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Identification of Dmrt2a downstream genes during zebrafish early development using a timely controlled approach

BACKGROUND: Dmrt2a is a zinc finger like transcription factor with several roles during zebrafish early development: left-right asymmetry, synchronisation of the somite clock genes and fast muscle differentiation. Despite the described functions, Dmrt2a mechanism of action is unknown. Therefore, wit...

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Autores principales: Pinto, Rita Alexandra, Almeida-Santos, José, Lourenço, Raquel, Saúde, Leonor
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6006574/
https://www.ncbi.nlm.nih.gov/pubmed/29914374
http://dx.doi.org/10.1186/s12861-018-0173-5
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author Pinto, Rita Alexandra
Almeida-Santos, José
Lourenço, Raquel
Saúde, Leonor
author_facet Pinto, Rita Alexandra
Almeida-Santos, José
Lourenço, Raquel
Saúde, Leonor
author_sort Pinto, Rita Alexandra
collection PubMed
description BACKGROUND: Dmrt2a is a zinc finger like transcription factor with several roles during zebrafish early development: left-right asymmetry, synchronisation of the somite clock genes and fast muscle differentiation. Despite the described functions, Dmrt2a mechanism of action is unknown. Therefore, with this work, we propose to identify Dmrt2a downstream genes during zebrafish early development. RESULTS: We generated and validated a heat-shock inducible transgenic line, to timely control dmrt2a overexpression, and dmrt2a mutant lines. We characterised dmrt2a overexpression phenotype and verified that it was very similar to the one described after knockdown of this gene, with left-right asymmetry defects and desynchronisation of somite clock genes. Additionally, we identified a new phenotype of somite border malformation. We generated several dmrt2a mutant lines, but we only detected a weak to negligible phenotype. As dmrt2a has a paralog gene, dmrt2b, with similar functions and expression pattern, we evaluated the possibility of redundancy. We found that dmrt2b does not seem to compensate the lack of dmrt2a. Furthermore, we took advantage of one of our mutant lines to confirm dmrt2a morpholino specificity, which was previously shown to be a robust knockdown tool in two independent studies. Using the described genetic tools to perform and validate a microarray, we were able to identify six genes downstream of Dmrt2a: foxj1b, pxdc1b, cxcl12b, etv2, foxc1b and cyp1a. CONCLUSIONS: In this work, we generated and validated several genetic tools for dmrt2a and identified six genes downstream of this transcription factor. The identified genes will be crucial to the future understanding of Dmrt2a mechanism of action in zebrafish. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12861-018-0173-5) contains supplementary material, which is available to authorized users.
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spelling pubmed-60065742018-06-26 Identification of Dmrt2a downstream genes during zebrafish early development using a timely controlled approach Pinto, Rita Alexandra Almeida-Santos, José Lourenço, Raquel Saúde, Leonor BMC Dev Biol Research Article BACKGROUND: Dmrt2a is a zinc finger like transcription factor with several roles during zebrafish early development: left-right asymmetry, synchronisation of the somite clock genes and fast muscle differentiation. Despite the described functions, Dmrt2a mechanism of action is unknown. Therefore, with this work, we propose to identify Dmrt2a downstream genes during zebrafish early development. RESULTS: We generated and validated a heat-shock inducible transgenic line, to timely control dmrt2a overexpression, and dmrt2a mutant lines. We characterised dmrt2a overexpression phenotype and verified that it was very similar to the one described after knockdown of this gene, with left-right asymmetry defects and desynchronisation of somite clock genes. Additionally, we identified a new phenotype of somite border malformation. We generated several dmrt2a mutant lines, but we only detected a weak to negligible phenotype. As dmrt2a has a paralog gene, dmrt2b, with similar functions and expression pattern, we evaluated the possibility of redundancy. We found that dmrt2b does not seem to compensate the lack of dmrt2a. Furthermore, we took advantage of one of our mutant lines to confirm dmrt2a morpholino specificity, which was previously shown to be a robust knockdown tool in two independent studies. Using the described genetic tools to perform and validate a microarray, we were able to identify six genes downstream of Dmrt2a: foxj1b, pxdc1b, cxcl12b, etv2, foxc1b and cyp1a. CONCLUSIONS: In this work, we generated and validated several genetic tools for dmrt2a and identified six genes downstream of this transcription factor. The identified genes will be crucial to the future understanding of Dmrt2a mechanism of action in zebrafish. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12861-018-0173-5) contains supplementary material, which is available to authorized users. BioMed Central 2018-06-19 /pmc/articles/PMC6006574/ /pubmed/29914374 http://dx.doi.org/10.1186/s12861-018-0173-5 Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Pinto, Rita Alexandra
Almeida-Santos, José
Lourenço, Raquel
Saúde, Leonor
Identification of Dmrt2a downstream genes during zebrafish early development using a timely controlled approach
title Identification of Dmrt2a downstream genes during zebrafish early development using a timely controlled approach
title_full Identification of Dmrt2a downstream genes during zebrafish early development using a timely controlled approach
title_fullStr Identification of Dmrt2a downstream genes during zebrafish early development using a timely controlled approach
title_full_unstemmed Identification of Dmrt2a downstream genes during zebrafish early development using a timely controlled approach
title_short Identification of Dmrt2a downstream genes during zebrafish early development using a timely controlled approach
title_sort identification of dmrt2a downstream genes during zebrafish early development using a timely controlled approach
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6006574/
https://www.ncbi.nlm.nih.gov/pubmed/29914374
http://dx.doi.org/10.1186/s12861-018-0173-5
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