓肯检验。统计显着性被认为是在p &lt;0.05。 当使用如上描述的方法，将袖带注入导致的同侧异常性疼痛如示于图1，一旦鼠标习惯于在测试过程中，在的von Frey测试缩爪阈值随时间保持稳定，并且不受手术操作本身 ，如图深水动物。然而它应该是注意的是，短暂的手术后异常性疼痛通常可以在深水小鼠中观察到。当这样的异常性疼痛存在时，缩爪反应，几天后手术后恢复到基线。在袖口小鼠，同侧异常性疼痛是已经存在于第一天手术后和下保持2个月以上（参见9和图1，F8，344 = 29.5，P &lt;0.001）。当它是由的von Frey试验，如上所述测量袖带诱发的异常性疼痛同侧保持在C57BL/6J小鼠，但是在其它条件异常性疼痛的对侧足一个存在，也可观察到8。对于基线的绝对值通常是4和6克之间在C57BL/6J小鼠，但是测试协议可能会影响这些值。 三环类抗抑郁药都属于临床一线治疗神经性疼痛。在这个模型中，三环类抗抑郁药去甲替林（5毫克/千克，腹膜内，每天两次）Relieves的神经性异常性疼痛后约2周的治疗， 如图2（F7，91 = 15.3，P &lt;0.001; 事后：（CuffNor =深）&gt; CuffSal在p &lt;0.001天29 - 34）。在这个剂量的抗抑郁药无急性镇痛作用观察16,17。模仿持久的疼痛缓解是存在于服用这样的药物的患者中，小鼠可以在早晨药物给药而不是之后才进行试验。这种程序允许的长效作用由治疗前几天催芽评估。在这种情况下，它需要1到2周的治疗，观察持久救济的神经性异常性疼痛的。当处理中断时，复发时在3至4天18通常观察到的。除了一些抗抑郁药，gabapentinoids是其他首选治疗神经性疼痛。加巴喷丁在该模型16中的急性和短暂的镇痛作用，但它也显示延迟和lon<html> G-持久的每一天给药前测试的动物（ 图3，P &lt;0.001）时的止痛作用。这个动作是比抗抑郁药物更快。 <img alt="图1" fo:content-width="5in" fo:src="/files/ftp_upload/51608/51608fig1highres.jpg" src="/files/ftp_upload/51608/51608fig1.jpg" /> 在小鼠神经病理性疼痛的袖口模型如图1所示。机械缩足阈值。成年雄性C57BL/6J小鼠习惯于冯弗雷过程，直到一个稳定的基线，获得（基准线表示在图上点0）。两个爪​​进行了测试。袖口小鼠显示患侧机械痛为表现的降低缩足阈值（N = 10每组）。 tp_upload/51608/51608fig2.jpg“/&gt; 图2。延迟止痛三环类抗抑郁药的作用。后2周手术后，小鼠接受腹腔治疗，一天两次（早晚各一次）与任何0.9％氯化钠或5毫克/公斤盐酸去甲替林（N = 5或6元组）。冯弗雷测试是早上治疗前完成。有了这个过程中，去甲替林的延迟的止痛作用是观察，其中需要治疗的大约12天。 <img alt="图3" fo:content-width="5in" fo:src="/files/ftp_upload/51608/51608fig3highres.jpg" src="/files/ftp_upload/51608/51608fig3.jpg" /> 加巴喷丁的图3。抗痛敏作用。后三周手术后，小鼠接受腹腔治疗，一天两次（早晚各一次）与任何0.9％NaCl或10毫克/千克加巴喷丁（每组N = 5）。冯弗雷测试的早晨，以前做过换货。有了这个过程中，加巴喷丁的延迟和持久的止痛作用是观察。数据开始治疗前和治疗第6天出现。

Watch this Scientific Journal Video about The Sciatic Nerve Cuffing Model of Neuropathic Pain in Mice at JoVE.com

The Sciatic Nerve Cuffing Model of Neuropathic Pain in Mice

神经性疼痛是一种病变或疾病影响的体感系统的结果。在“袖口模式”对小鼠神经病理性疼痛是由周围的坐骨神经的主要分支聚乙烯袖口的植入。机械痛是使用von Frey细丝测试。

神经病理性疼痛小鼠坐骨神经掴型号

Neuropathic pain arises as a consequence of a lesion or a disease affecting the somatosensory system. This syndrome results from maladaptive changes in ...

the-sciatic-nerve-cuffing-model-of-neuropathic-pain-in-mice

Research

JoVE Journal

Medicine

47.8K 次观看.  Centre National de la Recherche Scientifique. 神经性疼痛是一种病变或疾病影响的体感系统的结果。在“袖口模式”对小鼠神经病理性疼痛是由周围的坐骨神经的主要分支聚乙烯袖口的植入。机械痛是使用von Frey细丝测试。 

视频: The Sciatic Nerve Cuffing Model of Neuropathic Pain in Mice

The Spared Nerve Injury (SNI) Model of Induced Mechanical Allodynia in Mice

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared nerve injury (SNI) model induces symptoms of neuropathic pain such as mechanical allodynia i.e. pain due to tactile stimuli that do not normally provoke a painful response [1]. 
The SNI mouse model involves ligation of two of the three branches of the sciatic nerve (the tibial nerve and the common peroneal nerve), while the sural nerve is left intact [2]. The lesion results in marked hypersensitivity in the lateral area of the paw, which is innervated by the spared sural nerve. The non-operated side of the mouse can be used as a control. The advantages of the SNI model are the robustness of the response and that it doesn&#x2019;t require expert microsurgical skills.
The threshold for mechanical pain response is determined by testing with von Frey filaments of increasing bending force, which are repetitively pressed against the lateral area of the paw [3], [4]. A positive pain reaction is defined as sudden paw withdrawal, flinching and/or paw licking induced by the filament. A positive response in three out of five repetitive stimuli is defined as the pain threshold. 
As demonstrated in the video protocol, C57BL/6 mice experience profound allodynia as early as the day following surgery and maintain this for several weeks.

The Spared Nerve Injury SNI Model of Induced Mechanical Allodynia in Mice

The Spared Nerve Injury animal model is described here as a mouse model of peripheral neuropathic pain following partial denervation of the sciatic nerve by lesioning the tibial and common peroneal nerve branches, leaving the remaining sural nerve intact. Behavioral modification resulting from mechanical allodynia is quantified by von Frey filaments.

幸免神经损伤（SNI）模型引起的机械触感小鼠

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared ...

科研

医学

Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats

Chronic neuropathic pain, resulting from damage to the central or peripheral nervous system, is a prevalent and debilitating condition, affecting 7-18% of the population1,2. Symptoms include spontaneous (tingling, burning, electric-shock like) pain, dysaesthesia, paraesthesia, allodynia (pain resulting from normally non-painful stimuli) and hyperalgesia (an increased response to painful stimuli). The sensory symptoms are co-morbid with behavioural disabilities, such as insomnia and depression. To study chronic neuropathic pain several animal models mimicking peripheral nerve injury have been developed, one of the most widely used is Bennett and Xie's (1988) unilateral sciatic nerve chronic constriction injury (CCI)3 (Figure 1). Here we present a method for performing CCI and testing pain hypersensitivity.
CCI is performed under anaesthesia, with the sciatic nerve on one side exposed by making a skin incision, and cutting through the connective tissue between the gluteus superficialis and biceps femoris muscles. Four chromic gut ligatures are tied loosely around the sciatic nerve at 1 mm intervals, to just occlude but not arrest epineural blood flow. The wound is closed with sutures in the muscle and staples in the skin. The animal is then allowed to recover from surgery for 24 hrs before pain hypersensitivity testing begins.
For behavioural testing, rats are placed into the testing apparatus and are allowed to habituate to the testing procedure. The area tested is the mid-plantar surface of the hindpaw (Figure 2), which falls within the sciatic nerve distribution. Mechanical withdrawal threshold is assessed by mechanically stimulating both injured and uninjured hindpaws using an electronic dynamic plantar von Frey aesthesiometer or manual von Frey hairs4. The mechanical withdrawal threshold is the maximum pressure exerted (in grams) that triggers paw withdrawal. For measurement of thermal withdrawal latency, first described by Hargreaves et al (1988), the hindpaw is exposed to a beam of radiant heat through a transparent glass surface using a plantar analgesia meter5,6. The withdrawal latency to the heat stimulus is recorded as the time for paw withdrawal in both injured and uninjured hindpaws. Following CCI, mechanical withdrawal threshold, as well as thermal withdrawal latency in the injured paw are both significantly reduced, compared to baseline measurements and the uninjured paw (Figure 3). The CCI model of peripheral nerve injury combined with pain hypersensitivity testing provides a model system to investigate the effectiveness of potential therapeutic agents to modify chronic neuropathic pain. In our laboratory, we utilise CCI alongside thermal and mechanical sensitivity of the hindpaws to investigate the role of neuro-immune interactions in the pathogenesis and treatment of neuropathic pain.

Due to the simplicity of surgery and the robust behavioural outcome, chronic constriction of the sciatic nerve is one of the pre-eminent animal models of neuropathic pain. Within 24 hrs following surgery, pain hypersensitivity is established and can be quantitatively measured using a von Frey aesthesiometer (mechanical test) and plantar analgesia meter (thermal test).

慢性缩窄大鼠坐骨神经疼痛过敏测试

Chronic neuropathic pain, resulting from damage to the central or peripheral nervous system, is a prevalent and debilitating condition, affecting 7-18% of ...

Breathing-controlled Electrical Stimulation (BreEStim) for Management of Neuropathic Pain and Spasticity

Electrical stimulation (EStim) refers to the application of electrical current to muscles or nerves in order to achieve functional and therapeutic goals. It has been extensively used in various clinical settings. Based upon recent discoveries related to the systemic effects of voluntary breathing and intrinsic physiological interactions among systems during voluntary breathing, a new EStim protocol, Breathing-controlled Electrical Stimulation (BreEStim), has been developed to augment the effects of electrical stimulation. In BreEStim, a single-pulse electrical stimulus is triggered and delivered to the target area when the airflow rate of an isolated voluntary inspiration reaches the threshold. BreEStim integrates intrinsic physiological interactions that are activated during voluntary breathing and has demonstrated excellent clinical efficacy. Two representative applications of BreEStim are reported with detailed protocols: management of post-stroke finger flexor spasticity and neuropathic pain in spinal cord injury.

Breathing-controlled Electrical Stimulation BreEStim for Management of Neuropathic Pain and Spasticity

The purpose is to present a new method, breathing-control electrical stimulation (BreEStim) for management of neuropathic pain and spasticity.

呼吸控制电气的刺激（BreEStim）的神经性疼痛和痉挛的管理

Electrical stimulation (EStim) refers to the application of electrical current to muscles or nerves in order to achieve functional and therapeutic goals. ...

Tibial Nerve Transection - A Standardized Model for Denervation-induced Skeletal Muscle Atrophy in Mice

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. Although originally developed and used extensively in the rat due to its larger size, the tibial nerve in mice is big enough that it can be easily manipulated with either crush or transection, leaving the peroneal and sural nerve branches of the sciatic nerve intact and thereby preserving their target muscles. Thus, this model offers the advantages of inducing less morbidity and impediment of ambulation than the sciatic nerve transection model and also allows investigators to study the physiologic, cellular and molecular biologic mechanisms regulating the process of muscle atrophy in genetically engineered mice. The tibial nerve supplies the gastrocnemius, soleus and plantaris muscles, so its transection permits the study of denervated skeletal muscle composed of fast twitch type II fibers and/or slow twitch type I fibers. Here we demonstrate the tibial nerve transection model in the C57Black6 mouse. We assess the atrophy of the gastrocnemius muscle, as a representative muscle, at 1, 2, and 4 weeks post-denervation by measuring muscle weights and fiber type specific cross-sectional area on paraffin-embedded histologic sections immunostained for fast twitch myosin.

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of skeletal muscle atrophy. The model surgical protocol is described and demonstrated in C57Black6 mice. &#160;

胫骨神经切断术 - 一个标准化的模型失神经诱导小鼠骨骼肌

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. ...

Urinary Bladder Distention Evoked Visceromotor Responses as a Model for Bladder Pain in Mice

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized by increased urgency and frequency of urination, as well as nocturia and general pelvic pain, especially upon bladder filling or voiding. Despite years of research, the cause of IC/BPS remains elusive and treatment strategies are unable to provide complete relief to patients. In order to study nervous system contributions to the condition, many animal models have been developed to mimic the pain and symptoms associated with IC/BPS. One such murine model is urinary bladder distension (UBD). In this model, compressed air of a specific pressure is delivered to the bladder of a lightly anesthetized animal over a set period of time. Throughout the procedure, wires in the superior oblique abdominal muscles record electrical activity from the muscle. This activity is known as the visceromotor response (VMR) and is a reliable and reproducible measure of nociception. Here, we describe the steps necessary to perform this technique in mice including surgical manipulations, physiological recording, and data analysis. With the use of this model, the coordination between primary sensory neurons, spinal cord secondary afferents, and higher central nervous system areas involved in bladder pain can be unraveled. This basic science knowledge can then be clinically translated to treat patients suffering from IC/BPS.

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition characterized in part by pelvic pain. In order to study nervous system contributions to the condition, a physiological model of urinary bladder pain is used in mice and rats.

膀胱胀诱发内脏运动反应为膀胱疼痛小鼠模型

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition ...

Facial Nerve Axotomy in Mice: A Model to Study Motoneuron Response to Injury

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and either cut or crush it to induce peripheral nerve injury. Advantages of this surgery are its simplicity, high reproducibility, and the lack of effect on vital functions or mobility from the subsequent facial paralysis, thus resulting in a relatively mild surgical outcome compared to other nerve injury models. A major advantage of using a cranial nerve injury model is that the motoneurons reside in a relatively homogenous population in the facial motor nucleus in the pons, simplifying the study of the motoneuron cell bodies. Because of the symmetrical nature of facial nerve innervation and the lack of crosstalk between the facial motor nuclei, the operation can be performed unilaterally with the unaxotomized side serving as a paired internal control. A variety of analyses can be performed postoperatively to assess the physiologic response, details of which are beyond the scope of this article. For example, recovery of muscle function can serve as a behavioral marker for reinnervation, or the motoneurons can be quantified to measure cell survival. Additionally, the motoneurons can be accurately captured using laser microdissection for molecular analysis. Because the facial nerve axotomy is minimally invasive and well tolerated, it can be utilized on a wide variety of genetically modified mice. Also, this surgery model can be used to analyze the effectiveness of peripheral nerve injury treatments. Facial nerve injury provides a means for investigating not only motoneurons, but also the responses of the central and peripheral glial microenvironment, immune system, and target musculature. The facial nerve injury model is a widely accepted peripheral nerve injury model that serves as a powerful tool for studying nerve injury and regeneration.

We present a surgical protocol detailing how to perform a cut or crush axotomy on the facial nerve in the mouse. The facial nerve axotomy can be employed to study the physiological response to nerve injury and test therapeutic techniques.

面神经干切断小鼠：模型，研究运动神经元损伤反应

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and ...

Chronic Constriction Injury of the Rat's Infraorbital Nerve (IoN-CCI) to Study Trigeminal Neuropathic Pain

Animal models are important tools to study the pathophysiology and pharmacology of neuropathic pain. This manuscript describes the surgical and behavioral procedures to study trigeminal neuropathic pain in rats. To meet the specificity of trigeminal neuropathic pain syndromes, the infraorbital nerve (IoN) is subjected to a chronic constriction injury (CCI) by loosely ligating the nerve. An intra-orbital approach is presented here to expose and ligate the IoN in the orbital cavity. After IoN ligation, rats exhibit changes in spontaneous behavior and in response to von Frey hair stimulation that are indicative of persistent pain and mechanical allodynia. Two phases can be defined in the development of the behavioral changes. During the first week following IoN-CCI (phase 1), rats show an increased and asymmetric face grooming activity, i.e., with face wash strokes primarily directed to the nerve-injured IoN territory. A distinction is made between face grooming behavior that is part of a more general body grooming behavior, which remains largely unaffected by IoN-CCI, and face grooming that is neither preceded nor followed by body grooming, which is significantly increased after IoN-CCI. During this period, responsiveness to mechanical stimulation of the IoN territory is reduced. This hyporesponsiveness is abruptly replaced by an extreme hyperresponsiveness whereby even very weak stimulus intensities provoke nocifensive behavior (phase 2). The phenomenological similarities between these behavioral alterations and reported signs of facial pain (i.e., responses to noxious stimulation of the face) suggest the presence of dysesthesia/paresthesia and mechanical allodynia in the ligated IoN territory.

Chronic Constriction Injury of the Rat&#039;s Infraorbital Nerve IoN-CCI to Study Trigeminal Neuropathic Pain

This manuscript describes a rodent model to study trigeminal neuropathic pain. The methods described include surgical procedures to perform a chronic constriction injury of the rat&#8217;s infraorbital nerve and the post-surgical behavioral tests to measure the changes in spontaneous and evoked behavior that are indicative of persistent pain and allodynia.

老鼠的眶下神经（ION-CCI）研究三叉神经性疼痛性损伤

Animal models are important tools to study the pathophysiology and pharmacology of neuropathic pain. This manuscript describes the surgical and behavioral ...

The Monoiodoacetate Model of Osteoarthritis Pain in the Mouse

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 current treatments for OA pain such as NSAIDS or opiates are neither sufficiently effective nor devoid of detrimental side effects. Animal models of OA are being developed to improve our understanding of OA-related pain mechanisms and define novel pharmacological targets for therapy. Currently available models of OA in rodents include surgical and chemical interventions into one knee joint. The monoiodoacetate (MIA) model has become a standard for modelling joint disruption in OA in both rats and mice. The model, which is easier to perform in the rat, involves injection of MIA into a knee joint that induces rapid pain-like responses in the ipsilateral limb, the level of which can be controlled by injection of different doses. Intra-articular injection of MIA disrupts chondrocyte glycolysis by inhibiting glyceraldehyde-3-phosphatase dehydrogenase and results in chondrocyte death, neovascularization, subchondral bone necrosis and collapse, as well as inflammation. The morphological changes of the articular cartilage and bone disruption are reflective of some aspects of patient pathology. Along with joint damage, MIA injection induces referred mechanical sensitivity in the ipsilateral hind paw and weight bearing deficits that are measurable and quantifiable. These behavioral changes resemble some of the symptoms reported by the patient population, thereby validating the MIA injection in the knee as a useful and relevant pre-clinical model of OA pain.
The aim of this article is to describe the methodology of intra-articular injections of MIA and the behavioral recordings of the associated development of hypersensitivity with a mind to highlight the necessary steps to give consistent and reliable recordings.

Osteoarthritis (OA), or degenerative joint disease, is a debilitating condition associated with pain that remains only partially controlled by available analgesics. Animal models are being developed to improve our understanding of OA-related pain mechanisms. Here we describe the methodology for the monoiodoacetate model of OA pain in the mouse.

骨关节炎疼痛的鼠标Monoiodoacetate模式

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 ...

Subcutaneous Trigeminal Nerve Field Stimulation for Refractory Facial Pain

Chronic or neuropathic trigeminal facial pain can be challenging to treat. Neurosurgical procedures should be applied when conservative treatment fails. Neuromodulation techniques for chronic facial pain include deep brain stimulation and motor cortex stimulation, which are complex to perform. Subcutaneous nerve field stimulation is certified for chronic back pain and is the least invasive form of neuromodulation. We applied this technique to treat chronic and neuropathic trigeminal pain as an individual therapy concept. First, trial stimulation is performed. Subcutaneous leads are placed in the painful trigeminal dermatome under local anesthesia. The leads are connected to an external neurostimulator that applies constant stimulation. Patients undergo a 12 day outpatient trial to assess the effect of the stimulation. Electrodes are removed after the trial. If the patient reports pain reduction of at least 50% in intensity and/or attack frequency, a reduction in medication or increase in quality of life, permanent implantation is scheduled. New electrodes are implanted under general anesthesia and are subcutaneously tunneled to an infraclavicular internal pulse generator. Patients are able to turn stimulation on and off and to increase or decrease the stimulation amplitude as needed.
This technique represents a minimal invasive alternative to other more invasive means of neuromodulation for trigeminal pain such as motor cortex stimulation or deep brain stimulation.

Treating chronic or neuropathic facial pain can be challenging when medical or standard treatment fails. Subcutaneous nerve field stimulation is the least invasive form of neuromodulation and is used for chronic back pain. We applied this technology to treat chronic and neuropathic trigeminal facial pain.

皮下三叉神经场刺激难治性面部疼痛

Chronic or neuropathic trigeminal facial pain can be challenging to treat. Neurosurgical procedures should be applied when conservative treatment fails. ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...