A novel high-throughput assay for islet respiration reveals uncoupling of rodent and human islets

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dc.contributor.authorWikstrom, J.
dc.contributor.authorSereda, S.
dc.contributor.authorStiles, L.
dc.contributor.authorElorza, A.
dc.contributor.authorAllister, E.
dc.contributor.authorNeilson, A.
dc.contributor.authorFerrick, D.
dc.contributor.authorWheeler, M.
dc.contributor.authorShirihai, O.
dc.date.accessioned2023-06-23T20:14:08Z
dc.date.available2023-06-23T20:14:08Z
dc.date.issued2012-05
dc.descriptionIndexación: Scopus.es
dc.description.abstractThe pancreatic beta cell is unique in its response to nutrient by increased fuel oxidation. Recent studies have demonstrated that oxygen consumption rate (OCR) may be a valuable predictor of islet quality and long term nutrient responsiveness. To date, high-throughput and user-friendly assays for islet respiration are lacking. The aim of this study was to develop such an assay and to examine bioenergetic efficiency of rodent and human islets. Methodology/Principal Findings: The XF24 respirometer platform was adapted to islets by the development of a 24-well plate specifically designed to confine islets. The islet plate generated data with low inter-well variability and enabled stable measurement of oxygen consumption for hours. The F1F0 ATP synthase blocker oligomycin was used to assess uncoupling while rotenone together with myxothiazol/antimycin was used to measure the level of non-mitochondrial respiration. The use of oligomycin in islets was validated by reversing its effect in the presence of the uncoupler FCCP. Respiratory leak averaged to 59% and 49% of basal OCR in islets from C57Bl6/J and FVB/N mice, respectively. In comparison, respiratory leak of INS-1 cells and C2C12 myotubes was measured to 38% and 23% respectively. Islets from a cohort of human donors showed a respiratory leak of 38%, significantly lower than mouse islets. Conclusions/Significance: The assay for islet respiration presented here provides a novel tool that can be used to study islet mitochondrial function in a relatively high-throughput manner. The data obtained in this study shows that rodent islets are less bioenergetically efficient than human islets as well as INS1 cells.es
dc.description.urihttps://journals.plos.org/plosone/article?id=10.1371/journal.pone.0033023#ack
dc.identifier.citationPLoS ONE, Volume 7, Issue 514, May 2012, Article number e33023es
dc.identifier.doi10.1371/journal.pone.0033023
dc.identifier.issn1932-6203
dc.identifier.urihttps://repositorio.unab.cl/xmlui/handle/ria/51047
dc.language.isoenes
dc.publisherPublic Library of Science (PLoS)es
dc.rights.licenseAttribution 2.0 Generic (CC BY 2.0)
dc.rights.urihttps://plos.org/open-science/open-access/
dc.subjectPancreas Isletes
dc.subjectPancreatectomyes
dc.subjectAutograftinges
dc.titleA novel high-throughput assay for islet respiration reveals uncoupling of rodent and human isletses
dc.typeArtículoes
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