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Conditional gene targeting in mouse pancreatic ß-Cells: analysis of ectopic Cre transgene expression in the brain.

OBJECTIVE: Conditional gene targeting has been extensively used for in vivo analysis of gene function in β-cell biology. The objective of this study was to examine whether mouse transgenic Cre lines, used to mediate β-cell- or pancreas-specific recombination, also drive Cre expression in the brain. RESEARCH DESIGN AND METHODS: Transgenic Cre lines driven by Ins1, Ins2, and Pdx1 promoters were bred to R26R reporter strains. Cre activity was assessed by β-galactosidase or yellow fluorescent protein expression in the pancreas and the brain. Endogenous Pdx1 gene expression was monitored using Pdx1(tm1Cvw) lacZ knock-in mice. Cre expression in β-cells and co-localization of Cre activity with orexin-expressing and leptin-responsive neurons within the brain was assessed by immunohistochemistry. RESULTS: All transgenic Cre lines examined that used the Ins2 promoter to drive Cre expression showed widespread Cre activity in the brain, whereas Cre lines that used Pdx1 promoter fragments showed more restricted Cre activity primarily within the hypothalamus. Immunohistochemical analysis of the hypothalamus from Tg(Pdx1-cre)(89.1Dam) mice revealed Cre activity in neurons expressing orexin and in neurons activated by leptin. Tg(Ins1-Cre/ERT)(1Lphi) mice were the only line that lacked Cre activity in the brain. CONCLUSIONS: Cre-mediated gene manipulation using transgenic lines that express Cre under the control of the Ins2 and Pdx1 promoters are likely to alter gene expression in nutrient-sensing neurons. Therefore, data arising from the use of these transgenic Cre lines must be interpreted carefully to assess whether the resultant phenotype is solely attributable to alterations in the islet β-cells.

Pubmed ID: 20802254

Authors

  • Wicksteed B
  • Brissova M
  • Yan W
  • Opland DM
  • Plank JL
  • Reinert RB
  • Dickson LM
  • Tamarina NA
  • Philipson LH
  • Shostak A
  • Bernal-Mizrachi E
  • Elghazi L
  • Roe MW
  • Labosky PA
  • Myers MG
  • Gannon M
  • Powers AC
  • Dempsey PJ

Journal

Diabetes

Publication Data

December 30, 2010

Associated Grants

  • Agency: NIDDK NIH HHS, Id: DK057768
  • Agency: NIDDK NIH HHS, Id: DK065131
  • Agency: NIDDK NIH HHS, Id: DK071052
  • Agency: NIDDK NIH HHS, Id: DK072473
  • Agency: NIDDK NIH HHS, Id: DK073716
  • Agency: NIDDK NIH HHS, Id: DK074966
  • Agency: NIDDK NIH HHS, Id: DK59637
  • Agency: NIDDK NIH HHS, Id: DK63363
  • Agency: NIDDK NIH HHS, Id: DK66636
  • Agency: NIDDK NIH HHS, Id: DK69603
  • Agency: NICHD NIH HHS, Id: HD36720
  • Agency: BLRD VA, Id: I01 BX000666
  • Agency: NIDDK NIH HHS, Id: P60 DK020572
  • Agency: NIDDK NIH HHS, Id: P60 DK020593
  • Agency: NIDDK NIH HHS, Id: P60 DK020593
  • Agency: NIDDK NIH HHS, Id: P60 DK020595
  • Agency: NIDDK NIH HHS, Id: P60 DK20595
  • Agency: NIDDK NIH HHS, Id: R01 DK068764
  • Agency: NIDDK NIH HHS, Id: R01 DK069603
  • Agency: NIDDK NIH HHS, Id: R01 DK073716
  • Agency: NIDDK NIH HHS, Id: R01 DK084236
  • Agency: NIDDK NIH HHS, Id: R33 DK066636
  • Agency: NIDDK NIH HHS, Id: T32DK07563
  • Agency: NIDDK NIH HHS, Id: U01 DK089572

Mesh Terms

  • Animals
  • Brain
  • Crosses, Genetic
  • Estrogen Antagonists
  • Female
  • Galactosides
  • Gene Targeting
  • Genes, Reporter
  • Immunoglobulin G
  • Immunohistochemistry
  • Insulin
  • Insulin-Secreting Cells
  • Integrases
  • Leptin
  • Male
  • Mice
  • Mice, Transgenic
  • Reverse Transcriptase Polymerase Chain Reaction
  • Swine
  • Tamoxifen