Lee Seul Ki, Schmidt Tiffany M
Department of Neurobiology, Northwestern University, Evanston, IL, USA.
Methods Mol Biol. 2018;1753:275-287. doi: 10.1007/978-1-4939-7720-8_19.
Melanopsin-expressing, intrinsically photosensitive retinal ganglion cells (ipRGCs) are a relatively recently discovered class of photoreceptor. ipRGCs can be subdivided into at least five subtypes (M1-M5), each of which has a distinct complement of morphological and physiological properties. ipRGC subtypes can be identified morphologically based on a combination of dendritic morphology and immunostaining for a cell-type specific marker. In this chapter, we describe methods for conclusively identifying each of the five ipRGC subtypes through a combination of patch clamp electrophysiology, Neurobiotin filling, visualization of ipRGC dendrites, and immunostaining for the marker SMI-32.
表达黑视蛋白的内在光敏性视网膜神经节细胞(ipRGCs)是一类相对较新发现的光感受器。ipRGCs可至少细分为五种亚型(M1 - M5),每种亚型都具有独特的形态和生理特性组合。基于树突形态和针对细胞类型特异性标志物的免疫染色相结合,可从形态学上识别ipRGC亚型。在本章中,我们描述了通过膜片钳电生理学、Neurobiotin填充、ipRGC树突可视化以及针对标志物SMI - 32的免疫染色相结合的方法来最终鉴定五种ipRGC亚型中的每一种。