Differential mitochondrial and cellular responses between H vs. J mtDNA haplogroup-containing human RPE transmitochondrial cybrid cells
| Autor: | Ana Rubin Panvini, Anzor Gvritishvili, Hannah Galvan, Sonali Nashine, Shari R. Atilano, M. Cristina Kenney, Joyce Tombran-Tink |
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| Rok vydání: | 2022 |
| Předmět: |
1.1 Normal biological development and functioning
mtDNA J haplogroup AMD Neurodegenerative Medical Biochemistry and Metabolomics Eye Ophthalmology & Optometry DNA Mitochondrial Article Macular Degeneration Cellular and Molecular Neuroscience Underpinning research Opthalmology and Optometry Genetics 2.1 Biological and endogenous factors Humans Aetiology Eye Disease and Disorders of Vision Age-related macular degeneration Neurosciences DNA Hydrogen Peroxide mtDNA H haplogroup Sensory Systems Mitochondrial Mitochondria Ophthalmology Calcium Mitochondrial DNA haplogroups |
| Zdroj: | Exp Eye Res |
| ISSN: | 0014-4835 |
| DOI: | 10.1016/j.exer.2022.109013 |
| Popis: | Mitochondrial dysfunction is associated with several retinal degenerative diseases including Age-related Macular Degeneration (AMD). Human mitochondrial DNA (mtDNA) haplogroups are inherited from a common ancestral clan and are defined by specific sets of genetic differences. The purpose of this study was to determine and compare the effects of mtDNA haplogroups H and J on transcriptome regulation and cellular resilience to oxidative stress in human RPE cytoplasmic hybrid (cybrid) cell lines in vitro. ARPE-19 cybrid cell lines containing mtDNA haplogroups H and J were created by fusing platelets obtained from normal individuals containing H and J haplogroups with mitochondria-deficient (Rho0) ARPE-19cell lines. These cybrids were exposed to oxidative stress using 300μM hydrogen peroxide (H2O2), following which mitochondrial structural dynamics was studied at varying time points using the mitochondrial markers - TOMM20 (Translocase of Outer Mitochondrial Membrane 20) and Mitotracker. To evaluate mitochondrial function, levels of ROS, ΔΨm and [Ca2+]m were measured using flow cytometry, and ATP levels were measured using luminescence. The H and J cybrid cell transcriptomes were compared using RNAseq to determine how changes in mtDNA regulate gene expression. Inflammatory and angiogenic markers were measured using Luminex assay to understand how these mtDNAs influenced cellular response to oxidative stress. Actin filaments' morphology was examined using confocal microscopy. Following exposure to H2O2 stress, the J cybrids showed increased mitochondrial swelling and perinuclear localization, disturbed fission and fusion, increased calcium uptake (p |
| Databáze: | OpenAIRE |
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