Chapter 16
In Vivo Analysis of the Mesenchymal-to-Epithelial Transition
During Chick Secondary Neurulation
Elena Gonzalez-Gobartt, Guillaume Allio, Bertrand Be ´ naze ´ raf,
and Elisa Martı ´
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.
Key words Mesenchymal-to-epithelial transition, Secondary neurulation, Neural tube formation,
Chick embryo, In ovo electroporation, In vivo time-lapse imaging, Cell dynamics
1 Introduction
During development, the entire vertebrate peripheral nervous system (PNS) is formed through the epithelial-to-mesenchymal transition (EMT) of neuroepithelial cells, which generates migratory
neural crest cells, one of the best studied examples of physiological
EMT [1–3]. By contrast, the mesenchymal-to-epithelial transition
(MET) is the reverse process and plays an important role during
organogenesis, as well as in the elongation of the caudal nervous
system, a process known as secondary neurulation (SN). Indeed,
the formation of the vertebrate neural tube (NT) involves two
different morphogenetic events, primary (PN) and secondary
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_16,
© 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_16) contains supplementary material, which is available to authorized users.
183
In Vivo Analysis of the Mesenchymal-to-Epithelial Transition
During Chick Secondary Neurulation
Elena Gonzalez-Gobartt, Guillaume Allio, Bertrand Be ´ naze ´ raf,
and Elisa Martı ´
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.
Key words Mesenchymal-to-epithelial transition, Secondary neurulation, Neural tube formation,
Chick embryo, In ovo electroporation, In vivo time-lapse imaging, Cell dynamics
1 Introduction
During development, the entire vertebrate peripheral nervous system (PNS) is formed through the epithelial-to-mesenchymal transition (EMT) of neuroepithelial cells, which generates migratory
neural crest cells, one of the best studied examples of physiological
EMT [1–3]. By contrast, the mesenchymal-to-epithelial transition
(MET) is the reverse process and plays an important role during
organogenesis, as well as in the elongation of the caudal nervous
system, a process known as secondary neurulation (SN). Indeed,
the formation of the vertebrate neural tube (NT) involves two
different morphogenetic events, primary (PN) and secondary
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_16,
© 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_16) contains supplementary material, which is available to authorized users.
183