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Development of a Rapid Insulin Assay by Homogenous Time-Resolved Fluorescence

Direct measurement of insulin is critical for basic and clinical studies of insulin secretion. However, current methods are expensive and time-consuming. We developed an insulin assay based on homogenous time-resolved fluorescence that is significantly more rapid and cost-effective than current comm...

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Autores principales: Farino, Zachary J., Morgenstern, Travis J., Vallaghe, Julie, Gregor, Nathalie, Donthamsetti, Prashant, Harris, Paul E., Pierre, Nicolas, Freyberg, Robin, Charrier-Savournin, Fabienne, Javitch, Jonathan A., Freyberg, Zachary
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4743966/
https://www.ncbi.nlm.nih.gov/pubmed/26849707
http://dx.doi.org/10.1371/journal.pone.0148684
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author Farino, Zachary J.
Morgenstern, Travis J.
Vallaghe, Julie
Gregor, Nathalie
Donthamsetti, Prashant
Harris, Paul E.
Pierre, Nicolas
Freyberg, Robin
Charrier-Savournin, Fabienne
Javitch, Jonathan A.
Freyberg, Zachary
author_facet Farino, Zachary J.
Morgenstern, Travis J.
Vallaghe, Julie
Gregor, Nathalie
Donthamsetti, Prashant
Harris, Paul E.
Pierre, Nicolas
Freyberg, Robin
Charrier-Savournin, Fabienne
Javitch, Jonathan A.
Freyberg, Zachary
author_sort Farino, Zachary J.
collection PubMed
description Direct measurement of insulin is critical for basic and clinical studies of insulin secretion. However, current methods are expensive and time-consuming. We developed an insulin assay based on homogenous time-resolved fluorescence that is significantly more rapid and cost-effective than current commonly used approaches. This assay was applied effectively to an insulin secreting cell line, INS-1E cells, as well as pancreatic islets, allowing us to validate the assay by elucidating mechanisms by which dopamine regulates insulin release. We found that dopamine functioned as a significant negative modulator of glucose-stimulated insulin secretion. Further, we showed that bromocriptine, a known dopamine D2/D3 receptor agonist and newly approved drug used for treatment of type II diabetes mellitus, also decreased glucose-stimulated insulin secretion in islets to levels comparable to those caused by dopamine treatment.
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spelling pubmed-47439662016-02-11 Development of a Rapid Insulin Assay by Homogenous Time-Resolved Fluorescence Farino, Zachary J. Morgenstern, Travis J. Vallaghe, Julie Gregor, Nathalie Donthamsetti, Prashant Harris, Paul E. Pierre, Nicolas Freyberg, Robin Charrier-Savournin, Fabienne Javitch, Jonathan A. Freyberg, Zachary PLoS One Research Article Direct measurement of insulin is critical for basic and clinical studies of insulin secretion. However, current methods are expensive and time-consuming. We developed an insulin assay based on homogenous time-resolved fluorescence that is significantly more rapid and cost-effective than current commonly used approaches. This assay was applied effectively to an insulin secreting cell line, INS-1E cells, as well as pancreatic islets, allowing us to validate the assay by elucidating mechanisms by which dopamine regulates insulin release. We found that dopamine functioned as a significant negative modulator of glucose-stimulated insulin secretion. Further, we showed that bromocriptine, a known dopamine D2/D3 receptor agonist and newly approved drug used for treatment of type II diabetes mellitus, also decreased glucose-stimulated insulin secretion in islets to levels comparable to those caused by dopamine treatment. Public Library of Science 2016-02-05 /pmc/articles/PMC4743966/ /pubmed/26849707 http://dx.doi.org/10.1371/journal.pone.0148684 Text en © 2016 Farino et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Farino, Zachary J.
Morgenstern, Travis J.
Vallaghe, Julie
Gregor, Nathalie
Donthamsetti, Prashant
Harris, Paul E.
Pierre, Nicolas
Freyberg, Robin
Charrier-Savournin, Fabienne
Javitch, Jonathan A.
Freyberg, Zachary
Development of a Rapid Insulin Assay by Homogenous Time-Resolved Fluorescence
title Development of a Rapid Insulin Assay by Homogenous Time-Resolved Fluorescence
title_full Development of a Rapid Insulin Assay by Homogenous Time-Resolved Fluorescence
title_fullStr Development of a Rapid Insulin Assay by Homogenous Time-Resolved Fluorescence
title_full_unstemmed Development of a Rapid Insulin Assay by Homogenous Time-Resolved Fluorescence
title_short Development of a Rapid Insulin Assay by Homogenous Time-Resolved Fluorescence
title_sort development of a rapid insulin assay by homogenous time-resolved fluorescence
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4743966/
https://www.ncbi.nlm.nih.gov/pubmed/26849707
http://dx.doi.org/10.1371/journal.pone.0148684
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