Tao Wenjing, Sun Lina, Shi Hongjuan, Cheng Yunying, Jiang Dongneng, Fu Beide, Conte Matthew A, Gammerdinger William J, Kocher Thomas D, Wang Deshou
Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, 400715, P. R. China.
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, P. R. China.
BMC Genomics. 2016 May 4;17:328. doi: 10.1186/s12864-016-2636-z.
MicroRNAs (miRNAs) represent a second regulatory network that has important effects on gene expression and protein translation during biological process. However, the possible role of miRNAs in the early stages of fish sex differentiation is not well understood. In this study, we carried an integrated analysis of miRNA and mRNA expression profiles to explore their possibly regulatory patterns at the critical stage of sex differentiation in tilapia.
We identified 279 pre-miRNA genes in tilapia genome, which were highly conserved in other fish species. Based on small RNA library sequencing, we identified 635 mature miRNAs in tilapia gonads, in which 62 and 49 miRNAs showed higher expression in XX and XY gonads, respectively. The predicted targets of these sex-biased miRNAs (e.g., miR-9, miR-21, miR-30a, miR-96, miR-200b, miR-212 and miR-7977) included genes encoding key enzymes in steroidogenic pathways (Cyp11a1, Hsd3b, Cyp19a1a, Hsd11b) and key molecules involved in vertebrate sex differentiation (Foxl2, Amh, Star1, Sf1, Dmrt1, and Gsdf). These genes also showed sex-biased expression in tilapia gonads at 5 dah. Some miRNAs (e.g., miR-96 and miR-737) targeted multiple genes involved in steroid synthesis, suggesting a complex miRNA regulatory network during early sex differentiation in this fish.
The sequence and expression patterns of most miRNAs in tilapia are conserved in fishes, indicating the basic functions of vertebrate miRNAs might share a common evolutionary origin. This comprehensive analysis of miRNA and mRNA at the early stage of molecular sex differentiation in tilapia XX and XY gonads lead to the discovery of differentially expressed miRNAs and their putative targets, which will facilitate studies of the regulatory network of molecular sex determination and differentiation in fishes.
微小RNA(miRNA)代表了第二个调控网络,在生物过程中对基因表达和蛋白质翻译具有重要影响。然而,miRNA在鱼类性别分化早期阶段的可能作用尚未得到充分了解。在本研究中,我们对miRNA和mRNA表达谱进行了综合分析,以探索它们在罗非鱼性别分化关键阶段的可能调控模式。
我们在罗非鱼基因组中鉴定出279个前体miRNA基因,这些基因在其他鱼类物种中高度保守。基于小RNA文库测序,我们在罗非鱼性腺中鉴定出635个成熟miRNA,其中分别有62个和49个miRNA在XX和XY性腺中表达较高。这些性别偏向性miRNA(例如miR-9、miR-21、miR-30a、miR-96、miR-200b、miR-212和miR-7977)的预测靶标包括编码类固醇生成途径关键酶(Cyp11a1、Hsd3b、Cyp19a1a、Hsd11b)的基因以及参与脊椎动物性别分化的关键分子(Foxl2、Amh、Star1、Sf1、Dmrt1和Gsdf)。这些基因在5日龄罗非鱼性腺中也表现出性别偏向性表达。一些miRNA(例如miR-96和miR-737)靶向多个参与类固醇合成基因,表明该鱼类早期性别分化过程中存在复杂的miRNA调控网络。
罗非鱼中大多数miRNA的序列和表达模式在鱼类中是保守的,这表明脊椎动物miRNA的基本功能可能具有共同的进化起源。对罗非鱼XX和XY性腺分子性别分化早期阶段的miRNA和mRNA进行的综合分析,发现了差异表达的miRNA及其推定靶标,这将有助于研究鱼类分子性别决定和分化的调控网络。