Analysis of Neuro-Neuronal Synapses Using Embryonic Chick Ciliary Ganglion via Single-Axon Tracing, Electrophysiology, and Optogenetic Techniques.
Analysis of Neuro-Neuronal Synapses Using Embryonic Chick Ciliary Ganglion via Single-Axon Tracing, Electrophysiology, and Optogenetic Techniques.
复制标题
通过单轴突追踪、电生理学和光遗传学技术，使用胚胎鸡睫状神经节分析神经神经元突触。
DOI：
10.1002/cpns.64
复制
发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Egawa R. Yawo H.
中科院分区：
文献类型：
--
作者：
Egawa R. Yawo H.
萼型突触是一种巨大的突触结构，其中突触前末梢以一对一的方式缠绕突触后神经元。由于其简单性和生理记录方法的高可及性，它已被用作突触的实验模型系统数十年。特别是，胚胎鸡睫状神经节（CG）的花萼具有巨大的潜力，突触科学，因为更灵活的遗传操作相比，其他突触。在这里，我们描述了使用CG和尖端分子工具研究突触前形态、生理和发育的方法。我们概述了使用卵内电穿孔进行突触前基因操作的分步方案，分离的CG的制备，透明CG中单轴突追踪的3D成像，突触前末端的电生理学，以及使用光遗传学分子试剂的全光学方法。这些方法将有助于今后对突触和神经元回路的研究。John Wiley & Sons，Inc._关于我们
The calyx‐type synapse is a giant synaptic structure in which a presynaptic terminal wraps around a postsynaptic neuron in a one‐to‐one manner. It has been used for decades as an experimental model system of the synapse due to its simplicity and high accessibility in physiological recording methods. In particular, the calyx of the embryonic chick ciliary ganglion (CG) has enormous potential for synapse science because more flexible genetic manipulations are available compared with other synapses. Here, we describe methods to study presynaptic morphology, physiology, and development using CGs and cutting‐edge molecular tools. We outline step‐by‐step protocols for presynaptic gene manipulation using in ovo electroporation, preparation of isolated CGs, 3‐D imaging for single‐axon tracing in transparent CGs, electrophysiology of the presynaptic terminal, and an all‐optical approach using optogenetic molecular reagents. These methods will facilitate studies of the synapse and neuronal circuits in the future. © 2019 by John Wiley & Sons, Inc.