Chapter 20
Using Xenopus Neural Crest Explants to Study
Epithelial-Mesenchymal Transition
Nade ` ge Gouignard, Christian Rouvie ` re, and Eric Theveneau
Abstract
The epithelial-mesenchymal transition (EMT) converts coherent epithelial structures into single cells. EMT
is a dynamic cellular process that is not systematically completed (not all EMTs lead to single cells) and
reversible (cells can re-epithelialize). EMT is orchestrated at multiple levels from transcription, to posttranslational modifications, to protein turnover. It involves remodeling of polarity and adhesion and
enhances migratory capabilities. During physiological events such as embryogenesis or wound healing
EMT is used to initiate cell migration, but EMT can also occur in pathological settings. In particular, EMT
has been linked to fibrosis and cancer. Neural crest (NC) cells, an embryonic stem cell population whose
behavior recapitulates the main steps of carcinoma progression, are a great model to study EMT. In this
chapter, we provide a fully detailed protocol to extract NC cells from Xenopus embryos and culture them to
study the dynamics of cell–cell adhesion, cell motility, and dispersion.
Key words Neural crest, Epithelial-mesenchymal transition, Cell migration, Adhesion, Polarity,
Dispersion
1 Introduction
In the 1960s, during chick embryo gastrulation, epiblast cells were
shown to dissociate from the epithelium, lose their epithelial morphology, and become mesoderm and endoderm cells [1, 2]. The
concept of Epithelial-Mesenchymal Transformation was then introduced and defined as a complete loss of epithelial traits, including
apicobasal polarity and cell–cell adhesion, accompanied by total
acquisition of mesenchymal characteristics (e.g., front-back polarity, cell motility) [1, 2]. E-Cadherin, a cell–cell junction protein,
and vimentin, an intermediate filament protein, were soon proposed as makers for epithelial and mesenchymal cells, respectively.
Kyra Campbell and Eric Theveneau (eds.), The Epithelial-to Mesenchymal Transition: Methods and Protocols,
Methods in Molecular Biology, vol. 2179, https://doi.org/10.1007/978-1-0716-0779-4_20,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
Electronic supplementary material: The online version of this chapter (https://doi.org/10.1007/978-1-07160779-4_20) contains supplementary material, which is available to authorized users.
257
Using Xenopus Neural Crest Explants to Study
Epithelial-Mesenchymal Transition
Nade ` ge Gouignard, Christian Rouvie ` re, and Eric Theveneau
Abstract
The epithelial-mesenchymal transition (EMT) converts coherent epithelial structures into single cells. EMT
is a dynamic cellular process that is not systematically completed (not all EMTs lead to single cells) and
reversible (cells can re-epithelialize). EMT is orchestrated at multiple levels from transcription, to posttranslational modifications, to protein turnover. It involves remodeling of polarity and adhesion and
enhances migratory capabilities. During physiological events such as embryogenesis or wound healing
EMT is used to initiate cell migration, but EMT can also occur in pathological settings. In particular, EMT
has been linked to fibrosis and cancer. Neural crest (NC) cells, an embryonic stem cell population whose
behavior recapitulates the main steps of carcinoma progression, are a great model to study EMT. In this
chapter, we provide a fully detailed protocol to extract NC cells from Xenopus embryos and culture them to
study the dynamics of cell–cell adhesion, cell motility, and dispersion.
Key words Neural crest, Epithelial-mesenchymal transition, Cell migration, Adhesion, Polarity,
Dispersion
1 Introduction
In the 1960s, during chick embryo gastrulation, epiblast cells were
shown to dissociate from the epithelium, lose their epithelial morphology, and become mesoderm and endoderm cells [1, 2]. The
concept of Epithelial-Mesenchymal Transformation was then introduced and defined as a complete loss of epithelial traits, including
apicobasal polarity and cell–cell adhesion, accompanied by total
acquisition of mesenchymal characteristics (e.g., front-back polarity, cell motility) [1, 2]. E-Cadherin, a cell–cell junction protein,
and vimentin, an intermediate filament protein, were soon proposed as makers for epithelial and mesenchymal cells, respectively.
Kyra Campbell and Eric Theveneau (eds.), The Epithelial-to Mesenchymal Transition: Methods and Protocols,
Methods in Molecular Biology, vol. 2179, https://doi.org/10.1007/978-1-0716-0779-4_20,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
Electronic supplementary material: The online version of this chapter (https://doi.org/10.1007/978-1-07160779-4_20) contains supplementary material, which is available to authorized users.
257