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Pluripotent state transitions coordinate morphogenesis in mouse and human embryos

Shahbazi, Marta N. and Scialdone, Antonio and Skorupska, Natalia and Weberling, Antonia and Recher, Gaelle and Zhu, Meng and Jedrusik, Agnieszka and Devito, Liani G. and Noli, Laila and Macaulay, Iain C. and Buecker, Christa and Khalaf, Yakoub and Ilic, Dusko and Voet, Thierry and Marioni, John C. and Zernicka-Goetz, Magdalena (2017) Pluripotent state transitions coordinate morphogenesis in mouse and human embryos. Nature, 552 (7684). pp. 239-243. ISSN 0028-0836. PMCID PMC5768241. https://resolver.caltech.edu/CaltechAUTHORS:20190408-111046314

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[img] Image (JPEG) (Extended Data Figure 1 : Epiblast gene expression patterns at peri-implantation stages) - Supplemental Material
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[img] Image (JPEG) (Extended Data Figure 2 : Dynamics of polarization, lumenogenesis and naive pluripotency exit in mES cells) - Supplemental Material
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[img] Image (JPEG) (Extended Data Figure 3 : mES cells initiate polarization in naive conditions in a 3D Matrigel culture) - Supplemental Material
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[img] Image (JPEG) (Extended Data Figure 4 : 2i/LIF inhibits lumen formation in mES cells) - Supplemental Material
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[img] Image (JPEG) (Extended Data Figure 5 : Exit from naive pluripotency is required for lumenogenesis in mES cells) - Supplemental Material
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[img] Image (JPEG) (Extended Data Figure 6 : Sialomucins are required for mES cell lumenogenesis) - Supplemental Material
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[img] Image (JPEG) (Extended Data Figure 7 : Otx2 and Oct4 induce Podxl expression upon naive pluripotency exit) - Supplemental Material
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[img] Image (JPEG) (Extended Data Figure 8 : Cgn is induced upon naive pluripotency exit and mediates Rab11 vesicle tethering to the apical membrane) - Supplemental Material
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[img] Image (JPEG) (Extended Data Figure 9 : Characterization of pluripotency gene expression and epiblast morphogenesis in post-implantation human embryos) - Supplemental Material
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[img] Image (JPEG) (Extended Data Figure 10 : Exit from naive pluripotency is required for lumenogenesis in hES cells) - Supplemental Material
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[img] PDF (Supplementary Table 2. This file contains antibodies used in this study) - Supplemental Material
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[img] PDF (Supplementary Table 3. This file contains a list of RT-PCR primers used in this study) - Supplemental Material
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[img] MS Excel (Supplementary Table 1. This file contains genes expressed in the mouse epiblast at peri-implantation stages) - Supplemental Material
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[img] Video (AVI) (ΔPE-Oct4-GFP mESCs cultured in 3D matrigel without 2i/LIF and imaged every 30 minutes) - Supplemental Material
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[img] Video (AVI) (Rex1::GFPd2 mESCs cultured in 3D matrigel without 2i/LIF and imaged every 30 minutes) - Supplemental Material
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[img] Video (AVI) (Nanog-YFP mTmG mESCs cultured in 3D matrigel without 2i/LIF and imaged every 30 minutes) - Supplemental Material
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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.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1038/nature24675DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5768241/PubMed CentralArticle
https://rdcu.be/bvrtbPublisherFree ReadCube access -- Article
https://doi.org/10.1038/nature25995DOIErratum
https://rdcu.be/bvrvxPublisherFree ReadCube access -- Erratum
ORCID:
AuthorORCID
Shahbazi, Marta N.0000-0002-1599-5747
Recher, Gaelle0000-0001-9004-0118
Zhu, Meng0000-0001-6157-8840
Zernicka-Goetz, Magdalena0000-0002-7004-2471
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.
Funders:
Funding AgencyGrant Number
Wellcome Trust098287/Z/12/Z
European Research Council (ERC)669198
Katholieke Universiteit LeuvenPFV/10/016
European Molecular Biology LaboratoryUNSPECIFIED
Cancer Research UKUNSPECIFIED
Fundación Ramón ArecesUNSPECIFIED
European Molecular Biology Organization (EMBO)UNSPECIFIED
Wellcome Trust105031/D/14/Z
Royal SocietyUNSPECIFIED
Issue or Number:7684
PubMed Central ID:PMC5768241
Record Number:CaltechAUTHORS:20190408-111046314
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20190408-111046314
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:94567
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:09 Apr 2019 15:34
Last Modified:09 Mar 2020 13:19

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