Palahnuk Samantha B, Abdala Jonathan A, Gospodarev Vadim V, Wilson Christopher G
Center for Perinatal Biology, Department of Basic Sciences, Loma Linda University; Department of Biology, The College of New Jersey.
Center for Perinatal Biology, Department of Basic Sciences, Loma Linda University.
J Vis Exp. 2019 Mar 23(145). doi: 10.3791/58870.
Mammalian inspiratory rhythm is generated from a neuronal network in a region of the medulla called the preBötzinger complex (pBC), which produces a signal driving the rhythmic contraction of inspiratory muscles. Rhythmic neural activity generated in the pBC and carried to other neuronal pools to drive the musculature of breathing may be studied using various approaches, including en bloc nerve recordings and transverse slice recordings. However, previously published methods have not extensively described the brainstem-spinal cord dissection process in a transparent and reproducible manner for future studies. Here, we present a comprehensive overview of a method used to reproducibly cut rhythmically-active brainstem slices containing the necessary and sufficient neuronal circuitry for generating and transmitting inspiratory drive. This work builds upon previous brainstem-spinal cord electrophysiology protocols to enhance the likelihood of reliably obtaining viable and rhythmically-active slices for recording neuronal output from the pBC, hypoglossal premotor neurons (XII pMN), and hypoglossal motor neurons (XII MN). The work presented expands upon previous published methods by providing detailed, step-by-step illustrations of the dissection, from whole rat pup, to in vitro slice containing the XII rootlets.
哺乳动物的吸气节律由延髓中一个称为前包钦格复合体（pBC）的区域中的神经网络产生，该网络产生驱动吸气肌节律性收缩的信号。可以使用多种方法研究在pBC中产生并传递到其他神经元池以驱动呼吸肌肉组织的节律性神经活动，包括整体神经记录和横向切片记录。然而，先前发表的方法并未以透明且可重复的方式广泛描述用于未来研究的脑干-脊髓解剖过程。在此，我们全面概述了一种用于可重复切割包含产生和传递吸气驱动所需且足够的神经元回路的有节律活动的脑干切片的方法。这项工作建立在先前的脑干-脊髓电生理协议基础上，以提高可靠获得用于记录来自pBC、舌下运动前神经元（XII pMN）和舌下运动神经元（XII MN）的神经元输出的存活且有节律活动切片的可能性。通过提供从整个幼鼠到包含舌下神经根的体外切片的详细、逐步解剖图示，本文所呈现的工作在先前发表的方法基础上进行了扩展。
