Tremblay K D, Dunn N R, Robertson E J
Molecular and Cellular Biology, Harvard University, 16 Divinity Avenue, Cambridge, MA 02138, USA.
Development. 2001 Sep;128(18):3609-21. doi: 10.1242/dev.128.18.3609.
The Smad proteins are important intracellular mediators of the transforming growth factor beta (TGFbeta) family of secreted growth factors. Smad1 is an effector of signals provided by the bone morphogenetic protein (BMP) sub-group of TGFbeta molecules. To understand the role of Smad1 in mouse development, we have generated a Smad1 loss-of-function allele using homologous recombination in ES cells. Smad1-/- embryos die by 10.5 dpc because they fail to connect to the placenta. Mutant embryos are first recognizable by 7.0 dpc, owing to a characteristic localized outpocketing of the visceral endoderm at the posterior embryonic/extra-embryonic junction, accompanied by a dramatic twisting of the epiblast and nascent mesoderm. Chimera analysis reveals that these two defects are attributable to a requirement for Smad1 in the extra-embryonic tissues. By 7.5 dpc, Smad1-deficient embryos show a marked impairment in allantois formation. By contrast, the chorion overproliferates, is erratically folded within the extra-embryonic space and is impeded in proximal migration. BMP signals are known to be essential for the specification and proliferation of primordial germ cells. We find a drastic reduction of primordial germ cells in Smad1-deficient embryos, suggesting an essential role for Smad1-dependent signals in primordial germ cell specification. Surprisingly, despite the key involvement of BMP signaling in tissues of the embryo proper, Smad1-deficient embryos develop remarkably normally. An examination of the expression domains of Smad1, Smad5 and Smad8 in early mouse embryos show that, while Smad1 is uniquely expressed in the visceral endoderm at 6.5 dpc, in other tissues Smad1 is co-expressed with Smad5 and/or Smad8. Collectively, these data have uncovered a unique function for Smad1 signaling in coordinating the growth of extra-embryonic structures necessary to support development within the uterine environment.
Smad蛋白是分泌型生长因子转化生长因子β(TGFβ)家族重要的细胞内介质。Smad1是TGFβ分子中骨形态发生蛋白(BMP)亚组所提供信号的效应器。为了解Smad1在小鼠发育中的作用,我们利用胚胎干细胞中的同源重组产生了一个功能缺失的Smad1等位基因。Smad1基因敲除胚胎在胚胎期第10.5天死亡,因为它们无法与胎盘相连。突变胚胎在胚胎期第7.0天首次可被识别,其特征是在内脏内胚层与胚胎后/胚外连接处出现局部向外突出,同时伴有外胚层和新生中胚层的剧烈扭曲。嵌合体分析表明,这两个缺陷归因于胚外组织中对Smad1的需求。到胚胎期第7.5天,Smad1缺陷胚胎的尿囊形成明显受损。相比之下,绒毛膜过度增殖,在胚外空间内折叠不规则,近端迁移受阻。已知BMP信号对于原始生殖细胞的特化和增殖至关重要。我们发现Smad1缺陷胚胎中的原始生殖细胞急剧减少,这表明Smad1依赖性信号在原始生殖细胞特化中起重要作用。令人惊讶的是,尽管BMP信号在胚胎自身组织中起关键作用,但Smad1缺陷胚胎发育基本正常。对早期小鼠胚胎中Smad1、Smad5和Smad8表达域的检查表明,虽然Smad1在胚胎期第6.5天在内脏内胚层中独特表达,但在其他组织中Smad1与Smad5和/或Smad8共同表达。总体而言,这些数据揭示了Smad1信号在协调子宫环境中支持发育所需的胚外结构生长方面的独特功能。