Morales A V, de la Rosa E J, de Pablo F
Department of Cell and Developmental Biology, CSIC, Madrid, Spain.
Dev Dyn. 1996 Aug;206(4):343-53. doi: 10.1002/(SICI)1097-0177(199608)206:4<343::AID-AJA1>3.0.CO;2-I.
cCdx-B (formerly cHox-cad 2) is a chick homeobox-containing gene related to the Drosophila caudal. Compared with other caudal homologues, its similarity is highest with the murine Cdx-4. In the present study, we characterize the localization of cCdx-B transcripts to the caudal region of the embryo by using reverse transcription-polymerase chain reaction (RT-PCR) and, in detail, by using in situ hybridization. Chick embryos from gastrulation to early organogenesis were hybridized with digoxigenin-labeled riboprobes, and the pattern of expression of cCdx-B mRNA was analyzed in wholemount embryos and in tissue sections. In the early gastrula, transcripts were localized in a gradient through the caudal half of the embryo, in the epiblast and the mesoderm cells, but not including Hensen's node. During neurulation, cCdx-B transcripts were found more rostrally, with high levels localized in Hensen's node and the posterior neural plate. Expression was also high in paraxial mesoderm, with a rostral limit in the most recently formed somite. There was no expression in definitive endoderm. During late neurulation and tail bud formation, cCdx-B mRNA expression regressed posteriorly and was finally confined to the tail bud region. This pattern of expression of cCdx-B, regulated in time and space, is different from that of the other known chick caudal homologue, cCdx-A. Both genes may play a coordinated role in the posterior axial patterning of the chick embryo, whereas cCdx-B may specify further the identity of the tail region.
cCdx-B(以前称为cHox-cad 2)是一种含有鸡同源异型框的基因,与果蝇的尾端基因相关。与其他尾端同源基因相比,它与小鼠的Cdx-4相似度最高。在本研究中,我们通过逆转录聚合酶链反应(RT-PCR),并详细地通过原位杂交来确定cCdx-B转录本在胚胎尾端区域的定位。用洋地黄毒苷标记的核糖探针与从原肠胚形成到早期器官发生阶段的鸡胚胎进行杂交,并在整装胚胎和组织切片中分析cCdx-B mRNA的表达模式。在早期原肠胚中,转录本以梯度形式定位在胚胎尾端的后半部分,存在于上胚层和中胚层细胞中,但不包括亨氏结。在神经胚形成过程中,cCdx-B转录本在更靠前的位置被发现,高水平表达定位于亨氏结和后神经板。在近轴中胚层中表达也很高,在最近形成的体节处有一个前端界限。在确定的内胚层中没有表达。在神经胚形成后期和尾芽形成过程中,cCdx-B mRNA表达向后退缩,最终局限于尾芽区域。cCdx-B这种在时间和空间上受到调控的表达模式,与另一个已知的鸡尾端同源基因cCdx-A不同。这两个基因可能在鸡胚胎的后轴模式形成中发挥协同作用,而cCdx-B可能进一步确定尾区的特征。