Cellular ROS imaging with hydro-Cy3 dye is strongly influenced by mitochondrial membrane potential

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dc.contributor.author Zhdanov, Alexander V.
dc.contributor.author Aviello, Gabriella
dc.contributor.author Knaus, Ulla G.
dc.contributor.author Papkovsky, Dmitri B.
dc.date.accessioned 2016-12-09T14:40:27Z
dc.date.available 2016-12-09T14:40:27Z
dc.date.issued 2016-11-04
dc.identifier.citation Zhdanov, A. V., Aviello, G., Knaus, U. G. and Papkovsky, D. B. (2017) 'Cellular ROS imaging with hydro-Cy3 dye is strongly influenced by mitochondrial membrane potential', Biochimica et Biophysica Acta (BBA) - General Subjects, 1861(2), pp. 198-204. doi:10.1016/j.bbagen.2016.10.023 en
dc.identifier.volume 1861 en
dc.identifier.issued 22 en
dc.identifier.startpage 198 en
dc.identifier.endpage 204 en
dc.identifier.issn 0304-4165
dc.identifier.uri http://hdl.handle.net/10468/3368
dc.identifier.doi 10.1016/j.bbagen.2016.10.023
dc.description.abstract Background: Hydrocyanines are widely used as fluorogenic probes to monitor reactive oxygen species (ROS) generation in cells. Their brightness, stability to autoxidation and photobleaching, large signal change upon oxidation, pH independence and red/near infrared emission are particularly attractive for imaging ROS in live tissue. Methods: Using confocal fluorescence microscopy we have examined an interference of mitochondrial membrane potential (ΔΨm) with fluorescence intensity and localisation of a commercial hydro-Cy3 probe in respiring and non-respiring colon carcinoma HCT116 cells. Results: We found that the oxidised (fluorescent) form of hydro-Cy3 is highly homologous to the common ΔΨm-sensitive probe JC-1, which accumulates and aggregates only in ‘energised’ negatively charged mitochondrial matrix. Therefore, hydro-Cy3 oxidised by hydroxyl and superoxide radicals tends to accumulate in mitochondrial matrix, but dissipates and loses brightness as soon as ΔΨm is compromised. Experiments with mitochondrial inhibitor oligomycin and uncoupler FCCP, as well as a common ROS producer paraquat demonstrated that signals of the oxidised hydro-Cy3 probe rapidly and strongly decrease upon mitochondrial depolarisation, regardless of the rate of cellular ROS production. Conclusions: While analysing ROS-derived fluorescence of commercial hydrocyanine probes, an accurate control of ΔΨm is required. General significance: If not accounted for, non-specific effect of mitochondrial polarisation state on the behaviour of oxidised hydrocyanines can cause artefacts and data misinterpretation in ROS studies. en
dc.description.sponsorship Science Foundation Ireland (SFI grant 12/RC/2276) en
dc.format.mimetype application/pdf en
dc.language.iso en en
dc.publisher Elsevier en
dc.rights © 2016, Elsevier. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ en
dc.rights.uri http://creativecommons.org/licenses/by-nc-nd/4.0/ en
dc.subject ROS probes en
dc.subject Hydrocyanines en
dc.subject Hydro-Cy3 en
dc.subject Mitochondrial membrane potential en
dc.subject Live cell imaging en
dc.title Cellular ROS imaging with hydro-Cy3 dye is strongly influenced by mitochondrial membrane potential en
dc.type Article (peer-reviewed) en
dc.internal.authorcontactother Dmitri Papkovsky, School Of Biochemistry & Cell Biology, University College Cork, Cork, Ireland. +353-21-490-3000 Email: d.papkovsky@ucc.ie en
dc.internal.availability Full text available en
dc.check.info Access to this item is restricted until 12 months after publication by the request of the publisher. en
dc.check.date 2017-11-04
dc.date.updated 2016-12-09T14:31:09Z
dc.description.version Accepted Version en
dc.internal.rssid 375019528
dc.contributor.funder Science Foundation Ireland en
dc.description.status Peer reviewed en
dc.identifier.journaltitle Biochimica et Biophysica Acta (BBA) - General Subjects en
dc.internal.copyrightchecked No !!CORA!! en
dc.internal.licenseacceptance Yes en
dc.internal.IRISemailaddress d.papkovsky@ucc.ie en


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© 2016, Elsevier. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ Except where otherwise noted, this item's license is described as © 2016, Elsevier. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
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