Inducible gene expression in transgenic Xenopus embryos

Curr Biol. 2000 Jul 13;10(14):849-52. doi: 10.1016/s0960-9822(00)00596-0.

Abstract

The amphibian Xenopus laevis has been successfully used for many years as a model system for studying vertebrate development. Because of technical limitations, however, molecular investigations have mainly concentrated on early stages. We have developed a straightforward method for stage-specific induction of gene expression in transgenic Xenopus embryos [1] [2]. This method is based on the Xenopus heat shock protein 70 (Xhsp70 [3]) promoter driving the expression of desired gene products. We found that ubiquitous expression of the transgene is induced upon relatively mild heat treatment. Green fluorescent protein (GFP) was used as a marker to monitor successful induction of gene expression in transgenic embryos. We used this method to study the stage specificity of Wnt signalling function. Transient ectopic Wnt-8 expression during early neurulation was sufficient to repress anterior head development and this capacity was restricted to early stages of neurulation. By transient over-expression at different stages of development, we show that frizzled-7 disrupted morphogenesis sequentially from anterior to posterior along the dorsal axis as development proceeds. These results demonstrate that this method for inducible gene expression in transgenic Xenopus embryos will be a very powerful tool for temporal analysis of gene function and for studying molecular mechanisms of vertebrate organogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Cytoskeletal Proteins
  • Gene Expression Regulation, Developmental
  • Genes, Reporter
  • Green Fluorescent Proteins
  • HSP70 Heat-Shock Proteins / genetics
  • Luminescent Proteins / genetics
  • Phenotype
  • Promoter Regions, Genetic
  • Proteins / genetics
  • Recombinant Fusion Proteins / genetics
  • Signal Transduction
  • Wnt Proteins
  • Xenopus laevis / embryology*
  • Xenopus laevis / genetics*
  • Zebrafish Proteins

Substances

  • Cytoskeletal Proteins
  • HSP70 Heat-Shock Proteins
  • Luminescent Proteins
  • Proteins
  • Recombinant Fusion Proteins
  • Wnt Proteins
  • Zebrafish Proteins
  • wnt8a protein, zebrafish
  • Green Fluorescent Proteins