Abstract
Analysis of canonical Wnt signaling during vertebrate development by means of knock-out or transgenic approaches is often hampered by functional redundancy as well as pathway bifurcations downstream of the manipulated components. We report the design of an optimized chimera capable of blocking transcriptional activation of Lef1/Tcf-beta-catenin target genes, thus enabling intervention with the canonical Wnt pathway at its nuclear end point. This construct was made hormone-inducible, both functionally and transcriptionally, and was transgenically integrated in Xenopus embryos. Down-regulation of target genes was clearly observed upon treatment of these embryos with dexamethasone. In addition, exposure of variously aged transgenic embryos to dexamethasone caused complex phenotypes with many new but also several recognizable features stemming from inhibition of canonical Wnt signaling. At least in some tissues, a significant reduction in cell proliferation and an increase in programmed cell death appeared to underlie these phenotypes. Our inducible transgenic system can serve a broad range of experimental settings designed to unveil new functional aspects of Lef1/Tcf-beta-catenin signaling during vertebrate embryogenesis.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Animals
-
Animals, Genetically Modified
-
Antineoplastic Agents, Hormonal / pharmacology
-
Apoptosis
-
Blotting, Western
-
Bromodeoxyuridine / pharmacology
-
Cell Nucleus / metabolism*
-
Cell Proliferation
-
DNA-Binding Proteins / antagonists & inhibitors
-
DNA-Binding Proteins / genetics*
-
DNA-Binding Proteins / metabolism*
-
Dexamethasone / pharmacology
-
Down-Regulation
-
Drosophila
-
Embryo, Nonmammalian / metabolism*
-
Gene Expression Regulation, Developmental*
-
Genes, Reporter
-
Genetic Techniques*
-
Genetic Vectors
-
Homeodomain Proteins / genetics
-
Humans
-
In Situ Nick-End Labeling
-
Lymphoid Enhancer-Binding Factor 1
-
Mice
-
Microscopy, Fluorescence
-
Phenotype
-
Plasmids / metabolism
-
Promoter Regions, Genetic
-
Protein Structure, Tertiary
-
Proto-Oncogene Proteins / metabolism*
-
RNA / chemistry
-
RNA / metabolism
-
Receptors, Glucocorticoid / genetics
-
Reverse Transcriptase Polymerase Chain Reaction
-
Signal Transduction
-
Transcription Factors / antagonists & inhibitors
-
Transcription Factors / genetics*
-
Transcription Factors / metabolism*
-
Transcription, Genetic
-
Transcriptional Activation
-
Transgenes
-
Wnt Proteins
-
Xenopus laevis
Substances
-
Antineoplastic Agents, Hormonal
-
DNA-Binding Proteins
-
Homeodomain Proteins
-
LEF1 protein, human
-
Lef1 protein, mouse
-
Lymphoid Enhancer-Binding Factor 1
-
Proto-Oncogene Proteins
-
Receptors, Glucocorticoid
-
Transcription Factors
-
Wnt Proteins
-
engrailed homeobox proteins
-
RNA
-
Dexamethasone
-
Bromodeoxyuridine