The increase in maternal expression of axin1 and axin2 contribute to the zebrafish mutant ichabod ventralized phenotype

J Cell Biochem. 2015 Mar;116(3):418-30. doi: 10.1002/jcb.24993.

Abstract

β-Catenin is a central effector of the Wnt pathway and one of the players in Ca(+)-dependent cell-cell adhesion. While many wnts are present and expressed in vertebrates, only one β-catenin exists in the majority of the organisms. One intriguing exception is zebrafish that carries two genes for β-catenin. The maternal recessive mutation ichabod presents very low levels of β-catenin2 that in turn affects dorsal axis formation, suggesting that β-catenin1 is incapable to compensate for β-catenin2 loss and raising the question of whether these two β-catenins may have differential roles during early axis specification. Here we identify a specific antibody that can discriminate selectively for β-catenin1. By confocal co-immunofluorescent analysis and low concentration gain-of-function experiments, we show that β-catenin1 and 2 behave in similar modes in dorsal axis induction and cellular localization. Surprisingly, we also found that in the ich embryo the mRNAs of the components of β-catenin regulatory pathway, including β-catenin1, are more abundant than in the Wt embryo. Increased levels of β-catenin1 are found at the membrane level but not in the nuclei till high stage. Finally, we present evidence that β-catenin1 cannot revert the ich phenotype because it may be under the control of a GSK3β-independent mechanism that required Axin's RGS domain function.

Keywords: AXIS FORMATION; Axin2-RGS DOMAIN; BETA-CATENIN; Wnt signaling; ZEBRAFISH.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibody Specificity
  • Axin Protein / genetics
  • Axin Protein / metabolism*
  • Blastula / drug effects
  • Blastula / metabolism
  • Cell Nucleus / drug effects
  • Cell Nucleus / metabolism
  • Embryonic Development / drug effects
  • Embryonic Development / genetics
  • Female
  • Gene Expression Regulation, Developmental / drug effects
  • Genes, Dominant
  • Immunohistochemistry
  • Lithium Chloride / pharmacology
  • Mutation / genetics*
  • Phenotype
  • Protein Stability / drug effects
  • Protein Transport
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Signal Transduction / drug effects
  • Up-Regulation / drug effects
  • Up-Regulation / genetics
  • Zebrafish / embryology
  • Zebrafish / genetics*
  • Zebrafish Proteins / genetics
  • Zebrafish Proteins / metabolism*
  • beta Catenin / metabolism

Substances

  • Axin Protein
  • Axin1 protein, zebrafish
  • RNA, Messenger
  • Zebrafish Proteins
  • axin2 protein, zebrafish
  • beta Catenin
  • Lithium Chloride