Li Mingyou, Zhu Feng, Li Zhendong, Hong Ni, Hong Yunhan
Ministry of Education Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, College of Fisheries and Life Sciences, Shanghai Ocean University, 999 Hucheng Huan Road, Shanghai 201306, China.
Department of Biological Sciences, National University of Singapore, Science Drive 4, Singapore 117543, Singapore.
Sci Rep. 2016 Jun 22;6:28317. doi: 10.1038/srep28317.
The DAZ family genes boule, daz and dazl have conserved functions in primordial germ cell (PGC) migration, germ stem cell proliferation, differentiation and meiosis progression. It has remained unknown whether this family is required for PGC formation in developing embryos. Our recent study in the fish medaka (Oryzias latipes) has defined dnd as the critical PGC specifier and predicted the presence of additional factors essential for PGC formation. Here we report that dazl is a second key player for medaka PGC formation. Dazl knockdown did not prevent PGC formation even in the absence of normal somatic structures. It turned out that a high level of Dazl protein was maternally supplied and persisted until gastrulation, and hardly affected by two antisense morpholino oligos targeting the dazl RNA translation. Importantly, microinjection of a Dazl antibody remarkably reduced the number of PGCs and even completely abolished PGC formation without causing detectable somatic abnormality. Therefore, medaka PGC formation requires the Dazl protein as maternal germ plasm component, offering first evidence that dazl is a critical player in PGC formation in vivo. Our results demonstrate that antibody neutralization is a powerful tool to study the roles of maternal protein factors in PGC development in vivo.
DAZ家族基因boule、daz和dazl在原始生殖细胞(PGC)迁移、生殖干细胞增殖、分化及减数分裂进程中具有保守功能。目前尚不清楚该家族在发育胚胎的PGC形成过程中是否是必需的。我们最近对青鳉(Oryzias latipes)的研究已将dnd定义为关键的PGC决定因子,并预测存在其他对PGC形成至关重要的因子。在此我们报告,dazl是青鳉PGC形成的第二个关键因子。即使在没有正常体细胞结构的情况下,敲低dazl也不会阻止PGC形成。结果发现,高水平的Dazl蛋白由母体提供并持续到原肠胚形成期,且几乎不受两种靶向dazl RNA翻译的反义吗啉代寡核苷酸的影响。重要的是,显微注射Dazl抗体显著减少了PGC的数量,甚至完全消除了PGC的形成,且未引起可检测到的体细胞异常。因此,青鳉PGC的形成需要Dazl蛋白作为母体生殖质成分,这首次证明dazl是体内PGC形成中的关键因子。我们的结果表明,抗体中和是研究母体蛋白因子在体内PGC发育中作用的有力工具。