Transgenic quails reveal dynamic TCF/β-catenin signaling during avian embryonic development

  1. Hila Barzilai-Tutsch
  2. Valerie Morin
  3. Gauthier Toulouse
  4. Oleksandr Chernyavskiy
  5. Stephen Firth
  6. Christophe Marcelle  Is a corresponding author
  7. Olivier Serralbo  Is a corresponding author
  1. Claude Bernard University Lyon 1, France
  2. Monash University, Australia
  3. CSIRO Health and Biosecurity, Australia

Abstract

The Wnt/bβ-catenin signaling pathway is highly conserved throughout evolution, playing crucial roles in several developmental and pathological processes. Wnt ligands can act at a considerable distance from their sources and it is therefore necessary to examine not only the Wnt-producing but also the Wnt-receiving cells and tissues to fully appreciate the many functions of this pathway. To monitor Wnt activity, multiple tools have been designed which consist of multimerized Wnt signaling response elements (TCF/LEF binding sites) driving the expression of fluorescent reporter proteins (e.g. GFP, RFP) or of LacZ. The high stability of those reporters leads to a considerable accumulation in cells activating the pathway, thereby making them easily detectable. However, this makes them unsuitable to follow temporal changes of the pathway's activity during dynamic biological events. Even though fluorescent transcriptional reporters can be destabilized to shorten their half-lives, this dramatically reduces signal intensities, particularly when applied in vivo. To alleviate these issues, we developed two transgenic quail lines in which high copy number (12x or 16x) of the TCF/LEF binding sites drive the expression of destabilized GFP variants. Translational enhancer sequences derived from viral mRNAs were used to increase signal intensity and specificity. This resulted in transgenic lines efficient for the characterisation of TCF/β-catenin transcriptional dynamic activities during embryogenesis, including using in vivo imaging. Our analyses demonstrate the use of this transcriptional reporter to unveil novel aspects of Wnt signaling, thus opening new routes of investigation into the role of this pathway during amniote embryonic development.

Data availability

Figure 1 - figure supplement 1-Source Data 1 and Figure 1 - figure supplement 2-Source Data 1 contain the numerical data used to generate the figures

Article and author information

Author details

  1. Hila Barzilai-Tutsch

    NeuroMyoGene Institute (INMG), Claude Bernard University Lyon 1, Lyon, France
    Competing interests
    The authors declare that no competing interests exist.
  2. Valerie Morin

    NeuroMyoGene Institute (INMG), Claude Bernard University Lyon 1, Lyon, France
    Competing interests
    The authors declare that no competing interests exist.
  3. Gauthier Toulouse

    NeuroMyoGene Institute (INMG), Claude Bernard University Lyon 1, Lyon, France
    Competing interests
    The authors declare that no competing interests exist.
  4. Oleksandr Chernyavskiy

    Monash University, Clayton, Australia
    Competing interests
    The authors declare that no competing interests exist.
  5. Stephen Firth

    Monash University, Clayton, Australia
    Competing interests
    The authors declare that no competing interests exist.
  6. Christophe Marcelle

    NeuroMyoGene Institute (INMG), Claude Bernard University Lyon 1, LYON, France
    For correspondence
    christophe.marcelle@univ-lyon1.fr
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9612-7609
  7. Olivier Serralbo

    CSIRO Health and Biosecurity, Geelong, Australia
    For correspondence
    olivier.serralbo@csiro.au
    Competing interests
    The authors declare that no competing interests exist.

Funding

No funding has supported this study.

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the Monash University. All of the animals were handled according to approved institutional animal care and use committee of Monash University (Research Ethics & Compliance numbers: ERM#27128 and ERM#18809)

Reviewing Editor

  1. Marianne E Bronner, California Institute of Technology, United States

Publication history

  1. Preprint posted: June 11, 2021 (view preprint)
  2. Received: August 4, 2021
  3. Accepted: July 13, 2022
  4. Accepted Manuscript published: July 14, 2022 (version 1)
  5. Version of Record published: August 19, 2022 (version 2)

Copyright

© 2022, Barzilai-Tutsch et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Hila Barzilai-Tutsch
  2. Valerie Morin
  3. Gauthier Toulouse
  4. Oleksandr Chernyavskiy
  5. Stephen Firth
  6. Christophe Marcelle
  7. Olivier Serralbo
(2022)
Transgenic quails reveal dynamic TCF/β-catenin signaling during avian embryonic development
eLife 11:e72098.
https://doi.org/10.7554/eLife.72098
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