Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, USA.
Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, USA.
Dev Biol. 2014 Aug 15;392(2):283-94. doi: 10.1016/j.ydbio.2014.05.017. Epub 2014 Jun 12.
The intestinal epithelium performs vital roles in organ function by absorbing nutrients and providing a protective barrier. The zinc-finger containing transcription factors GATA4 and GATA6 regulate enterocyte gene expression and control regional epithelial cell identity in the adult intestinal epithelium. Although GATA4 and GATA6 are expressed in the developing intestine, loss of either factor alone during the period of epithelial morphogenesis and cytodifferentiation fails to disrupt these processes. Therefore, we tested the hypothesis that GATA4 and GATA6 function redundantly to control these aspects of intestinal development. We used Villin-Cre, which deletes specifically in the intestinal epithelium during the period of villus development and epithelial cytodifferentiation, to generate Gata4Gata6 double conditional knockout embryos. Mice lacking GATA4 and GATA6 in the intestinal epithelium died within 24h of birth. At E18.5, intestinal villus architecture and epithelial cell populations were altered. Enterocytes were lost, and goblet cells were increased. Proliferation was also increased in GATA4-GATA6 deficient intestinal epithelium. Although villus morphology appeared normal at E16.5, the first time at which both Gata4 and Gata6 were efficiently reduced, changes in expression of markers of enterocytes, goblet cells, and proliferative cells were detected. Moreover, goblet cell number was increased at E16.5. Expression of the Notch ligand Dll1 and the Notch target Olfm4 were reduced in mutant tissue indicating decreased Notch signaling. Finally, we found that GATA4 occupies chromatin near the Dll1 transcription start site suggesting direct regulation of Dll1 by GATA4. We demonstrate that GATA4 and GATA6 play an essential role in maintaining proper intestinal epithelial structure and in regulating intestinal epithelial cytodifferentiation. Our data highlight a novel role for GATA factors in fine tuning Notch signaling during intestinal epithelial development to repress goblet cell differentiation.
肠上皮通过吸收营养物质和提供保护屏障来发挥重要作用。锌指转录因子 GATA4 和 GATA6 调节肠上皮细胞基因表达,并控制成年肠上皮细胞的区域上皮细胞身份。尽管 GATA4 和 GATA6 在发育中的肠道中表达,但在肠上皮形态发生和细胞分化期间单独丢失任一因子都不会破坏这些过程。因此,我们测试了 GATA4 和 GATA6 冗余地发挥作用以控制这些肠发育方面的假设。我们使用 Villin-Cre,其在绒毛发育和上皮细胞分化期间特异性地在肠上皮中缺失,以生成 Gata4Gata6 双条件性敲除胚胎。肠上皮中缺乏 GATA4 和 GATA6 的小鼠在出生后 24 小时内死亡。在 E18.5 时,肠绒毛结构和上皮细胞群体发生改变。肠上皮细胞丢失,杯状细胞增加。GATA4-GATA6 缺陷肠上皮中的增殖也增加。尽管在 E16.5 时绒毛形态似乎正常,这是 Gata4 和 Gata6 首次被有效降低的时候,已经检测到肠上皮中肠上皮细胞、杯状细胞和增殖细胞标记物的表达变化。此外,在 E16.5 时杯状细胞数量增加。突变组织中 Notch 配体 Dll1 和 Notch 靶标 Olfm4 的表达减少表明 Notch 信号降低。最后,我们发现 GATA4 占据 Dll1 转录起始位点附近的染色质,表明 GATA4 对 Dll1 的直接调节。我们证明 GATA4 和 GATA6 在维持适当的肠上皮结构和调节肠上皮细胞分化方面发挥着重要作用。我们的数据突出了 GATA 因子在精细调节肠上皮发育过程中的 Notch 信号以抑制杯状细胞分化中的新作用。