Gupta Rishikesh Kumar, Clark Natalie, Yeager Kaylee, Fisch Alexander, Tiriac Alexandre
Department of Biological Sciences, Vanderbilt University.
Department of Biological Sciences, Vanderbilt University; Vanderbilt Brain Institute, Vanderbilt University; Department of Ophthalmology and Visual Sciences, Vanderbilt University;
J Vis Exp. 2025 Jun 24(220). doi: 10.3791/68493.
Spontaneous retinal waves are a hallmark of retinal network activity during development, playing a crucial role in the formation of the visual system by influencing the refinement of axons, permeability of vasculature, and overall maturation of neural circuits. These waves are commonly studied in ex vivo retinal preparations using multielectrode arrays (MEAs), which enable electrophysiological recordings of large populations of retinal ganglion cell (RGC) activity. MEA-based electrophysiology has become a powerful tool due to its ease of use to rapidly collect high-throughput data, thus making it ideally suited to study retinal activity in a variety of experimental conditions. In this protocol, we outline the critical steps for preparing retinal tissue for the acquisition of electrophysiological data using a High-Density MEA (HD-MEA) on an electrophysiology platform. The process begins with the careful isolation of intact retinas from neonatal animals under physiological conditions. Once prepared, the retina is carefully mounted onto an HD-MEA chip, which consists of a grid of 26,400 electrodes capable of performing simultaneous extracellular recordings from at least 1,000 RGCs. Recordings can last up to several hours. Ultimately, this methodological approach offers valuable applications in investigating retinal development, disease, and potentially cross-species comparative studies, contributing to broader advancements in neuroscience and vision research.
自发视网膜波是视网膜网络在发育过程中的一个标志，通过影响轴突的精细化、血管的通透性以及神经回路的整体成熟，在视觉系统的形成中发挥着关键作用。这些波通常在离体视网膜标本中使用多电极阵列（MEA）进行研究，该阵列能够对大量视网膜神经节细胞（RGC）的活动进行电生理记录。基于MEA的电生理学因其易于快速收集高通量数据而成为一种强大的工具，因此非常适合在各种实验条件下研究视网膜活动。在本方案中，我们概述了在电生理平台上使用高密度MEA（HD-MEA）获取电生理数据时制备视网膜组织的关键步骤。该过程始于在生理条件下从新生动物中小心分离出完整的视网膜。一旦制备好，视网膜会被小心地安装到HD-MEA芯片上，该芯片由26400个电极组成的网格构成，能够同时对至少1000个RGC进行细胞外记录。记录可持续数小时。最终，这种方法在研究视网膜发育、疾病以及潜在的跨物种比较研究中具有重要应用价值，有助于神经科学和视觉研究取得更广泛的进展。
