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dc.contributor.authorVowinckel, Jakob
dc.contributor.authorHartl, Johannes
dc.contributor.authorButler, Richard
dc.contributor.authorRalser, Markus
dc.date.accessioned2015-07-14T11:29:18Z
dc.date.available2015-07-14T11:29:18Z
dc.date.issued2015-09
dc.identifier.citationVowinckel et al. Mitochondrion (2015) Vol. 24, pp. 77-86. doi:10.1016/j.mito.2015.07.001
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 in, 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 ∆fzo1, ∆ref2, and ∆dnm1 mutants, under oxidative stress and in cells lacking mtDNA, even when the network is fragmented to a similar extent. We find that mitochondrial fission and a decline in membrane potential do regularly, but not necessarily, co-occur. MitoLoc hence simplifies the measurement of mitochondrial membrane potential in parallel to detect morphological changes in mitochondrial networks. Marker plasmid open-source software as well as the mathematical procedures are made openly available.
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.languageEnglish
dc.language.isoen
dc.publisherElsevier BV
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 of mitochondrial network morphology and membrane potential in single cells.
dc.typeArticle
dc.description.versionThis is the final version. It was first published by Elsevier at http://www.sciencedirect.com/science/article/pii/S1567724915300088.
prism.endingPage86
prism.publicationDate2015
prism.publicationNameMitochondrion
prism.startingPage77
prism.volume24
dc.rioxxterms.funderWellcome Trust
dc.rioxxterms.funderERC
dc.rioxxterms.projectidRG 093735/Z/10/Z
dc.rioxxterms.projectid260809
dcterms.dateAccepted2015-07-07
rioxxterms.versionofrecord10.1016/j.mito.2015.07.001
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2015-07-13
dc.contributor.orcidButler, Richard [0000-0002-3885-1332]
dc.contributor.orcidRalser, Markus [0000-0001-9535-7413]
dc.identifier.eissn1872-8278
rioxxterms.typeJournal Article/Review
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