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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 753–761 (2011)Cite this article

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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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Figure 1: Characterization of β-catfl/fl and β-catΔ/Δ mESCs.
Figure 2: Self-renewal markers are not affected in β-catfl/fl and β-catΔ/Δ mESCs under different culture conditions.
Figure 3: Cell–cell adhesion defects in β-catfl/fl and β-catΔ/Δ mESCs in the presence and absence of Jup knockdown.
Figure 4: Morphological appearance and plakoglobin levels of β-catfl/fl, β-catΔ/Δ, β-catrescWT and β-catrescΔC embryoid bodies during differentiation.
Figure 5: Analyses of Tcf/Lef-mediated transcriptional activity.
Figure 6: Neuroectodermal differentiation potential of β-catfl/fl, β-catΔ/Δ, β-catrescWT and β-catrescΔC embryoid bodies.
Figure 7: Mesendodermal differentiation potential of β-catfl/fl, β-catΔ/Δ, β-catrescWT and β-catrescΔC embryoid bodies.

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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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  1. Domenico Migliorini

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

Authors and Affiliations

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

    Natalia Lyashenko, Markus Winter, Domenico Migliorini & Christine Hartmann

  2. Department of Pharmacology, Institute for Stem Cell and Regenerative Medicine, 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.

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Correspondence to Christine Hartmann.

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Lyashenko, N., Winter, M., Migliorini, D. et al. Differential requirement for the dual functions of β-catenin in embryonic stem cell self-renewal and germ layer formation. Nat Cell Biol 13, 753–761 (2011). https://doi.org/10.1038/ncb2260

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