Identification of Wnt Pathway Target Genes Regulating the Division and Differentiation of Larval Seam Cells and Vulval Precursor Cells in Caenorhabditis elegans.
Identification of Wnt Pathway Target Genes Regulating the Division and Differentiation of Larval Seam Cells and Vulval Precursor Cells in Caenorhabditis elegans.
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DOI：
10.1534/g3.115.017715
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发表时间：
2015-06-05
期刊：
影响因子：
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通讯作者：
Eisenmann DM
中科院分区：
文献类型：
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作者：
Gorrepati L;Krause MW;Chen W;Brodigan TM;Correa-Mendez M;Eisenmann DM
进化上保守的Wnt/β-catenin信号通路在后生动物发育过程中起着重要作用，调节许多过程，包括细胞命运特化、细胞迁移和干细胞更新。Wnt配体结合导致转录效应子β-连环蛋白的稳定和靶基因表达的上调以介导细胞应答。在秀丽隐杆线虫幼虫发育过程中，Wnt/β-catenin信号通路参与了腹侧外阴前体细胞（VPCs）和侧缝细胞的命运特化。由于对Wnt信号通路在幼虫VPC和缝细胞分化过程中作用的靶点知之甚少，我们试图鉴定这两种皮下细胞类型中受Wnt信号调节的基因。我们有条件地激活Wnt信号在幼虫动物，并进行细胞类型特异性的“mRNA标签”，以丰富VPC和接缝细胞特异性的mRNA，然后使用微阵列分析，以检查基因表达相比，对照动物。我们鉴定了239个响应Wnt信号而激活的基因，并进一步鉴定了50个基因。这些基因中的大多数在正常发育期间在接缝和/或外阴谱系中表达，并且9个基因的功能降低导致接缝细胞和外阴细胞的适当分裂、命运指定、命运执行或分化中的缺陷。因此，这些技术的组合成功地从少量细胞中鉴定了潜在的细胞类型特异性Wnt途径靶基因，并增加了我们对这些C.线虫幼虫的下皮细胞。
The evolutionarily conserved Wnt/β-catenin signaling pathway plays a fundamental role during metazoan development, regulating numerous processes including cell fate specification, cell migration, and stem cell renewal. Wnt ligand binding leads to stabilization of the transcriptional effector β-catenin and upregulation of target gene expression to mediate a cellular response. During larval development of the nematode Caenorhabditis elegans, Wnt/β-catenin pathways act in fate specification of two hypodermal cell types, the ventral vulval precursor cells (VPCs) and the lateral seam cells. Because little is known about targets of the Wnt signaling pathways acting during larval VPC and seam cell differentiation, we sought to identify genes regulated by Wnt signaling in these two hypodermal cell types. We conditionally activated Wnt signaling in larval animals and performed cell type–specific "mRNA tagging" to enrich for VPC and seam cell–specific mRNAs, and then used microarray analysis to examine gene expression compared to control animals. Two hundred thirty-nine genes activated in response to Wnt signaling were identified, and we characterized 50 genes further. The majority of these genes are expressed in seam and/or vulval lineages during normal development, and reduction of function for nine genes caused defects in the proper division, fate specification, fate execution, or differentiation of seam cells and vulval cells. Therefore, the combination of these techniques was successful at identifying potential cell type–specific Wnt pathway target genes from a small number of cells and at increasing our knowledge of the specification and behavior of these C. elegans larval hypodermal cells.