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RPE65 Leu450Met变异小鼠眼底成像光诱导视网膜退化与氧化应激和细胞凋亡相关

Zhong X, Aredo B, Ding Y, et al.

期刊名称:Investigative ophthalmology & visual science

卷期:2016年第57卷第13期

摘要

目的:氧化应激,部分由于光引起,在许多视网膜疾病,包括黄斑变性和视网膜营养不良中具有重要作用。 RPE65Leu450Met变体在C57BL / 6和许多基因修饰小鼠中表达。它对光诱导的视网膜变性(LIRD)具有明显抗性。我们的目标是研发一种有效的方法来诱导耐药小鼠产生LIRD,这将重现在已知的LIRD模型中观察到的机制。方法:使用一种鼠眼底照相机将C57BL / 6J小鼠视网膜暴露于光之下。使用两种方案(含和不含腹膜内荧光素)。光学相干断层扫描(OCT)有助于确定视网膜损伤的位置和程度。还进行了组织学,TUNEL检测,定量(qPCR和免疫组织化学检查。结果:两种方案均可导致C57BL / 6J小鼠发生LIRD。光学相干断层扫描和组织学表明视网膜损伤始于感光器/外视网膜水平,并且在上视网膜中更突出。眼底照相递送光诱导的视网膜变性(FCD-LIRD)与凋亡,视网膜下小胶质细胞/巨噬细胞,氧化应激反应基因表达增加,C3d沉积相关。结论:我们描述了光诱导视网膜变性的两个新模型的特点,这两种模型均对C57BL / 6J小鼠有效并且可以在严重程度方面进行调节。我们预期FCD-LIRD可用于探索耐药小鼠中LIRD机制,这对增加我们理解视网膜对光损伤和氧化应激反应来说很重要。

Purpose:Oxidative stress, partly due to light, has an important role in many retinal diseases, including macular degeneration and retinal dystrophies. The Leu450Met variant of RPE65 is expressed in C57BL/6 and in many genetically modified mice. It confers significant resistance to light induced retinal degeneration (LIRD). Our goal was to develop an effective and efficient method to induce LIRD in resistant mice that would recapitulate mechanisms seen in known models of LIRD.Methods:The retinas of C57BL/6J mice were exposed to light using a murine fundus camera. Two protocols (with and without intraperitoneal fluorescein) were used. Optical coherence tomography (OCT) helped determine the location and extent of retinal damage. Histology, TUNEL assay, quantitative (q) PCR, and immunohistochemistry were performed.Results:Both protocols consistently generated LIRD in C57BL/6J mice. Optical coherence tomography and histology demonstrated that retinal damage starts at the level of the photoreceptor/outer retina and is more prominent in the superior retina. Fundus camera-delivered light-induced retinal degeneration (FCD-LIRD) is associated with apoptosis, subretinal microglia/macrophages, increased expression of oxidative stress response genes, and C3d deposition.Conclusions:We characterize two new models of light-induced retinal degeneration that are effective in C57BL/6J mice, and can be modulated in terms of severity. We expect FCD-LIRD to be useful in exploring mechanisms of LIRD in resistant mice, which will be important in increasing our understanding of the retinal response to light damage and oxidative stress.


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