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Epigenetic restriction of embryonic cell lineage fate by methylation of Elf5

Nature Cell Biology volume 10, pages 1280–1290 (2008)Cite this article

Abstract

Mouse ES cells can differentiate into all three germ layers of the embryo but are generally excluded from the trophoblast lineage. Here we show that ES cells deficient in DNA methylation can differentiate efficiently into trophoblast derivatives. In a genome-wide screen we identified the transcription factor Elf5 as methylated and repressed in ES cells, and hypomethylated and expressed in TS and methylation-deficient ES cells. Elf5 creates a positive-feedback loop with the TS cell determinants Cdx2 and Eomes that is restricted to the trophoblast lineage by epigenetic regulation of Elf5. Importantly, the late-acting function of Elf5 allows initial plasticity and regulation in the early blastocyst. Thus, Elf5 functions as a gatekeeper, downstream of initial lineage determination, to reinforce commitment to the trophoblast lineage or to abort this pathway in epiblast cells. This epigenetic restriction of cell lineage fate provides a molecular mechanism for Waddington's concept of canalization of developmental pathways.

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Figure 1: Dnmt1-deficiency enables trophoblast differentiation from cells committed to the embryonic cell lineage.
Figure 2: Trophoblast differentiation is induced in wild-type ES cells upon inhibition of DNA methylation and is independent of Oct4 downregulation.
Figure 3: Dnmt1−/− ES cells transdifferentiate into functional trophoblast derivatives.
Figure 4: Global promoter methylation screen identifies Elf5 as the key gene that is methylated in ES cells and unmethylated in TS cells.
Figure 5: Elf5 is a TS cell transcription factor specific for proliferative trophoblasts.
Figure 6: Epigenetically controlled Elf5 expression acts in a positive-feedback loop to reinforce trophoblast identity.
Figure 7: Gatekeeper function of Elf5 in cell lineage specification.

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Acknowledgements

We would like to thank Fatima Santos and Annabelle Lewis for expert help with confocal microscopy and qPCR design, respectively. We also thank En Li for Dnmt1− mice and ES cells, and Haruhiko Koseki, Nick Gilbert and David Skalnik for ES cell lines. This work was supported by an MRC Career Development Award to M.H., by the Croucher Foundation Fellowship to R.K.N., and by BBSRC, MRC, EU NoE The Epigenome, CellCentric, and DIUS.

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Authors and Affiliations

  1. Laboratory of Developmental Genetics and Imprinting, The Babraham Institute, Cambridge, CB22 3AT, UK

    Ray Kit Ng, Wendy Dean, Claire Dawson, Diana Lucifero, Zofia Madeja, Wolf Reik & Myriam Hemberger

  2. Centre for Trophoblast Research, University of Cambridge, Cambridge, CB2 3EG, UK

    Ray Kit Ng, Zofia Madeja, Wolf Reik & Myriam Hemberger

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Contributions

R.K.N. and M.H. performed the main body of experiments; W.D. performed the chimaera and blastocyst outgrowth experiments; C.D. contributed to the qPCR analysis; D.L. and Z.M. performed parts of the bisulphite sequencing; R.K.N., W.D., W.R. and M.H. analysed and discussed the data and M.H. and W.R. wrote the manuscript.

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Correspondence to Myriam Hemberger.

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Ng, R., Dean, W., Dawson, C. et al. Epigenetic restriction of embryonic cell lineage fate by methylation of Elf5. Nat Cell Biol 10, 1280–1290 (2008). https://doi.org/10.1038/ncb1786

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