Mitochondrial event localiser (MEL) to quantitativelydescribe fission, fusion and depolarisation in the three-dimensional space

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dc.contributor.authorTheart, Rensu P.en_ZA
dc.contributor.authorKriel, Jurgenen_ZAe
dc.contributor.authorDu Toit, Andreen_ZA
dc.contributor.authorLoos, Benen_ZA
dc.contributor.authorNiesler, Thomas R.en_ZA
dc.date.accessioned2021-01-19T13:41:26Z
dc.date.available2021-01-19T13:41:26Z
dc.date.issued2020-12
dc.descriptionCITATION: Theart, R. P., et al. 2020. Mitochondrial event localiser (MEL) to quantitativelydescribe fission, fusion and depolarisation in the three-dimensional space. PLoS ONE, 15(12): e0229634, doi:10.1371/journal.pone.0229634
dc.descriptionThe original publication is available at https://journals.plos.org/plosone/
dc.descriptionPublication of this article was funded by the Stellenbosch University Open Access Fund
dc.description.abstractENGLISH ABSTRACT: Mitochondrial fission and fusion play an important role not only in maintaining mitochondrial homeostasis but also in preserving overall cellular viability. However, quantitative analysis based on the three-dimensional localisation of these highly dynamic mitochondrial events in the cellular context has not yet been accomplished. Moreover, it remains largely uncertain where in the mitochondrial network depolarisation is most likely to occur. We present the mitochondrial event localiser (MEL), a method that allows high-throughput, automated and deterministic localisation and quantification of mitochondrial fission, fusion and depolarisation events in large three-dimensional microscopy time-lapse sequences. In addition, MEL calculates the number of mitochondrial structures as well as their combined and average volume for each image frame in the time-lapse sequence. The mitochondrial event locations can subsequently be visualised by superposition over the fluorescence micrograph z-stack. We apply MEL to both control samples as well as to cells before and after treatment with hydrogen peroxide (H2O2). An average of 9.3/7.2/2.3 fusion/fission/depolarisation events per cell were observed respectively for every 10 sec in the control cells. With peroxide treatment, the rate initially shifted toward fusion with and average of 15/6/3 events per cell, before returning to a new equilibrium not far from that of the control cells, with an average of 6.2/6.4/3.4 events per cell. These MEL results indicate that both pre-treatment and control cells maintain a fission/fusion equilibrium, and that depolarisation is higher in the post-treatment cells. When individually validating mitochondrial events detected with MEL, for a representative cell for the control and treated samples, the true-positive events were 47%/49%/14% respectively for fusion/fission/depolarisation events. We conclude that MEL is a viable method of quantitative mitochondrial event analysis.en_ZA
dc.description.urihttps://journals.plos.org/plosone/article?id=10.1371/journal.pone.0229634
dc.description.versionPublishers' Version
dc.format.extent23 pages : illustrationsen_ZA
dc.identifier.citationTheart, R. P., et al. 2020. Mitochondrial event localiser (MEL) to quantitativelydescribe fission, fusion and depolarisation in the three-dimensional space. PLoS ONE, 15(12): e0229634, doi:10.1371/journal.pone.0229634
dc.identifier.issn1932-6203 (online)
dc.identifier.otherdoi:10.1371/journal.pone.0229634
dc.identifier.urihttp://hdl.handle.net/10019.1/108999
dc.language.isoen_ZAen_ZA
dc.publisherPublic Library of Scienceen_ZA
dc.rights.holderAuthors retain copyrighten_ZA
dc.subjectMitochondrial fissionen_ZA
dc.subjectFission (Biology)en_ZA
dc.subjectGene fusionen_ZA
dc.subjectHomeostasisen_ZA
dc.subjectImage processing -- Digital techniquesen_ZA
dc.subjectMitochondrial event localiseren_ZA
dc.titleMitochondrial event localiser (MEL) to quantitativelydescribe fission, fusion and depolarisation in the three-dimensional spaceen_ZA
dc.typeArticleen_ZA
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