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

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  • Additional Information
    • Source:
      Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE
    • Publication Information:
      Original Publication: San Francisco, CA : Public Library of Science
    • Subject Terms:
      Fluorescence*; Insulin/*analysis ; Insulin-Secreting Cells/*metabolism; Animals ; Bromocriptine/chemistry ; Bromocriptine/pharmacology ; Cell Line ; Insulin/metabolism ; Insulin Secretion ; Insulin-Secreting Cells/cytology ; Mice ; Rats ; Receptors, Dopamine D2/agonists ; Receptors, Dopamine D2/metabolism ; Receptors, Dopamine D3/agonists ; Receptors, Dopamine D3/metabolism
    • 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.
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    • Grant Information:
      K05 DA022413 United States DA NIDA NIH HHS; K08 DA031241 United States DA NIDA NIH HHS; R01 DK077493 United States DK NIDDK NIH HHS; R01 DK104740 United States DK NIDDK NIH HHS
    • Accession Number:
      0 (Insulin)
      0 (Receptors, Dopamine D2)
      0 (Receptors, Dopamine D3)
      3A64E3G5ZO (Bromocriptine)
    • Publication Date:
      Date Created: 20160206 Date Completed: 20160707 Latest Revision: 20190222
    • Publication Date:
      20240104
    • Accession Number:
      PMC4743966
    • Accession Number:
      10.1371/journal.pone.0148684
    • Accession Number:
      26849707