Abstract
Mitochondrial DNA (mtDNA) encodes a variety of rRNAs, tRNAs, and respiratory chain complex proteins. The integrity of mtDNA supports the mitochondrial functions and plays an essential role in numerous physiological and pathological processes. Mutations in mtDNA cause metabolic diseases and aging. The mtDNA within the human cells are packaged into hundreds of nucleoids within the mitochondrial matrix. Knowledge of how the nucleoids are dynamically distributed and organized within mitochondria is key to understanding mtDNA structure and functions. Therefore, visualizing the distribution and dynamics of mtDNA within mitochondria is a powerful approach to gain insights into the regulation of mtDNA replication and transcription. In this chapter, we describe the methods of observing mtDNA and its replication with fluorescence microscopy in both fixed and live cells using different labeling strategies.
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Acknowledgments
This work was supported by the funding from the National Key R&D Program of China (No.2017YFA0505300) and the National Science Foundation of China (21825401) to Y.S.; grants from the Australia National Health and Medical Council (NHMRC, APP1177374) and the Australia National Heart Foundation (NHF, 102592) to Q.P.S.; and the funding from China Scholarship Council (No. 201706170028) to X.D.
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Di, X., Qin, J., Sun, Y., Su, Q.P. (2023). Visualize the Distribution and Dynamics of Mitochondrial DNA (mtDNA) Nucleoids with Multiple Labeling Strategies. In: Nicholls, T.J., Uhler, J.P., Falkenberg, M. (eds) Mitochondrial DNA. Methods in Molecular Biology, vol 2615. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2922-2_6
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DOI: https://doi.org/10.1007/978-1-0716-2922-2_6
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2921-5
Online ISBN: 978-1-0716-2922-2
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