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Discordance between GLP-1R gene and protein expression in mouse pancreatic islet cells.

Publication ,  Journal Article
Gray, SM; Xin, Y; Ross, EC; Chazotte, BM; Capozzi, ME; El, K; Svendsen, B; Ravn, P; Sloop, KW; Tong, J; Gromada, J; Campbell, JE; D'Alessio, DA
Published in: J Biol Chem
August 14, 2020

The insulinotropic actions of glucagon-like peptide 1 receptor (GLP-1R) in β-cells have made it a useful target to manage type 2 diabetes. Metabolic stress reduces β-cell sensitivity to GLP-1, yet the underlying mechanisms are unknown. We hypothesized that Glp1r expression is heterogeneous among β-cells and that metabolic stress decreases the number of GLP-1R-positive β-cells. Here, analyses of publicly available single-cell RNA-Seq sequencing (scRNASeq) data from mouse and human β-cells indicated that significant populations of β-cells do not express the Glp1r gene, supporting heterogeneous GLP-1R expression. To check these results, we used complementary approaches employing FACS coupled with quantitative RT-PCR, a validated GLP-1R antibody, and flow cytometry to quantify GLP-1R promoter activity, gene expression, and protein expression in mouse α-, β-, and δ-cells. Experiments with Glp1r reporter mice and a validated GLP-1R antibody indicated that >90% of the β-cells are GLP-1R positive, contradicting the findings with the scRNASeq data. α-cells did not express Glp1r mRNA and δ-cells expressed Glp1r mRNA but not protein. We also examined the expression patterns of GLP-1R in mouse models of metabolic stress. Multiparous female mice had significantly decreased β-cell Glp1r expression, but no reduction in GLP-1R protein levels or GLP-1R-mediated insulin secretion. These findings suggest caution in interpreting the results of scRNASeq for low-abundance transcripts such as the incretin receptors and indicate that GLP-1R is widely expressed in β-cells, absent in α-cells, and expressed at the mRNA, but not protein, level in δ-cells.

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Published In

J Biol Chem

DOI

EISSN

1083-351X

Publication Date

August 14, 2020

Volume

295

Issue

33

Start / End Page

11529 / 11541

Location

United States

Related Subject Headings

  • Single-Cell Analysis
  • Mice, Inbred C57BL
  • Mice
  • Insulin-Secreting Cells
  • Humans
  • Glucagon-Like Peptide-1 Receptor
  • Gene Expression
  • Cells, Cultured
  • Biochemistry & Molecular Biology
  • Animals
 

Citation

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Gray, S. M., Xin, Y., Ross, E. C., Chazotte, B. M., Capozzi, M. E., El, K., … D’Alessio, D. A. (2020). Discordance between GLP-1R gene and protein expression in mouse pancreatic islet cells. J Biol Chem, 295(33), 11529–11541. https://doi.org/10.1074/jbc.RA120.014368
Gray, Sarah M., Yurong Xin, Elizabeth C. Ross, Bryanna M. Chazotte, Megan E. Capozzi, Kimberley El, Berit Svendsen, et al. “Discordance between GLP-1R gene and protein expression in mouse pancreatic islet cells.J Biol Chem 295, no. 33 (August 14, 2020): 11529–41. https://doi.org/10.1074/jbc.RA120.014368.
Gray SM, Xin Y, Ross EC, Chazotte BM, Capozzi ME, El K, et al. Discordance between GLP-1R gene and protein expression in mouse pancreatic islet cells. J Biol Chem. 2020 Aug 14;295(33):11529–41.
Gray, Sarah M., et al. “Discordance between GLP-1R gene and protein expression in mouse pancreatic islet cells.J Biol Chem, vol. 295, no. 33, Aug. 2020, pp. 11529–41. Pubmed, doi:10.1074/jbc.RA120.014368.
Gray SM, Xin Y, Ross EC, Chazotte BM, Capozzi ME, El K, Svendsen B, Ravn P, Sloop KW, Tong J, Gromada J, Campbell JE, D’Alessio DA. Discordance between GLP-1R gene and protein expression in mouse pancreatic islet cells. J Biol Chem. 2020 Aug 14;295(33):11529–11541.

Published In

J Biol Chem

DOI

EISSN

1083-351X

Publication Date

August 14, 2020

Volume

295

Issue

33

Start / End Page

11529 / 11541

Location

United States

Related Subject Headings

  • Single-Cell Analysis
  • Mice, Inbred C57BL
  • Mice
  • Insulin-Secreting Cells
  • Humans
  • Glucagon-Like Peptide-1 Receptor
  • Gene Expression
  • Cells, Cultured
  • Biochemistry & Molecular Biology
  • Animals