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• W2041767939 abstract "purpose. To measure changes in nuclear gene expression resulting from mitochondrial dysfunction in retinal pigment epithelial cells. methods. ARPE-19 retinal pigment epithelial cells were depleted of their mitochondrial (mt)DNA by passaging in a low concentration of ethidium bromide. Loss of mitochondrial DNA was determined by uridine auxotrophy and quantitative real-time polymerase chain reaction of isolated DNA. Loss of mitochondrial membrane potential was estimated by uptake of JC-1. Changes in nuclear gene expression were determined by quantitative real-time reverse transcription–polymerase chain reaction of isolated total RNA from ethidium-bromide–treated and untreated cells. Morphologic and phenotypic changes were determined by phase-contrast microscopy, sensitivity to the oxidant tert-butyl hydroperoxide (tBH), and invasion assay. results. ARPE-19 cells became auxotrophic for growth on uridine after eight passages in 50 ng/mL ethidium bromide. Quantitative PCR revealed almost complete loss of mitochondrial DNA (ρ0 cells). Uptake of JC-1 was reduced in the ρ0 cells, indicating reduction of mitochondrial membrane potential. Quantitative RT-PCR measured increased expression of genes coding for drusen components, lipid transport, extracellular matrix components, and responses to inflammation in the ρ0 cells. The ρ0 cells also exhibited an increased sensitivity to killing by tBH and increased migration and invasion through solubulized basement membrane–coated tissue culture inserts. conclusions. ARPE-19 cells respond to loss of mitochondrial function by changes in nuclear gene expression that resemble changes observed in age-related macular degeneration. The results lead to the hypothesis that loss of mitochondrial function with age and resultant changes in nuclear gene expression may explain some of the changes in the macula that are associated with the known clinical manifestations of age-related macular degeneration." @default.
• W2041767939 created "2016-06-24" @default.
• W2041767939 creator A5025480370 @default.
• W2041767939 creator A5069220558 @default.
• W2041767939 date "2005-05-01" @default.
• W2041767939 modified "2023-10-17" @default.
• W2041767939 title "Nuclear Gene Expression Changes Due to Mitochondrial Dysfunction in ARPE-19 Cells: Implications for Age-Related Macular Degeneration" @default.
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• W2041767939 doi "https://doi.org/10.1167/iovs.04-1327" @default.
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