Yuan Lihong, Li Linmiao, Zhang Xiujuan, Jiang Haiying, Chen Jinping
School of Life Sciences and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou, China.
Guangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Institute of Applied Biological Resources, Guangzhou, China.
PeerJ. 2019 Apr 12;7:e6709. doi: 10.7717/peerj.6709. eCollection 2019.
Sturgeons are considered living fossils, and have a very high conservation and economic value. Studies on the molecular mechanism of sturgeon gonadal development and sex differentiation would not only aid in understanding vertebrate sex determination but also benefit sturgeon aquaculture. Piwi-interacting RNAs (piRNAs) have been shown to function in germline or gonadal development. In this study, we performed small RNA deep sequencing and microarray hybridization to identify potential sturgeon piRNAs.
Male and female sturgeon gonads were collected and used for small RNA sequencing on an Illumina HiSeq platform with the validation of piRNA expression by microarray chip. The program Bowtie and -mer scheme were performed to filter small RNA reads and discover potential sturgeon piRNAs. A known piRNA database, the coding sequence (CDS), 5' and 3' untranslated region (UTR) database of the transcriptome, Gene Ontology (GO) database and KEGG pathway database were searched subsequently to analyze the potential bio-function of sturgeon piRNAs.
A total of 875,679 putative sturgeon piRNAs were obtained, including 93 homologous to known piRNAs and hundreds showing sex-specific and sex-biased expression. Further analysis showed that they are predominant in both the ovaries and testes and those with a sex-specific expression pattern are nearly equally distribution between sexes. This may imply a relevant role in sturgeon gonadal development. KEGG pathway and GO annotation analyses indicated that they may be related to sturgeon reproductive processes.
Our study provides the first insights into the gonadal piRNAs in a sturgeon species and should serve as a useful resource for further elucidation of the gene regulation involved in the sex differentiation of vertebrates. These results should also facilitate the technological development of early sex identification in sturgeon aquaculture.
鲟鱼被视为活化石,具有极高的保护和经济价值。对鲟鱼性腺发育和性别分化的分子机制进行研究,不仅有助于理解脊椎动物的性别决定,还对鲟鱼养殖有益。已有研究表明,Piwi相互作用RNA(piRNA)在生殖系或性腺发育中发挥作用。在本研究中,我们进行了小RNA深度测序和微阵列杂交,以鉴定潜在的鲟鱼piRNA。
采集雄性和雌性鲟鱼性腺,在Illumina HiSeq平台上进行小RNA测序,并通过微阵列芯片验证piRNA表达。使用Bowtie程序和-mer方案过滤小RNA读数,发现潜在的鲟鱼piRNA。随后搜索已知的piRNA数据库、转录组的编码序列(CDS)、5'和3'非翻译区(UTR)数据库、基因本体(GO)数据库和KEGG通路数据库,以分析鲟鱼piRNA的潜在生物学功能。
共获得875,679个推定的鲟鱼piRNA,其中93个与已知piRNA同源,数百个表现出性别特异性和性别偏向性表达。进一步分析表明,它们在卵巢和睾丸中均占主导地位,且具有性别特异性表达模式的piRNA在两性之间的分布几乎相等。这可能意味着其在鲟鱼性腺发育中发挥相关作用。KEGG通路和GO注释分析表明,它们可能与鲟鱼的生殖过程有关。
我们的研究首次揭示了鲟鱼性腺中的piRNA,应为进一步阐明脊椎动物性别分化所涉及的基因调控提供有用资源。这些结果也应有助于鲟鱼养殖中早期性别鉴定技术的发展。