Olmstead Deborah N, Mesnard-Hoaglin Nichole A, Batka Richard J, Haulcomb Melissa M, Miller Whitney M, Jones Kathryn J
Anatomy and Cell Biology, Indiana University School of Medicine; Research and Development Services, Richard L. Roudebush VA Medical Center.
Department of Anatomy and Cell Biology, University of Illinois, Chicago.
J Vis Exp. 2015 Feb 23(96):e52382. doi: 10.3791/52382.
The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and either cut or crush it to induce peripheral nerve injury. Advantages of this surgery are its simplicity, high reproducibility, and the lack of effect on vital functions or mobility from the subsequent facial paralysis, thus resulting in a relatively mild surgical outcome compared to other nerve injury models. A major advantage of using a cranial nerve injury model is that the motoneurons reside in a relatively homogenous population in the facial motor nucleus in the pons, simplifying the study of the motoneuron cell bodies. Because of the symmetrical nature of facial nerve innervation and the lack of crosstalk between the facial motor nuclei, the operation can be performed unilaterally with the unaxotomized side serving as a paired internal control. A variety of analyses can be performed postoperatively to assess the physiologic response, details of which are beyond the scope of this article. For example, recovery of muscle function can serve as a behavioral marker for reinnervation, or the motoneurons can be quantified to measure cell survival. Additionally, the motoneurons can be accurately captured using laser microdissection for molecular analysis. Because the facial nerve axotomy is minimally invasive and well tolerated, it can be utilized on a wide variety of genetically modified mice. Also, this surgery model can be used to analyze the effectiveness of peripheral nerve injury treatments. Facial nerve injury provides a means for investigating not only motoneurons, but also the responses of the central and peripheral glial microenvironment, immune system, and target musculature. The facial nerve injury model is a widely accepted peripheral nerve injury model that serves as a powerful tool for studying nerve injury and regeneration.
本手术方案的目的是在面神经从茎乳孔穿出时暴露支配面部肌肉组织的面神经,并将其切断或挤压以诱导周围神经损伤。该手术的优点包括操作简单、重复性高,以及后续面瘫对重要功能或活动能力无影响,因此与其他神经损伤模型相比,手术结果相对较轻。使用颅神经损伤模型的一个主要优点是运动神经元位于脑桥面神经运动核中相对同质的群体中,简化了对运动神经元细胞体的研究。由于面神经支配的对称性以及面神经运动核之间缺乏串扰,该手术可单侧进行,未切断轴突的一侧作为配对的内部对照。术后可进行多种分析以评估生理反应,其细节超出了本文的范围。例如,肌肉功能的恢复可作为再支配的行为标志物,或者可对运动神经元进行量化以测量细胞存活情况。此外,可使用激光显微切割精确捕获运动神经元以进行分子分析。由于面神经切断术微创且耐受性良好,它可用于多种基因修饰小鼠。而且,该手术模型可用于分析周围神经损伤治疗的有效性。面神经损伤不仅为研究运动神经元,还为研究中枢和外周神经胶质微环境、免疫系统及靶肌肉组织的反应提供了一种手段。面神经损伤模型是一种广泛接受的周围神经损伤模型,是研究神经损伤和再生的有力工具。