The Fugu tyrp1 promoter directs specific GFP expression in zebrafish:: tools to study the RPE and the neural crest-derived melanophores
The Fugu tyrp1 promoter directs specific GFP expression in zebrafish:: tools to study the RPE and the neural crest-derived melanophores
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DOI：
10.1111/j.1600-0749.2006.00349.x
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发表时间：
2006-12-01
期刊：
影响因子：
--
通讯作者：
Wei, Xiangyun
中科院分区：
文献类型：
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作者：
Zou, Jian;Beermann, Friedrich;Wei, Xiangyun
在脊椎动物中，色素细胞只占细胞总数的一小部分，它们与其他类型的细胞混合在一起。这使得很难将它们分离出来，以分析它们在发展背景下的功能。为了缓解这一困难，我们在1 kb的Fugu tyrp1启动子的控制下，产生了两个稳定的转基因斑马鱼系pt101和pt102，它们在黑色素细胞中表达绿色荧光蛋白（GFP）。在pt101中，GFP在视网膜色素上皮细胞（RPE）和神经嵴源性黑色素细胞（NCDM）中均有表达，而在pt102中，GFP主要在NCDM中表达。我们的研究结果表明，河豚tyrp1启动子可以以细胞类型特异性的方式指导斑马鱼的转基因表达。此外，我们的研究结果为斑马鱼RPE细胞和NCDM中黑色素合成基因的差异调控提供了证据。利用这些鱼中不同的GFP表达水平，我们通过流式细胞术分离了黑素细胞，并揭示了从RPE细胞中分选NCDM的能力。因此，这些转基因系是研究斑马鱼黑色素细胞的有用工具。
In vertebrates, pigment cells account for a small percentage of the total cell population and they intermingle with other cell types. This makes it difficult to isolate them for analyzes of their functions in the context of development. To alleviate such difficulty, we generated two stable transgenic zebrafish lines (pt101 and pt102) that express green fluorescent protein (GFP) in melanophores under the control of the 1 kb Fugu tyrp1 promoter. In pt101, GFP is expressed in both retinal pigment epithelium (RPE) cells and the neural crest-derived melanophores (NCDM), whereas in pt102, GFP is predominately expressed in the NCDM. Our results indicate that the Fugu tyrp1 promoter can direct transgene expression in a cell-type-specific manner in zebrafish. In addition, our findings provide evidence supporting differential regulations of melanin-synthesizing genes in RPE cells and the NCDM in zebrafish. Utilizing the varying GFP expression levels in these fish, we have isolated melanophores via flow cytometry and revealed the capability of sorting the NCDM from RPE cells as well. Thus, these transgenic lines are useful tools to study melanophores in zebrafish.