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Published January 28, 2022 | Supplemental Material + Submitted + Published
Journal Article Open

Single-cell atlas of early chick development reveals gradual segregation of neural crest lineage from the neural plate border during neurulation

Abstract

The epiblast of vertebrate embryos is comprised of neural and non-neural ectoderm, with the border territory at their intersection harboring neural crest and cranial placode progenitors. Here, we a generate single-cell atlas of the developing chick epiblast from late gastrulation through early neurulation stages to define transcriptional changes in the emerging 'neural plate border' as well as other regions of the epiblast. Focusing on the border territory, the results reveal gradual establishment of heterogeneous neural plate border signatures, including novel genes that we validate by fluorescent in situ hybridization. Developmental trajectory analysis infers that segregation of neural plate border lineages only commences at early neurulation, rather than at gastrulation as previously predicted. We find that cells expressing the prospective neural crest marker Pax7 contribute to multiple lineages, and a subset of premigratory neural crest cells shares a transcriptional signature with their border precursors. Together, our results suggest that cells at the neural plate border remain heterogeneous until early neurulation, at which time progenitors become progressively allocated toward defined neural crest and placode lineages. The data also can be mined to reveal changes throughout the developing epiblast.

Additional Information

© 2022 Williams et. al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited. Preprinted: 08 August 2021; Received: 05 October 2021; Accepted: 01 December 2021; Published: 28 January 2022. Fluorescence activated cell sorting was performed at California Institute of Technology Flow Cytometry Facility using BD Biosciences FACSAria Cell Sorter with Patrick Cannon. 10 X libraries were prepared in the Thomson lab at California Institute of Technology with assistance from Jeff Park. Illumina sequencing was performed at the Millard and Muriel Jacob at California Institute of Technology with Igor Antoshechkin. HH4 10 X library was constructed at MRC Weatherall Institute of Molecular Medicine, University of Oxford with assistance from Kevin Clark (FACS facility), Dr Neil Ashley (single-cell facility) and Tim Rostron (NGS sequencing facility). Confocal microscopy was performed within the Biological Imaging Facility at the Beckman Institute, California Institute of Technology with assistance from Dr Giada Spigolon. We thank members of the Bronner and Sauka-Spengler labs for their support and helpful discussions. This work was funded by NIH R01DE027538 to MEB/RMW, Wellcome Trust Senior Research Fellowship (215615/Z/19/Z) to TSS/RMW and Radcliffe Department of Medicine Scholarship and MRC DTP Supplementary Funding to ML. Author contributions: Ruth M Williams, Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Validation, Visualization, Writing – original draft; Martyna Lukoseviciute, Formal analysis, Writing – review and editing; Tatjana Sauka-Spengler, Formal analysis, Funding acquisition, Supervision, Writing – review and editing; Marianne E Bronner, Conceptualization, Funding acquisition, Project administration, Resources, Supervision, Writing – review and editing. Competing interests: Marianne E Bronner: Senior editor, eLife. The other authors declare that no competing interests exist. Data availability: Sequencing data have been deposited in GEO under accession codes GSE181577.

Attached Files

Published - elife-74464-v1.pdf

Submitted - 2021.08.08.455573v1.full.pdf

Supplemental Material - elife-74464-supp1-v1.xlsx

Supplemental Material - elife-74464-transrepform1-v1.docx

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Additional details

Created:
August 20, 2023
Modified:
December 22, 2023