Mitochondrial Ceramide Effects on the Retinal Pigment Epithelium in Diabetes
Abstract
:1. Introduction
2. Results
2.1. Diabetes Results in Retinal Mitochondrial Ceramide Accumulation
2.2. Diabetes Results in Pro-Inflammatory Changes in Human Retinal Pigment Epithelium (RPE) Cells
2.3. Diabetes Results in Mitochondrial Fragmentation in Human RPE Cells
2.4. Diabetes Induces Acid Sphingomyelinase (ASM)-Mediated Changes in Mitochondrial Function of Human RPE Cells
2.5. Mitochondrial ASM Contributes to Impaired Mitochondrial Function In Vitro
3. Discussion
4. Materials and Methods
4.1. Rodents
4.2. Cell Culture
4.3. Mitochondrial Isolation
4.4. Mass Spectrometry
4.5. Immunocytochemistry and Mitochondrial Morphology
4.6. Quantitative Real-Time Polymerase Chain Reaction
4.7. Western Blot Analysis
4.8. Citrate Synthase Activity
4.9. Microrespirometry
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
CCCP | Carbonyl cyanide m-chlorophenylhydrazone |
DR | Diabetic retinopathy |
ICAM-1 | Intercellular adhesion molecule 1 |
VDAC | Voltage dependent anion channel |
LAMP-1 | Lysosome associated membrane protein 1 |
ASM | Acid sphingomyelinase |
RCR | Respiratory control ratio |
RPE | Retinal pigment epithelium |
ROS | Reactive oxygen species |
STZ | Streptozotocin |
VEGF | Vascular endothelial growth factor |
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Levitsky, Y.; Hammer, S.S.; Fisher, K.P.; Huang, C.; Gentles, T.L.; Pegouske, D.J.; Xi, C.; Lydic, T.A.; Busik, J.V.; Proshlyakov, D.A. Mitochondrial Ceramide Effects on the Retinal Pigment Epithelium in Diabetes. Int. J. Mol. Sci. 2020, 21, 3830. https://doi.org/10.3390/ijms21113830
Levitsky Y, Hammer SS, Fisher KP, Huang C, Gentles TL, Pegouske DJ, Xi C, Lydic TA, Busik JV, Proshlyakov DA. Mitochondrial Ceramide Effects on the Retinal Pigment Epithelium in Diabetes. International Journal of Molecular Sciences. 2020; 21(11):3830. https://doi.org/10.3390/ijms21113830
Chicago/Turabian StyleLevitsky, Yan, Sandra S. Hammer, Kiera P. Fisher, Chao Huang, Travan L. Gentles, David J. Pegouske, Caimin Xi, Todd A. Lydic, Julia V. Busik, and Denis A. Proshlyakov. 2020. "Mitochondrial Ceramide Effects on the Retinal Pigment Epithelium in Diabetes" International Journal of Molecular Sciences 21, no. 11: 3830. https://doi.org/10.3390/ijms21113830