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MitoLoc: A method for the simultaneous quantification of mitochondrial network morphology and membrane potential in single cells

Journal article published in 2015 by Jakob Vowinckel, Johannes Hartl, Richard Butler, Markus Ralser ORCID
This paper is available in a repository.
This paper is available in a repository.

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Preprint: policy unknown
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Postprint: policy unknown
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Abstract

This is the final version. It was first published by Elsevier at http://www.sciencedirect.com/science/article/pii/S1567724915300088. ; 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 ?fzo1, ?ref2, and ?dnm1 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.