Name: a neonatal mouse spinal cord compression injury model
Uploaded: 2016-03-27T14:00:00.000+00:00
Duration: 13 min 31 s
Description: Watch this Scientific Journal Video about A Neonatal Mouse Spinal Cord Compression Injury Model at JoVE.com

This work has been supported by grants from the South-Eastern Norway Regional Health Authority (JLB, 2014119; JCG, project numbers 2015045 and 2012065), by the Norwegian Research Council (JCG, project number 23 00 00) and the University of Oslo.

该实验方案已通过国家动物研究的权威机构挪威（Forsøksdyrutvalget，局部实验批准文号12.4591）符合欧盟动物保护条例（联邦欧洲实验动物科学协会）。作出了努力，以尽量减少使用动物和他们的痛苦的数量。在这篇文章产后（P）的第1天的野生型ICR（印迹控制区）小鼠（Jackson，USA）中所使用的程序进行说明，但同样的方法也可以在稍后阶段使用。 1.新生小鼠构建气体麻醉系统（图1） <ol><li>建立从注射器的前端的鼻面罩。连接这对塑料管（ 图1 -红色油管和图2A1）的3路阀。 </li><li>钻一个小孔的鼻罩的一侧，这个连接的塑料管从去除气体溢出面具。结束该管道或者在真空泵轻微的负压设定，或者在通风橱（ 图1 -亮绿色管路）。 </li><li>从150毫米×25毫米的塑料培养皿（ 图2A2）做一个麻醉室。 <ol><li>在一侧上，使大到足以容纳鼠标的头部和鼻罩的孔。 </li><li>在相反侧，使两个较小的孔，通过该塑料管，并从鼻罩可插入（ 图1 -红色和亮绿色管道，分别）。 </li><li>使在盖的顶部的第三孔和连接到该第三塑料管，在真空泵（ 图1 -暗绿色管路）结束。该第三管的目的是确保尚未通过从鼻罩的出口捕获的任何过量的气体被除去。 </li></ol></li><li>在任何类型的培养皿的底部凿洞建立一个睡眠室是足够大到contain鼠标和具有光滑的和连缘（盘的开口必须位于与表以防止气体泄漏平齐）。在腔室连接孔与塑料管（ 图1 -棕色管）的3路阀。放置睡眠室通风橱下。 </li><li>一个3通旋塞阀连接到从蒸发器（ 图1 -黄色油管和图2A3）的出口管。 </li><li>蒸发器的入口连接到氧气供给（ 图1 -蓝色管）。 </li></ol> 2.变形例一Yasargil临时瘤迷你夹创建压缩工具的（图2和表1） <ol><li>牢牢贴上剪辑用卡子的立场。使用视觉控制双目放大镜，每个剪辑叶片的尖端的外表面的文件下降到使用磨石大约150μm的最终厚度安装在钻头（ 图2B和C）。 </LI&gt; <li>使该夹子止动件通过用微型刀（ 表1）在立体显微镜下切聚乙烯毛细管（ 表1）的一段短，并把这个在叶片中的一个（ 图2A4及图2B和C）。这可以防止剪辑的全封闭，并创建标准化的压缩尺寸。当夹子闭合时叶片间的距离为约230微米。使每个实验作为聚乙烯材料在使用过程中可能会压缩，这会改变叶片间空间的新塞子。 注意:剪辑的弹簧张力减小随着时间的推移，使得后约80按压夹具不再完全关闭到塞子并需要更换。 </li></ol> 3.准备手术前<ol><li>放置在睡眠室（ 图1）的鼠标，并开始用4％异氟烷麻醉（ 图2A5 </strong&gt;）蒸发在纯氧，使用蒸发器（ 图2A3和表1）。 </li><li>用薄塑料镊子轻轻捏脚趾之间的皮肤上测试鼠标的撤回反射。认真执行此新出生的小鼠很容易受伤。捏在眼前青紫太难结果。在镇静的开头执行此测试触发反射，并提供所需要的力的量的良好指示。 </li><li>一旦反射被取消，从睡眠室移开鼠标，并将其放置在手术台上俯卧位置与插入所述鼻罩提供4％异氟烷在纯氧（ 图1）混合的连续供应的口鼻部。确保手术过程中暖垫被打开，并设置为37-38°C作为低温可能是致命的。 </li><li>实现完全镇痛，注入皮下50μl的局部麻醉剂布比卡因（2.5毫克/毫升， <strong&gt;的图2A6）在手术位点（在这里报导的实验，这是在胸水平（T）的9-T11）。使用胰岛素注射器（300微升，30 G， 图2A7和表1），以进行注射。 </li><li>减少传递到鼻罩到1-2％的异氟烷​​浓度。 </li></ol> 4.背椎板切除术<ol><li>微观控制下进行手术。 </li><li>用葡萄糖酸洗必泰（ 表1＃19）清洗手术区至少30秒后，使在T9-T11使用microknife（ 图2A8）一个1-2毫米的横向的皮肤切口。 注意:在ICR新生小鼠胃的喙部，可见当它含有乳，是面向椎骨水平T12-T13（ 图3）。另一个具有里程碑意义的是胸部的皮下脂肪组织聚集，在大约T8-9结束的延髓部分。这一里程碑式的是切开皮肤后，才可见。 </li><l我&gt;使用镊子（ 图2A9和A10）通过拉动皮肤轻轻吻侧和尾侧（皮肤泪容易，创造一个平直伤口），以扩大在横向方向上的皮肤开口至8-9毫米。这提供了到脊柱足够的横向连接。 </li><li>通过插入止血明胶海绵的无菌片（ 图2A11和表1）皮下喙和尾部的切口缩回从底层结构皮肤切口的边缘。这扩大了开口并防止皮肤缩回和手术过程中遮蔽面积。止血明胶海绵并不需要在使用前盐水浸泡。 </li><li>以暴露脊柱，解剖用薄剪刀（ 图2A12，和表1）椎旁的肌肉。切肌肉的附 ​​着物脊柱和暴露椎板（ 图4A）。不E也认为在这个阶段的棘突不发达。 </li><li>确定中线和两个薄片（其在此阶段是软骨）薄剪刀（ 图4B）之间横向切割。小心地将椎板和硬膜（ 图4C）之间的薄镊子一个刀片，用抓钳椎板，小心地将其提起，直到一块打破了，留下硬脑膜完好（ 图4D）。重复此2-3次，以获得1-2段长椎板切除术。 </li><li>使用薄镊子作为骨钳，取出关节突关节部位两侧以获得足够的空间来放置椎管内的剪辑。清洁手术区和控制小块止血明胶海绵止血。 </li></ol> 5.脊髓压迫损伤<ol><li>打开剪辑持有人修改后的动脉瘤迷你夹子（ 图2A13和图2B）和地点次在脊髓中的面连接的空间，且电源线任一侧ë叶片。确保被插入深深足以影响脊髓的腹侧部分的叶片。如果这是不可能的，去除更多的小面关节的。 </li><li>迅速释放微型夹，用夹子夹持器保持在适当位置，以防止其滑动。保持15秒的压缩。 </li><li>迅速打开迷你夹子将其取出。实现了对称的压缩，反向微型夹的方向，并使用由所述出血性水肿从第一压缩作为引导取得的容易看到标记，重新定位在相反方向的夹子的第二15秒压缩（现有实验表明，这种产生对称的组织学和生理缺陷，而单 ​​按压不要1）。硬脑膜不应由压缩而损坏。 </li><li>清洁区域并保持与止血明胶海绵件止血。</li><li>除去在手术开始被放置在皮肤切口的边缘根据该止血明胶海绵的碎片并用无菌6.0缝合和针保持器（ 图2A14和15）关闭皮肤切口。 </li><li>注入皮下0.75毫克/公斤体重丁丙诺啡使用胰岛素注射器（300微升，30克）在无菌PBS中稀释（图2A16）。 </li></ol> 6.术后护理<ol><li>从鼻罩移开鼠标和直到麻醉消退和鼠标变得警报（1-3小时通常是足够的）并将其放置在温度控制的腔室组在30℃下。 </li><li>注入地西泮（ 图2B17）腹膜内进入母体（8克/公斤体重）。这将创建减少食人过程中的第一个晚上的风险，当这种风险是最高的一个麻木。 </li><li>返回操作鼠标的垃圾。 </li><li>如果垃圾为lARGE（&gt; 12幼仔），去除一些不操作幼仔的，优选当它们在大小不同的较大的动物，减少牛奶竞争。产妇护理的操作幼崽是最好的ICR行，如果产仔数约为9幼崽。 </li><li>对疼痛管理，在术后第一天施用丁丙诺啡（0.75毫克/公斤体重）皮下注射，每天一次，使用胰岛素注射器（300微升，30克）。用于皮下注射适当体积为30-50微升。在新生小鼠发声和鼓动是疼痛的良好指标。 </li><li>执行使用评分表，以评估营养，体重，脱水，疼痛，伤口愈合，尿潴留和感染状态的损伤小鼠的每日检查。根据所获得的评分，提供特别的照顾，如在异常营养的情况下的无菌儿科营养溶液（ 表1＃18）的注射。评分表还0;人性化的定义终点指标。不拒绝受伤的幼仔母亲是最好的照顾者。 </li><li>在膀胱功能障碍的异常的情况下，直到恢复功能执行膀胱按摩一天两次。这是通过将小鼠以仰卧姿势，一手用指尖在rostro - 尾方向轻轻按摩小腹完成。 </li></ol>

The authors have nothing to disclose.

在这篇文章中被描述为一个剪辑生成的SCC损伤P1小鼠的程序。相同的程序也可以在稍后阶段进行。压缩伤害物在P5，P7，P9和P12（Züchner， 等人 ，在制备手稿）成功地进行。在所有产后阶段，是用异氟烷在纯氧气化获得全身麻醉，但麻醉结果在很大程度上取决于年龄。在P1〜P4的初步尝试，前局部麻醉引入协议，难以得到深且长时间镇静由于镇静和过量不足之间的窄的剂量效应窗口。另外，有关在新生动物异氟烷的神经毒性作用的关注已经提出27-30。异氟醚的组合和局部麻醉剂布比卡因导致更深的和更稳定的麻醉，同时允许通过的2-3倍的异氟烷减少剂量。不同类型的anestheSIA已为新生儿啮齿类动物中描述，包括cryoanesthesia 31,32，但cryoanesthesia的一个潜在的不便是其神经保护作用（由33,34综述），这可能是有效的和可再现的损伤的产生复杂化。基于巴比妥麻醉被认为具有在新生小鼠低效率，由于较低的血清白蛋白和身体脂肪的比成人35,36。 虽然相当侵入性和创伤性，一旦该过程手术期间建立的死亡率是低的。但是，存在需要特别注意以提高手术小鼠的恢复和存活的过程中的关键步骤。一个重要的问题是要选择具有生存的手术的最佳机会幼仔。当垃圾较大的个体幼崽的营养状况而变化。除了手术过程中发生的不可避免的出血，操作幼仔花小时■从母亲走，他们往往不会在第二天早上之前喝牛奶。因此，这是选择已经具有在胃中一定量的牛奶幼仔的优点。这是通过腹部皮肤从P0到P7随时可见。 在第一天晚上的工作是小狗在由母亲被蚕食很大的风险。在这种模式的初始发展以上被操作的小鼠的一半以下早晨被丢失，在保持架的血液明显迹象。 Necrophagy，吃人肉和杀婴啮齿类动物中已经研究了几十年37-40。在这项研究中，食人只看到过一次，但被认为比necrophagy一个更可能的解释是因为被送回笼子里的小狗都一般在这样良好的状态，通过在夜间自然原因死亡似乎是不可能的。这促使使用可逆药理剂如地西泮，以减少焦虑和侵略性i的想法n中的母亲（41审查）。地西泮的腹腔注射大大提高的情况下，从超过60％在第一天晚上死亡率下降到不足20％。 通过剔除，扰乱垃圾尽可能少，术后如下回报减少产仔数是可以惠及手术动物额外的元素。然而，只留下操作幼仔与母亲并无益处。操作/未手术幼仔的最佳平衡可根据线变化，但对于ICR和SCID-ICR小鼠离去4-5操作幼仔（伤害或假）3-4无操作幼仔一起得到最佳的结果。 在一般意义上，这新生儿脊髓损伤模型的主要限制是，在新生儿脊髓在许多方面不同于成人脊髓，并且因此可以不提供比得上从成人脊髓损伤模型获得的那些实验结果。这种差异包括总体尺寸和脊髓体积，细胞数，代表不足的特定细胞类型，例如少突胶质细胞，未成熟的免疫应答和未成熟的神经元回路的。从实验在这个模型中得出的结论，因此必须慎重考虑。另一方面，该模型是有关用于儿科的SCI的相对较少研究情形。此外，相对于成人脊髓损伤模型的明显缺点是还因为它可以允许的可塑性机制，虽然在成人脊髓微创现存如果恢复，可能代表治疗性基板的澄清的电位强度。可以想到的新生儿或甚至胚胎条件恢复可以通过较不发达的细胞或组织的植入或通过与产生与早期发育特征成人组织试剂处理来实现。使用酶来消除perineuronal网是后一种方法42,43的一个例子。 例如 ，切断，半切，撞击，气囊压缩，钳挤压，静重压缩等就影响设备，在这个方向上的努力在导致脊髓损伤模型的一个重要方面成年啮齿类动物，其中的影响的多个参数，例如速度，力和持续时间可以被操纵（由44中综述）。另一种方法中，涉及设备少，采用了克尔洛希德动脉瘤夹45,46的变形例。这2方法是互补的冲击模仿挫伤，而夹模仿一个压迫性损伤具有一定程度的并发缺血。由于大量的大小限制和新生小鼠的更大的脆弱性，病死率较高较长的手术以及DEVEL的相关费用展中规模较小的设备，它被选定为开发剪辑生成的压缩，而不是一个撞击产生的挫伤的方法。这是通过调整市售瘤迷你夹以容纳新生小鼠1的脊柱的大小来实现的。添加塞子确保标准化的压缩宽度和只要剪辑的张力压缩到塞子的限制，在压缩的过程中的静态相位以最小的宽度的力应该变化不大。什么是不规范是在其动态相位压缩的速度，因为这会在其生命周期剪辑张力的变化而变化。作为压缩的静止阶段持续比动态相位长得多，而且很少有表明脊髓组织施加多对迷你夹叶片的反作用力的，它很可能是损伤的严重程度是最依赖静态阶段。然而，这仍然是进行测试。伤严重程度可能取决于多种因素，其中包括静态压缩力和持续时间，压缩和减压的速度，小型剪辑的位置，并在同一部位进行按压的次数。因此，在这些参数的组合变化可能导致受伤的严重程度由弱到严重的频谱的产生。尽管可变性的潜力，在我们之前发表的研究报告1中，我们获得在组织学，生理和行为水平相一致的结果，所以几乎没有表明接受标准化是很难达到的。我们注意到，在该研究中，我们使用的验证的多个方法中的每一级，包括行为测试如空气步， 如图5。 在此新生脊髓损伤模型的损伤备件轴突一定比例的，从而提供了一种情况有利于通过重新modelin引发自适应可塑性幸免连接g且新电路的形成。另外，由于新生小鼠是非常适合用于通过许多实验方法调查，有可能使用该模型来研究功能恢复和自适应可塑性与一种综合方法，包括行为测试，逆行并顺行轴突追踪，免疫组织化学，电和高-throughput光学记录1。作为一个例子，我们采取了这种综合方法的优点在于脑干体外 wholemount制剂和脊髓损伤1使用高通量钙成像具体递减输入的水平来证明网络重新建模。这可以进一步通过使用神经光遗传学和光遗传学药理工具来评估脊髓神经元的特定亚群之间的突触连接的重塑推动。

During the last decade, increasing evidence obtained from different spinal cord injury (SCI) models has shown that spinal networks can reorganize spontaneously to contribute to functional recovery1-9. Adaptive plasticity has as a consequence become an important topic in SCI research. It has been shown that plasticity encompasses regrowth of spared axons, sprouting of new axon collaterals and the formation of novel synaptic connections. Much of this knowledge has been obtained from behavioral or anatomical studies in adult animals. An important limitation of adult spinal cord studies is the difficulty of performing high-throughput physiological assessment, which is easier in neonatal preparations1. One major difference is that wholemount ex vivo preparations of the adult brainstem and spinal cord have low viability. Another is that adult spinal tissue is more opaque to light because it is thicker and myelinated. Although recent advances in in vivo imaging (see for example, 10-12) may partially overcome these problems, the possibility of performing high throughput imaging at any desired dorsoventral depth at multiple sites along a given brainstem-spinal cord preparation is currently only feasible in neonates. The immature state of axon myelination in the neonatal spinal cord facilitates high-throughput ex vivo optical recording, thus permitting a dynamic assessment of functional synaptic connections13-17. Combined with genetically encoded calcium reporters and optogenetic stimulation and pharmacology tools, optical approaches can contribute to a deeper understanding of the mechanisms underlying adaptive plasticity.
It is estimated that between 1-10% of all spinal cord injuries affect infants and children18-22. In contrast to adult SCI the pathogenesis and potential for spontaneous recovery in pediatric SCI is less studied. Using a neonatal SCI model can therefore provide more insight into pediatric SCI and contribute to a better understanding of the pathogenetic and recovery mechanisms involved. Moreover, post-SCI plasticity supporting functional recovery in the adult spinal cord is believed to involve at least in part the same mechanisms that govern the development of the central nervous system such as axon growth, branching and formation of new synapses23-26. Thus, using a neonatal SCI model could provide important insights into mechanisms that are also operative in the adult spinal cord, or that could potentially be reinstated in the adult spinal cord (for example by implantation of fetal cells or tissue or of tissue constructed de novo from pluripotent stem cells) to facilitate recovery.
The neonatal mouse thus provides a platform for an integrative, multi-methodological approach to investigating adaptive plasticity following spinal cord injury, in which a combination of behavioral, physiological, anatomical, molecular and genetic methods can be readily employed. Establishing standardized neonatal injury models is an important step in implementing such studies.

<table><tbody><tr><td>Plastic syringe (30 or 50&amp;nbsp;ml)</td><td></td><td></td><td></td></tr><tr><td>Plastic Petri dish (150 x 25&amp;nbsp;mm)</td><td></td><td></td><td></td></tr><tr><td>Fortec isoflurane vaporizer</td><td>Cyprane</td><td></td><td>We use and old device out of production, check the link for newer device</td></tr><tr><td>Yasargil temporary aneurysm mini-clip</td><td>&amp;AElig;sculap</td><td>FE681K</td><td></td></tr><tr><td>Fine-Bore Polyethylene tubing ID 0.58&amp;nbsp;mm, OD 0.96&amp;nbsp;mm</td><td>Smiths Medical</td><td>800/100/200</td><td></td></tr><tr><td>Isoflurane (Forene)</td><td>Abbott GmbH &amp;amp; Co. KG</td><td></td><td></td></tr><tr><td>Marcain (Bupivacain)</td><td>AstraZeneca</td><td></td><td></td></tr><tr><td>Insuline seyringe 0.3&amp;nbsp;ml 30&amp;nbsp;G x 8&amp;nbsp;mm</td><td>VWR</td><td>80086-442</td><td></td></tr><tr><td>Ultra Fine Micro Knife 5&amp;nbsp;mm cutting edge</td><td>Fine Science Tools</td><td>10315-12</td><td></td></tr><tr><td>Extra Fine Graefe Forceps &amp;ndash; 0.5&amp;nbsp;mm Tip</td><td>Fine Science Tools</td><td>1153-10</td><td>Not really necessary, often the teeth are too large</td></tr><tr><td>Forceps SuperGrip Straight</td><td>Fine Science Tools</td><td>00632-11</td><td>Two forceps are necessary</td></tr><tr><td>Spongostan Special 70 x 50 x 1&amp;nbsp;mm</td><td>Ferrosan</td><td></td><td></td></tr><tr><td>Vannas Spring Scissors &amp;ndash; 2&amp;nbsp;mm Blades Straight</td><td>Fine Science Tools</td><td>15000-03</td><td></td></tr><tr><td>Vario Clip Applying Forceps</td><td>Aesculap</td><td>FE502T</td><td></td></tr><tr><td>Vicryl 6&amp;ndash;0 (Ethicon)</td><td>Johnson and Johnson</td><td>J105G</td><td></td></tr><tr><td>Diethrich micro needle holder</td><td></td><td>11-510-20</td><td></td></tr><tr><td>Temgesic (buprenorphine)</td><td>Schering-Plough</td><td></td><td></td></tr><tr><td>Stesolid (diazepam)</td><td>Actavis</td><td></td><td>Also known as Valium</td></tr><tr><td>Pedamix</td><td>Fresenius Kabi</td><td></td><td></td></tr><tr><td>Klorhexidinsprit (chlorhexidine gluconate)</td><td>Fresenius Kabi</td><td>D08A C02</td><td></td></tr></tbody></table>

a neonatal mouse spinal cord compression injury model

脊髓损伤（SCI）通常会导致破坏性的神经功能障碍，特别是通过对纤维从大脑降至脊髓的损伤。研究的主要领域当前的重点是自适应的可塑性背后脊髓损伤后自发或诱发功能恢复的机制。自发性功能恢复据报道，在生活中更大的早期，提高对如何适应可塑性变化脊髓开发有趣的问题。为了促进这种动态的调查，我们已经开发出在新生儿鼠标脊髓损伤模型。该模型具有儿科SCI，这是太少了研究的相关性。因为在成人神经可塑性涉及到一些相同的机制，在生命早期1神经可塑性，这种模式可能会潜在地具有一定的相关性也为成人脊髓损伤。在这里，我们描述了新生小鼠产生可再生的脊髓压迫（SCC）伤害了整个过程早在出生后（P），每日1次的SCC是通过在给定的脊髓水平执行椎板切除术来实现，然后（在胸椎水平9-11此处描述）使用改良的Yasargil动脉瘤迷你夹迅速压缩和解压缩的脊髓。如先前所描述的，受伤新生小鼠可以为行为缺陷被测试或牺牲用于使用电和高通量的光记录技术1 体外突触连接的生理分析。早期的和正在进行的研究使用行为和生理评估表明后肢运动的戏剧性，急性损伤后2周内完整的功能恢复，并在功能电路的变化在下降鉴定突触连接1级的第一个证据。

脊髓压迫性损伤和功能丧失 如前所述，通过优化的术前，手术中和术后的程序，在新生小鼠可再现压缩的SCI模型可以得到1。放置在夹子（ 图2B和C）中的一个叶片的聚乙烯塞子防止夹子的全封闭，并在约230微米保持刀片间的距离一致。扭转夹在这两个按压结果之间的取向以对称损伤，通过组织学后遗症如判断（ 图5A和1）。微型夹取出后，立即将压缩脊髓组织变得因失血性挫伤和水肿暗。染色曙红和苏木已经有一天，一个受伤的脊髓连续切片观察接近病灶震中（ 图5A）时压脚提升损伤揭示的组织的逐渐恶化。椎管内空腔的存在或血液中的病变是不寻常的。 行为评估，例如通过手术后在非重跟踪后肢轨迹轴承条件几个小时，显示后肢运动在SCC损伤小鼠急剧损害相比，其中仅执行椎板切除术假的对照小鼠（ 图5B和1） 。此测试可以重复直到鼠标能够执行需要承载其自身的重量1其他行为测试。 死亡率和手术后恢复 术中的死亡率的主要原因是呼吸暂停和引起的达到足够anesthesi所需的高浓度的异氟烷心脏停搏一个。介绍麻药布比卡因到手术协议允许降低异氟醚的浓度，从而减少显著死亡率。在最近的一系列实验包括超过20只，术中死亡率为零。与此相反，手术后存活主要由他们的母亲在接受手术小鼠的影响。一个显著的改善发生时减少了返回操作小鼠垃圾1日前交付地西泮（IP为8g / kg体重）母亲单注焦虑和侵略性。所操作的小鼠接受和术后恢复可以通过牛奶在胃中存在进行监控。一个P1-P7鼠标有醉酒牛奶的胃是通过腹部皮肤（ 图3）明明白白的可见。馈送的在操作时，假控制和未手术的小鼠的比较是评估损伤的营养状况有用ð老鼠。评估的操作与未手术的小鼠的生长表明，尽管在第一术后天稍微体重减轻，手术小鼠的生长曲线此后迅速恢复正常（ 图6）。涉及膀胱功能障碍或感染的死亡率从未在研究了只要7周小鼠甚至观察到。 <table border="1"><tbody><tr><td> 数图。 2 </td><td> 名称 </td><td> 制造商/供应商 </td><td> 参考＃ </td><td colspan="3"> 链接 </td><td> 评论 </td></tr><tr><td> 1 </td><td>塑料注射器（30或50毫升） </td><td></td><td></td><td colspan="3"></td><td></td></tr><tr><td> 2 </td><td>塑料培养皿（150×25毫米） </td><td></td><td></td><td colspan="3"></td><td></td></tr><tr><td> 3 </td><td> FORTEC异氟醚蒸发</td><td> CypraNE </td><td></td><td colspan="3"> http://www.mssmedical.co.uk/products/new-vaporisers/ </td><td>我们使用老设备生产出来的，检查链接更新的设备</td></tr><tr><td> 4A </td><td> Yasargil临时瘤迷你夹</td><td> AESCULAP </td><td> FE681K </td><td colspan="3"> http://www.aesculapusa.com/assets/base/doc/DOC697_Rev_C-Yasargil_Aneurysm_Clip.pdf </td><td></td></tr><tr><td> 4B </td><td>细孔聚乙烯毛细管ID0.58毫米，外径0.96毫米</td><td>史密斯医疗</td><td> 800/100/200 </td><td colspan="3"> http://www.smiths-medical.com/industrialproducts/8/39/ </td><td></td></tr><tr><td>五</td><td>异氟醚（Forene） </td><td>雅培GMBH＆Co. KG的</td><td></td><td colspan="3"> http://www.life-sciences-europe.com/product/forene-abbott-gmbh-wiesbaden-group-narcotic-germany-west-2001-1858.html </td><td></td></tr><tr><td> 6 </td><td> Marcain（布比卡因） </td><td>阿斯利康</td><td></td><td colspan="3"> http://www.astrazeneca.co.uk/medicines01/neuroscience/Product/marcaine </td><td></td></tr><tr><td> 7 </td><td>胰岛素注射器0.3毫升30国祥8毫米</td><td> VWR </td><td> 80086-442 </td><td colspan="3"> https://us.vwr.com/store/catalog/product.jsp?product_id=4646138 </td><td></td></tr><tr><td> 8 </td><td>超细微型刀5毫米前沿</td><td>精细的科学工具</td><td> 10315-12 </td><td colspan="3"> http://www.finescience.de/katalog_ansicht.asp?Suchtyp= 吉＆suchkatalog = 0019900000＆reloadmenu = 1 </td><td></td></tr><tr><td> 9 </td><td>特细镊格雷夫 - 0.5毫米提示</td><td>精细的科学工具</td><td> 1153年至1110年</td><td colspan="3"> http://www.finescience.de/katalog_ansicht.asp?Suchtyp= 吉＆suchkatalog = 0055700000＆reloadmenu = 1 </td><td>不是真的有必要，往往牙齿过大</td></tr><tr> &lt;TD&gt; 10 </td><td>镊SuperGrip直</td><td>精细的科学工具</td><td> 00632-11 </td><td colspan="3"> http://www.finescience.de/katalog_ansicht.asp?Suchtyp= 吉＆suchkatalog = 0053500000＆reloadmenu = 1 </td><td>两个镊子是必要的</td></tr><tr><td> 11 </td><td> Spongostan特别70×50×1毫米</td><td>费诺森</td><td></td><td colspan="3"></td><td></td></tr><tr><td> 12 </td><td> Vannas弹簧剪刀 - 2毫米直叶片</td><td>精细的科学工具</td><td> 15000-03 </td><td colspan="3"> http://www.finescience.de/katalog_ansicht.asp?Suchtyp= 吉＆suchkatalog = 0012800000＆reloadmenu = 1 </td><td></td></tr><tr><td> 13 </td><td> VARIO夹镊应用</td><td>蛇牌</td><td> FE502T </td><td colspan="3"> http://www.aesculapusa.com/assets/base/doc/DOC697_Rev_C-Yasargil_Aneurysm_Clip.pdf </td><td></td></tr><tr><td> 14 </td><td>薇乔60（爱惜康） </td><td>强生公司</td><td> J105G </td><td colspan="3"></td><td></td></tr><tr><td> 15 </td><td> Diethrich微持针器</td><td></td><td> 11-510-20 </td><td colspan="3"> http://trimed-ltd.com/Products/Suture-Instruments/Micro-Needle-Holders-With-Tungsten-Carbide-Inserts/Ref-11-29.html </td><td></td></tr><tr><td> 16 </td><td> Temgesic（丁丙诺啡） </td><td>先灵葆雅</td><td></td><td colspan="3"></td><td></td></tr><tr><td> 17 </td><td> Stesolid（地西泮） </td><td>阿特维斯</td><td></td><td colspan="3"></td><td>又称安定</td></tr><tr><td> 18 </td><td> Pedamix </td><td>费森尤斯卡比</td><td></td><td colspan="3"> http://www.helsebiblioteket.no/retningslinjer/pediatri/mage-tarm-lever-ern%C3%A6ring/parenteral-ern%C3%A6ring </td><td></td></tr><tr><td> 19 </td><td> Klorhexidinsprit（洗必泰） </td><td> FresenIUS卡比</td><td> D08A C02 </td><td colspan="3"> http://www.felleskatalogen.no/medisin/klorhexidinsprit-fresenius-kabi-klorhexidinsprit-farget-fresenius-kabi-fresenius-kabi-560639 </td><td></td></tr></tbody></table> 表1列出的工具和设备在新生小鼠产生剪辑驱动脊髓压迫损伤。 <img alt="图1" src="/files/ftp_upload/53498/53498fig1.jpg" /> 图1.麻醉安装示意图，该示意图介绍了麻醉设置专为新生小鼠，与睡眠室初始麻醉和手术过程中持续麻醉鼻罩装置。 <img alt="图2" src="/files/ftp_upload/53498/53498fig2.jpg" /> 图2.主要工具和压缩剪辑。（A）在手术过程中使用的工具。所述编号对应于表1中 。（B和C）一种Yasargil临时瘤迷你夹与每个叶片手动下调到约150微米厚的前端中使用的注释。由聚乙烯管（ 表1）的切片的止动件放置在叶片中的一个，以防止夹子的全封闭。比例尺:2毫米。应用:夹子涂药（＃12 A）; ST:塞<a href="https://www.jove.com/files/ftp_upload/53498/53498fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/53498/53498fig3.jpg" /> 图3为地标新生ICR小鼠脊髓水平的术前评估。（A）与奶白色的个P1 ICR小鼠侧面观<html> tomach。胃的延髓部分对应于T12-T13脊髓水平。 （B）的麻醉下P1 ICR小鼠以俯卧位。虽然越来越多的困难比在（A）的可视化，充满了牛奶肚子辨认。胃的延髓部分表示T12-T13脊髓水平。比例尺:0.5厘米<img alt="图4" src="/files/ftp_upload/53498/53498fig4.jpg" /> 图4.背椎板切除术。（A）椎旁肌解剖。请注意，在这个年龄段的棘突不发达。 （B）的薄剪刀椎板横向切片。椎板和硬膜之间的薄镊子的一个叶片的（C）的介绍。入口点由箭头示出。 （D）椎板切除。比例尺:2毫米。 文件/ ftp_upload / 53498 / 53498fig5.jpg"/&gt; 图5.组织学和P1脊髓压迫损伤后的行为结果。（A）在离震中受伤的距离不同脊髓节曙红和苏木素染色从受伤的小鼠（1天伤后）。 （B）前肢及后肢运动轨迹的代表痕迹损伤后或假控制椎板切除术后观察6小时。在顶部迹线代表从该动物的侧视图观察轨迹。在底部迹线代表从动物的腹侧观察轨迹。也1见。比例尺:250微米。 DH:背角; L，左; R:右; SCC:脊髓压迫; VH:腹角<a href="https://www.jove.com/files/ftp_upload/53498/53498fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <i毫克ALT ="图6"SRC ="/文件/ ftp_upload / 53498 / 53498fig6.jpg"/&gt; 图6.比较生长曲线。直方图显示体重增加腭裂和SCC损伤小鼠从出生后1天〜9日龄。

Watch this Scientific Journal Video about A Neonatal Mouse Spinal Cord Compression Injury Model at JoVE.com

A Neonatal Mouse Spinal Cord Compression Injury Model

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

新生儿小鼠脊髓压迫损伤模型

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

a-neonatal-mouse-spinal-cord-compression-injury-model

Research

JoVE Journal

Medicine

12.4K 次观看.  Oslo University Hospital. This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery. 

视频: A Neonatal Mouse Spinal Cord Compression Injury Model

Neural Stem Cell Transplantation in Experimental Contusive Model of Spinal Cord Injury

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can be used both to investigate the biological responses to injury and to test potential therapies. Contusion or compression injury delivered to the surgically exposed spinal cord are the most widely used models of the pathology. In this report the experimental contusion is performed by using the Infinite Horizon (IH) Impactor device, which allows the creation of a reproducible injury animal model through definition of specific injury parameters. Stem cell transplantation is commonly considered a potentially useful strategy for curing this debilitating condition. Numerous studies have evaluated the effects of transplanting a variety of stem cells. Here we demonstrate an adapted method for spinal cord injury followed by tail vein injection of cells in CD1 mice. In short, we provide procedures for: i) cell labeling with a vital tracer, ii) pre-operative care of mice, iii) execution of a contusive spinal cord injury, and iv) intravenous administration of post mortem neural precursors. This contusion model can be utilized to evaluate the efficacy and safety of stem cell transplantation in a regenerative medicine approach.

Spinal cord injury is a traumatic condition that causes severe morbidity and high mortality. In this work we describe in detail a contusion model of spinal cord injury in mice followed by a transplantation of neural stem cells.

在脊髓损伤的实验挫伤型号神经干细胞移植

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can ...

科研

医学

Calibrated Forceps Model of Spinal Cord Compression Injury

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The calibrated forceps model of compression injury is a convenient, low cost, and very reproducible animal model for SCI. We used a pair of modified forceps in accordance with the method published by Plemel et al. (2008) to laterally compress the spinal cord to a distance of 0.35 mm. In this video, we will demonstrate a dorsal laminectomy to expose the spinal cord, followed by compression of the spinal cord with the modified forceps. In the video, we will also address issues related to the care of paraplegic laboratory animals. This injury model produces mice that exhibit impairment in sensation, as well as impaired hindlimb locomotor function. Furthermore, this method of injury produces consistent aberrations in the pathology of the SCI, as determined by immunohistochemical methods. After watching this video, viewers should be able to determine the necessary supplies and methods for producing SCI of various severities in the mouse for studies on SCI and/or treatments designed to mitigate impairment after injury.

Spinal cord injury models should be highly reproducible. We demonstrate that the calibrated forceps compression model of spinal cord injury is an easy to use surgical method for generating reproducible injuries to the murine spinal cord.

脊髓压迫损伤的校准钳型

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The ...

A Tissue Displacement-based Contusive Spinal Cord Injury Model in Mice

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between experimental animals. Several contusive SCI models have been developed to produce injuries using different mechanisms. The severity of the SCI is based on the height that a given weight is dropped, the injury force, or the spinal cord displacement. In the current study, we introduce a novel mouse contusive SCI device, the Louisville Injury System Apparatus (LISA) impactor, which can create a displacement-based SCI with high injury velocity and accuracy. This system utilizes laser distance sensors combined with advanced software to produce graded and highly-reproducible injuries. We performed a contusive SCI at the 10th thoracic vertebral (T10) level in mice to demonstrate the step-by-step procedure. The model can also be applied to the cervical and lumbar spinal levels.

We introduce a tissue displacement-based contusive spinal cord injury model that can produce a consistent contusive spinal cord injury in adult mice.

基于组织位移的小鼠脊髓损伤模型

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between ...

Induction of Complete Transection-Type Spinal Cord Injury in Mice

Spinal cord injury (SCI) largely leads to irreversible and permanent loss of function, most commonly as a result of trauma. Several treatment options, such as cell transplantation methods, are being researched to overcome the debilitating disabilities arising from SCI. Most pre-clinical animal trials are conducted in rodent models of SCI. While rat models of SCI have been widely used, mouse models have received less attention, even though mouse models can have significant advantages over rat models. The small size of mice equates to lower animal maintenance costs than for rats, and the availability of numerous transgenic mouse models is advantageous for many types of studies. Inducing repeatable and precise injury in the animals is the primary challenge for SCI research, which in small rodents requires high-precision surgery. The transection-type injury model has been a commonly used injury model over the last decade for transplantation-based therapeutic research, however a standardized method for inducing a complete transection-type injury in mice does not exist. We have developed a surgical protocol for inducing a complete transection type injury in C57BL/6 mice at thoracic vertebral level 10 (T10). The procedure uses a small tip drill instead of rongeurs to precisely remove the lamina, after which a thin blade with rounded cutting edge is used to induce the spinal cord transection. This method leads to reproducible transection-type injury in small rodents with minimal collateral muscle and bone damage and therefore minimizes confounding factors, specifically where behavioral functional outcomes are analyzed.

This protocol describes how to create a precise laminectomy for induction of stable transection-type spinal cord injury in the mouse model, with minimal collateral damage for spinal cord injury research.

小鼠完全转型脊髓损伤的诱导

Spinal cord injury (SCI) largely leads to irreversible and permanent loss of function, most commonly as a result of trauma. Several treatment options, ...

神经科学

A Neuronal Apoptosis Model induced by Spinal Cord Compression in Rat

As a severe progressive degenerative disease, cervical spondylotic myelopathy (CSM) has a poor prognosis and is associated with physical pain, stiffness, motor or sensory dysfunction, and a high risk of spinal cord injury and acroparalysis. Thus, therapeutic strategies that promote efficient spinal cord regeneration in this chronic and progressive disease are urgently needed. Effective and reproducible animal spinal cord compression models are required to understand the complex biological mechanism underlying CSM. Most spinal cord injury models reflect acute and structural destructive conditions, whereas animal models of CSM present a chronic compression in the spinal cord. This paper presents a protocol to generate a rat spinal cord compression model, which was further evaluated by assessing the behavioral score and observing the compressed spinal cord region. The behavioral assessments showed decreased monitor motor disability, including joint movements, stepping ability, coordination, trunk stability, and limb muscle strength. Hematoxylin and eosin (H&#38;E) staining and immunostaining revealed considerable neuronal apoptosis in the compressed region of the spinal cord.

Here, we present a protocol to generate a rat spinal cord compression model, assess its behavioral score, and observe the compressed spinal cord region. The behavioral assessments showed decreased monitor motor disability. Hematoxylin and eosin staining and immunostaining revealed considerable neuronal apoptosis in the compressed region of the spinal cord.

As a severe progressive degenerative disease, cervical spondylotic myelopathy (CSM) has a poor prognosis and is associated with physical pain, stiffness, ...

Establishing a Mouse Contusion Spinal Cord Injury Model Based on a Minimally Invasive Technique

Using minimally invasive methods to model spinal cord injury (SCI) can minimize behavioral and histological differences between experimental animals, thereby improving the reproducibility of the experiments.
These methods need two requirements to be fulfilled: clarity of the surgical anatomical pathway and simplicity and convenience of the laboratory device. Crucially for the operator, a clear anatomical pathway provides minimally invasive exposure, which avoids additional damage to the experimental animal during the surgical procedures and allows the animal to maintain a consistent and stable anatomical morphology during the experiment.
In this study, the use of a novel integrated platform called the SCI coaxial platform for spinal cord injury in small animals to expose the T9 level spinal cord in a minimally invasive way and stabilize and immobilize the vertebra of mice using a vertebral stabilizer is researched, and, finally, a coaxial gravity impactor is used to contuse the spinal cord of mice to approach different degrees of T9 spinal cord injury. Finally, histological results are provided as a reference for the readers.

Minimally invasive techniques and a simple laboratory device improve the reproducibility of the spinal cord injury model by reducing operative damage to the experimental animals and allowing anatomical morphology maintenance. The method is worthwhile because the reliable results and reproducible procedure facilitate investigations of the mechanisms of disease reparation.

基于微创技术的小鼠挫伤脊髓损伤模型的建立

Using minimally invasive methods to model spinal cord injury (SCI) can minimize behavioral and histological differences between experimental animals, ...

Establishment of Central Cord Syndrome Model in C57BL/6J Mouse

Animal models of central cord syndrome (CCS) could substantially benefit preclinical research. Identifiable anatomical pathways can give minimally invasive exposure approaches and reduce extra injury to experimental animals during operation, enabling the maintenance of consistent and stable anatomical morphology during experiments to minimize behavioral and histological differences between individuals to improve the reproducibility of experiments. In this study, the C6 level spinal cord was exposed using a spinal cord injury coaxial platform (SCICP) and combination with a minimally invasive technique. With the assistance of a vertebral stabilizator, we fixed the vertebrae and compressed the spinal cords of C57BL/6J mice with 5 g/mm2 and 10 g/mm2 weights with SCICP to induce different degrees of C6 spinal cord injury. In line with the previous description of CCS, the results reveal that the lesion in this model is concentrated in the gray matter around the central cord, enabling further research into CCS. Finally, histological results are provided as a reference for the readers.

The present protocol simulating central cord syndrome (CCS) in mice has improved repeatability and minimized operation damage to the experimental animals, avoiding disrupting the anatomical structure excessively. The strategy in this study is advantageous because it allows for research into injury mechanisms by producing consistent results.

C57BL/6J小鼠中索综合征模型的建立

Animal models of central cord syndrome (CCS) could substantially benefit preclinical research. Identifiable anatomical pathways can give minimally ...

用于精确小鼠模型的脊髓损伤挫伤装置

Automated Impactor for Contusive Spinal Cord Injury Model in Mice

Spinal cord injury (SCI) due to traumatic injuries such as car accidents and falls is associated with permanent spinal cord dysfunction. Creation of contusion models of spinal cord injury by impacting the spinal cord results in similar pathologies to most spinal cord injuries in clinical practice. Accurate, reproducible, and convenient animal models of spinal cord injury are essential for studying spinal cord injury. We present a novel automated spinal cord injury contusion device for mice, the Guangzhou Jinan University smart spinal cord injury system, that can produce spinal cord injury contusion models with accuracy, reproducibility, and convenience. The system accurately produces models of varying degrees of spinal cord injury via laser distance sensors combined with an automated mobile platform and advanced software. We used this system to create three levels of spinal cord injury mice models, determined their Basso mouse scale (BMS) scores, and performed behavioral as well as staining assays to demonstrate its accuracy and reproducibility. We show each step of the development of the injury models using this device, forming a standardized procedure. This method produces reproducible spinal cord injury contusion mice models and reduces human manipulation factors via convenient handling procedures. The developed animal model is reliable for studying spinal cord injury mechanisms and associated treatment approaches.

作者聚焦：深入了解脊髓损伤研究的创新

Presented here is a novel automated spinal cord injury contusion device for mice, which can accurately produce spinal cord injury contusion models with varying degrees.

用于小鼠挫伤脊髓损伤模型的自动撞击器

Spinal cord injury (SCI) due to traumatic injuries such as car accidents and falls is associated with permanent spinal cord dysfunction. Creation of ...

新生儿小鼠脊髓压迫损伤模型

摘要

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此视频中的章节

在脊髓损伤的实验挫伤型号神经干细胞移植

脊髓压迫损伤的校准钳型

基于组织位移的小鼠脊髓损伤模型

小鼠完全转型脊髓损伤的诱导

A Neuronal Apoptosis Model induced by Spinal Cord Compression in Rat

基于微创技术的小鼠挫伤脊髓损伤模型的建立

C57BL/6J小鼠中索综合征模型的建立

用于小鼠挫伤脊髓损伤模型的自动撞击器

用于小鼠挫伤脊髓损伤模型的自动撞击器

用于小鼠挫伤脊髓损伤模型的自动撞击器