Single cell transcriptional analysis of regenerating parietal cells (PCs) in murine gastric epithelium recovering from Diphtheria toxin injury

Highly specialized, acid-secreting PCs are one of many epithelial lineages that line the gastric epithelium. Even though PCs are functionally and structurally distinct from other lineages in the stomach, the molecular mechanisms that govern their specification and maturation from the gastric stem cells are relatively unknown. We used single cell RNA sequencing (scRNA-seq) to construct a transcriptional profile for regenerating PCs and identify candidate regulators of PC fate specification and maturation. Overall design: Transgenic Atp4b-DTR mice were treated daily with diphtheria toxin for 3 days to induce reversible PC ablation. The gastric corpus was harvested at 36, 48, or 72 hours after the final injection for the experimental groups and from vehicle treated mice for the uninjured control sample. The corpus was dissociated to the single cell level using a combination of mechanical and chemical techniques. Three mice were used per sample group/condition.