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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
要旨

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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Sigma-Aldrich
ラミニン EHS肉腫基底膜由来, 1-2 mg/mL in Tris-buffered saline, 0.2 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
アンチマイシンA 放線菌由来
Sigma-Aldrich
インスリン ヒト, recombinant, expressed in yeast (proprietary host)
Sigma-Aldrich
オリゴマイシンA, ≥99% (HPLC)
Sigma-Aldrich
デフェロキサミン メシル酸塩, powder, ≥92.5% (TLC)
Sigma-Aldrich
カルボニルシアニド 3-クロロフェニルヒドラゾン, ≥97% (TLC), powder
Sigma-Aldrich
HEPES ナトリウム塩, BioPerformance Certified, suitable for cell culture, ≥99.0%
Sigma-Aldrich
スタウロスポリン 放線菌由来, ≥98% (HPLC), film
Sigma-Aldrich
抗Brn-3a抗体 POU-ドメインタンパク質 クローン5A3.2, culture supernatant, clone 5A3.2, Chemicon®
Sigma-Aldrich
CORM-A1, ≥95% (NMR)
Sigma-Aldrich
1,2-ジリノレオイル-3-パルミトイル-rac-グリセリン, ≥95% (TLC), liquid