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Assessment of cellular reduced glutathione content in Pseudokirchneriella subcapitata using monochlorobimane

2012Journal article Open access
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dc.contributor.authorMachado, Manuela D.
dc.contributor.authorSoares, Eduardo V.
dc.date.accessioned2017-09-14T13:29:41Z
dc.date.available2017-09-14T13:29:41Z
dc.date.issued2012
dc.description.abstractThe green alga Pseudokirchneriella subcapitata has been extensively used for the assessment of adverse impacts of pollutants. Glutathione is involved in antioxidant defence and drug detoxification. Intracellular reduced glutathione (GSH) concentration can be used as an indicator of the health of cells. This work describes a simple and fast fluorescent cell-based assay for the evaluation of intracellular GSH in the alga P. subcapitata, using monochlorobimane (mBCl). Metabolically active algal cells incubated with 50 μmol L−1 mBCl form fluorescent bimane–glutathione (B-SG) adducts that can be measured fluorometrically. The distribution of GSH (B-SG adducts) in whole cells can be observed by epifluorescence microscopy, in the form of blue fluorescent spots. Depletion of cellular GSH with iodoacetamide, inhibition of glutathione S-transferase with ethacrynic acid or heat-induced death of the cells inhibited the formation of fluorescent adducts in the presence of mBCl. The fluorometric assay, using the 96-well microplate format, was able to detect GSH depletion in algal cells. This cell-based assay can be used to evaluate decreases in GSH content due to exposure to toxicants. This assay is amenable to automation and may be useful in high-throughput toxicity screening using the alga P. subcapitata.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1007/s10811-012-9811-7pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.22/10261
dc.language.isoengpt_PT
dc.publisherSpringer Verlagpt_PT
dc.relation.ispartofseriesJournal of Applied Phycology;Vol. 24, Issue 6
dc.relation.publisherversionhttps://link.springer.com/article/10.1007/s10811-012-9811-7pt_PT
dc.subjectFluorescence microscopypt_PT
dc.subjectGlutathionept_PT
dc.subjectMicroplate assaypt_PT
dc.subjectMonochlorobimanept_PT
dc.subjectSelenastrum capricornutumpt_PT
dc.subjectToxicity assaypt_PT
dc.titleAssessment of cellular reduced glutathione content in Pseudokirchneriella subcapitata using monochlorobimanept_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.titleJournal of Applied Phycologypt_PT
person.familyNameSoares
person.givenNameEduardo
person.identifier93637
person.identifier.ciencia-idD612-0D0E-4984
person.identifier.orcid0000-0003-2280-5291
person.identifier.ridO-5908-2015
person.identifier.scopus-author-id7102060173
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublicationb3ebdee1-84fa-4183-a1d8-c3cdcaaef219
relation.isAuthorOfPublication.latestForDiscoveryb3ebdee1-84fa-4183-a1d8-c3cdcaaef219

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