Purified Wnt-5a increases differentiation of midbrain dopaminergic cells and dishevelled phosphorylation

J Neurochem. 2005 Mar;92(6):1550-3. doi: 10.1111/j.1471-4159.2004.03022.x.


The Wnt family of lipoproteins regulates several aspects of the development of the nervous system. Recently, we reported that Wnt-3a enhances the proliferation of midbrain dopaminergic precursors and that Wnt-5a promotes their differentiation into dopaminergic neurones. Here we report the purification of hemagglutinin-tagged Wnt-5a using a three-step purification method similar to that previously described for Wnt-3a. Haemagglutinin-tagged Wnt-5a was biologically active and induced the differentiation of immature primary midbrain precursors into tyrosine hydroxylase-positive dopaminergic neurones. Using a substantia nigra-derived dopaminergic cell line (SN4741), we found that Wnt-5a, unlike Wnt-3a, did not promote beta-catenin phosphorylation or stabilization. However, both Wnt-5a and Wnt-3a activated dishevelled, as assessed by a phosphorylation-dependent mobility shift. Moreover, the activity of Wnt-5a on dishevelled was blocked by pre-treatment with acyl protein thioesterase-1, indicating that palmitoylation of Wnt-5a is necessary for its function. Thus, our results suggest that Wnt-3a and Wnt-5a, respectively, activate canonical and non-canonical Wnt signalling pathways in ventral midbrain dopaminergic cells. Furthermore, we identify dishevelled as a key player in transducing both Wnt canonical and non-canonical signals in dopaminergic cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Animals
  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology*
  • Cell Line
  • Cells, Cultured
  • Dishevelled Proteins
  • Dopamine / metabolism*
  • Hemagglutinins / metabolism
  • Mice
  • Neurons / cytology
  • Neurons / metabolism*
  • Palmitic Acid / metabolism
  • Phosphoproteins
  • Phosphorylation / drug effects
  • Proteins / drug effects
  • Proteins / isolation & purification
  • Proteins / metabolism*
  • Proteins / pharmacology*
  • Rats
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction / physiology
  • Stem Cells / cytology
  • Stem Cells / metabolism
  • Substantia Nigra / cytology
  • Substantia Nigra / embryology*
  • Substantia Nigra / metabolism
  • Thiolester Hydrolases / pharmacology
  • Wnt Proteins
  • Wnt-5a Protein
  • Wnt3 Protein
  • Wnt3A Protein


  • Adaptor Proteins, Signal Transducing
  • Dishevelled Proteins
  • Hemagglutinins
  • Phosphoproteins
  • Proteins
  • Recombinant Fusion Proteins
  • WNT3A protein, human
  • Wnt Proteins
  • Wnt-5a Protein
  • Wnt3 Protein
  • Wnt3A Protein
  • Wnt3a protein, mouse
  • Wnt5a protein, rat
  • Palmitic Acid
  • LYPLA1 protein, human
  • Thiolester Hydrolases
  • Dopamine