Pluripotent state transitions coordinate morphogenesis in mouse and human embryos
Abstract
The foundations of mammalian development lie in a cluster of embryonic epiblast stem cells. In response to extracellular matrix signalling, these cells undergo epithelialization and create an apical surface in contact with a cavity, a fundamental event for all subsequent development. Concomitantly, epiblast cells transit through distinct pluripotent states, before lineage commitment at gastrulation. These pluripotent states have been characterized at the molecular level5, but their biological importance remains unclear. Here we show that exit from an unrestricted naive pluripotent state is required for epiblast epithelialization and generation of the pro-amniotic cavity in mouse embryos. Embryonic stem cells locked in the naive state are able to initiate polarization but fail to undergo lumenogenesis. Mechanistically, exit from naive pluripotency activates an Oct4-governed transcriptional program that results in expression of glycosylated sialomucin proteins and the vesicle tethering and fusion events of lumenogenesis. Similarly, exit of epiblasts from naive pluripotency in cultured human post-implantation embryos triggers amniotic cavity formation and developmental progression. Our results add tissue-level architecture as a new criterion for the characterization of different pluripotent states, and show the relevance of transitions between these states during development of the mammalian embryo.
Additional Information
© 2017 Springer Nature Publishing AG. received 19 April; accepted 2 November 2017. Published online 29 November 2017. We thank K. McNagny, J. Hanna and R. Jaenisch for reagents and discussions; F. Martin-Belmonte, D. Glover, C. Lynch, M. Serrano, A. Hupalowska, F. Antonica and M. Petruzzelli for feedback; J. N. Skepper for help with electron microscopy; W. Mansfield for help with embryo transfer. This work was supported by Wellcome Trust (098287/Z/12/Z) and ERC (669198) grants to M.Z.-G. Work in the laboratory of T.V. was supported by Wellcome Trust and KU Leuven (SymBioSys PFV/10/016). Work in the laboratory of J.C.M. was supported by EMBL and Cancer Research UK. M.N.S. was supported by Ramon Areces and EMBO postdoctoral fellowships; A.S. by a Wellcome Trust strategic award (105031/D/14/Z) and G.R. by a Newton fellowship. Contributions M.N.S. designed, performed and analysed most of the experiments. A.S. analysed the sequencing data. N.S. and A.W. performed experiments in Fig. 4g and Extended Data Figs 5–7. M.Z. and G.R. helped with embryo experiments and image analysis. A.J., L.G.D. and L.N. helped with human embryo cultures. I.C.M. prepared cDNA libraries. C.B. generated and analysed ChIP–seq data. D.I. and Y.K. supervised the human embryo experiments. T.V. supervised the cDNA library preparation. J.C.M. supervised the computational analyses of the sequencing data. M.Z.-G. supervised the study. M.N.S. and M.Z.-G. conceived the project and wrote the manuscript. Code availability: Code used for this study is available from the corresponding author upon reasonable request. Data availability: RNA sequencing data are available at Array Express (https://www.ebi.ac.uk/arrayexpress/) under accession number E-MTAB-5147. The immunofluorescence and time-lapse data that support the findings of this study are available from the corresponding author upon reasonable request. Source Data for RT–PCR experiments and quantifications of the immunofluorescence data are provided with the paper. The authors declare no competing financial interests. 28 February 2018 -- Please see accompanying Erratum (http://doi.org/10.1038/nature25995). Extended Data Fig. 4 has been replaced, to correct the missing colours in the key to panels c, h, k, n and q, and to correct the missing colours of the graph in panel k.Errata
In this Letter, owing to an error during the production process, the keys to the bar charts in Extended Data Fig. 4c, h, k, n and q all appeared as open white boxes, instead of displaying the different colours used in graphs (for example, to denote the 'polarized' and 'non-polarized' sections, or the 'lumen', 'rosette' and 'disorganized' sections). In addition, in Extended Data Fig. 4k, the colours in the graph (to denote the 'lumen', 'rosette' and 'disorganised') were also missing. This figure has been corrected online.Attached Files
Accepted Version - emss-74765.pdf
Supplemental Material - nature24675-s1.pdf
Supplemental Material - nature24675-s2.xls
Supplemental Material - nature24675-s3.pdf
Supplemental Material - nature24675-s4.pdf
Supplemental Material - nature24675-sf1.jpg
Supplemental Material - nature24675-sf10.jpg
Supplemental Material - nature24675-sf2.jpg
Supplemental Material - nature24675-sf3.jpg
Supplemental Material - nature24675-sf4.jpg
Supplemental Material - nature24675-sf5.jpg
Supplemental Material - nature24675-sf6.jpg
Supplemental Material - nature24675-sf7.jpg
Supplemental Material - nature24675-sf8.jpg
Supplemental Material - nature24675-sf9.jpg
Supplemental Material - nature24675-sv1.avi
Supplemental Material - nature24675-sv2.avi
Supplemental Material - nature24675-sv3.avi
Files
Name | Size | Download all |
---|---|---|
md5:35ced4590b8d872c7a15006c53ab180a
|
165.4 kB | Preview Download |
md5:3cd7fa176e8d00d2f2e7f662bfa882bf
|
151.3 kB | Preview Download |
md5:03e085395ce74c5075cfb6fb723f7f6d
|
162.4 kB | Preview Download |
md5:8fbee7463a55af1eb88659f689deea3c
|
156.7 kB | Preview Download |
md5:bbd417ec0d66f7ccc77cbb4954f69ae7
|
16.0 MB | Download |
md5:76107f9d64356ca2a932055ee7ac1366
|
156.8 kB | Preview Download |
md5:6af94a82a1306d7b2c6419d2eb691585
|
124.4 kB | Preview Download |
md5:b56d82ba68096873ab0afe79e7a4f9c8
|
131.6 kB | Preview Download |
md5:cf84748499a175c0bd9282e426e34e86
|
89.1 kB | Preview Download |
md5:802ce9225250ccb74cf8d49904c26984
|
68.4 kB | Preview Download |
md5:b0a2f78625d6da2655e239e46788cf7e
|
11.5 MB | Download |
md5:051914cc6e3bde4f2753a771b754be07
|
8.7 MB | Preview Download |
md5:3c6c6dfe32aebc1cbcc67a9159d68e54
|
6.7 MB | Download |
md5:bacf5dd8c2df6bc5f4a07158f77363ed
|
162.3 kB | Preview Download |
md5:82e4e7b8fd8432b592e5ca2adfacc14c
|
75.7 kB | Preview Download |
md5:ab06fdfa34acea9f1e8d583bc5de0335
|
162.9 kB | Preview Download |
md5:2d7377de1ac6a98c428f6243bc8a2e52
|
140.4 kB | Preview Download |
md5:ba8ac4bc336f9e00a12b0b9f8e287268
|
8.1 MB | Download |
Additional details
- PMCID
- PMC5768241
- Eprint ID
- 94567
- Resolver ID
- CaltechAUTHORS:20190408-111046314
- 098287/Z/12/Z
- Wellcome Trust
- 669198
- European Research Council (ERC)
- PFV/10/016
- Katholieke Universiteit Leuven
- European Molecular Biology Laboratory
- Cancer Research UK
- Fundación Ramón Areces
- European Molecular Biology Organization (EMBO)
- 105031/D/14/Z
- Wellcome Trust
- Royal Society
- Created
-
2019-04-09Created from EPrint's datestamp field
- Updated
-
2023-06-01Created from EPrint's last_modified field