MitoLoc: A method for the simultaneous quantification mitochondrial network morphology and membrane potential in single cells
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Vowinckel, J., Hartl, J., Butler, R., & Ralser, M. (2015). MitoLoc: A method for the simultaneous quantification mitochondrial network morphology and membrane potential in single cells. Mitochondrion, 24 77-86. https://doi.org/10.1016/j.mito.2015.07.001
Mitochondria assemble into flexible networks. Here we present a simple method for the simultaneous quantification of mitochondrial membrane potential and network morphology that is based on computational co-localisation analysis of differentially imported fluorescent marker proteins. Established, but not restricted to, Saccharomyces cerevisiae, MitoLoc reproducibly measures changes in membrane potential induced by the uncoupling agent CCCP, by oxidative stress, in respiratory deficient cells, and in Δfzo1, Δref2, and Δdnm1 mutants that possess fission and fusion defects. In combination with super-resolution images, MitoLoc uses 3D reconstruction to calculate six geometrical classifiers which differentiate network morphologies in fission/fusion mutants, under oxidative stress and in cells lacking mtDNA. We find that mitochondrial fission and a decline in membrane potential do regularly, but not facultatively, co-occur. We make markers, open-source software as well as the mathematical procedures openly available. MitoLoc simplifies the measurement of mitochondrial membrane potential in parallel to detect morphological changes in mitochondrial networks.
This work was supported by funding from the Wellcome Trust (RG 093735/Z/10/Z) and the ERC (Starting grant 260809). M.R. is a Wellcome Trust Research Career Development and Wellcome-Beit prize fellow.
Wellcome Trust (093735/Z/10/Z)
European Research Council (260809)
External DOI: https://doi.org/10.1016/j.mito.2015.07.001
This record's URL: https://www.repository.cam.ac.uk/handle/1810/248930
Attribution 2.0 UK: England & Wales
Licence URL: http://creativecommons.org/licenses/by/2.0/uk/
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