A crystal-clear zebrafish for in vivo imaging.
A crystal-clear zebrafish for in vivo imaging.
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DOI：
10.1038/srep29490
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发表时间：
2016-07-06
影响因子：
4.6
通讯作者：
Hindges R
中科院分区：
文献类型：
--
作者：
Antinucci P;Hindges R
斑马鱼（Danio rerio）幼鱼是研究亚细胞、细胞和系统水平生物学现象的理想动物模型。然而，即使在这种动物模型中，高度色素沉着组织（如眼睛）的光学可及性也是有限的。提高斑马鱼幼体透明度的典型策略需要使用高毒性化合物（例如1-苯基-2-硫脲，PTU）或色素沉着突变株（例如casper突变株）。迄今为止，这些策略都没有产生身体和眼睛都透明的正常行为的幼虫。在这里，我们提出了晶体，一个光学透明的斑马鱼突变体，通过结合不同的可行的突变影响皮肤色素沉着。与先前描述的组合突变体casper相比，晶体突变体在视网膜色素上皮中也缺乏色素沉着，因此能够光学进入眼睛。与PTU处理的动物不同，水晶幼虫能够像野生型幼虫一样有效地执行视觉引导行为，例如视动反应。为了验证晶体幼虫在体内的应用，我们进行了全脑光片成像和视网膜神经活动的双光子钙成像。总之，这种新的组合色素沉着突变体是一种理想的脊椎动物工具，完全通畅的结构和功能的生物过程中的体内调查，特别是当成像组织内或之间的眼睛。
The larval zebrafish (Danio rerio) is an excellent vertebrate model for in vivo imaging of biological phenomena at subcellular, cellular and systems levels. However, the optical accessibility of highly pigmented tissues, like the eyes, is limited even in this animal model. Typical strategies to improve the transparency of zebrafish larvae require the use of either highly toxic chemical compounds (e.g. 1-phenyl-2-thiourea, PTU) or pigmentation mutant strains (e.g. casper mutant). To date none of these strategies produce normally behaving larvae that are transparent in both the body and the eyes. Here we present crystal, an optically clear zebrafish mutant obtained by combining different viable mutations affecting skin pigmentation. Compared to the previously described combinatorial mutant casper, the crystal mutant lacks pigmentation also in the retinal pigment epithelium, therefore enabling optical access to the eyes. Unlike PTU-treated animals, crystal larvae are able to perform visually guided behaviours, such as the optomotor response, as efficiently as wild type larvae. To validate the in vivo application of crystal larvae, we performed whole-brain light-sheet imaging and two-photon calcium imaging of neural activity in the retina. In conclusion, this novel combinatorial pigmentation mutant represents an ideal vertebrate tool for completely unobstructed structural and functional in vivo investigations of biological processes, particularly when imaging tissues inside or between the eyes.