Hashimoto-Partyka Minako K, Yuge Masahiro, Cho Ken W Y
Department of Developmental and Cell Biology, University of California, Irvine, California 92697-2300, USA.
Dev Biol. 2003 Jan 1;253(1):125-38. doi: 10.1006/dbio.2002.0867.
The classical three-signal model of amphibian mesoderm induction and more recent modifications together propose that an activin-like signaling activity is uniformly distributed across the vegetal half of the Xenopus blastula and that this activity contributes to mesoderm induction. In support of this, we have previously shown that the activin-response element (DE) of the goosecoid promoter is uniformly activated across the vegetal half of midgastrula-stage embryos. Here, we further examine the nature of this activity by measuring DE activation by endogenous signals over time. We find that the spatiotemporal pattern of DE activation is much more dynamic than was previously appreciated and also conclude that DE(6X)Luc activity reflects endogenous nodal signaling in the embryo. Using both the DE(6X)Luc construct and endogenous Xbra and Xgsc expression as read-outs for nodal activity, and the cleavage-mutant version of Xnr2 (CmXnr2) to regionally suppress endogenous nodal activity, we demonstrate that nodal signals act cell-autonomously in Xenopus gastrulae. Nodal-expressing cells are unable to rescue either reporter gene activation or target gene expression in distant nodal-deficient cells, suggesting that nodals function at short range in this context. Finally, we show that DE activation by endogenous signals occurs in the absence of dorsal beta-catenin-mediated signaling, but that the timing of dorsal initiation is altered. We conclude that nodal signals in Xenopus gastrulae function cell autonomously at short ranges and that the spatiotemporal pattern of this signaling along the dorsoventral axis is regulated by maternal Wnt-like signaling.
两栖动物中胚层诱导的经典三信号模型以及最近的修正共同表明,一种激活素样信号活性在非洲爪蟾囊胚的植物半球均匀分布,且这种活性有助于中胚层诱导。为此,我们之前已表明,鹅膏蕈氨酸启动子的激活素反应元件(DE)在原肠胚中期胚胎的植物半球被均匀激活。在此,我们通过测量内源性信号随时间对DE的激活作用,进一步研究这种活性的本质。我们发现,DE激活的时空模式比之前所认识到的更加动态,并且还得出结论,DE(6X)Luc活性反映了胚胎中的内源性节点信号。使用DE(6X)Luc构建体以及内源性Xbra和Xgsc表达作为节点活性的读出指标,并使用Xnr2的切割突变体版本(CmXnr2)来局部抑制内源性节点活性,我们证明节点信号在非洲爪蟾原肠胚中以细胞自主方式起作用。表达节点的细胞无法挽救远处缺乏节点的细胞中的报告基因激活或靶基因表达,这表明在这种情况下,节点在短距离内发挥作用。最后,我们表明内源性信号对DE的激活在没有背侧β-连环蛋白介导的信号传导的情况下发生,但背侧起始的时间发生了改变。我们得出结论,非洲爪蟾原肠胚中的节点信号在短距离内以细胞自主方式起作用,并且这种信号沿背腹轴的时空模式受母体类Wnt信号传导调节。