Kemp Caroline R, Willems Erik, Wawrzak Danuta, Hendrickx Marijke, Agbor Agbor Terence, Leyns Luc
Vrije Universiteit Brussel, Lab for Cell Genetics, Brussels, Belgium.
Dev Dyn. 2007 Jul;236(7):2011-9. doi: 10.1002/dvdy.21198.
Wnt signaling has been shown to be important in the patterning of the gastrulating mouse embryo, especially in axis formation. To this date, there is no clear indication that the Wnt receptors, Frizzleds (Fzds), are involved in such early specification. Moreover, at the gastrulation stage, the only Fzd with a known characterized expression pattern is Fzd8, which is expressed in the anterior visceral endoderm (aVE) (Lu et al. [2004] Gene Expr Patterns 4:569-572). Following a real time RT-PCR study to evaluate Fzd expression in the gastrulating embryo, we used whole-mount in situ hybridization to reveal new expression domains for Fzd5, Fzd7, and Fzd10. Fzd5 is expressed in the aVE and Fzd7 expression is restricted to the epiblast of the gastrulating embryo. The expression pattern of Fzd10 in the primitive streak of the gastrula suggests it has a role in mesoderm induction. We also show that the purified, secreted forms of the extracellular cysteine-rich domains (CRDs) of FZD5, Fzd7, and Fzd8 can antagonize Wnt3a-induced beta-Catenin accumulation in L-cells, whereas in mouse embryonic stem cells, these CRDs can inhibit spontaneous mesoderm formation and promote neural differentiation. Our data demonstrate that Fzd5, Fzd7, and Fzd10 are expressed in distinct domains of the gastrulating embryo, and that the CRDs of FZD5, Fzd7, and Fzd8 can regulate Wnts, indicating that Fzds interpret Wnt signals during embryonic mesoderm and neural induction.
Wnt信号通路已被证明在原肠胚形成期的小鼠胚胎模式形成中很重要,尤其是在轴的形成过程中。迄今为止,尚无明确迹象表明Wnt受体卷曲蛋白(Fzds)参与了这种早期的细胞特化过程。此外,在原肠胚形成阶段,唯一具有已知特征性表达模式的Fzd是Fzd8,它在内脏内胚层前部(aVE)表达(Lu等人,[2004]《基因表达模式》4:569 - 572)。在进行实时RT-PCR研究以评估原肠胚形成期胚胎中Fzd的表达后,我们使用全胚胎原位杂交来揭示Fzd5、Fzd7和Fzd10的新表达域。Fzd5在aVE中表达,Fzd7的表达局限于原肠胚形成期胚胎的上胚层。Fzd10在原肠胚原条中的表达模式表明它在中胚层诱导中起作用。我们还表明,纯化的FZD5、Fzd7和Fzd8的富含半胱氨酸的细胞外结构域(CRDs)的分泌形式可以拮抗Wnt3a诱导的L细胞中β-连环蛋白的积累,而在小鼠胚胎干细胞中,这些CRDs可以抑制自发的中胚层形成并促进神经分化。我们的数据表明,Fzd5、Fzd7和Fzd10在原肠胚形成期胚胎的不同区域表达,并且FZD5、Fzd7和Fzd8的CRDs可以调节Wnt信号通路,这表明Fzds在胚胎中胚层和神经诱导过程中解读Wnt信号。