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用于原发性视网膜神经节细胞和实验性青光眼模型中定量噬黑素评估的-QC 报告器。

The -QC Reporter for Quantitative Mitophagy Assessment in Primary Retinal Ganglion Cells and Experimental Glaucoma Models.

机构信息

Department of Cellular and Molecular Biology, Centro de Investigaciones Biológicas Margarita Salas, CSIC, 28040 Madrid, Spain.

Experimental-Ophthalmo-Biology Group, Universidad del País Vasco, 48950 Leioa, Spain.

出版信息

Int J Mol Sci. 2020 Mar 10;21(5):1882. doi: 10.3390/ijms21051882.

Abstract

Mitochondrial damage plays a prominent role in glaucoma. The only way cells can degrade whole mitochondria is via autophagy, in a process called mitophagy. Thus, studying mitophagy in the context of glaucoma is essential to understand the disease. Up to date limited tools are available for analyzing mitophagy in vivo. We have taken advantage of the -QC reporter, a recently generated mouse model that allows an accurate mitophagy assessment to fill this gap. We used primary RGCs and retinal explants derived from -QC mice to quantify mitophagy activation in vitro and ex vivo. We also analyzed mitophagy in retinal ganglion cells (RGCs), in vivo, using different mitophagy inducers, as well as after optic nerve crush (ONC) in mice, a commonly used surgical procedure to model glaucoma. Using -QC reporter we quantified mitophagy induced by several known inducers in primary RGCs in vitro, ex vivo and in vivo. We also found that RGCs were rescued from some glaucoma relevant stress factors by incubation with the iron chelator deferiprone (DFP). Thus, the -QC reporter-based model is a valuable tool for accurately analyzing mitophagy in the context of glaucoma.

摘要

线粒体损伤在青光眼发病机制中起着重要作用。细胞降解整个线粒体的唯一途径是通过自噬,这一过程被称为线粒体自噬。因此,研究青光眼背景下的线粒体自噬对于理解该疾病至关重要。迄今为止,体内分析线粒体自噬的工具非常有限。我们利用最近生成的 -QC 报告小鼠模型来填补这一空白,该模型可用于准确评估线粒体自噬。我们使用来自 -QC 小鼠的原代 RGC 和视网膜外植体在体外和体内进行定量分析,以检测线粒体自噬的激活。我们还使用不同的线粒体自噬诱导剂,以及在小鼠视神经钳夹(ONC)后(一种常用的模拟青光眼的手术方法),在体内分析了视网膜神经节细胞(RGC)中的线粒体自噬。使用 -QC 报告基因,我们在体外、体内和体内检测了几种已知诱导剂诱导的原代 RGC 中的线粒体自噬。我们还发现,用铁螯合剂去铁酮(DFP)孵育可以挽救 RGC 免受一些与青光眼相关的应激因素的影响。因此,基于 -QC 报告基因的模型是一种准确分析青光眼背景下线粒体自噬的有价值的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/86b8/7084520/cb5133d743f8/ijms-21-01882-g001.jpg

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