大鼠模型中的面部神经外科研究同氧抑制与再生
Summary
该协议描述了大鼠模型中面部神经手术的可重复方法，包括各种可诱导损伤模式的描述。
Abstract
该协议描述了研究大鼠面部神经损伤模型中的六角再生和抑制的一致和可重复的方法。面部神经可以沿其整个长度进行操纵，从颅内部分到外时过程。用于再生特性实验研究的主要神经损伤类型有三种：神经挤压、截断和神经间隙。可能的干预范围很广，包括神经手术操作、神经活性试剂或细胞的输送，以及中央或终末期器官操作。该模型在研究神经再生方面的优点包括简单性、可重复性、物种间一致性、大鼠的可靠生存率以及相对于鼠型增加解剖尺寸。其局限性涉及与小鼠模型相比，基因操作更为有限，大鼠的再生能力更胜一等，因此面部神经科学家必须仔细评估恢复的时间点，以及是否将结果转化为高等动物和人类研究。面部神经损伤大鼠模型允许功能性、电生理和组形参数，用于神经再生的解释和比较。因此，它具有巨大的潜力，进一步理解和治疗面部神经损伤在人类患者的破坏性后果。
Introduction
头部和颈部部位的颅神经损伤可继发先天性、传染性、特发性、致源性、创伤性、神经学、肿瘤或全身病因1。颅神经VII，或面部神经，通常受到影响。面部神经功能障碍的发生率可能很大，因为它每年影响每10万人20至30人2。面部神经的主要运动分支是时态、酶、布卡、边缘性腺和颈椎分支;根据所涉及的分支，后果可能包括口腔无能或流口水、角膜干燥、视觉场阻塞继为普托西斯、肌张力障碍或面部不对称22、3。3长期发病包括同步性现象，或一个面部肌肉群的非自愿运动，试图自愿收缩一个独特的面部肌肉群。眼腔同步性是最常见的异常再生作为面部神经损伤的后遗症，导致功能障碍，尴尬，自尊下降，生活质量低下3。对单个分支的伤害决定了有选择地受损的功能。
面部神经损伤的临床治疗效果不达标，需要进一步研究，以改善效果。类固醇可以缓解急性面部神经肿胀， 而肉毒杆菌是有用的天生运动;但是，在医生的军备中，主要的重建选择包括通过神经修复、替代或恢复33、4、5、64,5,6进行手术干预。根据面部神经损伤的类型，面部神经外科医生可能使用多种选择。对于简单的截断，神经再抗是有用的，而电缆移植修复更适合神经缺陷;为了恢复功能，外科医生可以选择静态或动态面部恢复手术。在许多面部神经损伤和随后的修复的情况下，即使在有经验的面部神经外科医生手中，最好的结果仍然导致持续的面部不对称和功能损害7。
这些不理想的结果刺激了对面部神经再生的广泛研究。广泛的关注主题包括完善和创新神经修复技术，确定各种神经再生因素的影响，以及评估特定神经抑制剂的潜力，以帮助对抗合成酶88，9，10，119,10,11的长期结果。虽然体外模型可用于评估一些有利于生长或抑制因子的特征，但最好通过可翻译的动物模型完成关于这一主题的真正转化研究。
决定使用哪种动物模型可能具有挑战性，因为研究人员已经利用了大型动物，如绵羊和小动物模型，如老鼠12，13。12,虽然大型动物模型提供理想的解剖可视化效果，但使用它们需要专用设备和人员，不易或容易获得。此外，推动一项研究来证明效果可能很高，而且可能不属于许多科学中心的可行范围。因此，小型动物模型被最常用的。小鼠模型可用于评估与面部神经手术相关的一些结果;然而，神经长度的有限会限制科学家模拟某些模式的能力，如大间隙损伤14。
因此，大鼠鼠原型已成为科学家可以执行创新外科手术或利用抑制或促进生长因子并评估各种结果参数效果的主力模型。大鼠面部神经解剖学是可预测和容易接近在一个可重复的方式。与鼠标模型相比，其规模更大，允许对各种手术缺陷进行建模，范围从简单的截面到5毫米的间隙15，16。15,这进一步允许在缺陷部位应用复杂的干预，包括局部因子的放置、因子的神经内注入，以及等体移植或桥17、18、19、20、21、22、23。17,18,19,20,21,22,23
大鼠的温顺性质，其可靠的解剖学，其有效神经再生的倾向，允许收集许多结果措施，以回应上述手术模式的伤害24。通过大鼠模型，面部神经科学家能够通过免疫组化学评估对损伤、神经和肌肉组学结果的电生理反应，通过跟踪颤垫的运动和评估眼闭合的功能结果，以及通过荧光或共聚焦显微镜，通过荧光或,共聚焦显微镜，包括11、22、23、25、26、27、28、29,27,28等29微微和宏观变化。11,22,2325,26因此，以下协议将概述对大鼠面部神经的手术方法以及可诱发的损伤模式。
Protocol
所有干预措施均严格按照国家卫生研究院（NIH）准则进行。该实验协议在实施前得到了密歇根大学机构动物护理和使用委员会（IACUC）的批准。使用了十周大的成年雌性斯普拉格-道利大鼠。 1. 手术前 确保手术前有适当的消毒手术器械、镇痛药、麻醉药物和氧气。有关完整列表，请参阅材料表。 2. 术前设置 确保有足够的工…
Representative Results
在最初的外科手术后，有两种主要的结果测量：活体动物的连续测量和需要牺牲动物的测量。连续测量的例子包括电生理测定，如复合肌肉动作势谱测量30，通过激光辅助或摄像手段9评估面部肌肉运动，甚至重复现场成像荧光转基因动物面部神经的再生31，32。,32图1演示…
Discussion
大鼠面部神经损伤模型已成为最通用的系统，评估神经营养因子，由于其手术可及性，分支模式，和生理意义27，29，33，34，35，36。27,29,33,34,35,36视频演示与转基因动物数据应用的结合，为神经再生现象的科学研究提供了新的可能。该模型允许对创?…
Declarações
The authors have nothing to disclose.
Acknowledgements
S.A.A.由美国面部整形和重建外科学会莱斯利·伯恩斯坦助学金计划资助。
Materials
| 1.8% isoflurane | VetOne | 13985-030-40 | |
| 11-0 nylon microsutures | AROSuture | TK-117038 | |
| 4-0 monocryl suture | VWR | 75982-084 | |
| Buprenorphine SR | ZooPharm | MIF 900-006 | |
| Carprofen | Sigma-Aldrich | MFCD00079028 | |
| Chlorhexidine | VWR | IC19135805 | |
| Jeweler forceps | VWR | 21909-458 | |
| Micro Weitlaner retractor | VWR | 82030-146 | |
| Micro-scissors | VWR | 100492-348 | |
| Mini tenotomy scissors | VWR | 89023-522 | |
| Number 15 scalpel blade | VWR | 102097-834 | |
| Operating microscope | Leica | ||
| Petrolatum eye gel | Pharmaderm | B002LUWBEK | |
| Sterile water | VWR | 89125-834 | |
| Tissue adhesive | Vetbond, 3M | NC9259532 | |
| Water conductor pad | Aqua Relief System | ARS2000B |
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Ali, S. A., Stebbins, A. W., Hanks, J. E., Kupfer, R. A., Hogikyan, N. D., Feldman, E. L., Brenner, M. J. Facial Nerve Surgery in the Rat Model to Study Axonal Inhibition and Regeneration. J. Vis. Exp. (159), e59224, doi:10.3791/59224 (2020).