Whole embryo culture, transcriptomics and RNA interference identify TBX1 and FGF11 as novel regulators of limb development in the mouse
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...
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| Veröffentlicht in: | Scientific reports 2020-02, Vol.10 (1), p.3597-3597, Article 3597 |
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| creator | Tejedor, Gautier Laplace-Builhé, Béryl Luz-Crawford, Patricia Assou, Said Barthelaix, Audrey Mathieu, Marc Kissa, Karima Jorgensen, Christian Collignon, Jérôme Chuchana, Paul Djouad, Farida |
| description | 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. |
| doi_str_mv | 10.1038/s41598-020-60217-w |
| format | Article |
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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
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Tbx1
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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. 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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.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>32107392</pmid><doi>10.1038/s41598-020-60217-w</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0001-6448-7995</orcidid><orcidid>https://orcid.org/0000-0002-7683-3661</orcidid><orcidid>https://orcid.org/0000-0002-1212-4006</orcidid><orcidid>https://orcid.org/0000-0002-1437-587X</orcidid><orcidid>https://orcid.org/0000-0001-8248-6822</orcidid><orcidid>https://orcid.org/0000-0001-6947-8094</orcidid><oa>free_for_read</oa></addata></record> |
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| ispartof | Scientific reports, 2020-02, Vol.10 (1), p.3597-3597, Article 3597 |
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| source | MEDLINE; Nature Free; DOAJ Directory of Open Access Journals; EZB-FREE-00999 freely available EZB journals; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry; Springer Nature OA Free Journals |
| subjects | 13/31 14/19 42/89 631/136/1455 631/80/82/23 64/60 Animals Arrays Biochemistry, Molecular Biology Cells, Cultured Cellular Biology Computational Biology Development Biology Embryo, Mammalian Embryology and Organogenesis Embryos Fibroblast Growth Factors - genetics Fibroblast Growth Factors - metabolism Forelimb - physiology Gene expression Gene Expression Profiling Gene Expression Regulation, Developmental Genomics Humanities and Social Sciences Laboratory animals Life Sciences Mice Molecular biology multidisciplinary Organogenesis - genetics RNA Interference RNA, Small Interfering - genetics RNA-mediated interference Science Science (multidisciplinary) Software T-Box Domain Proteins - genetics T-Box Domain Proteins - metabolism Tbx1 protein Transcription Transcriptome Transcriptomics Viviparity |
| title | Whole embryo culture, transcriptomics and RNA interference identify TBX1 and FGF11 as novel regulators of limb development in the mouse |
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