Developmental Biology Institute of Marseille Luminy (IBDML), CNRS UMR 6216, Université de la Méditerranée, Campus de Luminy, Marseille, France.
Dev Dyn. 2010 Jan;239(1):346-53. doi: 10.1002/dvdy.22174.
A critical mediator of cell-cell signaling events during embryogenesis is the highly conserved Wnt family of secreted proteins. Reporter constructs containing multimerized TCF DNA binding sites have been used to detect Wnt beta-catenin dependent activity during animal development. In this report, we have constructed and compared several TCF green fluorescent protein (GFP) reporter constructs. They contained 3, 8, or 12 TCF binding sites upstream of a minimal promoter driving native or destabilized enhanced GFP (EGFP). We have used the electroporation of somites in the chick embryo as a paradigm to test them in vivo. We have verified that they all respond to Wnt signaling in vivo. We have then assessed their efficiency at reflecting the activity of the Wnt pathway. Using destabilized EGFP reporter constructs, we show that somite cells dynamically regulate Wnt/beta-catenin-dependent signaling, a finding that was confirmed by performing time-lapse video confocal observation of electroporated embryos.
在胚胎发生过程中，细胞间信号事件的关键介质是高度保守的 Wnt 家族分泌蛋白。含有多聚化 TCF DNA 结合位点的报告构建体已被用于检测动物发育过程中 Wnt β-连环蛋白依赖性活性。在本报告中，我们构建并比较了几种 TCF 绿色荧光蛋白 (GFP) 报告构建体。它们在驱动天然或不稳定增强 GFP（EGFP）的最小启动子上游含有 3、8 或 12 个 TCF 结合位点。我们使用鸡胚体节的电穿孔作为范例在体内对它们进行了测试。我们已经验证了它们都可以在体内响应 Wnt 信号。然后，我们评估了它们反映 Wnt 途径活性的效率。使用不稳定的 EGFP 报告构建体，我们表明体节细胞动态调节 Wnt/β-连环蛋白依赖性信号，这一发现通过对电穿孔胚胎进行延时视频共聚焦观察得到了证实。
