Department of Biological Sciences, University of Maryland Baltimore County, Baltimore, Maryland 21250.
G3 (Bethesda). 2014 Apr 16;4(4):733-47. doi: 10.1534/g3.113.009522.
The Wnt signaling pathway plays a fundamental role during metazoan development, where it regulates diverse processes, including cell fate specification, cell migration, and stem cell renewal. Activation of the beta-catenin-dependent/canonical Wnt pathway up-regulates expression of Wnt target genes to mediate a cellular response. In the nematode Caenorhabditis elegans, a canonical Wnt signaling pathway regulates several processes during larval development; however, few target genes of this pathway have been identified. To address this deficit, we used a novel approach of conditionally activated Wnt signaling during a defined stage of larval life by overexpressing an activated beta-catenin protein, then used microarray analysis to identify genes showing altered expression compared with control animals. We identified 166 differentially expressed genes, of which 104 were up-regulated. A subset of the up-regulated genes was shown to have altered expression in mutants with decreased or increased Wnt signaling; we consider these genes to be bona fide C. elegans Wnt pathway targets. Among these was a group of six genes, including the cuticular collagen genes, bli-1 col-38, col-49, and col-71. These genes show a peak of expression in the mid L4 stage during normal development, suggesting a role in adult cuticle formation. Consistent with this finding, reduction of function for several of the genes causes phenotypes suggestive of defects in cuticle function or integrity. Therefore, this work has identified a large number of putative Wnt pathway target genes during larval life, including a small subset of Wnt-regulated collagen genes that may function in synthesis of the adult cuticle.
Wnt 信号通路在后生动物发育过程中发挥着基础性作用,它调节着包括细胞命运特化、细胞迁移和干细胞更新在内的多种过程。β-连环蛋白依赖性/经典 Wnt 信号通路的激活上调 Wnt 靶基因的表达,从而介导细胞反应。在秀丽隐杆线虫中,经典 Wnt 信号通路调节幼虫发育过程中的几个过程;然而,这个途径的几个靶基因尚未被鉴定。为了解决这一缺陷,我们采用了一种新的方法,即在幼虫发育的特定阶段通过过表达激活的β-连环蛋白蛋白来条件性激活 Wnt 信号,然后使用微阵列分析来鉴定与对照动物相比表达发生改变的基因。我们鉴定出 166 个差异表达基因,其中 104 个上调。上调基因的一部分在 Wnt 信号减少或增加的突变体中表现出表达改变;我们认为这些基因是真正的秀丽隐杆线虫 Wnt 途径靶基因。其中包括一组六个基因,包括表皮胶原基因 bli-1 col-38、col-49 和 col-71。这些基因在正常发育过程中 L4 中期表达峰值最高,表明它们在成虫表皮形成中具有作用。这一发现与功能降低几个基因的表型一致,提示在表皮功能或完整性方面存在缺陷。因此,这项工作在幼虫期鉴定出了大量潜在的 Wnt 途径靶基因,包括一小部分 Wnt 调节的胶原基因,它们可能在合成成虫表皮中发挥作用。