Pownall M E, Isaacs H V, Slack J M
Developmental Biology Programme, School of Biology and Biochemistry, University of Bath, UK.
Curr Biol. 1998 May 21;8(11):673-6. doi: 10.1016/s0960-9822(98)70257-x.
We have shown previously that fibroblast growth factor (FGF) signalling in posterior regions of the Xenopus embryo is required for the development of the trunk and tail via a molecular pathway that includes the caudal-related gene Xcad3 and the posterior Hox genes [1]. These results have been contested by the work of Kroll and Amaya [2], which shows that Xenopus embryos transgenic for a dominant-negative form of the FGF receptor (FGF-RI) express posterior Hox genes normally, leading these authors to suggest that the FGFs are not required for anteroposterior (A-P) patterning of the dorsal axis. In order to investigate the apparent discrepancy between these studies, we have produced Xenopus embryos transgenic for two inhibitors of the FGF/Caudal pathway: a kinase-deficient dominant-negative FGF receptor (XFD) [3]; and a domain-swapped form of Xcad3 (Xcad-EnR) in which the activation domain of Xcad3 is replaced by the repression domain of the Drosophila Engrailed protein. Both of these were introduced as fusions with the green fluorescent protein (GFP), which allows identification of non-mosaic transgenic embryos at early gastrula stages by simply looking for GFP fluorescence. Analysis of gene expression in embryos transgenic for these constructs indicated that the activation of posterior Hox genes during early neurula stages absolutely requires FGF signalling and transcriptional activation by Xcad3, while the maintenance of Hox gene expression in the trunk and tail during later development is independent of both FGF and Xcad.
我们之前已经表明,非洲爪蟾胚胎后部区域的成纤维细胞生长因子(FGF)信号传导对于躯干和尾巴的发育是必需的,其通过一条分子途径实现,该途径包括与尾相关的基因Xcad3和后部Hox基因[1]。这些结果受到了Kroll和Amaya[2]研究工作的质疑,他们的研究表明,转染了显性负性形式FGF受体(FGF-RI)的非洲爪蟾胚胎正常表达后部Hox基因,这使得他们认为FGF对于背轴的前后(A-P)模式形成并非必需。为了研究这些研究之间明显的差异,我们构建了转染FGF/尾途径两种抑制剂的非洲爪蟾胚胎:一种激酶缺陷型显性负性FGF受体(XFD)[3];以及一种Xcad3的结构域交换形式(Xcad-EnR),其中Xcad3的激活结构域被果蝇Engrailed蛋白的抑制结构域所取代。这两种抑制剂都与绿色荧光蛋白(GFP)融合引入,这使得在原肠胚早期阶段,通过简单观察GFP荧光就能鉴定非嵌合转基因胚胎。对转染这些构建体的胚胎中的基因表达分析表明,在神经胚早期阶段后部Hox基因的激活绝对需要FGF信号传导和Xcad3的转录激活,而在后期发育过程中躯干和尾巴中Hox基因表达的维持则独立于FGF和Xcad。