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Am J Physiol Endocrinol Metab. 2018 Dec 11. doi: 10.1152/ajpendo.00229.2018. [Epub ahead of print]

Transgene-associated hGH expression in pancreatic β-cells impairs identification of sex-based gene expression differences.

Author information

1
Cell and Developmental Biology, Vanderbilt University School of Medicine.
2
Molecular Physiology and Biophysics, Vanderbilt University School of Medicine.
3
Center for Stem Cell Biology, Vanderbilt University School of Medicine.
4
Center for Stem Cell Biology, Vanderbilt University School of Medicine, United States.

Abstract

Fluorescence protein reporter genes are widely used to identify and sort murine pancreatic ß-cells. In this study we compared use of the MIP-GFP transgene, which exhibits aberrant expression of human growth hormone (hGH), with a newly derived Ins2Apple allele that lacks hGH expression on the expression of sex-specific genes. ß-cells from MIP-GFP transgenic mice exhibit changes in the expression of 7,733 genes, or greater than half of their transcriptome, when compared to ß-cells from Ins2Apple/+ mice. To determine how these differences might affect a typical differential gene expression study, we analyzed the effect of sex on gene expression using both reporter lines. 657 differentially-expressed genes were identified between male and female ß-cells containing the Ins2Apple allele. Female ß-cells exhibit higher expression of Xist, Tmed9, Arpc3, Eml2 and several islet-enriched transcription factors, including Nkx2-2 and Hnf4a, whereas male ß-cells exhibited a generally higher expression of genes involved in cell cycle regulation. In marked contrast, the same male versus female comparison of ß-cells containing the MIP-GFP transgene revealed only 115 differentially-expressed genes, and comparison of the two lists of differentially expressed genes revealed only 17 that were common to both analyses. These results indicate 1) that male and female ß-cells differ in their expression of key transcription factors and cell cycle regulators, and 2) that the MIP-GFP transgene may attenuate sex-specific differences that distinguish male and female ß-cells, thereby impairing the identification of sex-specific variations.

KEYWORDS:

Pancreatic β-cells; RNA-seq; gene expression; growth hormone; sex

PMID:
30532991
DOI:
10.1152/ajpendo.00229.2018

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