Gastrulation EMT Is Independent of P-Cadherin Downregulation.
Gastrulation EMT Is Independent of P-Cadherin Downregulation.
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DOI：
10.1371/journal.pone.0153591
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Antin PB
中科院分区：
文献类型：
--
作者：
Moly PK;Cooley JR;Zeltzer SL;Yatskievych TA;Antin PB
上皮间质转化（EMT）是一个进化上保守的过程，在此过程中细胞失去上皮特征并获得迁移表型。尽管 Snail 和其他转录抑制因子对上皮钙粘蛋白的下调通常被认为是 EMT 的先决条件，但最近的研究对这一观点提出了挑战。在这里，我们研究了鸡胚原肠胚形成过程中 E-钙粘蛋白和 P-钙粘蛋白表达和定位、Snail 功能和 EMT 之间的关系。表达分析表明，虽然在外胚层中检测到 E-钙粘蛋白转录物，但在原条或中胚层中未检测到，但 P-钙粘蛋白 mRNA 和蛋白质存在于外胚层、原条和中胚层中。目前还没有特异性识别 E-钙粘蛋白的抗体。在 EMT 过程中，P-钙粘蛋白从上皮外胚层细胞的侧面重新定位到新生中胚层细胞的圆周分布。用 E-钙粘蛋白表达构建体电穿孔的细胞经历 EMT 并迁移到中胚层。对 Snail 功能的检查表明，使用吗啉降低 Slug (SNAI2) 蛋白水平不能抑制 EMT，并且外胚层细胞中人或鸡 Snail 的表达不能诱导 EMT。相比之下，表达 Rho 抑制肽 C3 的细胞快速退出外胚层，而不激活 Slug 或中胚层标记物 N-钙粘蛋白。总之，这些实验表明外胚层细胞在保留 P-钙粘蛋白的同时经历 EMT，并提出了关于禽类原肠胚形成过程中 EMT 调节机制的问题。
Epithelial-mesenchymal transition (EMT) is an evolutionarily conserved process during which cells lose epithelial characteristics and gain a migratory phenotype. Although downregulation of epithelial cadherins by Snail and other transcriptional repressors is generally considered a prerequisite for EMT, recent studies have challenged this view. Here we investigate the relationship between E-cadherin and P-cadherin expression and localization, Snail function and EMT during gastrulation in chicken embryos. Expression analyses show that while E-cadherin transcripts are detected in the epiblast but not in the primitive streak or mesoderm, P-cadherin mRNA and protein are present in the epiblast, primitive and mesoderm. Antibodies that specifically recognize E-cadherin are not presently available. During EMT, P-cadherin relocalizes from the lateral surfaces of epithelial epiblast cells to a circumferential distribution in emerging mesodermal cells. Cells electroporated with an E-cadherin expression construct undergo EMT and migrate into the mesoderm. An examination of Snail function showed that reduction of Slug (SNAI2) protein levels using a morpholino fails to inhibit EMT, and expression of human or chicken Snail in epiblast cells fails to induce EMT. In contrast, cells expressing the Rho inhibitor peptide C3 rapidly exit the epiblast without activating Slug or the mesoderm marker N-cadherin. Together, these experiments show that epiblast cells undergo EMT while retaining P-cadherin, and raise questions about the mechanisms of EMT regulation during avian gastrulation.