Abstract
BackgroundDmrt2a 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.ResultsWe 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.ConclusionsIn 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.
Highlights
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
We induced a heat-shock (Fig. 1b) at bud-stage given that this time-point coincides with dmrt2a early expression (Fig. 1c), marks the beginning of somitogenesis [11] and precedes the formation of zebrafish Left-Right Organiser (LRO) [12]
Using our transgenic line we described for the first time dmrt2a overexpression phenotype
Summary
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. The DMRT (Doublesex and Mab Related Transcription factors) family of zinc finger like transcription factors has been classically associated with sexual determination and differentiation. These functions remain fairly conserved in the different animals studied so far, during evolution different members of this family acquired different functions, such as nervous system development, somitogenesis and left-right asymmetry establishment [1]. A fish-specific paralog gene – dmrt2b – was identified with similar
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