Online monitoring of stimulus-induced gene expression in pancreatic β-cells

Tilo Moede, Barbara Leibiger, Per Olof Berggren, Ingo B. Leibiger

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13 Scopus citations

Abstract

Fluorescent proteins have been extensively used as protein "tags" to study the subcellular localization of proteins and/or their translocation upon stimulation or as markers for transfection in transient and stable expression systems. However, they have not been frequently used as reporter genes to monitor stimulus-induced gene expression in mammalian cells. Here we demonstrate the use of fluorescent proteins to study stimulus-induced gene transcription. The general applicability of the approach is exemplified by doxycyclin-(Tet-On) and phorbol 12-myristate 13-acetate-induced (c-fos) promoter activation, with green fluorescent protein (GFP) and red fluorescent protein (DsRed) as semiquantitative and immediate reporters, of transcription activation. Under the control of β-cell-specific promoters, such as the rat insulin 1 promoter or the rat upstream glucokinase promoter, this approach allowed us to monitor online glucose-induced gene transcription in primary β-cells at the single-cell level as well as in the context of the islet of Langerhans. Applying discretely detectable fluorescent proteins, for example GFP and DsRed, enabled us to simultaneously monitor stimulus-induced transcription by two different promoters in the same cell.

Original languageEnglish (US)
Pages (from-to)S15-S19
JournalDiabetes
Volume50
Issue numberSUPPL. 1
DOIs
StatePublished - Jan 1 2001

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ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism

Cite this

Moede, T., Leibiger, B., Berggren, P. O., & Leibiger, I. B. (2001). Online monitoring of stimulus-induced gene expression in pancreatic β-cells. Diabetes, 50(SUPPL. 1), S15-S19. https://doi.org/10.2337/diabetes.50.2007.s15