Microarray 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 microarray analysis to interrogate transcriptional changes that occur in regenerating PCs and to identify candidate regulators of PC fate specification and maturation. Transgenic Atp4b-DTR mice were treated daily with diphtheria toxin for 3 days to induce reversible PC ablation. The gastric corpus was harvested at 24, 36, 48, or 72 hours after the final injection for the experimental groups and from vehicle treated mice for the uninjured control sample. Two independent samples were collected for 48 and72h timepoints; all others had 1 sample. The corpus was collected, snap frozen, then RNA isolated using Qiagen RNeasy Micro Kit and qulality assesed with Nanodrop.

高度特化的泌酸壁细胞（Parietal Cells, PCs）是构成胃上皮的多种上皮细胞谱系之一。尽管壁细胞在功能与结构上均与胃内其他细胞谱系存在显著差异，但调控其从胃干细胞定向分化并成熟的分子机制仍未完全明晰。本研究通过基因芯片（microarray）分析探究了再生状态下壁细胞的转录组变化，并筛选得到调控壁细胞命运定向与成熟过程的候选因子。我们对转Atp4b-DTR基因的小鼠每日给予白喉毒素处理，持续3天，以诱导可逆的壁细胞清除模型。实验组小鼠于末次注射后24、36、48或72小时收取胃体组织，未损伤对照组小鼠则收取溶剂处理后的组织样本。48小时与72小时时间点各收取两份独立样本，其余时间点各收取一份样本。收取胃体组织后进行快速冷冻，随后使用Qiagen RNeasy微量试剂盒提取RNA，并通过Nanodrop评估RNA质量。