This work is funded by NIH RO1 NS 40433 (K.J.J.).

执行所有的程序都是由印第安纳大学医学院实验动物管理和使用委员会的批准，并按照卫生指引研究院。 1.手术技巧<ol><li>这个过程通过使用无菌手套，仪器，并根据美国国立卫生研究院指南25的无菌外科领域中保持无菌技术。消毒高压灭菌他们开始在手术前的工具（ 见表特定试剂/设备的完整列表）。使用玻璃珠灭菌的操作过程中，以消毒的工具。 </li></ol> 2.麻醉和准备<ol><li>麻醉小鼠在麻醉箱以0.9升/分钟的氧气和2.5％的异氟醚用兽医异氟烷蒸发器系统的混合物。确保鼠标并没有改变身体姿势从包装盒中取出前作出回应。 </li><li>应用眼药膏谅解备忘录SE的眼睛以保护他们免受干燥。 </li><li>从箱气流切换到鼻锥。将鼠标正视在其左侧上的加热垫覆盖有手术垫和吸收长凳纸与它的鼻子和嘴的锥体内。连续监测小鼠的呼吸节奏和速​​度，并调整异氟烷水平根据需要（2.5之间 - 3％异氟醚）以维持麻醉的适当水平，并且使用脚趾捏反射来确认总镇静。 </li></ol> 3.手术方法<ol><li>对齐并与外科领域关注的实体镜。调整鼻锥和磁带下来，以便它位于沿视野的边缘。 </li><li>用鼠标卧在其左侧，用胶带将右耳到鼻锥体的边缘，在那里露出的切口将作出在耳后的区域。确保耳后静脉水平会穿过耳朵。需要注意的是T的正确位置他的动物和耳朵的录像都是为了快速找到面神经的关键。 </li><li>润湿毛发上，并用70％乙醇在耳朵后面，刮胡子使用剃刀或手术刀刀片的外科手术部位。预湿的皮草，使剃须在这个解剖位置比较容易。 </li><li>清洁与碘溶液，如聚维酮碘的外科擦洗（7.5％聚维酮碘），随后用70％乙醇的皮肤。重复该清洗2次以上，以彻底消毒的区域。 </li><li>确定在何处作出切口，从耳尾端跟踪耳后静脉的区域后方的耳突起。使用弹簧剪刀，制作4毫米的切口2 - 3毫米后的突起。 </li><li>通过解剖用钝性分离皮下脂肪和筋膜。避免直接切削用剪刀因为血管或肌肉组织很容易损坏。 </li><li>如果发生出血，将压力施加到手术部位用无菌棉签为至少30秒。如果发生显著流体损失，以达到的用25或27G的针的无菌0.9％盐溶液0.5毫升腹膜内注入小鼠。 </li><li>使用几个键标，副神经，耳道，和前二腹肌（下面描述），以定位面神经。剖析围绕这些地标，直到面神经的分支可视化。当它被发现的神经将显示为显著固体白色组织和一层筋膜附着到下面的结构。 <ol><li>发现副神经，从头骨的尾部部分，其行进到支配斜方肌，一旦皮下脂肪和筋膜已解剖。面神经是深副神经。 </li><li>发现软骨耳道看起来珍珠白，并且可以看出延髓面神经。 </li><li>发现前二腹肌的肌腹的位于顶部和caudal面神经。 </li></ol></li><li>如果面神经的主要分支的可视化，跟踪他们背来找到自己的出身，从茎乳孔。用细尖杜蒙钳＃四十五分之五保持手术部位的开放，推进下面的神经路径的春天剪刀的提示，然​​后将镊子背侧，以保持新的先进地区开放。 </li><li>可视面神经与颧骨，颊的躯干，并且在这一点上下颌缘分支。 注:颞支会发现接近孔。边际下颌神经分支到其上，下部分更靠近钳口，从而那些神经分支将不会在这个水平可见的。 <ol><li>如果执行神经切断，轻轻稳定神经的精尖钳和切割​​神经与弹簧剪刀。避免将过多的牵引神经与血管钳，以防止avulsing从脑干神经。推树桩相互远离，或切割并移除远端神经的一部分，以确保没有重新连接可以发生。 </li><li>如果执行一个挤压伤，利用蒙特＃四十五分之五镊子压缩神经为30秒，使用恒定压力以切断所有轴突，则在垂直于第一粉碎部位的第二角度重复此压碎。避免将压力可变数量的30秒美眉期间，否则伤害将是动物之间的不一致。 </li></ol></li></ol> 4.关闭和恢复<ol><li>重新定位的脂肪和肌肉在底层结构。 </li><li>近似​​切口的边缘，并使用7.5毫米伤口夹闭合伤口。缝线或胶水还可以接受的伤口闭合。手术后止痛药可以在此时提供。 </li><li>取下鼠标的耳朵磁带。关掉异氟烷流动，并允许小鼠呼吸纯氧为30秒至1分钟。 PL王牌鼠标在一个空笼没有被褥从麻醉中恢复。 </li><li>当鼠标被回收，检查其面瘫的验证标志的行为。胡须就会瘫痪和角度背对着脸颊，鼻子就会偏离，并且眼睛也不会因空气的粉扑闪烁。 </li><li>房子动物联合手术后，如果他们是女性。避免住房雄性小鼠联合，因为他们更积极，而且往往强行删除其cagemate的伤口剪辑，从而导致感染。如有必要，提供手术后止痛剂，此时，。 </li><li>监测小鼠，每天一次，连续几天的操作后，以确保没有感染或其他并发症发生术后。手术后10天，如果他们没有倒下了自己 - 删除伤口片段7。 </li><li>适用于润滑眼膏，以患眼每天以防止角膜并发症，无论是直到眨眼反射是重覆盖或直到安乐死。 </li></ol>

<ol><li>Kaufman, M., Bard, J. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aKaufman%2C+M+%22The+Anatomical+Basis+of+Mouse+Development%22">The Anatomical Basis of Mouse Development</a>. , Elsevier. New York, NY. (1999).</li>
<li>Ashwell, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+adult+mouse+facial+nerve+nucleus%3A+morphology+and+musculotopic+organization%5BTitle%5D)+AND+%22Journal+of+Anatomy%22%5BJournal%5D)">The adult mouse facial nerve nucleus: morphology and musculotopic organization.</a> Journal of Anatomy. 135, 531-538 (1982).</li>
<li>Sunderland, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+classification+of+peripheral+nerve+injuries+producing+loss+of+function%5BTitle%5D)+AND+%22Brain+%3A+A+Journal+Of+Neurology%22%5BJournal%5D)">A classification of peripheral nerve injuries producing loss of function.</a> Brain : A Journal Of Neurology. 74, 491-516 (1951).</li>
<li>Beahrs, T., Tanzer, L., Sanders, V. M., Jones, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Functional+recovery+and+facial+motoneuron+survival+are+influenced+by+immunodeficiency+in+crush-axotomized+mice%5BTitle%5D)+AND+%22Experimental+Neurology%22%5BJournal%5D)">Functional recovery and facial motoneuron survival are influenced by immunodeficiency in crush-axotomized mice.</a> Experimental Neurology. 221, 225-230 (2010).</li>
<li>Mesnard, N. A., Haulcomb, M. M., Tanzer, L., Sanders, V., Jones, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Delayed+functional+recovery+in+presymptomatic+mSOD1G93A+mice+following+facial+nerve+crush+axotomy%5BTitle%5D)+AND+%22Journal+of+Neurodegeneration+%26+Regeneration%22%5BJournal%5D)">Delayed functional recovery in presymptomatic mSOD1G93A mice following facial nerve crush axotomy.</a> Journal of Neurodegeneration & Regeneration. 4, 21-25 (2013).</li>
<li>Komiyama, M., Shibata, H., Suzuki, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Somatotopic+representation+of+facial+muscles+within+the+facial+nucleus+of+the+mouse.+A+study+using+the+retrograde+horseradish+peroxidase+and+cell+degeneration+techniques%5BTitle%5D)+AND+%22Brain+Behav+Evol%22%5BJournal%5D)">Somatotopic representation of facial muscles within the facial nucleus of the mouse. A study using the retrograde horseradish peroxidase and cell degeneration techniques.</a> Brain Behav Evol. 24, 144-151 (1984).</li>
<li>Canh, M. Y., Serpe, C. J., Sanders, V., Jones, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((CD4%28%2B%29+T+cell-mediated+facial+motoneuron+survival+after+injury%3A+Distribution+pattern+of+cell+death+and+rescue+throughout+the+extent+of+the+facial+motor+nucleus%5BTitle%5D)+AND+%22Journal+of+Neuroimmunology%22%5BJournal%5D)">CD4(+) T cell-mediated facial motoneuron survival after injury: Distribution pattern of cell death and rescue throughout the extent of the facial motor nucleus.</a> Journal of Neuroimmunology. 181, 93-99 (2006).</li>
<li>Isokawa-Akesson, M., Komisaruk, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Difference+in+projections+to+the+lateral+and+medial+facial+nucleus%3A+anatomically+separate+pathways+for+rhythmical+vibrissa+movement+in+rats%5BTitle%5D)+AND+%22Exp+Brain+Res%22%5BJournal%5D)">Difference in projections to the lateral and medial facial nucleus: anatomically separate pathways for rhythmical vibrissa movement in rats.</a> Exp Brain Res. 65, 385-398 (1987).</li>
<li>Streit, W., Kreutzberg, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Response+of+endogenous+glial+cells+to+motor+neuron+degeneration+induced+by+toxic+ricin%5BTitle%5D)+AND+%22The+Journal+of+Comparative+Neurology%22%5BJournal%5D)">Response of endogenous glial cells to motor neuron degeneration induced by toxic ricin.</a> The Journal of Comparative Neurology. 268, 248-263 (1988).</li>
<li>Serpe, C. J., Tetzlaff, J. E., Coers, S., Sanders, V., Jones, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Functional+recovery+after+facial+nerve+crush+is+delayed+in+severe+combined+immunodeficient+mice%5BTitle%5D)+AND+%22Brain%2C+Behavior%2C+And+Immunity%22%5BJournal%5D)">Functional recovery after facial nerve crush is delayed in severe combined immunodeficient mice.</a> Brain, Behavior, And Immunity. 16, 808-812 (2002).</li>
<li>Lal, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Electrical+stimulation+facilitates+rat+facial+nerve+recovery+from+a+crush+injury.+Otolaryngology--Head+And+Neck+Surgery%5BTitle%5D)+AND+%22Official+Journal+Of+American+Academy+Of+Otolaryngology-Head+And+Neck+Surgery%22%5BJournal%5D)">Electrical stimulation facilitates rat facial nerve recovery from a crush injury. Otolaryngology--Head And Neck Surgery.</a> Official Journal Of American Academy Of Otolaryngology-Head And Neck Surgery. 139, 68-73 (2008).</li>
<li>Tomov, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+Example+of+Neural+Plasticity+Evoked+by+Putative+Behavioral+Demand+and+Early+Use+of+Vibrissal+Hairs+after+Facial+Nerve+Transection%5BTitle%5D)+AND+%22Experimental+Neurology%22%5BJournal%5D)">An Example of Neural Plasticity Evoked by Putative Behavioral Demand and Early Use of Vibrissal Hairs after Facial Nerve Transection.</a> Experimental Neurology. 178, 207-218 (2002).</li>
<li>Skouras, E., Angelov, D. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Experimental+studies+on+post-transectional+facial+nerve+regrowth+and+functional+recovery+of+paralyzed+muscles+of+the+face+in+rats+and+mice%5BTitle%5D)+AND+%22Anatomy+%28International+Journal+of+Experimental+and+Clinical+Anatomy%29%22%5BJournal%5D)">Experimental studies on post-transectional facial nerve regrowth and functional recovery of paralyzed muscles of the face in rats and mice.</a> Anatomy (International Journal of Experimental and Clinical Anatomy). 4, 1-27 (2010).</li>
<li>Xin, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((IL-10+within+the+CNS+is+necessary+for+CD4%2B+T+cells+to+mediate+neuroprotection%29%5BTitle%5D)+AND+%22Brain%2C+Behavior%2C+And+Immunity%22%5BJournal%5D)">IL-10 within the CNS is necessary for CD4+ T cells to mediate neuroprotection).</a> Brain, Behavior, And Immunity. 25, 820-829 (2011).</li>
<li>Mesnard, N. A., Sanders, V. M., Jones, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Differential+gene+expression+in+the+axotomized+facial+motor+nucleus+of+presymptomatic+SOD1+mice%5BTitle%5D)+AND+%22The+Journal+of+Comparative+Neurology%22%5BJournal%5D)">Differential gene expression in the axotomized facial motor nucleus of presymptomatic SOD1 mice.</a> The Journal of Comparative Neurology. 519, 3488-3506 (2011).</li>
<li>Mesnard, N. A., Alexander, T. D., Sanders, V. M., Jones, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Use+of+laser+microdissection+in+the+investigation+of+facial+motoneuron+and+neuropil+molecular+phenotypes+after+peripheral+axotomy%5BTitle%5D)+AND+%22Experimental+Neurology%22%5BJournal%5D)">Use of laser microdissection in the investigation of facial motoneuron and neuropil molecular phenotypes after peripheral axotomy.</a> Experimental Neurology. 225, 94-103 (2010).</li>
<li>Franchi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Changes+in+motor+representation+related+to+facial+nerve+damage+and+regeneration+in+adult+rats%5BTitle%5D)+AND+%22Experimental+Brain+Research%22%5BJournal%5D)">Changes in motor representation related to facial nerve damage and regeneration in adult rats.</a> Experimental Brain Research. 135, 53-65 (2000).</li>
<li>Munera, A., Cuestas, D. M., Troncoso, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Peripheral+facial+nerve+lesions+induce+changes+in+the+firing+properties+of+primary+motor+cortex+layer+5+pyramidal+cells%5BTitle%5D)+AND+%22Neuroscience%22%5BJournal%5D)">Peripheral facial nerve lesions induce changes in the firing properties of primary motor cortex layer 5 pyramidal cells.</a> Neuroscience. 223, 140-151 (2012).</li>
<li>Liu, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Hereditary+absence+of+complement+C5+in+adult+mice+influences+Wallerian+degeneration%2C+but+not+retrograde+responses%2C+following+injury+to+peripheral+nerve%5BTitle%5D)+AND+%22Journal+of+the+Peripheral+Nervous+System%22%5BJournal%5D)">Hereditary absence of complement C5 in adult mice influences Wallerian degeneration, but not retrograde responses, following injury to peripheral nerve.</a> Journal of the Peripheral Nervous System. 4, 123-133 (1999).</li>
<li>Ferri, C., Moore, F., Bisby, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+facial+nerve+injury+on+mouse+motoneurons+lacking+the+p75+low-affinity+neurotrophin+receptor%5BTitle%5D)+AND+%22Journal+of+Neurobiology%22%5BJournal%5D)">Effects of facial nerve injury on mouse motoneurons lacking the p75 low-affinity neurotrophin receptor.</a> Journal of Neurobiology. 34, 1-9 (1997).</li>
<li>Zhou, R. Y., Xu, J., Chi, F. L., Chen, L. H., Li, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Differences+in+sensitivity+to+rocuronium+among+orbicularis+oris+muscles+innervated+by+normal+or+damaged+facial+nerves+and+gastrocnemius+muscle+innervated+by+somatic+nerve+in+rats%3A+combined+morphological+and+functional+analyses%5BTitle%5D)+AND+%22The+Laryngoscope%22%5BJournal%5D)">Differences in sensitivity to rocuronium among orbicularis oris muscles innervated by normal or damaged facial nerves and gastrocnemius muscle innervated by somatic nerve in rats: combined morphological and functional analyses.</a> The Laryngoscope. 122, 1831-1837 (2012).</li>
<li>Jinno, S., Yamada, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Using+comparative+anatomy+in+the+axotomy+model+to+identify+distinct+roles+for+microglia+and+astrocytes+in+synaptic+stripping%5BTitle%5D)+AND+%22Neuron+Glia+Biology%22%5BJournal%5D)">Using comparative anatomy in the axotomy model to identify distinct roles for microglia and astrocytes in synaptic stripping.</a> Neuron Glia Biology. 7, 55-66 (2011).</li>
<li>Jones, K. J., Serpe, C. J., Byram, S. C., Deboy, C. A., Sanders, V. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Role+of+the+immune+system+in+the+maintenance+of+mouse+facial+motoneuron+viability+after+nerve+injury%5BTitle%5D)+AND+%22Brain%2C+Behavior%2C+And+Immunity%22%5BJournal%5D)">Role of the immune system in the maintenance of mouse facial motoneuron viability after nerve injury.</a> Brain, Behavior, And Immunity. 19, 12-19 (2005).</li>
<li>Moran, L. B., Graeber, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+facial+nerve+axotomy+model%5BTitle%5D)+AND+%22Brain+research.+Brain+research%22%5BJournal%5D)">The facial nerve axotomy model.</a> Brain research. Brain research. 44, 154-178 (2004).</li>
<li>Council, N. R. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aCouncil%2C+NR+%22Guide+for+the+Care+and+Use+of+Laboratory+Animals%3A+Eighth+Edition%22">Guide for the Care and Use of Laboratory Animals: Eighth Edition</a>. , The National Academies Press. New York, NY. (2011).</li>
<li>Serpe, C. J., Kohm, A. P., Huppenbauer, C. B., Sanders, V., Jones, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Exacerbation+of+Facial+Motoneuron+Loss+after+facial+nerve+transection+in+severe+combined+immunodeficient+%28scid%29+mice%5BTitle%5D)+AND+%22Neuroscience%22%5BJournal%5D)">Exacerbation of Facial Motoneuron Loss after facial nerve transection in severe combined immunodeficient (scid) mice.</a> Neuroscience. 19, (1999).</li>
<li>Mesnard-Hoaglin, N. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((SOD1%28G93A%29+transgenic+mouse+CD4%28%2B%29+T+cells+mediate+neuroprotection+after+facial+nerve+axotomy+when+removed+from+a+suppressive+peripheral+microenvironment%5BTitle%5D)+AND+%22Brain%2C+Behavior%2C+And+Immunity%22%5BJournal%5D)">SOD1(G93A) transgenic mouse CD4(+) T cells mediate neuroprotection after facial nerve axotomy when removed from a suppressive peripheral microenvironment.</a> Brain, Behavior, And Immunity. 40, 55-60 (2014).</li>
<li>Wang, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Establishment+and+assessment+of+the+perinatal+mouse+facial+nerve+axotomy+model+via+a+subauricular+incision+approach%5BTitle%5D)+AND+%22Experimental+Biology+And+Medicine%22%5BJournal%5D)">Establishment and assessment of the perinatal mouse facial nerve axotomy model via a subauricular incision approach.</a> Experimental Biology And Medicine. 237, 1249-1255 (2012).</li>
<li>Sharma, N., Moeller, C. W., Marzo, S. J., Jones, K. J., Foecking, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Combinatorial+treatments+enhance+recovery+following+facial+nerve+crush%5BTitle%5D)+AND+%22The+Laryngoscope%22%5BJournal%5D)">Combinatorial treatments enhance recovery following facial nerve crush.</a> The Laryngoscope. 120, 1523-1530 (2010).</li>
<li>Lieberman, D. M., Jan, T. A., Ahmad, S. O., Most, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+corticosteroids+on+functional+recovery+and+neuron+survival+after+facial+nerve+injury+in+mice%5BTitle%5D)+AND+%22Archives+of+Facial+Plastic+Surgery%22%5BJournal%5D)">Effects of corticosteroids on functional recovery and neuron survival after facial nerve injury in mice.</a> Archives of Facial Plastic Surgery. 13, 117-124 (2011).</li>
<li>Serpe, C. J., Coers, S., Sanders, V. M., Jones, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((CD4%2B+T%2C+but+not+CD8%2B+or+B%2C+lymphocytes+mediate+facial+motoneuron+survival+after+facial+nerve+transection%5BTitle%5D)+AND+%22Brain%2C+Behavior%2C+And+Immunity%22%5BJournal%5D)">CD4+ T, but not CD8+ or B, lymphocytes mediate facial motoneuron survival after facial nerve transection.</a> Brain, Behavior, And Immunity. 17, 393-402 (2003).</li>
<li>Haulcomb, M. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Axotomy-induced+target+disconnection+promotes+an+additional+death+mechanism+involved+in+motoneuron+degeneration+in+ALS+transgenic+mice%5BTitle%5D)+AND+%22The+Journal+of+Comparative+Neurology%22%5BJournal%5D)">Axotomy-induced target disconnection promotes an additional death mechanism involved in motoneuron degeneration in ALS transgenic mice.</a> The Journal of Comparative Neurology. , (2014).</li>
<li>Bauder, A. R., Ferguson, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Reproducible+mouse+sciatic+nerve+crush+and+subsequent+assessment+of+regeneration+by+whole+mount+muscle+analysis%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments+%3A+JoVE%22%5BJournal%5D)">Reproducible mouse sciatic nerve crush and subsequent assessment of regeneration by whole mount muscle analysis.</a> Journal of Visualized Experiments : JoVE. (60), (2012).</li>
<li>Richner, M., Bjerrum, O. J., Nykjaer, A., Vaegter, C. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+spared+nerve+injury+%28SNI%29+model+of+induced+mechanical+allodynia+in+mice%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments+%3A+JoVE%22%5BJournal%5D)">The spared nerve injury (SNI) model of induced mechanical allodynia in mice.</a> Journal of Visualized Experiments : JoVE. (54), (2011).</li>
</ol>

The authors have nothing to disclose.

The critical step for this protocol is positioning the mouse properly before surgery is begun. If the mouse is not lying flat on its side, the ear is not taped at the correct angle, or the incision is made in an incorrect location, then finding the facial nerve becomes much more difficult. When this technique is mastered, surgeries will take only minutes per mouse. 
Either sutures, glue, or wound clips can be used to close the wound. Wound clips are preferred because of the small size of incis...

许多周围神经损伤模型存在，但一个突出的运动神经元的研究是面神经干切断模型。面神经，也被称为脑神经七，起源于脑桥和支配的面部表情1,2的肌肉。在此手术协议，面神经在其出口处从茎乳孔露出，或者切断或粉碎。神经损伤的严重性可以分类以下桑德兰3的分类，其中区分基于所述轴突，神经内膜，神经束膜和外膜的完整性的损伤，这是结缔组织层，其依次绕轴突束包裹。在挤压伤（axonotmesis），轴突被切断，但神经束膜和外膜被保留。从面神经压榨完整的功能恢复发生在11天左右，因为完整的神经鞘作为管道内其轴突继续生长4,5。在另一方面，在该切伤（神经断裂），轴突和所有3结缔组织层被切断，并且整个远端神经必须重新生长，恢复肌肉的神经支配。往往是在人类患者神经切断伤进行神经外膜手术重新连接，但是复苏的结果是很少最佳。进一步的研究是需要理解为什么神经未能重新生长到它的目标和什么疗法可用于改善和加速再生处理。 有许多优势，使用面神经干切断模型研究神经损伤。一，面神经干切断过程快速，简便，具有很强的可重复性;和面部肌肉的合成瘫痪不会影响重要功能，是深受动物的耐受性。因为这是一个脑神经损伤模型，研究了运动神经元细胞体被简化，因为运动神经元驻留在一个相对同质种群在第Ë面部运动核在脑桥。人口不基于面部运动核中的亚核图案不同，因为有七个亚核每个特定于支配肌肉的特定组，因此响应于干切断亚核的差异可能会影响结果-2,6,7-。 面神经损伤模型的一个主要好处是，unaxotomized方可以作为一个配对的内部控制，因为神经支配是非常对称的，还有就是面部运动核8之间没有串扰。使用这种手术方法的另一个优点是，缺乏直接创伤至CNS或血脑屏障9中断。并发症，如出血过多和感染是罕见的这一程序。 各种分析可以被执行以评估对神经损伤的生理反应。眼眨眼反射和晶须活性的恢复可以作为一个行为测量功能恢复10,11。触须活动的录像是目前最有效的方法检测面神经支配12,13的恢复。安乐死后，可以在运动神经元胞体的面部运动核内进行脑干组织学分析。面部运动核被细分成七个亚核，每个特定的某些面部肌肉，从而为应对不同检查损伤2,6。面部运动神经元进行计数来定量细胞存活，或免疫组织化学可用于鉴定生物标志物和特定细胞群14。面部运动核可以利用激光捕获到神经损伤15,16的细胞反应的分子分析准确显微切割。面神经干切断的影响，可以在运动皮层17,18进行分析。此外，神经可以解剖研究沃勒变性或19轴突再生20和肌肉能够被去除以研究神经肌肉接头21。面神经切断，也可以用来研究所附中枢和外周神经胶质细胞22，目标肌肉21，以及免疫系统反应23。虽然取得了显着的研究面神经切断模型24，是必需的外周神经损伤的进一步研究，因为神经损伤是为患者一个显著问题和目前的治疗不能产生最佳的效果。该模型是一种强大的工具，用于检查到神经损伤的生理反应和分析神经再生疗法的有效性。

<table><tbody><tr><td>Stereo microscope</td><td>Leica</td><td>M60</td><td></td></tr><tr><td>Labeling tape</td><td>Fisher Scientific</td><td>15-952</td><td></td></tr><tr><td>Vannas-T&amp;uuml;bingen spring scissors - straight/sharp/8.5 cm/5 mm cutting edge</td><td>Fine Science Tools</td><td>15003-08</td><td>Sterilize before use</td></tr><tr><td>Dumont #5/45 forceps - standard tips/angled 45&amp;deg;/Dumoxel/11 cm</td><td>Fine Science Tools</td><td>11251-35</td><td>Sterilize before use</td></tr><tr><td>Michel suture clips - 7.5 mm x 1.75 mm</td><td>Fine Science Tools</td><td>12040-01</td><td>Described as &quot;wound clip&quot; in protocol, sterilize before use</td></tr><tr><td>Hagenbarth cross action wound clip applier 5&quot;</td><td>George Tiemann &amp;amp; Co</td><td>160-910</td><td>Used to apply wound clip, sterilize before use</td></tr><tr><td>Michel suture clip applicator &amp;amp; remover - For 7.5 mm clips</td><td>Fine Science Tools</td><td>12029-12</td><td>Used to remove wound clip</td></tr><tr><td>0.9% Sodium chloride injection, USP</td><td>Hospira</td><td>0409-4888-10</td><td></td></tr><tr><td>Betadine, 16 oz, with dispenser</td><td>Fisher Scientific</td><td>19-027132</td><td></td></tr><tr><td>70% Ethanol</td><td></td><td></td><td></td></tr><tr><td>Glass bead sterilizer</td><td></td><td></td><td></td></tr></tbody></table>

facial nerve axotomy in mice: a model to study motoneuron response to injury

这种手术协议的目标是从茎乳孔露出面神经，它支配的面部肌肉，在其出口处，要么切割或粉碎它来诱导外周神经损伤。这种手术的优点是它的简单性，高再现性，以及缺乏对从后续面神经麻痹生命功能或流动性的效果，因此导致相对温和的手术结果相比其他神经损伤的模型。使用颅神经损伤模型的一个主要优点是，运动神经元驻留在一个相对同质种群的面部运动核在脑桥，简化了运动神经元细胞体的研究。因为面部神经支配的对称性质和缺乏面部运动核之间的串扰的，操作可以单方面与unaxotomized侧作为配对的内部控制进行的。各种分析术后可以进行以驴S中的生理反应，其中的细节超出了本文的范围。例如，肌肉功能的恢复可以作为行为标记神经支配，或运动神经元可以被量化以测量细胞的存活。此外，运动神经元能够精确地使用激光显微切割进行分子分析捕获。因为面部神经切断是微创和良好的耐受性，它可以用于在多种转基因小鼠。另外，该手术模型可用于分析的外周神经损伤的治疗的有效性。面神经损伤提供了一种装置，用于调查不仅运动神经元，但中枢和外周神经胶质的微环境，免疫系统，和目标肌肉组织也响应。面神经损伤模型是被广泛接受的外周神经损伤模型充当一个强有力的工具用于研究神经损伤和再生。

之后进行的面部神经切断，运动神经元的损失发生的损伤的结果。损伤后运动神经元的生存取决于许多变量，如性别，年龄的动物在手术时间，并在其中运动神经元计数完成的时间点，与莫兰和Graeber回顾24和 ​​神野与山田回顾22既总结了运动神经元存活的数据。通常情况下，约86％的运动神经元的28天后干切断14,15,26生存。运动神经元损失的动力学在Serpe 等人进行?...

Watch this Scientific Journal Video about Facial Nerve Axotomy in Mice: A Model to Study Motoneuron Response to Injury at JoVE.com

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

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 ...

facial-nerve-axotomy-mice-model-to-study-motoneuron-response-to

Research

JoVE Journal

Medicine

12.8K 次观看.  Indiana University School of Medicine. 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. 

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

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. ...

科研

医学

Motor Nerve Transection and Time-lapse Imaging of Glial Cell Behaviors in Live Zebrafish

The nervous system is often described as a hard-wired component of the body even though it is a considerably fluid organ system that reacts to external stimuli in a consistent, stereotyped manner, while maintaining incredible flexibility and plasticity. Unlike the central nervous system (CNS), the peripheral nervous system (PNS) is capable of significant repair, but we have only just begun to understand the cellular and molecular mechanisms that govern this phenomenon. Using zebrafish as a model system, we have the unprecedented opportunity to couple regenerative studies with in vivo imaging and genetic manipulation. Peripheral nerves are composed of axons surrounded by layers of glia and connective tissue. Axons are ensheathed by myelinating or non-myelinating Schwann cells, which are in turn wrapped into a fascicle by a cellular sheath called the perineurium. Following an injury, adult peripheral nerves have the remarkable capacity to remove damaged axonal debris and re-innervate targets. To investigate the roles of all peripheral glia in PNS regeneration, we describe here an axon transection assay that uses a commercially available nitrogen-pumped dye laser to axotomize motor nerves in live transgenic zebrafish. We further describe the methods to couple these experiments to time-lapse imaging of injured and control nerves. This experimental paradigm can be used to not only assess the role that glia play in nerve regeneration, but can also be the platform for elucidating the molecular mechanisms that govern nervous system repair.

Although the peripheral nervous system (PNS) is capable of significant repair after injury, little is known about the cellular and molecular mechanisms that govern this phenomenon. Using live, transgenic zebrafish and a reproducible nerve transection assay, we can study dynamic glial cell behaviors during nerve degeneration and regeneration.

电机神经切断和时间推移成像胶质细胞行为在现场斑马鱼

The  nervous system is often described as a hard-wired component of the body even  though it is a considerably fluid organ system that reacts to external ...

神经科学

The Fibular Nerve Injury Method: A Reliable Assay to Identify and Test Factors That Repair Neuromuscular Junctions

The neuromuscular junction (NMJ) undergoes deleterious structural and functional changes as a result of aging, injury and disease. Thus, it is imperative to understand the cellular and molecular changes involved in maintaining and repairing NMJs. For this purpose, we have developed a method to reliably and consistently examine regenerating NMJs in mice. This nerve injury method involves crushing the common fibular nerve as it passes over the lateral head of the gastrocnemius muscle tendon near the knee. Using 70 day old female mice, we demonstrate that motor axons begin to reinnervate previous postsynaptic targets within 7 days post-crush. They completely reoccupy their previous synaptic areas by 12 days. To determine the reliability of this injury method, we compared reinnervation rates between individual 70 day old female mice. We found that the number of reinnervated postsynaptic sites was similar between mice at 7, 9, and 12 days post-crush. To determine if this injury assay can also be used to compare molecular changes in muscles, we examined levels of the gamma-subunit of the muscle nicotinic receptor (gamma-AChR) and the muscle-specific kinase (MuSK). The gamma-AChR subunit and MuSK to are highly upregulated following denervation and return to normal levels following reinnervation of NMJs. We found a close relationship between transcript levels for these genes and innervation status of muscles. We believe that this method will accelerate our understanding of the cellular and molecular changes involved in repairing the NMJ and other synapses.

We have developed a nerve injury method to reliably examine muscle reinnervation, and thus regeneration of neuromuscular junctions in mice. This technique involves injuring the common fibular nerve via a simple and highly reproducible surgery. Muscle reinnervation in then assessed by whole-mounting the extensor digitorum longus muscle.

腓神经损伤方法:可靠检测，以确定和测试因素修复神经肌肉接头

The neuromuscular junction (NMJ) undergoes deleterious structural and functional changes as a result of aging, injury and disease. Thus, it is imperative ...

Facial Nerve Surgery in the Rat Model to Study Axonal Inhibition and Regeneration

This protocol describes consistent and reproducible methods to study axonal regeneration and inhibition in a rat facial nerve injury model. The facial nerve can be manipulated along its entire length, from its intracranial segment to its extratemporal course. There are three primary types of nerve injury used for the experimental study of regenerative properties: nerve crush, transection, and nerve gap. The range of possible interventions is vast, including surgical manipulation of the nerve, delivery of neuroactive reagents or cells, and either central or end-organ manipulations. Advantages of this model for studying nerve regeneration include simplicity, reproducibility, interspecies consistency, reliable survival rates of the rat, and an increased anatomic size relative to murine models. Its limitations involve a more limited genetic manipulation versus the mouse model and the superlative regenerative capability of the rat, such that the facial nerve scientist must carefully assess time points for recovery and whether to translate results to higher animals and human studies. The rat model for facial nerve injury allows for functional, electrophysiological, and histomorphometric parameters for the interpretation and comparison of nerve regeneration. It thereby boasts tremendous potential toward furthering the understanding and treatment of the devastating consequences of facial nerve injury in human patients.

This protocol describes a reproducible approach to facial nerve surgery in the rat model, including descriptions of various inducible patterns of injury.

大鼠模型中的面部神经外科研究同氧抑制与再生

This protocol describes consistent and reproducible methods to study axonal regeneration and inhibition in a rat facial nerve injury model. The facial ...

<meta charset="utf8"></head><body>使用大鼠模型评估颅面 VML 损伤中的生物材料

Rodent Model of Masseter Volumetric Muscle Loss for Studying Bioengineering Materials

Craniofacial volumetric muscle loss (VML) injuries can occur as a result of severe trauma, surgical excision, inflammation, and congenital or other acquired conditions. Treatment of craniofacial VML involves surgical, functional muscle transfer. However, these procedures are unable to restore normal function, sensation, or expression, and more commonly, these conditions go untreated. Very little research has been conducted on skeletal muscle regeneration in animal models of craniofacial VML. This manuscript describes a rat model for the study of craniofacial VML injury and a protocol for the histological evaluation of biomaterials in the treatment of these injuries. Liquid hydrogel and freeze-dried scaffolds are applied at the time of surgical VML creation, and masseters are excised at terminal time points up to 12 weeks with high retention rates and negligible complications. Hematoxylin and eosin (HE), Masson's Trichrome, and immunohistochemical analysis are used to evaluate parameters of skeletal muscle regeneration as well as biocompatibility and immunomodulation. While we demonstrate the study of a hyaluronic-acid-based hydrogel, this model provides a means for evaluating subsequent iterations of materials in VML injuries.

<meta charset="utf8"></head><body>用于研究生物工程材料的咬肌体积损失的啮齿动物模型

Here, we describe a protocol for the surgical creation of a volumetric muscle loss (VML) injury in the rat masseter, providing a reproducible and accessible model for the study of craniofacial muscle injuries and their treatment using biomaterials such as the novel hydrogel.

用于研究生物工程材料的咬肌体积损失的啮齿动物模型

Craniofacial volumetric muscle loss (VML) injuries can occur as a result of severe trauma, surgical excision, inflammation, and congenital or other ...

生物工程

<meta charset="utf8"></head><body>小鼠耳长提肌电穿孔和神经损伤

Combined In Vivo Electroporation and Short&#45;Term Reinnervation of the Cranial Levator Auris Longus Skeletal Muscle

The neuromuscular junction (NMJ) is the peripheral synapse controlling the contraction of skeletal muscle fibers to allow the coordinated movement of many organisms. At the NMJ, a presynaptic motor axon terminal contacts a muscle postsynaptic domain and is covered by terminal Schwann cells. The integrity and function of the NMJ is compromised under several conditions, including aging, neuromuscular diseases, and traumatic injuries. To analyze the potential contribution of muscle-derived proteins to NMJ maintenance and regeneration, an in vivo gene transfer strategy has been combined with the denervation of the cranial levator auris longus (LAL) muscle after mechanical nerve injury. Previous findings showed that the forced expression of control fluorescent proteins does not alter NMJ organization or neurotransmission. This procedure aims to describe a detailed method of in vivo electroporation of the LAL muscle followed by transection or crushing of the specific branch of the facial nerve innervating cranial muscles, leading to NMJ denervation and reinnervation, respectively. The combination of these experimental strategies in the convenient LAL muscle constitutes an efficient method to study the potential contribution of muscle protein overexpression or silencing in the context of short-term NMJ reinnervation.

<meta charset="utf8"></head><body>作者聚焦：肌肉蛋白在神经损伤后神经肌肉接头的再生作用

Here, we present a protocol that combines in vivo electroporation and denervation of the cranial levator auris longus (LAL) muscle. This procedure enables the study of the potential role of muscle-derived proteins in the regeneration of the neuromuscular synapse.

颅提肌耳长骨骼肌的体内电穿孔和短期神经再支配相结合

The neuromuscular junction (NMJ) is the peripheral synapse controlling the contraction of skeletal muscle fibers to allow the coordinated movement of many ...

面神经干切断小鼠:模型，研究运动神经元损伤反应

摘要

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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

电机神经切断和时间推移成像胶质细胞行为在现场斑马鱼

腓神经损伤方法:可靠检测，以确定和测试因素修复神经肌肉接头

大鼠模型中的面部神经外科研究同氧抑制与再生

用于研究生物工程材料的咬肌体积损失的啮齿动物模型

用于研究生物工程材料的咬肌体积损失的啮齿动物模型

用于研究生物工程材料的咬肌体积损失的啮齿动物模型

颅提肌耳长骨骼肌的体内电穿孔和短期神经再支配相结合

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颅提肌耳长骨骼肌的体内电穿孔和短期神经再支配相结合