Cargando…
Transgenic quails reveal dynamic TCF/β-catenin signaling during avian embryonic development
The Wnt/β-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...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9395189/ https://www.ncbi.nlm.nih.gov/pubmed/35833630 http://dx.doi.org/10.7554/eLife.72098 |
_version_ | 1784771633971462144 |
---|---|
author | Barzilai-Tutsch, Hila Morin, Valerie Toulouse, Gauthier Chernyavskiy, Oleksandr Firth, Stephen Marcelle, Christophe Serralbo, Olivier |
author_facet | Barzilai-Tutsch, Hila Morin, Valerie Toulouse, Gauthier Chernyavskiy, Oleksandr Firth, Stephen Marcelle, Christophe Serralbo, Olivier |
author_sort | Barzilai-Tutsch, Hila |
collection | PubMed |
description | The Wnt/β-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 (12× or 16×) 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 characterization 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. |
format | Online Article Text |
id | pubmed-9395189 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-93951892022-08-23 Transgenic quails reveal dynamic TCF/β-catenin signaling during avian embryonic development Barzilai-Tutsch, Hila Morin, Valerie Toulouse, Gauthier Chernyavskiy, Oleksandr Firth, Stephen Marcelle, Christophe Serralbo, Olivier eLife Developmental Biology The Wnt/β-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 (12× or 16×) 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 characterization 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. eLife Sciences Publications, Ltd 2022-07-14 /pmc/articles/PMC9395189/ /pubmed/35833630 http://dx.doi.org/10.7554/eLife.72098 Text en © 2022, Barzilai-Tutsch et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Developmental Biology Barzilai-Tutsch, Hila Morin, Valerie Toulouse, Gauthier Chernyavskiy, Oleksandr Firth, Stephen Marcelle, Christophe Serralbo, Olivier Transgenic quails reveal dynamic TCF/β-catenin signaling during avian embryonic development |
title | Transgenic quails reveal dynamic TCF/β-catenin signaling during avian embryonic development |
title_full | Transgenic quails reveal dynamic TCF/β-catenin signaling during avian embryonic development |
title_fullStr | Transgenic quails reveal dynamic TCF/β-catenin signaling during avian embryonic development |
title_full_unstemmed | Transgenic quails reveal dynamic TCF/β-catenin signaling during avian embryonic development |
title_short | Transgenic quails reveal dynamic TCF/β-catenin signaling during avian embryonic development |
title_sort | transgenic quails reveal dynamic tcf/β-catenin signaling during avian embryonic development |
topic | Developmental Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9395189/ https://www.ncbi.nlm.nih.gov/pubmed/35833630 http://dx.doi.org/10.7554/eLife.72098 |
work_keys_str_mv | AT barzilaitutschhila transgenicquailsrevealdynamictcfbcateninsignalingduringavianembryonicdevelopment AT morinvalerie transgenicquailsrevealdynamictcfbcateninsignalingduringavianembryonicdevelopment AT toulousegauthier transgenicquailsrevealdynamictcfbcateninsignalingduringavianembryonicdevelopment AT chernyavskiyoleksandr transgenicquailsrevealdynamictcfbcateninsignalingduringavianembryonicdevelopment AT firthstephen transgenicquailsrevealdynamictcfbcateninsignalingduringavianembryonicdevelopment AT marcellechristophe transgenicquailsrevealdynamictcfbcateninsignalingduringavianembryonicdevelopment AT serralboolivier transgenicquailsrevealdynamictcfbcateninsignalingduringavianembryonicdevelopment |