Thomas Jennifer L, Thummel Ryan
Department of Anatomy and Cell Biology, Wayne State University School of Medicine.
J Vis Exp. 2013 Oct 24(80):e51017. doi: 10.3791/51017.
Light-induced retinal degeneration (LIRD) is commonly used in both rodents and zebrafish to damage rod and cone photoreceptors. In adult zebrafish, photoreceptor degeneration triggers Müller glial cells to re-enter the cell cycle and produce transient-amplifying progenitors. These progenitors continue to proliferate as they migrate to the damaged area, where they ultimately give rise to new photoreceptors. Currently, there are two widely-used LIRD paradigms, each of which results in varying degrees of photoreceptor loss and corresponding differences in the regeneration response. As more genetic and pharmacological tools are available to test the role of individual genes of interest during regeneration, there is a need to develop a robust LIRD paradigm. Here we describe a LIRD protocol that results in widespread and consistent loss of both rod and cone photoreceptors in which we have combined the use of two previously established LIRD techniques. Furthermore, this protocol can be extended for use in pigmented animals, which eliminates the need to maintain transgenic lines of interest on the albino background for LIRD studies.
光诱导视网膜变性(LIRD)常用于啮齿动物和斑马鱼,以损伤视杆和视锥光感受器。在成年斑马鱼中,光感受器变性会触发穆勒胶质细胞重新进入细胞周期并产生瞬时扩增祖细胞。这些祖细胞在迁移到受损区域时继续增殖,最终在那里产生新的光感受器。目前,有两种广泛使用的LIRD模式,每种模式都会导致不同程度的光感受器损失以及再生反应的相应差异。随着越来越多的遗传和药理学工具可用于测试再生过程中单个感兴趣基因的作用,需要开发一种强大的LIRD模式。在这里,我们描述了一种LIRD方案,该方案会导致视杆和视锥光感受器广泛且一致地丧失,我们在其中结合了两种先前建立的LIRD技术。此外,该方案可扩展用于有色动物,这消除了在白化背景上维持感兴趣的转基因品系以进行LIRD研究的需要。