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Live Image Profiling of Neural Crest Lineages in Zebrafish Transgenic Lines

机译:斑马鱼转基因品系中神经C谱系的实时图像分析

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摘要

Zebrafish transgenic lines are important experimental tools for lineage tracing and imaging studies. It is crucial to precisely characterize the cell lineages labeled in transgenic lines to understand their limitations and thus properly interpret the data obtained from their use; only then can we confidently select a line appropriate for our particular research objectives. Here we profiled the cell lineages labeled in the closely related neural crest transgenic lines Tg(foxd3:GFP), Tg(sox10:eGFP) and Tg(sox10:mRFP). These fish were crossed to generate embryos, in which foxd3 and sox10 transgenic neural crest labeling could be directly compared at the cellular level using live confocal imaging. We have identified key differences in the cell lineages labeled in each line during early neural crest development and demonstrated that the most anterior cranial neural crest cells initially migrating out of neural tube at the level of forebrain and anterior midbrain express sox10: eGFP and sox10:mRFP, but not foxd3:GFP. This differential profile was robustly maintained in the differentiating progeny of the neural crest lineages until 3.5dpf. Our data will enable researchers to make an informed choice in selecting transgenic lines for future neural crest research.
机译:斑马鱼转基因品系是谱系追踪和成像研究的重要实验工具。准确表征转基因品系中标记的细胞谱系以了解其局限性,从而正确解释从其使用中获得的数据至关重要。只有这样,我们才能自信地选择适合我们特定研究目标的生产线。在这里,我们介绍了密切相关的神经c转基因系Tg(foxd3:GFP),Tg(sox10:eGFP)和Tg(sox10:mRFP)中标记的细胞谱系。将这些鱼杂交产生胚胎,在其中可以使用实时共聚焦成像在细胞水平上直接比较foxd3和sox10转基因神经c标记。我们已经确定了在早期神经c发育过程中各行中标记的细胞谱系的关键差异,并证明了最前颅神经neural细胞最初以前脑和中脑的水平移出神经管,并表达了sox10:eGFP和sox10:mRFP ,但不是foxd3:GFP。在神经profile谱系的分化后代中稳固地维持了这种差异特征,直至3.5dpf。我们的数据将使研究人员能够在选择转基因品系以进行未来神经c研究时做出明智的选择。

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