Generating transparent zebrafish: A refined method to improve detection of gene expression during embryonic development
Generating transparent zebrafish: A refined method to improve detection of gene expression during embryonic development
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DOI：
10.1007/s1012601-0053-4
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发表时间：
2001-11-01
影响因子：
3
通讯作者：
Olsson, PE
中科院分区：
文献类型：
--
作者：
Karlsson, J;von Hofsten, J;Olsson, PE
在斑马鱼（Danio rerio）中，色素沉着在胚胎发生期间开始，并开始于视网膜上皮和黑色素细胞中。色素细胞发育迅速，在几个小时内，它们就构成了胚胎的显著特征。为了通过整体原位杂交、共聚焦显微镜或GFP表达来改善信号检测，可以在胚胎发生期间用1-苯基2-硫脲（PTU）处理胚胎。PTU通过阻断黑色素途径中的所有酪氨酸酶依赖性步骤来抑制黑色素生成，但在高浓度下可能是有毒的。只要继续PTU处理，胚胎就保持透明。然而，PTU治疗必须在初始色素沉着之前开始，因为它不能去除已经形成的色素。在这里，我们提供了一个协议，产生透明的斑马鱼，同时避免毒性和致畸作用的PTU治疗。
In zebrafish (Danio rerio) pigmentation is initiated during embryogenesis and begins in the retinal epithelium and in the melanophores. The pigment cells develop rapidly, and within hours they constitute a prominent feature of the embryo. In order to improve signal detection by whole mount in situ hybridization, confocal microscopy, or expression of GFP, embryos may be treated with 1-phenyl 2-thiourea (PTU) during embryogenesis. PTU inhibits melanogenesis by blocking all tyrosinase-dependent steps in the melanin pathway but can be toxic at high concentrations. The embryos remain transparent as long as the PTU treatment is continued. However, PTU treatment must be initiated before the initial pigmentation because it does not remove already formed pigment. Here we provide a protocol for generating transparent zebrafish while avoiding the toxic and teratogenic effects of PTU treatment.