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

Author

Listed:
  • Zachary J Farino
  • Travis J Morgenstern
  • Julie Vallaghe
  • Nathalie Gregor
  • Prashant Donthamsetti
  • Paul E Harris
  • Nicolas Pierre
  • Robin Freyberg
  • Fabienne Charrier-Savournin
  • Jonathan A Javitch
  • Zachary Freyberg

Abstract

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.

Suggested Citation

  • Zachary J Farino & Travis J Morgenstern & Julie Vallaghe & Nathalie Gregor & Prashant Donthamsetti & Paul E Harris & Nicolas Pierre & Robin Freyberg & Fabienne Charrier-Savournin & Jonathan A Javitch , 2016. "Development of a Rapid Insulin Assay by Homogenous Time-Resolved Fluorescence," PLOS ONE, Public Library of Science, vol. 11(2), pages 1-17, February.
    • Handle: RePEc:plo:pone00:0148684
    DOI: 10.1371/journal.pone.0148684
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    References listed on IDEAS

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    1. Steven E. Kahn & Rebecca L. Hull & Kristina M. Utzschneider, 2006. "Mechanisms linking obesity to insulin resistance and type 2 diabetes," Nature, Nature, vol. 444(7121), pages 840-846, December.
    2. Carol Kilkenny & William J Browne & Innes C Cuthill & Michael Emerson & Douglas G Altman, 2010. "Improving Bioscience Research Reporting: The ARRIVE Guidelines for Reporting Animal Research," PLOS Biology, Public Library of Science, vol. 8(6), pages 1-5, June.
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