Department of Zoology and Physiology, and Program in Neuroscience, University of Wyoming, Laramie, Wyoming 82071, USA
Cold Spring Harb Protoc. 2021 Nov 1;2021(11):pdb.prot106898. doi: 10.1101/pdb.prot106898.
The tadpole retinotectal projection is the main component of the amphibian visual system. It comprises the retinal ganglion cells (RGCs) in the eye, which project an axon to synapse onto tectal neurons in the optic tectum. There are many attributes of this relatively simple projection that render it uniquely well-suited for studying the functional development of neural circuits. One major experimental advantage of this circuit is that it can be genetically or pharmacologically altered and then assessed at high resolution via whole-cell electrophysiological recordings using an ex vivo isolated brain preparation. This protocol provides instructions for performing such electrophysiological investigations using the ex-vivo-isolated brain preparation. It allows one to measure many different aspects of synaptic transmission between the RGC axons and individual postsynaptic tectal neurons, including AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) to NMDA (-methyl-d-aspartate) ratios, strength of individual RGC axons, paired pulse facilitation, and strength of individual synapses.
蝌蚪视网膜-顶盖投射是两栖类视觉系统的主要组成部分。它包括眼睛中的视网膜神经节细胞（RGCs），其轴突投射到视顶盖中的顶盖神经元上形成突触。这个相对简单的投射有许多属性，使其非常适合研究神经回路的功能发育。该回路的一个主要实验优势是，它可以通过遗传或药理学方法进行改变，然后使用离体脑切片进行高分辨率的全细胞膜片钳记录进行评估。本方案提供了使用离体脑切片进行这种电生理研究的说明。它可以测量 RGC 轴突和单个突触后顶盖神经元之间的突触传递的许多不同方面，包括 AMPA（α-氨基-3-羟基-5-甲基-4-异恶唑丙酸）与 NMDA（N-甲基-D-天冬氨酸）的比率、单个 RGC 轴突的强度、成对脉冲易化和单个突触的强度。
