Impact of Mitochondrial A3243G Heteroplasmy on Mitochondrial Bioenergetics and Dynamics of Directly Reprogrammed MELAS Neurons
Abstract
:1. Introduction
2. Material and Methods
2.1. Establish of Primary Skin Fibroblasts
2.2. Measurement of Mutant Heteroplasmy
2.3. Direct Reprogramming of Primary Skin Fibroblasts to iNs
2.4. Western Blotting
2.5. Detection of Mitochondrial Membrane Potential (ΔΨm)
2.6. Measurement of Mitochondrial Reactive Oxygen Species
2.7. Cellular Bioenergetics
2.8. Mitochondria Morphology and Network
2.9. Statistical Analysis
3. Results
3.1. Generation of iNs
3.2. iNs Retained Stable m.A3243G Heteroplasmy
3.3. RC Complexes Deficiency in MELAS iNs
3.4. High Heteroplasmy Increased ROS Production
3.5. Reduction in ΔΨm in iNs with High Heteroplasmy
3.6. Impact of Heteroplasmy on the Bioenergetics
3.7. Impact of Heteroplasmy on Mitochondria Dynamics
4. Discussion
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Lin, D.-S.; Huang, Y.-W.; Ho, C.-S.; Huang, T.-S.; Lee, T.-H.; Wu, T.-Y.; Huang, Z.-D.; Wang, T.-J. Impact of Mitochondrial A3243G Heteroplasmy on Mitochondrial Bioenergetics and Dynamics of Directly Reprogrammed MELAS Neurons. Cells 2023, 12, 15. https://doi.org/10.3390/cells12010015
Lin D-S, Huang Y-W, Ho C-S, Huang T-S, Lee T-H, Wu T-Y, Huang Z-D, Wang T-J. Impact of Mitochondrial A3243G Heteroplasmy on Mitochondrial Bioenergetics and Dynamics of Directly Reprogrammed MELAS Neurons. Cells. 2023; 12(1):15. https://doi.org/10.3390/cells12010015
Chicago/Turabian StyleLin, Dar-Shong, Yu-Wen Huang, Che-Sheng Ho, Tung-Sun Huang, Tsung-Han Lee, Tsu-Yen Wu, Zon-Darr Huang, and Tuan-Jen Wang. 2023. "Impact of Mitochondrial A3243G Heteroplasmy on Mitochondrial Bioenergetics and Dynamics of Directly Reprogrammed MELAS Neurons" Cells 12, no. 1: 15. https://doi.org/10.3390/cells12010015