Franco Joy A, Kloefkorn Heidi E, Hochman Shawn, Wilkinson Katherine A
Department of Mechanical Engineering, San José State University.
J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida.
J Vis Exp. 2014 Sep 24(91):51948. doi: 10.3791/51948.
Muscle sensory neurons innervating muscle spindles and Golgi tendon organs encode length and force changes essential to proprioception. Additional afferent fibers monitor other characteristics of the muscle environment, including metabolite buildup, temperature, and nociceptive stimuli. Overall, abnormal activation of sensory neurons can lead to movement disorders or chronic pain syndromes. We describe the isolation of the extensor digitorum longus (EDL) muscle and nerve for in vitro study of stretch-evoked afferent responses in the adult mouse. Sensory activity is recorded from the nerve with a suction electrode and individual afferents can be analyzed using spike sorting software. In vitro preparations allow for well controlled studies on sensory afferents without the potential confounds of anesthesia or altered muscle perfusion. Here we describe a protocol to identify and test the response of muscle spindle afferents to stretch. Importantly, this preparation also supports the study of other subtypes of muscle afferents, response properties following drug application and the incorporation of powerful genetic approaches and disease models in mice.
支配肌梭和高尔基腱器官的肌肉感觉神经元编码本体感觉所必需的长度和力量变化。其他传入纤维监测肌肉环境的其他特征，包括代谢产物积累、温度和伤害性刺激。总体而言，感觉神经元的异常激活可导致运动障碍或慢性疼痛综合征。我们描述了分离成年小鼠趾长伸肌（EDL）及其神经，用于体外研究牵张诱发的传入反应。用吸电极从神经记录感觉活动，并可使用尖峰分类软件分析单个传入纤维。体外制备允许对感觉传入进行良好控制的研究，而没有麻醉或改变肌肉灌注的潜在干扰因素。在这里，我们描述了一种识别和测试肌梭传入纤维对牵张反应的方案。重要的是，这种制备方法还支持对其他肌肉传入纤维亚型、药物应用后的反应特性以及在小鼠中采用强大的基因方法和疾病模型的研究。
