Wnt/β-Catenin Signaling Blockade Promotes Neuronal Induction and Dopaminergic Differentiation in Embryonic Stem Cells§

Authors

  • Lukás̆ C̆ajánek,

    1. Laboratory of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden
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  • Diogo Ribeiro,

    1. Laboratory of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden
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  • Isabel Liste,

    1. Laboratory of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden
    2. Center of Molecular Biology “Severo Ochoa” (CSIC-UAM), Autonomous University of Madrid, Madrid, Spain
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  • Clare L. Parish,

    1. Laboratory of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden
    2. Howard Florey Institute, University of Melbourne, Parkville, Victoria, Australia
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  • Vítĕzslav Bryja,

    Corresponding author
    1. Laboratory of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden
    2. Department of Cytokinetics, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic, and Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic
    • Institute of Experimental Biology, Faculty of Science, Masaryk University, CZ-611 37 Brno, Czech Republic
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    • Telephone: +420-532146226; Fax: +420-541211214

  • Ernest Arenas

    Corresponding author
    1. Laboratory of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden
    • Laboratory of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, S-171 77, Stockholm, Sweden
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    • Telephone: +46-8-52487663; Fax: +46-8-34 1960


  • Author contributions: LC: conception and design, collection and/or assembly of data, data analyses and interpretation, manuscript writing, final approval of manuscript; DR: collection and/or assembly of data, data analyses and interpretation; IL: collection and/or assembly of data, data analyses and interpretation; CLP: collection and/or assembly of data, data analyses and interpretation; VB: conception and design, data analyses and interpretation, manuscript writing; EA: conception and design, fund raising, data analyses and interpretation, manuscript writing, final approval of manuscript.

  • First published online in STEM CELLS EXPRESS September 1, 2009.

  • §

    Disclosure of potential conflicts of interest is found at the end of this article.

Abstract

Embryonic stem cells (ESCs) represent not only a promising source of cells for cell replacement therapy, but also a tool to study the molecular mechanisms underlying cellular signaling and dopaminergic (DA) neuron development. One of the main regulators of DA neuron development is Wnt signaling. Here we used mouse ESCs (mESCs) lacking Wnt1 or the low-density lipoprotein receptor-related protein 6 (LRP6) to decipher the action of Wnt/β-catenin signaling on DA neuron development in mESCs. We provide evidence that the absence of LRP6 abrogates responsiveness of mESCs to Wnt ligand stimulation. Using two differentiation protocols, we show that the loss of Wnt1 or LRP6 increases neuroectodermal differentiation and the number of mESC-derived DA neurons. These effects were similar to those observed following treatment of mESCs with the Wnt/β-catenin pathway inhibitor Dickkopf1 (Dkk1). Combined, our results show that decreases in Wnt/β-catenin signaling enhance neuronal and DA differentiation of mESCs. These findings suggest that: 1) Wnt1 or LRP6 are not strictly required for the DA differentiation of mESCs in vitro, 2) the levels of morphogens and their activity in ESC cultures need to be optimized to improve DA differentiation, and 3) by enhancing the differentiation and number of ESC-derived DA neurons with Dkk1, the application of ESCs for cell replacement therapy in Parkinson's disease may be improved. STEM CELLS 2009;27:2917–2927

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