title: Novel transgenic mice for inducible gene overexpression in pancreatic cells define glucocorticoid receptor-mediated regulations of beta cells.
abstract: Conditional gene deletion in specific cell populations has helped the understanding of pancreas development. Using this approach, we have shown that deleting the glucocorticoid receptor (GR) gene in pancreatic precursor cells leads to a doubled beta-cell mass. Here, we provide genetic tools that permit a temporally and spatially controlled expression of target genes in pancreatic cells using the Tetracycline inducible system. To efficiently target the Tetracycline transactivator (tTA) in specific cell populations, we generated Bacterial Artificial Chromosomes (BAC) transgenic mice expressing the improved Tetracycline transactivator (itTA) either in pancreatic progenitor cells expressing the transcription factor Pdx1 (BAC-Pdx1-itTA), or in beta cells expressing the insulin1 gene (BAC-Ins1-itTA). In the two transgenic models, itTA-mediated activation of reporter genes was efficient and subject to regulation by Doxycycline (Dox). The analysis of a tetracycline-regulated LacZ reporter gene shows that in BAC-Pdx1-itTA mice, itTA is expressed from embryonic (E) day 11.5 in all pancreatic precursor cells. In the adult pancreas, itTA is active in mature beta, delta cells and in few acinar cells. In BAC-Ins1-itTA mice tTA is active from E13.5 and is restricted to beta cells in fetal and adult pancreas. In both lines, tTA activity was suppressed by Dox treatment and re-induced after Dox removal. Using these transgenic lines, we overexpressed the GR in selective pancreatic cell populations and found that overexpression in precursor cells altered adult beta-cell fraction but not glucose tolerance. In contrast, GR overexpression in mature beta cells did not alter beta-cell fraction but impaired glucose tolerance with insufficient insulin secretion. In conclusion, these new itTA mouse models will allow fine-tuning of gene expression to investigate gene function in pancreatic biology and help us understand how glucocorticoid signaling affects on the long-term distinct aspects of beta-cell biology.
authors.affiliation: INSERM UMR-S 872, Centre de Recherches des Cordeliers, Paris, France. Bertrand.blondeau@crc.jussieu.fr
authors.forename: Bertrand
authors.initials: B
authors.lastname: Blondeau
authors.forename: Iman
authors.initials: I
authors.lastname: Sahly
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Massouridès
authors.forename: Amrit
authors.initials: A
authors.lastname: Singh-Estivalet
authors.forename: Bérengère
authors.initials: B
authors.lastname: Valtat
authors.forename: Delphine
authors.initials: D
authors.lastname: Dorchene
authors.forename: Frédéric
authors.initials: F
authors.lastname: Jaisser
authors.forename: Bernadette
authors.initials: B
authors.lastname: Bréant
authors.forename: Francois
authors.initials: F
authors.lastname: Tronche
mesh_terms: D000818:Animals; D004318:Doxycycline / Q000494:pharmacology; D005333:Fetus / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D005947:Glucose / Q000378:metabolism; D018398:Homeodomain Proteins / Q000378:metabolism; D006706:Homeostasis / Q000187:drug effects; D007328:Insulin / Q000235:genetics; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000378:metabolism*; D007763:Lac Operon; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology; D015534:Trans-Activators / Q000378:metabolism
chemical_list: D018398:Homeodomain Proteins; D007328:Insulin; D011965:Receptors, Glucocorticoid; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D013752:Tetracycline; D005947:Glucose; D004318:Doxycycline