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

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

International journal of molecular sciences (2020-03-14)
Ines Rosignol, Beatriz Villarejo-Zori, Petra Teresak, Elena Sierra-Filardi, Xandra Pereiro, Natalia Rodríguez-Muela, Elena Vecino, Helena L A Vieira, Katharina Bell, Patricia Boya
摘要

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 mito-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 mito-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 mito-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 mito-QC reporter-based model is a valuable tool for accurately analyzing mitophagy in the context of glaucoma.

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层粘连蛋白 来源于 Engelbreth-Holm-Swarm 小鼠肉瘤基底膜, 1-2 mg/mL in Tris-buffered saline, 0.2 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
胰岛素 人, recombinant, expressed in yeast (proprietary host)
Sigma-Aldrich
去铁胺 甲磺酸盐, powder, ≥92.5% (TLC)
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氰化羰基-3-氯苯腙, ≥97% (TLC), powder
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HEPES钠盐 钠盐, BioPerformance Certified, suitable for cell culture, ≥99.0%
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抗Brn-3a抗体,POU域蛋白,克隆5A3.2, culture supernatant, clone 5A3.2, Chemicon®
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CORM-A1, ≥95% (NMR)
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1,2-二亚油酰-3-棕榈酰基-外消旋-甘油, ≥95% (TLC), liquid