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dc.contributor.authorVowinckel, Jakoben
dc.contributor.authorHartl, Johannesen
dc.contributor.authorButler, Richarden
dc.contributor.authorRalser, Markusen
dc.date.accessioned2015-07-14T11:29:18Z
dc.date.available2015-07-14T11:29:18Z
dc.date.issued2015-07-13en
dc.identifier.citationVowinckel et al. Mitochondrion (2015) Vol. 24, pp. 77-86. doi:10.1016/j.mito.2015.07.001en
dc.identifier.issn1567-7249
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/248930
dc.description.abstractMitochondria 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.
dc.description.sponsorshipThis 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.
dc.languageEnglishen
dc.language.isoenen
dc.publisherElsevier
dc.rightsAttribution 2.0 UK: England & Wales*
dc.rights.urihttp://creativecommons.org/licenses/by/2.0/uk/*
dc.titleMitoLoc: A method for the simultaneous quantification mitochondrial network morphology and membrane potential in single cellsen
dc.typeArticle
dc.description.versionThis is the final version. It was first published by Elsevier at http://www.sciencedirect.com/science/article/pii/S1567724915300088.en
prism.endingPage86
prism.publicationDate2015en
prism.publicationNameMitochondrionen
prism.startingPage77
prism.volume24en
dc.rioxxterms.funderWellcome Trust
dc.rioxxterms.funderERC
dc.rioxxterms.projectidRG 093735/Z/10/Z
dc.rioxxterms.projectid260809
dcterms.dateAccepted2015-07-07en
rioxxterms.versionofrecord10.1016/j.mito.2015.07.001en
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2015-07-13en
dc.contributor.orcidButler, Richard [0000-0002-3885-1332]
dc.contributor.orcidRalser, Markus [0000-0001-9535-7413]
dc.identifier.eissn1872-8278
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idWellcome Trust (093735/Z/10/Z)
pubs.funder-project-idEuropean Research Council (260809)


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Attribution 2.0 UK: England & Wales
Except where otherwise noted, this item's licence is described as Attribution 2.0 UK: England & Wales