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Differential requirement for the dual functions of β-catenin in embryonic stem cell self-renewal and germ layer formation

Nature Cell Biology volume 13, pages 753761 (2011) | Download Citation

Abstract

Canonical Wnt signalling has been implicated in mouse and human embryonic stem cell (ESC) maintenance; however, its requirement is controversial. β-catenin is the key component in this highly conserved Wnt pathway, acting as a transcriptional transactivator. However, β-catenin has additional roles at the plasma membrane regulating cell–cell adhesion, complicating the analyses of cells/tissues lacking β-catenin. We report here the generation of a Ctnnb1 (β-catenin)-deficient mouse ESC (mESC) line and show that self-renewal is maintained in the absence of β-catenin. Cell adhesion is partially rescued by plakoglobin upregulation, but fails to be maintained during differentiation. When differentiated as aggregates, wild-type mESCs form descendants of all three germ layers, whereas mesendodermal germ layer formation and neuronal differentiation are defective in Ctnnb1-deficient mESCs. A Tcf/Lef-signalling-defective β-catenin variant, which re-establishes cadherin-mediated cell adhesion, rescues definitive endoderm and neuroepithelial formation, indicating that the β-catenin cell-adhesion function is more important than its signalling function for these processes.

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Acknowledgements

We thank W. Birchmeier (MDC, Berlin, Germany), R. Grosschedl (MPI, Freiburg, Germany) and A. Wutz (CSCR, Cambridge, UK) for reagents, R. Latham for Wnt3aCM, R. Peachey, G. Resch, P. Pasierbeck, G. Stengl, V. Komnenovic and M. Zeba for help and technical advice, M. Radolf and H. Scheuch for help with microarray studies, C. Theussl and J. Wojciechowski for blastocyst injections, and A. Smith for helpful comments. Research in the laboratory of C.H. has been supported by Boehringer Ingelheim, the project has been funded in part by the NoE Cells into Organs (LSHM-CT-2003-504468), N.L. was supported by the Austrian Science Fund (FWF Grant No. P19281-B16) and M.W. is supported by a grant from the Arthritis Research Campaign (ARC Grant No. 18075). R.T.M. is an investigator of the HHMI and T.B. was supported by NIH grant R01 GM081619-01.

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Author notes

    • Domenico Migliorini

    Present address: Laboratory of Molecular Cancer Biology, University of Leuven, Herestraat 49, 3000 Leuven, Belgium

Affiliations

  1. Research Institute of Molecular Pathology, Dr. Bohrgasse 7, 1030 Vienna, Austria

    • Natalia Lyashenko
    • , Markus Winter
    • , Domenico Migliorini
    •  & Christine Hartmann
  2. Institute for Stem Cell and Regenerative Medicine, Department of Pharmacology, Howard Hughes Medical Institute, University of Washington School of Medicine, Campus Box 358056, Seattle, Washington 98109, USA

    • Travis Biechele
    •  & Randall T. Moon

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Contributions

N.L. carried out, analysed and interpreted experiments. M.W. and D.M. carried out pulldown studies. T.B. and R.T.M. generated and provided BAR/fuBAR lentiviruses, and commented on the paper. C.H. supervised the study and wrote the paper together with N.L.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Christine Hartmann.

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DOI

https://doi.org/10.1038/ncb2260

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