Identification and characterization of microRNAs in the gonad of Trachinotus ovatus using Solexa sequencing
Identification and characterization of microRNAs in the gonad of Trachinotus ovatus using Solexa sequencing
复制标题
使用 Solexa 测序对卵形鲳鲹性腺中的 microRNA 进行鉴定和表征
DOI：
10.1016/j.cbd.2019.03.010
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发表时间：
2019-06-01
影响因子：
3
通讯作者：
Peng, Jinxia
中科院分区：
文献类型：
--
作者：
He, Pingping;Wei, Pinyuan;Peng, Jinxia
卵形Trachinotus ovatus（T.卵圆的卵圆的卵为了研究性分化的潜在调控机制，我们对T.卵形的总共13，423，691和11，734，953个原始读段，分别代表从卵巢和睾丸获得的91，883，908和86，879，726个18-35 nt长度的独特序列。映射到T. ovatus转录组参考序列（未发表数据），并与miRBase数据库比较，鉴定出179个已知成熟的miRNAs和100个新的miRNAs。表达分析显示，165个miRNAs在卵巢和睾丸中差异表达，为了验证Illumina的结果，对9个差异表达最大的miRNAs的表达模式进行了分析。（dre-miR-7a、dre-miR-7b、dre-miR-153a-3p、dre-miR-144-3p、dre-miR-301a、dre-miR-92a-3p、dre-miR-727-5p、Novel 124和Novel 190，以及可能与性腺发育相关的5种miRNA（dre-miR-143、dre-miR-101 a、dre-miR-202- 5 p、dre-let-7 c-5 p和dre-miR-181 a-5 p）的那些。RT-qPCR验证的所有miRNA的表达趋势与下一代测序数据一致。本研究通过对卵巢和精巢组织中差异表达的miRNAs进行鉴定和分析，为进一步研究卵巢和精巢组织中差异表达的miRNAs奠定了基础。ovatus将增加我们对miRNAs在性腺分化中的作用的理解。这些数据也有助于研究miRNA对硬骨鱼生殖的调控。
The Trachinotus ovatus (T. ovatus) reach sexual maturity as late as 4-5 years, and there are no obvious morphology differences between males and females, even at maturity, making the selection of male and female parents for selective breeding difficult. To examine the potential regulatory mechanism of sexual differentiation, we conducted a microRNAs (miRNAs) analysis on the ovaries and testes of T. ovatus. A total of 13,423,691 and 11,734,953 raw reads, representing 91,883,908 and 86,879,726 unique sequences of 18-35 nt length obtained from the ovaries and testes, respectively. After mapping to the T. ovatus transcriptome reference sequence (unpublished data), and comparing the miRNA sequences with the miRBase database, 179 known mature miRNAs and 100 novel miRNAs were identified. Expression analysis showed that, 165 miRNAs were differentially expressed between the ovary and testis.To validate the Illumina results, the expression patterns of the nine most differently expressed miRNAs (dre-miR-7a, dre-miR-7b, dre-miR-153a-3p, dre-miR-144-3p, dre-miR-301a, dre-miR-92a-3p, dre-miR-727-5p, Novel 124, and Novel 190, as well as those of five miRNAs that may relate to gonad development (dre-miR-143, dre-miR-101a, dre-miR-202-5p, dre-let-7c-5p, and dre-miR-181a-5p), were assayed with RT-qPCR. The expression trends for all the miRNAs validated with RT-qPCR were consistent with the next-generation sequencing data. The identification and characterization of the miRNAs differentially expressed between the ovary and the testis of the T. ovatus will increase our understanding of the role of miRNAs in gonad differentiation. These data also facilitate studies on miRNA regulation of teleost reproduction.