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Real‐time observation of Wnt β‐catenin signaling in the chick embryo
Author(s) -
Rios Anne C.,
Denans Nicolas,
Marcelle Christophe
Publication year - 2010
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.22174
Subject(s) - wnt signaling pathway , biology , green fluorescent protein , somite , wnt3a , microbiology and biotechnology , electroporation , embryo , reporter gene , beta catenin , lrp6 , live cell imaging , lrp5 , signal transduction , embryogenesis , cell , genetics , gene , gene expression
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 β‐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/β‐catenin–dependent signaling, a finding that was confirmed by performing time‐lapse video confocal observation of electroporated embryos. Developmental Dynamics 239:346–353, 2010. © 2009 Wiley‐Liss, Inc.