Xanthos J B, Kofron M, Wylie C, Heasman J
Department of Genetics, Cell Biology, and Development, University of Minnesota, 321 Church Street SE, Minneapolis, MN 55455, USA.
Development. 2001 Jan;128(2):167-80. doi: 10.1242/dev.128.2.167.
During cleavage stages, maternal VegT mRNA and protein are localized to the Xenopus embryo's vegetal region from which the endoderm will arise and where several zygotic gene transcripts will be localized. Previous loss-of-function experiments on this T-box transcription factor suggested a role for VegT in Xenopus endoderm formation. Here, we test whether VegT is required to initiate endoderm formation using a loss of function approach. We find that the endodermal genes, Bix1, Bix3, Bix4, Milk (Bix2), Mix.1, Mix.2, Mixer, Xsox17 alpha, Gata4, Gata5, Gata6 and endodermin, as well as the anterior endodermal genes Xhex and cerberus, and the organizer specific gene, Xlim1, are downstream of maternal VegT. We also find that the TGF beta s, Xnr1, Xnr2, Xnr4 and derrière rescue expression of these genes, supporting the idea that cell interactions are critical for proper endoderm formation. Additionally, inhibitory forms of Xnr2 and Derrière blocked the ability of VegT mRNA injection to rescue VegT-depleted embryos. Furthermore, a subset of endodermal genes was rescued in VegT-depleted vegetal masses by induction from an uninjected vegetal mass. Finally, we begin to establish a gene hierarchy downstream of VegT by testing the ability of Mixer and Gata5 to rescue the expression of other endodermal genes. These results identify VegT as the maternal regulator of endoderm initiation and illustrate the complexity of zygotic pathways activated by VegT in the embryo's vegetal region.
在卵裂阶段,母体VegT mRNA和蛋白质定位于非洲爪蟾胚胎的植物区域,内胚层将从该区域产生,并且一些合子基因转录本也将定位于此。先前针对这种T盒转录因子的功能丧失实验表明,VegT在非洲爪蟾内胚层形成中发挥作用。在此,我们使用功能丧失方法来测试VegT是否是启动内胚层形成所必需的。我们发现,内胚层基因Bix1、Bix3、Bix4、Milk(Bix2)、Mix.1、Mix.2、Mixer、Xsox17α、Gata4、Gata5、Gata6和内胚层决定蛋白,以及前内胚层基因Xhex和Cerberus,还有组织者特异性基因Xlim1,都是母体VegT的下游基因。我们还发现,TGFβ家族成员Xnr1、Xnr2、Xnr4和Derrière可挽救这些基因的表达,这支持了细胞间相互作用对于正确的内胚层形成至关重要的观点。此外,Xnr2和Derrière的抑制形式阻断了VegT mRNA注射挽救VegT缺失胚胎的能力。此外,通过未注射的植物块诱导,在VegT缺失的植物块中挽救了一部分内胚层基因。最后,我们通过测试Mixer和Gata5挽救其他内胚层基因表达的能力,开始建立VegT下游的基因层级关系。这些结果确定VegT是内胚层起始的母体调节因子,并阐明了VegT在胚胎植物区域激活的合子途径的复杂性。