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

The insulinotropic actions of glucagon-like peptide 1 receptor (GLP-1R) in beta-cells have made it a useful target to manage type 2 diabetes. Metabolic stress reduces beta-cell sensitivity to GLP-1, yet the underlying mechanisms are unknown. We hypothesized thatGlp1rexpression is heterogeneous among beta-cells and that metabolic stress decreases the number of GLP-1R-positive beta-cells. Here, analyses of publicly available single-cell RNA-Seq sequencing (scRNASeq) data from mouse and human beta-cells indicated that significant populations of beta-cells do not express theGlp1rgene, 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 alpha-, beta-, and delta-cells. Experiments withGlp1rreporter mice and a validated GLP-1R antibody indicated that >90% of the beta-cells are GLP-1R positive, contradicting the findings with the scRNASeq data. alpha-cells did not expressGlp1rmRNA and delta-cells expressedGlp1rmRNA but not protein. We also examined the expression patterns of GLP-1R in mouse models of metabolic stress. Multiparous female mice had significantly decreased beta-cellGlp1rexpression, 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 beta-cells, absent in alpha-cells, and expressed at the mRNA, but not protein, level in delta-cells.