Stromal DLK1 promotes proliferation and inhibits differentiation of the intestinal epithelium during development
Stromal DLK1 promotes proliferation and inhibits differentiation of the intestinal epithelium during development
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DOI：
10.1152/ajpgi.00445.2020
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发表时间：
2021-04-01
影响因子：
4.5
通讯作者：
Woods, Susan L.
中科院分区：
文献类型：
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作者：
Ichinose, Mari;Suzuki, Nobumi;Woods, Susan L.
发育中的肠的干/祖细胞在生物学上与它们的成年对应物不同。在这里，我们研究了调节胚胎干/祖细胞群体的微环境线索，重点是Notch途径因子δ样蛋白-1（DLK 1）的作用。使用从胚胎第14.5天（E14.5）或成年IMC收集的肠间充质细胞（IMC）的mRNA-seq分析以及与E14.5肠上皮类器官的新型共培养系统。加入重组DLK 1（rDLK）或Dlk 1 siRNA（siDlk 1）后，使用成像、再铺板效率测定、qPCR和免疫细胞化学比较上皮特征。采用免疫组织化学方法比较同窝Dlk 1（+）（/+）和Dlk 1（-/-）小鼠的肠道表型。使用转录组学分析，我们鉴定了来自胚胎间充质的可能调节发育中的上皮细胞的形态发生素，重点是Notch家族候选者DLK 1。免疫组化结果表明，DLK 1只在E14.5的肠间质中表达，位于新生绒毛的顶部，出生后下降，成年后转移到肠上皮。在共培养实验中，除了rDLK 1的成年IMC抑制类器官分化，而Dlk 1敲低胚胎IMC增加上皮细胞分化为分泌谱系细胞。与Dlk 1（+）（/+）胚胎相比，Dlk 1胚胎的Ki 67（+）细胞局限于绒毛基部，分泌细胞增多。间充质衍生的DLK 1在促进上皮干/前体扩张和防止发育中的肠道分化为分泌谱系方面发挥着重要作用。新&值得注意使用新型共培养系统、转录组学和转基因小鼠，我们研究了肠上皮之间的差异分子信号传导发育期间和成人中的间充质。我们发现Notch途径因子δ样蛋白-1（DLK 1）在发育过程中由基质产生，并揭示DLK 1在调节肠上皮干/前体扩张和分化为分泌谱系中的新作用。
The stem/progenitor cells of the developing intestine are biologically distinct from their adult counterparts. Here, we examine the microenvironmental cues that regulate the embryonic stem/progenitor population, focusing on the role of Notch pathway factor delta-like protein-1 (DLK1). mRNA-seq analyses of intestinal mesenchymal cells (IMCs) collected from embryonic day 14.5 (E14.5) or adult IMCs and a novel coculture system with E14.5 intestinal epithelial organoids were used. Following addition of recombinant DLK1 (rDLK) or Dlk1 siRNA (siDlk1), epithelial characteristics were compared using imaging, replating efficiency assays, qPCR, and immunocytochemistry. The intestinal phenotypes of littermate Dlk1(+)(/+) and Dlk1(-/-) mice were compared using immunohistochemistry. Using transcriptomic analyses, we identified morphogens derived from the embryonic mesenchyme that potentially regulate the developing epithelial cells, to focus on Notch family candidate DLK1. Immunohistochemistry indicated that DLK1 was expressed exclusively in the intestinal stroma at E14.5 at the top of emerging villi, decreased after birth, and shifted to the intestinal epithelium in adulthood. In coculture experiments, addition of rDLK1 to adult IMCs inhibited organoid differentiation, whereas Dlk1 knockdown in embryonic IMCs increased epithelial differentiation to secretory lineage cells. Dlk1 imice had restricted Ki67(+) cells in the villi base and increased secretory lineage cells compared with Dlk1(+)(/+) embryos. Mesenchyme-derived DLK1 plays an important role in the promotion of epithelial stem/precursor expansion and prevention of differentiation to secretory lineages in the developing intestine.NEW & NOTEWORTHY Using a novel coculture system, transcriptomics, and transgenic mice, we investigated differential molecular signaling between the intestinal epithelium and mesenchyme during development and in the adult. We show that the Notch pathway factor delta-like protein-1 (DLK1) is stromally produced during development and uncover a new role for DLK1 in the regulation of intestinal epithelial stem/precursor expansion and differentiation to secretory lineages.