A transcriptome-led exploration of molecular mechanisms regulating somatostatin-producing D-cells in the gastric epithelium
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Adriaenssens, A., Lam, B., Billing, L., Skeffington, K., Sewing, S., Reimann, F., & Gribble, F. (2015). A transcriptome-led exploration of molecular mechanisms regulating somatostatin-producing D-cells in the gastric epithelium. Endocrinology https://doi.org/10.1210/en.2015-1301
The stomach epithelium contains a myriad of enteroendocrine cells that modulate a range of physiological functions, including postprandial secretion of regulatory peptides, gastric motility, and nutrient absorption. Somatostatin (SST)-producing D-cells are present in the oxyntic and pyloric regions of the stomach, and provide a tonic inhibitory tone that regulates activity of neighboring enteroendocrine cells and gastric acid secretion. Cellular mechanisms underlying the effects of regulatory factors on gastric D-cells are poorly defined due to problems in identifying primary D-cells, and uncertainty remains about which stimuli influence D-cells directly. In this study, we introduce a transgenic mouse line, SST-Cre, which upon crossing with Cre reporter strains, facilitates the identification and purification of gastric D-cells, or cell-specific expression of genetically encoded calcium indicators. Populations of D-cells from the gastric antrum and corpus were isolated and analyzed by RNA sequencing and quantitative RT-PCR. The expression of hormones, hormone receptors, neurotransmitter receptors, and nutrient receptors was quantified. Pyy, Gipr, Chrm4, Calcrl, Taar1, and Casr were identified as genes that are highly enriched in D-cells compared with SST-negative cells. Hormone secretion assays performed in mixed gastric epithelial cultures confirmed that SST secretion is regulated by incretin hormones, cholecystokinin, acetylcholine, vasoactive intestinal polypeptide, calcitonin gene-related polypeptide, oligopetides, and trace amines. Cholecystokinin and oligopeptides elicited increases in intracellular calcium in single-cell imaging experiments performed using cultured D-cells. Our data provide the first transcriptomic analysis and functional characterization of gastric D-cells, and identify regulatory pathways that underlie the direct detection of stimuli by this cell type.
Gastric D-cells, somatostatin, enteroendocrine, RNA sequencing, G-protein coupled receptors
This work was supported by the Wellcome Trust (WT088357/Z/09/Z and WT084210/Z/07/Z) and the MRC Metabolic Diseases Unit (MRC_MC_UU_12012/3).
Medical Research Council (MC_UU_12012/5/B)
External DOI: https://doi.org/10.1210/en.2015-1301
This record's URL: https://www.repository.cam.ac.uk/handle/1810/250306
Creative Commons Attribution 4.0
Licence URL: http://creativecommons.org/licenses/by/4.0/