A functional Bucky ball-GFP transgene visualizes germ plasm in living zebrafish
A functional Bucky ball-GFP transgene visualizes germ plasm in living zebrafish
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DOI：
10.1016/j.gep.2015.05.003
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发表时间：
2015-05-01
影响因子：
1.2
通讯作者：
Dosch, Roland
中科院分区：
文献类型：
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作者：
Riemer, Stephan;Bontems, Franck;Dosch, Roland
在许多动物中，种系由称为种质的母体 RNA 颗粒指定。因此，胚胎发生过程中种质的正确定位对于生殖细胞的规范至关重要。在斑马鱼中，我们之前发现巴基球（Buc）是种质形成的关键调节因子。在这里，我们使用 Buc 抗体来描述其连续种质定位。此外，我们还生成了用于实时成像的转基因 Buc-GFP 系，该系可以可视化从卵子发生到幼虫阶段的种质组装。 Buc-GFP 的实时成像产生了令人惊叹的电影，因为它们突出了种质运动的动态细节。此外，我们发现受精后2天的原始生殖细胞中仍然检测到Buc。有趣的是，转基因拯救了 buc 突变体，从遗传学上证明了 Buc-GFP 融合蛋白是有功能的。这些结果表明，Buc-GFP 发挥了种系发育所必需的所有生化相互作用，并突出了该系分析种质形成分子调控的潜力。 (C) 2015 Elsevier B.V. 保留所有权利。
In many animals, the germline is specified by maternal RNA-granules termed germ plasm. The correct localization of germ plasm during embryogenesis is therefore crucial for the specification of germ cells. In zebrafish, we previously identified Bucky ball (Buc) as a key regulator of germ plasm formation. Here, we used a Buc antibody to describe its continuous germ plasm localization. Moreover, we generated a transgenic Buc-GFP line for live imaging, which visualizes germ plasm from its assembly during oogenesis up to the larval stages. Live imaging of Buc-GFP generated stunning movies, as they highlighted the dynamic details of germ plasm movements. Moreover, we discovered that Buc was still detected in primordial germ cells 2 days after fertilization. Interestingly, the transgene rescued buc mutants demonstrating genetically that the Buc-GFP fusion protein is functional. These results show that Buc-GFP exerts all biochemical interactions essential for germline development and highlight the potential of this line to analyze the molecular regulation of germ plasm formation. (C) 2015 Elsevier B.V. All rights reserved.