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斑马鱼转基因系中神经嵴谱系的实时成像。

Live image profiling of neural crest lineages in zebrafish transgenic lines.

机构信息

Department of Systems Immunology, Kangwon National University, Chuncheon 200-701, Korea.

出版信息

Mol Cells. 2013 Mar;35(3):255-60. doi: 10.1007/s10059-013-0001-5. Epub 2013 Feb 26.

Abstract

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.

摘要

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。精确描述转基因品系中标记的细胞谱系对于理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,我们才能有信心选择适合我们特定研究目标的品系。在这里,我们对密切相关的神经嵴转基因品系 Tg(foxd3:GFP)、Tg(sox10:eGFP)和 Tg(sox10:mRFP)中标记的细胞谱系进行了分析。这些鱼被杂交以产生胚胎,在这些胚胎中,使用活共聚焦成像可以在细胞水平上直接比较 foxd3 和 sox10 转基因神经嵴标记。我们已经确定了在早期神经嵴发育过程中每条品系标记的细胞谱系中的关键差异,并证明最初从脑神经管中迁移出来的最前颅神经嵴细胞在脑前部和中脑前部表达 sox10:eGFP 和 sox10:mRFP,但不表达 foxd3:GFP。这种差异表型在神经嵴谱系的分化后代中一直保持到 3.5dpf。我们的数据将使研究人员能够在选择未来神经嵴研究的转基因品系时做出明智的选择。

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