Abstract
Mitochondria are complex organelles with multifaceted roles in cell biology, acting as signaling hubs that implicate them in cellular physiology and pathology. Mitochondria are both the target and the origin of multiple signaling events, including redox processes and calcium signaling which are important for organellar function and homeostasis. One way to interrogate mitochondrial function is by live cell imaging. Elaborated approaches perform imaging of single mitochondrial dynamics in living cells and animals. Imaging mitochondrial signaling and function can be challenging due to the sheer number of mitochondria, and the speed, propagation, and potential short half-life of signals. Moreover, mitochondria are organized in functionally coupled interorganellar networks. Therefore, advanced analysis and postprocessing tools are needed to enable automated analysis to fully quantitate mitochondrial signaling events and decipher their complex spatiotemporal connectedness. Herein, we present a protocol for recording and automating analyses of signaling in neuronal mitochondrial networks.
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Acknowledgments
Experiments that form the basis of this protocol were performed in the laboratory of T. Misgeld (TU Munich) and Martin Kerschensteiner (LMU Munich). M.O.B. acknowledges helpful discussions with T. Dick (DKFZ Heidelberg) and M. Schwarzländer (University of Münster). M.O.B. and F.T.K. were supported by a physician-scientist fellowship of the Medical Faculty, University of Heidelberg and by the Hoffmann-Klose Foundation (University of Heidelberg). F.T.K. was supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation, KU 3555/1-1) and a research grant from Heidelberg University Hospital.
Author Contributions: F.T.K. and M.O.B. conceived the study. M.O.B. performed microscopy experiments. F.T.K. provided analytical tools. M.O.B. and F.T.K. performed image analysis of the data. F.T.K. and M.O.B. wrote the manuscript.
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Kurz, F.T., Breckwoldt, M.O. (2022). Automated Quantification and Network Analysis of Redox Dynamics in Neuronal Mitochondria. In: Cortassa, S., Aon, M.A. (eds) Computational Systems Biology in Medicine and Biotechnology. Methods in Molecular Biology, vol 2399. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1831-8_12
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DOI: https://doi.org/10.1007/978-1-0716-1831-8_12
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