这项工作是由欧盟第七框架计划（FP7/2007-2013）根据编号246556赠款协议（欧洲项目RBUCE-UP）的资助，HandiMedEx分配由法国公共投资委员会。马塞尔Bonay由Chancellerie德社区学院巴黎（腿华先生），全宗德Dotation RECHERCHE恩桑特Respiratoire和中心D&#39;援助Respiratoire住所德法兰西岛（CARDIF）支持

该方案通过卓越（Paris Sud酒店，编号246556赠款协议大学）和大学德凡尔赛圣昆廷 - 烯伊夫林省的RBUCE-UP椅子的伦理委员会。 无菌手术器械1。准备<ol><li>请用实验室的洗涤剂手术器械。 </li><li>高压灭菌前手术器械。 </li><li>在外科会话，通过将提示在热珠灭菌10分钟，在180℃下2次手术之间消毒的工具。 </li></ol> 2，准备药物<ol><li>准备2×1毫升注射器预麻醉药物的鸡尾酒及术后药物。 </li><li>根据大鼠的重量，制备的预麻醉药物的注射器:卡洛芬（5毫克/千克），丁丙诺啡（50微克/千克），拜有利（5毫克/千克）和美托咪啶（0.5毫克/千克）。完成音量，以1毫升乳酸林格液。 </li><li>在准备另一种注射器拨回，麻醉前用药:阿替美唑（500微克/公斤）。 </li></ol>鼠年3。麻醉<ol><li>管理皮下注射到动物的在步骤2.2中所述的预麻醉药物溶液。然后，将动物放回笼子里，等到镇静效果出现。 </li><li>将大鼠成一个封闭的腔充满了5％异氟醚在100％O 2，并等到呼吸节律减慢（大约30秒）。然后，从腔室取出大鼠，并将其放置到插管表。 </li></ol> 4，经口气管插管<ol><li>趴在他的背上的动物，然后通过将连接到他的门牙表表带固定头部。 </li><li>用光纤灯，照亮了整个胸腔的空间。然后，放置喉镜（或做了一个自定义的，柔等人 20节）在动物的嘴。可视化的声带。 </li><li>下滑e和放置一个导气管进气管（声带之间）。滑动经口气管（为16G导管的大小）上的导向。 </li><li>除去导向，并检查与喉癌镜放置在气管管子的端部的水分的存在，证实了该管的气管中，而不是在食道的适当位置。 </li><li>该管连接至一个啮齿动物呼吸机（683啮齿动物呼吸机，哈佛仪器）和调整异氟烷至2％的浓度（在100％O 2）。 </li><li>固定经口气管外科胶带。 </li></ol> 5，脊柱外科<ol><li>将动物在手术加热板腹卧位，用鼻子指着90°角的外科医生。围绕保持37.5°C体温整个手术。 </li><li>剃了头发与肩胛骨之间的快船，取下头发用纱布。 </li><li>请用优碘皮肤，然后用70％的酒精。重复此步骤3次。 </li><li>脚趾捏在手术开始前进行，以确保适当的麻醉深度。 &#160;然后，在肩胛骨之间剪刀进行横向切开皮肤rostro-尾端。 </li><li>通过下面的肌腱，以防止任何出血切开acromiotrapezius肌肉rostro-尾端。然后将分离菱形肌访问spinalis肌肉（围绕脊椎）。 </li><li>从C1收回spinalis肌肉到C3椎体。在C2椎体是一个具有突出的隆起。 </li><li>通过用无菌棉签清洁围绕脊椎的背侧部分的肌肉。 </li><li>开始小心地取出C2的隆起带咬骨钳。然后，继续认真直到脊髓背侧暴露。确保椎板切除术是一种背半椎板切除术。请特别注意包围脊髓，并在此区域附近动脉硬脑膜。 </li><li>用＃55镊子，解剖ROSTRO-尾端沿C2硬脑膜，接下来继续横向上的每个头端和尾端的一面。 </li><li>海绵立脑脊液。 </li><li>根据该颈脊神经后根号2与microscissors一个横截面。验证用微解剖刀，该病变的程度是足够接近到达脊髓（参见图2A为受伤的背视图）的中线。如果没有，那么另外一个切口可以作出来完成的伤害。如有出血，用消毒棉签。要小心，不要去对侧，否则动物将无法从伤病中恢复，将有一个呼吸衰竭。 </li><li>缝合肌肉作为保护层和背面缝合皮肤。请用优碘饱和的无菌纱布伤口。 </li><li>关闭异氟醚蒸发器，并注入了逆转​​药（阿替美唑[500微克/公斤，即时通讯]），检查体温。 </li><li>当动物开始呼吸对呼吸机，从呼吸机上拔下气管套管，然后取出经口气管。将动物在加热笼进行恢复。 </li></ol> 6，手术后护理手术后，动物被持续监控，以确保可能恢复的最佳环境。抗生素（拜有利，5毫克/千克），消炎（卡洛芬，5毫克/千克）和丁丙诺啡（50微克/公斤）的药物给予，每12小时进行第2天手术后，以防止感染，减少发生手术后的疼痛。大鼠获得自由采食，以软的食物和水（或胶化水为第一手术后一天）。皮下液体可以用来防止脱水的前几个术后天。体重和食物摄入量是每天监测。他们的环境是丰富整个​​实验过程和时间伤后（双壳，管道在他们的笼子）。

<ol>
	<li>Frankel, H. L., 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=Long-term+survival+in+spinal+cord+injury:+a+fifty+year+investigation.">Long-term survival in spinal cord injury: a fifty year investigation.</a> Spinal Cord. 36, 266-274 (1998).</li><li>Ramer, M. S., Harper, G. P., Bradbury, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progress+in+spinal+cord+research+-+a+refined+strategy+for+the+International+Spinal+Research+Trust.">Progress in spinal cord research - a refined strategy for the International Spinal Research Trust.</a> Spinal Cord. 38, 449-472 (2000).</li><li>Zimmer, M. B., Nantwi, K., Goshgarian, H. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+spinal+cord+injury+on+the+respiratory+system:+basic+research+and+current+clinical+treatment+options.">Effect of spinal cord injury on the respiratory system: basic research and current clinical treatment options.</a> J Spinal Cord Med. 30, 319-330 (2007).</li><li>Mantilla, C. B., Sieck, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuromuscular+adaptations+to+respiratory+muscle+inactivity.">Neuromuscular adaptations to respiratory muscle inactivity.</a> Respir Physiol Neurobiol. 169, 133-140 (2009).</li><li>Goshgarian, H. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+crossed+phrenic+phenomenon+and+recovery+of+function+following+spinal+cord+injury.">The crossed phrenic phenomenon and recovery of function following spinal cord injury.</a> Respir Physiol Neurobiol. 169, 85-93 (2009).</li><li>Nantwi, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recovery+of+respiratory+activity+after+C2+hemisection+(C2HS):+involvement+of+adenosinergic+mechanisms.">Recovery of respiratory activity after C2 hemisection (C2HS): involvement of adenosinergic mechanisms.</a> Respir Physiol Neurobiol. 169, 102-114 (2009).</li><li>Sandhu, M. S., 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=Respiratory+recovery+following+high+cervical+hemisection.">Respiratory recovery following high cervical hemisection.</a> Respir Physiol Neurobiol. 169, 94-101 (2009).</li><li>Lane, M. A., Lee, K. Z., Fuller, D. D., Reier, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+circuitry+and+respiratory+recovery+following+spinal+cord+injury.">Spinal circuitry and respiratory recovery following spinal cord injury.</a> Respir Physiol Neurobiol. 169, 123-132 (2009).</li><li>Seeds, N. W., Akison, L., Minor, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+plasminogen+activator+in+spinal+cord+remodeling+after+spinal+cord+injury.">Role of plasminogen activator in spinal cord remodeling after spinal cord injury.</a> Respir Physiol Neurobiol. 169, 141-149 (2009).</li><li>Alilain, W. J., Horn, K. P., Hu, H., Dick, T. E., Silver, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+regeneration+of+respiratory+pathways+after+spinal+cord+injury.">Functional regeneration of respiratory pathways after spinal cord injury.</a> Nature. 475, 196-200 (2011).</li><li>Vinit, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cervical+spinal+cord+injuries+and+respiratory+insufficiency:+a+revolutionary+treatment.">Cervical spinal cord injuries and respiratory insufficiency: a revolutionary treatment.</a> Med Sci (Paris. 28, 33-36 (2012).</li><li>Kastner, A., Gauthier, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Are+rodents+an+appropriate+pre-clinical+model+for+treating+spinal+cord+injury?+Examples+from+the+respiratory+system).">Are rodents an appropriate pre-clinical model for treating spinal cord injury? Examples from the respiratory system).</a> Exp Neurol. 213, 249-256 (2008).</li><li>Vinit, S., Lovett-Barr, M. R., Mitchell, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intermittent+hypoxia+induces+functional+recovery+following+cervical+spinal+injury.">Intermittent hypoxia induces functional recovery following cervical spinal injury.</a> Physiol Neurobiol. 169, 210-217 (2009).</li><li>Porter, W. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Path+of+the+Respiratory+Impulse+from+the+Bulb+to+the+Phrenic+Nuclei.">The Path of the Respiratory Impulse from the Bulb to the Phrenic Nuclei.</a> J Physiol. 17, 455-485 .</li><li>Nicaise, C., 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=Phrenic+motor+neuron+degeneration+compromises+phrenic+axonal+circuitry+and+diaphragm+activity+in+a+unilateral+cervical+contusion+model+of+spinal+cord+injury.">Phrenic motor neuron degeneration compromises phrenic axonal circuitry and diaphragm activity in a unilateral cervical contusion model of spinal cord injury.</a> Exp Neurol. 235, 539-552 (2012).</li><li>Goshgarian, H. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+crossed+phrenic+phenomenon:+a+model+for+plasticity+in+the+respiratory+pathways+following+spinal+cord+injury.">The crossed phrenic phenomenon: a model for plasticity in the respiratory pathways following spinal cord injury.</a> J Appl Physiol. 94, 795-810 (2003).</li><li>Vinit, S., Gauthier, P., Stamegna, J. C., Kastner, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+cervical+lateral+spinal+cord+injury+results+in+long-term+ipsilateral+hemidiaphragm+paralysis.">High cervical lateral spinal cord injury results in long-term ipsilateral hemidiaphragm paralysis.</a> J Neurotrauma. 23, 1137-1146 (2006).</li><li>Fuller, D. D., Johnson, S. M., Johnson, R. A., Mitchell, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+cervical+spinal+sensory+denervation+reveals+ineffective+spinal+pathways+to+phrenic+motoneurons+in+the+rat.">Chronic cervical spinal sensory denervation reveals ineffective spinal pathways to phrenic motoneurons in the rat.</a> Neurosci Lett. 323, 25-28 (2002).</li><li>Dougherty, B. J., Lee, K. Z., Lane, M. A., Reier, P. J., Fuller, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+the+spontaneous+crossed-phrenic+phenomenon+to+inspiratory+tidal+volume+in+spontaneously+breathing+rats.">Contribution of the spontaneous crossed-phrenic phenomenon to inspiratory tidal volume in spontaneously breathing rats.</a> J Appl Physiol. 112, 96-105 (2012).</li><li>Jou, I. M., 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=Simplified+rat+intubation+using+a+new+oropharyngeal+intubation+wedge.">Simplified rat intubation using a new oropharyngeal intubation wedge.</a> J Appl Physiol. 89, 1766-1770 (2000).</li><li>Fuller, D. D., 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=Graded+unilateral+cervical+spinal+cord+injury+and+respiratory+motor+recovery.">Graded unilateral cervical spinal cord injury and respiratory motor recovery.</a> Respir Physiol Neurobiol. 165, 245-253 (2009).</li><li>Vinit, S., Windelborn, J. A., Mitchell, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lipopolysaccharide+attenuates+phrenic+long-term+facilitation+following+acute+intermittent+hypoxia.">Lipopolysaccharide attenuates phrenic long-term facilitation following acute intermittent hypoxia.</a> Respir Physiol Neurobiol. 176, 130-135 (2011).</li><li>Ahmad, F., Wang, M. Y., Levi, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypothermia+for+Acute+Spinal+Cord+Injury-A+Review.">Hypothermia for Acute Spinal Cord Injury-A Review.</a> World Neurosurg. , (2013).</li><li>Lovett-Barr, M. 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=Repetitive+intermittent+hypoxia+induces+respiratory+and+somatic+motor+recovery+after+chronic+cervical+spinal+injury.">Repetitive intermittent hypoxia induces respiratory and somatic motor recovery after chronic cervical spinal injury.</a> J Neurosci. 32, 3591-3600 (2012).</li><li>Minor, K. H., Akison, L. K., Goshgarian, H. G., Seeds, N. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+injury-induced+plasticity+in+the+mouse--the+crossed+phrenic+phenomenon.">Spinal cord injury-induced plasticity in the mouse--the crossed phrenic phenomenon.</a> Exp Neurol. 200, 486-495 (2006).</li><li>Baussart, B., Stamegna, J. C., Polentes, J., Tadie, M., Gauthier, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+model+of+upper+cervical+spinal+contusion+inducing+a+persistent+unilateral+diaphragmatic+deficit+in+the+adult+rat.">A new model of upper cervical spinal contusion inducing a persistent unilateral diaphragmatic deficit in the adult rat.</a> Neurobiol Dis. 22, 562-574 (2006).</li><li>Golder, F. J., 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=Breathing+patterns+after+mid-cervical+spinal+contusion+in+rats.">Breathing patterns after mid-cervical spinal contusion in rats.</a> Exp Neurol. 231, 97-103 (2011).</li><li>Lane, M. A., 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=Respiratory+function+following+bilateral+mid-cervical+contusion+injury+in+the+adult+rat.">Respiratory function following bilateral mid-cervical contusion injury in the adult rat.</a> Exp Neurol. 235, 197-210 (2012).</li><li>Vinit, S., 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=Axotomized+bulbospinal+neurons+express+c-Jun+after+cervical+spinal+cord+injury.">Axotomized bulbospinal neurons express c-Jun after cervical spinal cord injury.</a> Neuroreport. 16, 1535-1539 (2005).</li><li>Guenther, C. H., Windelborn, J. A., Tubon, T. C., Yin, J. C., Mitchell, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+atypical+PKC+expression+and+activity+in+the+phrenic+motor+nucleus+following+cervical+spinal+injury.">Increased atypical PKC expression and activity in the phrenic motor nucleus following cervical spinal injury.</a> Exp Neurol. 234, 513-520 (2012).</li><li>Mantilla, C. B., Gransee, H. M., Zhan, W. Z., Sieck, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motoneuron+BDNF/TrkB+signaling+enhances+functional+recovery+after+cervical+spinal+cord+injury.">Motoneuron BDNF/TrkB signaling enhances functional recovery after cervical spinal cord injury.</a> Exp Neurol. 247, 101-109 (2013).</li><li>Vinit, S., Darlot, F., Aoulaiche, H., Boulenguez, P., Kastner, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+expression+of+c-Jun+and+HSP27+in+axotomized+and+spared+bulbospinal+neurons+after+cervical+spinal+cord+injury.">Distinct expression of c-Jun and HSP27 in axotomized and spared bulbospinal neurons after cervical spinal cord injury.</a> J Mol Neurosci. 45, 119-133 (2011).</li><li>Windelborn, J. A., Mitchell, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glial+activation+in+the+spinal+ventral+horn+caudal+to+cervical+injury.">Glial activation in the spinal ventral horn caudal to cervical injury.</a> Respir Physiol Neurobiol. 180, 61-68 (2012).</li><li>Vinit, S., Stamegna, J. C., Boulenguez, P., Gauthier, P., Kastner, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Restorative+respiratory+pathways+after+partial+cervical+spinal+cord+injury:+role+of+ipsilateral+phrenic+afferents.">Restorative respiratory pathways after partial cervical spinal cord injury: role of ipsilateral phrenic afferents.</a> Eur J Neurosci. 25, 3551-3560 (2007).</li><li>Dougherty, B. J., 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=Recovery+of+inspiratory+intercostal+muscle+activity+following+high+cervical+hemisection.">Recovery of inspiratory intercostal muscle activity following high cervical hemisection.</a> Respir Physiol Neurobiol. 183, 186-192 (2012).</li></ol>

作者宣称，他们有没有竞争的财务权益。

使得C2损伤模型的技术难点 该C2损伤小鼠模型是研究呼吸后皮损神经可塑性一个有趣的工具。然而，需要产生一个可重复的和可靠的模型的步骤多，而​​且每一个可能的研究的结果产生影响。例如，在插管过程中，非常小心的是要采取自气管管可产生气管的炎症，其可以导致多种并发症，如阻塞性呼吸衰竭，除了最初的呼吸功能不全，由于损伤本身。此外，在?...

脊髓损伤是在人群中观察到的与戏剧性的发生率，如永久性瘫痪一个常见的​​损​​伤。然而，损伤的严重程度取决于初始损伤的水平和程度。呼吸衰竭是病死率以下上颈段脊髓损伤（SCI）1的首要原因。目前，唯一的治疗治疗是将病人下辅助通气。由于少数患者可以断奶了辅助通气2，由于其发生后皮损延迟自然恢复，需要开发新的创新型非侵入性的治疗是当务之急3。拥有一个良好的标准化的临床前模型来观察颈椎脊髓损伤对呼吸功能不全的影响，因此，研究的推定治疗策略的应用，是必不可少的。 在本技术文章中，我们描述了一个特定的临床前小鼠模型Ø在C2级别F呼吸障碍引起的局部颈脊髓损伤。这种模式是目前使用世界各地的一些实验室（综述:4-13）。然而，在手术过程中的细微差别可以在不同的调查中可以观察到产生这个特殊的宫颈损伤小鼠模型。一个C2脊髓损伤对呼吸输出的影响最早是在1895年14波特描述。颈椎半切引起膈运动神经元的传入神经 ​​阻滞从他们的中央驱动（位于rVRG在脑干， 图1A）上受伤的同侧，导致无声膈神经活动和随后的膈肌麻痹。对侧保持不变，并允许动物生存。不同于位于一个较低的脊柱节段（例如，在C 4级15挫伤性损伤）不同SCI，在两侧膈运动神经元核的完整性被保存下来。一个CERV后iCal的C2伤害，一些自发的活动可以在同侧（膈和隔膜）可以观察到由于它越过脊柱中线在段水平C3-C6（交叉膈途径，CPP， 图1B）对侧沉默突触通路的激活。菲，这是激活，根据定义，一个C2半切结合对侧phrenicotomy从而诱发同侧局部膈神经恢复，可发生从几小时到几周损伤后16-18。这个CPP途径对呼吸复苏的真正有益作用是有限的19，并进一步调查处理应当制定提高自发恢复3的大小。 该协议提供了一个强大的类型的临床前小鼠模型来研究呼吸后皮损可塑性各级（从学前教育和膈运动神经元，的interneurons，分子和cellula呼吸生理r，则前肢为例）的运动，以及一个模型来测试旨在改善以下C2分型颈脊髓损伤的呼吸和运动恢复侵入性和非侵入性的治疗策略。

<table border="0" cellpadding="0" cellspacing="0" width="573">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;width:572px;">Animal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Male Sprague Dawley Rat</td>
			<td>Janvier</td>
			<td></td>
			<td>225-250g</td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;">Surgical Instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Student Dumont #5 forceps</td>
			<td>Fine Science Tool</td>
			<td>91150-20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Student Standard Pattern Forceps</td>
			<td>Fine Science Tool</td>
			<td>91100-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mayo-Stille Scissors</td>
			<td>Fine Science Tool</td>
			<td>14013-15</td>
			<td>Curved</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Student Vannas Spring Scissors</td>
			<td>Fine Science Tool</td>
			<td>91500-09</td>
			<td>Straight</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Spring Scissors - 8 mm Blades</td>
			<td>Fine Science Tool</td>
			<td>15025-10</td>
			<td>Straight Blunt/Blunt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Friedman Pearson Rongeur</td>
			<td>Fine Science Tool</td>
			<td>16121-14</td>
			<td>Curved</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dissecting Knife - Fine Tip</td>
			<td>Fine Science Tool</td>
			<td>10055-12</td>
			<td>Straight</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Olsen-Hegar Needle Holder</td>
			<td>Fine Science Tool</td>
			<td>12002-14</td>
			<td>Serrated</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Weitlaner-Locktite Retractor</td>
			<td>Fine Science Tool</td>
			<td>17012-11</td>
			<td>2x3 Blunt</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Absorbable surgical sutures</td>
			<td>Centravet</td>
			<td>BYO001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;">Equipment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hot Bead Steriliser</td>
			<td>Fine Science Tool</td>
			<td>18000-45</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Catheter&#160;</td>
			<td>Centravet</td>
			<td>CAT188</td>
			<td>16 gauge</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Laryngoscope</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Guide wire</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Laryngeal mirror</td>
			<td>Centravet</td>
			<td>MIR011</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lactated Ringers</td>
			<td>Centravet</td>
			<td>RIN020</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Syringe</td>
			<td>Centravet</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Needle</td>
			<td>Centravet</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">O2</td>
			<td>Air Liquid</td>
			<td>I1001M20R2A001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">683 RodentT Ventilator 115/230V</td>
			<td>Havard Apparatus</td>
			<td>55-0000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stand-Alone Vaporizer</td>
			<td>WPI</td>
			<td>EZ-155</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Thin line heated bed</td>
			<td>WPI</td>
			<td>EZ-211</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Air canister</td>
			<td>WPI</td>
			<td>EZ-258</td>
			<td></td>
		</tr>
		<tr height="21">
			<td colspan="4" height="21" style="height:21px;">Drugs</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Carprofen</td>
			<td>Centravet</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rimadyl</td>
			<td>Centravet</td>
			<td>RIM011</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Buprenorphine</td>
			<td>Centravet</td>
			<td>BUP001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Baytril</td>
			<td>Centravet</td>
			<td>BAY001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dexmedetomidine</td>
			<td>Centravet</td>
			<td>DEX010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Atipamezole</td>
			<td>Centravet</td>
			<td>ANT201</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Betadine Solution</td>
			<td>Centravet</td>
			<td>VET002</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane</td>
			<td>Centravet</td>
			<td>VET066</td>
			<td></td>
		</tr>
	</tbody>
</table>

a murine model of cervical spinal cord injury to study post-lesional respiratory neuroplasticity

颈椎脊髓损伤引起永久性瘫痪，并经常导致呼吸窘迫。迄今为止，还没有有效的治疗方法已发展到提高/改善呼吸衰竭以下高位颈脊髓损伤（SCI）。这里我们提出了很高的SCI在宫颈2（C2），同分异构的水平来研究不同的后皮损呼吸神经可塑性小鼠的临床前模型。该技术包括在C2级的外科局部损伤，这将导致隔膜的偏瘫，由于从位于脑干呼吸中枢膈运动神经元的传入神经阻滞。受伤的对侧保持不变，并允许动物恢复。不像影响运动功能（在胸椎和腰椎水平）等的SCI，呼吸功能并不需要动物的动机和赤字/恢复的量化可以容易地进行（隔膜和膈神经记录秒，全身通风）。此临床前C2脊髓损伤模型是一种功能强大，实用，可靠的临床前模型来研究各种呼吸道和非呼吸道神经可塑性的事件在不同层次（分子生理学），并测试不同的推定治疗策略，可以改善呼吸脊髓损伤患者。

损伤的程度 成功与这个特殊的实验模型的可重复性依赖于每个机械手/外科医师的经验。呼吸恢复（膈神经活动和膈肌活动）的继C2损伤的后续量相关，其余幸免腹外侧白质21。由于损伤是"手工"和需要一些实践从外科医生，每个损伤的程度要由组织学技术（组织用多聚甲醛4％，冰冻组织切片，甲酚紫染色的固定）进行检查，以确定准确的尺寸的受损组织?...

Watch this Scientific Journal Video about A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity at JoVE.com

A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

呼吸衰竭是死亡的跟随颈脊髓损伤的首要原因。有呼吸衰竭引起的局部损伤宫颈可重复的，可量化的，可靠的临床前动物模型将有助于了解后续的呼吸和非呼吸神经可塑性，并允许测试假定的维修策略。

颈脊髓损伤的小鼠模型研究后皮损呼吸神经可塑性

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed ...

a-murine-model-cervical-spinal-cord-injury-to-study-post-lesional

Research

JoVE Journal

Medicine

13.4K 次观看.  Université de Versailles Saint-Quentin-en-Yvelines. 呼吸衰竭是死亡的跟随颈脊髓损伤的首要原因。有呼吸衰竭引起的局部损伤宫颈可重复的，可量化的，可靠的临床前动物模型将有助于了解后续的呼吸和非呼吸神经可塑性，并允许测试假定的维修策略。 

视频: A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

在大鼠脊髓损伤后的急性和慢性触觉测试

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

科研

医学

A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of spinal cord injury (SCI) in which the thoracic spinal cord is injured. Additionally, because most human cord injuries occur as the result of blunt, non-penetrating trauma (e.g. motor vehicle accident, sporting injury) where the spinal cord is violently struck by displaced bone or soft tissues, the majority of SCI researchers are of the opinion that the most clinically relevant injury models are those in which the spinal cord is rapidly contused.1 Therefore, an important step in the preclinical evaluation of novel treatments on their way to human translation is an assessment of their efficacy in a model of contusion SCI within the cervical spinal cord. Here, we describe the technical aspects and resultant anatomical and behavioral outcomes of an unilateral contusive model of cervical SCI that employs the Infinite Horizon spinal cord injury impactor.
Sprague Dawley rats underwent a left-sided unilateral laminectomy at C5. To optimize the reproducibility of the biomechanical, functional, and histological outcomes of the injury model, we contused the spinal cords using an impact force of 150 kdyn, an impact trajectory of 22.5&deg; (animals rotated at 22.5&deg;), and an impact location off of midline of 1.4 mm. Functional recovery was assessed using the cylinder rearing test, horizontal ladder test, grooming test and modified Montoya's staircase test for up to 6 weeks, after which the spinal cords were evaluated histologically for white and grey matter sparing.
The injury model presented here imparts consistent and reproducible biomechanical forces to the spinal cord, an important feature of any experimental SCI model. This results in discrete histological damage to the lateral half of the spinal cord which is largely contained to the ipsilateral side of injury. The injury is well tolerated by the animals, but does result in functional deficits of the forelimb that are significant and sustained in the weeks following injury. The cervical unilateral injury model presented here may be a resource to researchers who wish to evaluate potentially promising therapies prior to human translation.

A reliable and repeatable way to produce a cervical unilateral spinal cord injury using the Infinite Horizon impactor is described. The method takes advantage of a custom designed frame and clamp to stabilize the spine. The standardized procedure and biomechanical injury parameters result in sufficient and sustained injuries.

单侧颈脊髓损伤的挫伤模型，利用无限的地平线冲击

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of ...

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

控制宫颈裂伤损伤小鼠

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

挫伤模型脊髓严重损伤的保护作用

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment (RMCA) in Individuals with Chronic Spinal Cord Injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination is disrupted by spinal cord injury (SCI), control of respiratory muscles innervated below the injury level is compromised1,2 leading to respiratory muscle dysfunction and pulmonary complications. These conditions are among the leading causes of death in patients with SCI3. Standard pulmonary function tests that assess respiratory motor function include spirometrical and maximum airway pressure outcomes: Forced Vital Capacity (FVC), Forced Expiratory Volume in one second (FEV1), Maximal Inspiratory Pressure (PImax) and Maximal Expiratory Pressure (PEmax)4,5. These values provide indirect measurements of respiratory muscle performance6. In clinical practice and research, a surface electromyography (sEMG) recorded from respiratory muscles can be used to assess respiratory motor function and help to diagnose neuromuscular pathology. However, variability in the sEMG amplitude inhibits efforts to develop objective and direct measures of respiratory motor function6. Based on a multi-muscle sEMG approach to characterize motor control of limb muscles7, known as the voluntary response index (VRI)8, we developed an analytical tool to characterize respiratory motor control directly from sEMG data recorded from multiple respiratory muscles during the voluntary respiratory tasks. We have termed this the Respiratory Motor Control Assessment (RMCA)9. This vector analysis method quantifies the amount and distribution of activity across muscles and presents it in the form of an index that relates the degree to which sEMG output within a test-subject resembles that from a group of healthy (non-injured) controls. The resulting index value has been shown to have high face validity, sensitivity and specificity9-11. We showed previously9 that the RMCA outcomes significantly correlate with levels of SCI and pulmonary function measures. We are presenting here the method to quantitatively compare post-spinal cord injury respiratory multi-muscle activation patterns to those of healthy individuals.

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment RMCA in Individuals with Chronic Spinal Cord Injury

The purpose of this publication is to present our original work on a multi-muscle surface electromyographic approach to quantitatively characterize respiratory muscle activation patterns in individuals with chronic spinal cord injury using vector-based analysis.

评价呼吸肌慢性脊髓损伤个人使用呼吸电机控制评估（RMCA）

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

控制皮层影响模型创伤性脑损伤

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter connected to a radio transmitter into the femoral artery of rats that underwent a T4 spinal cord transection with or without grafting of embryonic brainstem-derived neural stem cells expressing green fluorescent protein. Compared to other methods such as cannula insertion or tail-cuff, telemetry is advantageous to continuously monitor blood pressure and heart rate in freely moving animals. It is also capable of long term multiple data acquisitions. In spinal cord injured rats, basal cardiovascular data under unrestrained condition and autonomic dysreflexia in response to colorectal distension were successfully recorded. In addition, cardiovascular parameters before and after SCI can be compared in the same rat if a transmitter is implanted before a spinal cord transection. One limitation of the described telemetry procedure is that implantation in the femoral artery may influence the blood supply to the ipsilateral hindlimb.

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

无线电遥测系统，以监测大鼠脊髓横断和胚胎神经干细胞移植在心血管功能

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. Clinically, more than 50% of SCI affect the cervical spine1. As a consequence of the primary injury, a cascade of secondary mechanisms including inflammation, apoptosis, and demyelination occur finally leading to tissue scarring and development of intramedullary cavities2,3. Both represent physical and chemical barriers to cell transplantation, integration, and regeneration. Therefore, shaping the inhibitory environment and bridging cavities to create a supportive milieu for cell transplantation and regeneration is a promising therapeutic target4. Here, a contusion/compression model of cervical SCI using an aneurysm clip is described. This model is more clinically relevant than other experimental models, since complete transection or ruptures of the cord are rare. Also in comparison to the weight drop model, which in particular damage the dorsum columns, circumferential compression of the spinal cord appears advantageous. Clip closing force and duration can be adjusted to achieve different injury severity. A ring spring facilitates precise calibration and constancy of clip force. Under physiological conditions, synthetic self-assembling peptides (SAP) self-assemble into nanofibers and thus, are appealing for application in SCI5. They can be injected directly into the lesion minimizing damage to the cord. SAPs are biocompatible structures erecting scaffolds to bridge intramedullary cavities and thus, equip the damaged cord for regenerative treatments. K2(QL)6K2 (QL6) is a novel SAP introduced by Dong et al.6 In comparison to other peptides, QL6 self-assembles into &#946;-sheets at neutral pH6.14 days after SCI, after the acute stage, SAPs are injected into the center of the lesion and neural precursor cells (NPC) are injected into adjacent dorsal columns. In order to support cell survival, transplantation is combined with continuous subdural administration of growth factors by osmotic micro pumps for 7 days.

Treating cervical spinal cord injury with both self-assembling peptides (SAP) and neural precursor cells (NPC), together with growth factors, is a promising approach to promote regeneration and recovery. A contusion/compression aneurysm clip rat model of cervical SCI and combined treatment involving SAP injection and NPC transplantation is established.

在实验颈脊髓损伤协同使用神经前体细胞，并自组装肽的

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. ...

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 ...

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

Photothrombosis引起的局部脑缺血脊髓损伤的小鼠模型

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...