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In Vivo Analysis of the Mesenchymal-to-Epithelial Transition During Chick Secondary Neurulation

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The Epithelial-to Mesenchymal Transition

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2179))

Abstract

The neural tube in amniotic embryos forms as a result of two consecutive events along the anteroposterior axis, referred to as primary and secondary neurulation (PN and SN). While PN involves the invagination of a sheet of epithelial cells, SN shapes the caudal neural tube through the mesenchymal-to-epithelial transition (MET) of neuromesodermal progenitors, followed by cavitation of the medullary cord. The technical difficulties in studying SN mainly involve the challenge of labeling and manipulating SN cells in vivo. Here we describe a new method to follow MET during SN in the chick embryo, combining early in ovo chick electroporation with in vivo time-lapse imaging. This procedure allows the cells undergoing SN to be manipulated in order to investigate the MET process, permitting their cell dynamics to be followed in vivo.

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Author information

Authors and Affiliations

  1. Instituto de Biología Molecular de Barcelona, CSIC, Parc Científic de Barcelona, Barcelona, Spain

    Elena Gonzalez-Gobartt & Elisa Martí

  2. Centre de Biologie du Développement (CBD), Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse, France

    Guillaume Allio & Bertrand Bénazéraf

Authors
  1. Elena Gonzalez-Gobartt
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  2. Guillaume Allio
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  3. Bertrand Bénazéraf
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  4. Elisa Martí
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Corresponding author

Correspondence to Elisa Martí .

Editor information

Editors and Affiliations

  1. Department of Biomedical Science and Bateson Centre, University of Sheffield, Sheffield, UK

    Kyra Campbell

  2. Centre de Biologie du Développement (CBD) Centre de Biologie Intégrative (CBI), Université de Toulouse, Toulouse, France

    Eric Theveneau

1 Electronic Supplementary Material

Movie S1

In vivo time-lapse imaging of a WT chick embryo (5×) over 10 h showing normal tail bud elongation and blood vessel formation (hh:mm). Related to Fig. 4 (MOV 7547 kb)

Movie S2

In vivo time-lapse imaging of a pSox2:eGFP electroporated embryo (20×) over 10 h. Related to Fig. 5 (MOV 1509 kb)

Movie S3

In vivo time-lapse imaging of a pSox2:eGFP electroporated embryo showing the tracks obtained using the Manual Tracking plugin of the ImageJ software. Related to Fig. 5 (MOV 521 kb)

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Gonzalez-Gobartt, E., Allio, G., Bénazéraf, B., Martí, E. (2021). In Vivo Analysis of the Mesenchymal-to-Epithelial Transition During Chick Secondary Neurulation. In: Campbell, K., Theveneau, E. (eds) The Epithelial-to Mesenchymal Transition. Methods in Molecular Biology, vol 2179. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0779-4_16

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  • DOI: https://doi.org/10.1007/978-1-0716-0779-4_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0778-7

  • Online ISBN: 978-1-0716-0779-4

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