Conserved Mechanisms for Germ Cell-Specific Localization of nanos3 Transcripts in Teleost Species with Aquaculture Significance
Conserved Mechanisms for Germ Cell-Specific Localization of nanos3 Transcripts in Teleost Species with Aquaculture Significance
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DOI：
10.1007/s10126-013-9543-y
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发表时间：
2013-10
影响因子：
3
通讯作者：
Adrijana Skugor;Krasimir Slanchev;J. Torgersen;H. Tveiten;Ø. Andersen
中科院分区：
文献类型：
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作者：
Adrijana Skugor;Krasimir Slanchev;J. Torgersen;H. Tveiten;Ø. Andersen
随着全球鱼类数量的减少，水产养殖产量的重要性正在增加，但几个养殖物种的早性成熟会降低肌肉的生长和质量，逃逸可能会对野生种群产生负面影响。这些问题的一个可能的解决方案是通过去除胚胎原始生殖细胞(PGCs)来生产不育鱼，这是在斑马鱼中开发的一项技术。对mRNA稳定性的细胞特异性调控对于生殖细胞谱系的正确确定至关重要，通常涉及到microRNA(MiRNA)介导的体细胞靶向mRNAs的降解。本研究报道了在大西洋鳕鱼、大西洋鲑鱼和斑马鱼中发现的miRNA靶基因3的3‘非翻译区(UTR)的保守基序的功能作用。Codnanos3的3‘非编码区足以将绿色荧光蛋白(GFP)表达到这三个物种在系统发育上相距遥远的物种的注射胚胎中。通过将截短的3‘非编码区与GFP融合，然后在斑马鱼胚胎中注射，进一步研究了对PGC特异性表达具有重要意义的3’非编码区元件。GFP结构在PGCs和体细胞中的表达模式表明，近端的U富集区负责内源性Nanos3mRNA的PGC特异性稳定。吗啡介导的RNA结合蛋白Dead End(DND)的下调是PGC特异性的miRNA作用抑制物，取消了鳕鱼和斑马鱼胚胎中PGC的荧光，表明生殖细胞的存活和迁移存在保守的DND依赖机制。
The importance of the aquaculture production is increasing with the declining global fish stocks, but early sexual maturation in several farmed species reduces muscle growth and quality, and escapees could have a negative impact on wild populations. A possible solution to these problems is the production of sterile fish by ablation of the embryonic primordial germ cells (PGCs), a technique developed in zebrafish. Cell-specific regulation of mRNA stability is crucial for proper specification of the germ cell lineage and commonly involves microRNA (miRNA)-mediated degradation of targeted mRNAs in somatic cells. This study reports on the functional roles of conserved motifs in the 3′ untranslated region (UTR) of the miRNA target genenanos3identified in Atlantic cod, Atlantic salmon, and zebrafish. The 3′UTR of codnanos3was sufficient for targeting the expression of green fluorescent protein (GFP) to the presumptive PGCs in injected embryos of the three phylogenetically distant species. 3′UTR elements of importance for PGC-specific expression were further examined by fusing truncated 3′UTR variants of codnanos3to GFP followed by injections in zebrafish embryos. The expression patterns of the GFP constructs in PGCs and somatic cells suggested that the proximal U-rich region is responsible for the PGC-specific stabilization of the endogenousnanos3mRNA. Morpholino-mediated downregulation of the RNA-binding protein Dead end (DnD), a PGC-specific inhibitor of miRNA action, abolished the fluorescence of the PGCs in cod and zebrafish embryos, suggesting a conserved DnD-dependent mechanism for germ cell survival and migration.