Whole embryo culture, transcriptomics and RNA interference identify TBX1 and FGF11 as novel regulators of limb development in the mouse

Gautier Tejedor, Béryl Laplace-Builhé, Patricia Luz-Crawford, Said Assou, Audrey Barthelaix, Marc Mathieu, Karima Kissa, Christian Jorgensen, Jérôme Collignon, Paul Chuchana, Farida Djouad

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Identifying genes involved in vertebrate developmental processes and characterizing this involvement are daunting tasks, especially in the mouse where viviparity complicates investigations. Attempting to devise a streamlined approach for this type of study we focused on limb development. We cultured E10.5 and E12.5 embryos and performed transcriptional profiling to track molecular changes in the forelimb bud over a 6-hour time-window. The expression of certain genes was found to diverge rapidly from its normal path, possibly reflecting the activation of a stress-induced response. Others, however, maintained for up to 3 hours dynamic expression profiles similar to those seen in utero. Some of these resilient genes were known regulators of limb development. The implication of the others in this process was either unsuspected or unsubstantiated. The localized knockdown of two such genes, Fgf11 and Tbx1, hampered forelimb bud development, providing evidence of their implication. These results show that combining embryo culture, transcriptome analysis and RNA interference could speed up the identification of genes involved in a variety of developmental processes, and the validation of their implication.

Original languageEnglish
Article number3597
JournalScientific Reports
Volume10
Issue number1
DOIs
StatePublished - 1 Dec 2020

Bibliographical note

Funding Information:
We thank the MRI facilities for their assistance and SMARTY platform and Network of Animal facilities of Montpellier. This work was supported by Inserm, the University of Montpellier and grants from the Région Languedoc-Roussillon Chercheur(se)s d’Avenir. It contributes also to the Labex "Who am I?" #ANR-11-LABX-0071 and the Université de Paris Idex #ANR-18-IDEX-0001 funded by the french government.

Publisher Copyright:
© 2020, The Author(s).

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