Discordance between GLP-1R gene and protein expression in mouse pancreatic islet cells
| Autor: | Yurong Xin, Sarah M. Gray, David A. D'Alessio, Megan E. Capozzi, Elizabeth C. Ross, Kimberley El, Berit Svendsen, Jenny Tong, Jesper Gromada, Peter Ravn, Jonathan E. Campbell, Bryanna M. Chazotte, Kyle W. Sloop |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
endocrine system Incretin Gene Expression Biochemistry Glucagon-Like Peptide-1 Receptor Flow cytometry 03 medical and health sciences Mice Insulin-Secreting Cells Gene expression medicine Animals Humans Receptor Molecular Biology Gene Cells Cultured Messenger RNA 030102 biochemistry & molecular biology biology medicine.diagnostic_test digestive oral and skin physiology Cell Biology Cell sorting Cell biology Mice Inbred C57BL 030104 developmental biology Metabolism biology.protein Antibody Single-Cell Analysis hormones hormone substitutes and hormone antagonists |
| Zdroj: | J Biol Chem |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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