Weng Shi-Jun, Renna Jordan M, Chen Wei-Yi, Yang Xiong-Li
Department of Ophthalmology, State Key Laboratory of Medical Neurobiology and Institutes of Brain Science, Zhongshan Hospital, Fudan University, Shanghai, China.
Department of Biology, University of Akron, Akron, OH, USA.
Methods Mol Biol. 2018;1753:289-303. doi: 10.1007/978-1-4939-7720-8_20.
Intrinsically photosensitive retinal ganglion cells (ipRGCs) are a special subset of retinal output neurons capable of detecting and responding to light via a unique photopigment called melanopsin. Melanopsin activation is essential to a wide array of physiological functions, especially to those related to non-image-forming vision. Since ipRGCs only constitute a very small proportion of retinal ganglion cells, targeted recording of melanopsin-driven responses used to be a big challenge to vision researchers. Multielectrode array (MEA) recording provides a noninvasive, high throughput method to monitor melanopsin-driven responses. When synaptic inputs from rod/cone photoreceptors are silenced with glutamatergic blockers, extracellular electric signals derived from melanopsin activation can be recorded from multiple ipRGCs simultaneously by tens of microelectrodes aligned in an array. In this chapter we describe how our labs have approached MEA recording of melanopsin-driven light responses in adult mouse retinas. Instruments, tools and chemical reagents routinely used for setting up a successful MEA recording are listed, and a standard experimental procedure is provided. The implementation of this technique offers a useful paradigm that can be used to conduct functional assessments of ipRGCs and NIF vision.
内在光敏性视网膜神经节细胞（ipRGCs）是视网膜输出神经元的一个特殊子集，能够通过一种名为黑视蛋白的独特光色素检测光并对其作出反应。黑视蛋白的激活对于一系列生理功能至关重要，尤其是与非成像视觉相关的功能。由于ipRGCs仅占视网膜神经节细胞的极小比例，过去对视觉研究人员来说，靶向记录黑视蛋白驱动的反应是一项巨大挑战。多电极阵列（MEA）记录提供了一种非侵入性、高通量的方法来监测黑视蛋白驱动的反应。当用谷氨酸能阻滞剂使视杆/视锥光感受器的突触输入沉默时，由黑视蛋白激活产生的细胞外电信号可以通过排列成阵列的数十个微电极同时从多个ipRGCs记录下来。在本章中，我们描述了我们实验室如何对成年小鼠视网膜中黑视蛋白驱动的光反应进行MEA记录。列出了用于成功进行MEA记录的常规仪器、工具和化学试剂，并提供了标准实验程序。这项技术的实施提供了一个有用的范例，可用于对ipRGCs和非成像视觉进行功能评估。
