E-cadherin is required for cranial neural crest migration in Xenopus laevis
E-cadherin is required for cranial neural crest migration in Xenopus laevis
复制标题
DOI：
10.1016/j.ydbio.2016.02.007
复制
发表时间：
2016-03-15
影响因子：
2.7
通讯作者：
Kashef，Jubin
中科院分区：
文献类型：
--
作者：
Huang，Chaolie;Kratzer，Marie-Claire;Kashef，Jubin
颅神经脊是一种高度能动性和多潜能的胚胎细胞群，在整个胚胎中按一定的路线定向迁移，有助于面部结构的形成，包括软骨、骨和神经节。钙粘附素介导的细胞-细胞黏附在数控细胞的定向迁移中起着至关重要的作用。然而，迁移的NC共表达不同的钙粘附素亚型，它们各自的作用尚未得到充分的探索。在以前的研究中，单个钙粘附素亚型的表达已经用不同的方法分析，具有不同的敏感性，防止了表达水平的直接比较。在这里，我们首次全面和比较分析了六个钙粘附素超家族成员在非洲爪哇数控细胞迁移不同阶段的表达。通过应用定量RT-qPCR方法，我们可以确定每个表达的钙粘附素的拷贝数和丰度在不同的NC迁移阶段。利用这种方法，我们首次显示了E-钙粘蛋白和XB/C-钙粘蛋白在数控细胞中的表达，将它们添加为在数控细胞迁移过程中共表达的两个新的钙粘附素成员。在非洲爪蛙cnc迁移过程中，钙粘蛋白的共表达，尤其是E-钙粘蛋白的持续表达，与经典的上皮-间充质转化(EMT)模型中假定的钙粘蛋白表达的转换相矛盾。功能丧失实验进一步表明，E-钙粘附素对于体内正常的细胞迁移和体外细胞突起的形成都是必需的。E-钙粘蛋白的下调不能被其他经典钙粘蛋白联合注射挽救，这表明E-钙粘蛋白在介导数控细胞迁移中具有特定的功能。最后，通过对不同E-钙粘蛋白缺失突变体在E-钙粘蛋白形态变异胚胎中的重组实验，我们证明了E-钙粘蛋白的胞外区，而不是胞内区，足以在体内拯救CNC细胞的迁移。
The cranial neural crest (CNC) is a highly motile and multipotent embryonic cell population, which migrates directionally on defined routes throughout the embryo, contributing to facial structures including cartilage, bone and ganglia. Cadherin-mediated cell–cell adhesion is known to play a crucial role in the directional migration of CNC cells. However, migrating CNC co-express different cadherin subtypes, and their individual roles have yet to be fully explored. In previous studies, the expression of individual cadherin subtypes has been analysed using different methods with varying sensitivities, preventing the direct comparison of expression levels. Here, we provide the first comprehensive and comparative analysis of the expression of six cadherin superfamily members during different phases of CNC cell migration inXenopus. By applying a quantitative RT-qPCR approach, we can determine the copy number and abundance of each expressed cadherin through different phases of CNC migration. Using this approach, we show for the first time expression of E-cadherin and XB/C-cadherin in CNC cells, adding them as two new members of cadherins co-expressed during CNC migration. Cadherin co-expression during CNC migration inXenopus, in particular the constant expression of E-cadherin, contradicts the classical epithelial-mesenchymal transition (EMT) model postulating a switch in cadherin expression. Loss-of-function experiments further show that E-cadherin is required for proper CNC cell migrationin vivoand also for cell protrusion formationin vitro. Knockdown of E-cadherin is not rescued by co-injection of other classical cadherins, pointing to a specific function of E-cadherin in mediating CNC cell migration. Finally, through reconstitution experiments with different E-cadherin deletion mutants in E-cadherin morphant embryos, we demonstrate that the extracellular domain, but not the cytoplasmic domain, of E-cadherin is sufficient to rescue CNC cell migrationin vivo.