Dannhausen Katharina, Rashid Khalid, Langmann Thomas
Laboratory for Experimental Immunology of the Eye, Department of Ophthalmology, University of Cologne, Cologne, Germany.
Methods Mol Biol. 2018;1753:159-166. doi: 10.1007/978-1-4939-7720-8_10.
Microgliosis is a hallmark of degenerative processes in the retina. Reactive microglia migrate to the photoreceptor layer and the subretinal space during outer retinal degeneration. This process creates a toxic milieu where reactive microglia and dying photoreceptors recruit additional reactive phagocytes. This results in the release of a multitude of proinflammatory factors which accelerate photoreceptor demise. In this chapter, we outline in detail how to monitor microgliosis in the Fam161a-deficient mouse model of Retinitis Pigmentosa by performing immunohistochemical stainings of retinal cryosections and flat mounts using the marker Iba1. This protocol will serve as a guideline in evaluating microglia reactivity and localization in various mouse models of retinal degeneration.
小胶质细胞增生是视网膜退行性病变过程的一个标志。在视网膜外层变性期间,反应性小胶质细胞迁移至光感受器层和视网膜下间隙。这一过程会形成一个毒性环境,在此环境中,反应性小胶质细胞和濒死的光感受器会招募更多的反应性吞噬细胞。这导致大量促炎因子的释放,加速了光感受器的死亡。在本章中,我们详细概述了如何通过使用标记物离子钙结合衔接分子1(Iba1)对视网膜冰冻切片和平铺片进行免疫组织化学染色,来监测色素性视网膜炎的Fam161a基因缺陷小鼠模型中的小胶质细胞增生情况。本方案将作为评估各种视网膜变性小鼠模型中小胶质细胞反应性和定位的指导原则。