摘要
旨在解析circRNA在细粒棘球绦虫原头蚴、包囊壁和成虫的表达谱特性,为进一步阐明circRNA分子在细粒棘球绦虫生长发育中的作用提供理论依据。采集细粒棘球绦虫的原头蚴、包囊壁和成虫,提取总RNA,构建circRNA、miRNA以及mRNA文库,采用全转录组测序方法进行测序分析。通过生物信息学分析和功能预测筛选出差异表达的circRNA,并进行GO、KEGG以及ceRNA调控网络分析,采用实时荧光定量PCR(qRT-PCR)对其进行验证。结果在细粒棘球绦虫原头蚴、包囊壁和成虫共鉴定得到636个circRNA分子,其中差异表达的circRNA分子有140个,在原头蚴、包囊壁和成虫特异性表达的circRNA分子分别为44、2和0个。GO分析结果显示,成虫和原头蚴以及包囊壁相比,差异表达的circRNA分子主要富集于生物学过程的有性生殖、精子发生、雄性配子产生、轴系发生以及神经生成、分化等。KEGG分析结果显示,原头蚴和成虫相比,差异表达的circRNA的显著富集于Notch、三羧酸循环、磷脂酰肌醇、核黄素代谢通路以及脂肪酸合成和降解等信号通路。原头蚴和包囊壁相比,差异表达的circRNA显著富集于背腹轴形成、Notch、wnt以及FoxO信号通路等。成虫和包囊壁相比,差异表达的circRNA的显著富集于Notch、碳代谢、三羧酸循环以及糖酵解/糖异生通路。靶向预测circRNA-miRNA-mRNA调控网络结果显示,有24个差异表达circRNAs与14个miRNAs和54个mRNAs形成164个up-down-up的circRNA-miRNA-mRNA调控网路,有13个差异表达的circRNAs与7个miRNAs以及16个mRNAs构成36个down-up-down的circRNA-miRNA-mRNA调控网络。对部分circRNA分子的qRT-PCR验证结果表明,其mRNA转录水平与测序结果一致。综上所述,本研究首次系统解析了circRNA分子在细粒棘球绦虫原头蚴、包囊壁和成虫的差异表达谱特性,筛选了在原头蚴、包囊壁和成虫各阶段高表达及特异表达的circRNA分子,预测了差异表达circRNA潜在的circRNA-miRNA-mRNA调控网络,为进一步研究circRNA分子在细粒棘球绦虫生长发育中的作用于调控机制奠定了基础。
The purpose of this study was to analyze the expression profile of circRNA in protoscolex,hydatid cyst wall and adult of Echinococcus granulosus,and to provide theoretical basis for further elucidating the role of circRNA in the growth and development of Echinococcus granulosus.Total RNA was extracted from Echinococcus granulosus protoscoleces,hydatid cyst wall and adult worms,and the circRNA,miRNA and mRNA libraries were constructed.The differentially expressed circRNA were screened by bioinformatics analysis and functional prediction,and then were analyzed by the GO,KEGG and ceRNA regulatory networks.and part of t The differentially expressed circRNA were validated by real-time fluorescent quantitative PCR(qRT-PCR).In this study,636 circRNA molecules were identified from protoscolex,cyst wall and adult of Echinococcus Granulosus,of which 140 were differentially expressed.The specific expression of circRNA was 44,2 and 0 in protoscolex,cyst wall and adult worms,respectively.The results of the GO analysis showed that compared with protoscolices and cyst walls,the differentially expressed circRNA molecules in the adult worms were mainly concentrated in the biological processes of sexual reproduction,spermatogenesis,male gametogenesis,axogenesis,neurogenesis and differentiation.The results of KEGG analysis showed that the differentially expressed circRNA was significantly enriched in Notch,tricarboxylic acid cycle,phosphatidylinositol,riboflavin metabolic pathway and fatty acid synthesis and degradation pathway.Compared with Protoscoleces,t The differentially expressed circRNA was in cyst wall and protoscolices were significantly enriched in dorsal-ventral axis formation,Notch,wnt and FoxO signaling pathway.The differentially expressed circRNA was in cyst wall and adult worms were significantly enriched in Notch,carbon metabolism,tricarboxylic acid cycle and glycolysis/gluconeogenesis pathway.Targeted prediction of circRNA-miRNA-mRNA regulatory networks showed that 164 up-down-up circRNA-miRNA-mRNA regulatory networks were formed by 24 differentially expressed circRNAs,14 miRNAs and 54 mRNAs,36 down-up-down circRNA-miRNA-mRNA regulatory networks were composed of 13 differentially expressed circRNAs,7 miRNAs and 16 mRNAs.The results of qRT-PCR for some circRNA molecules showed that the mRNA transcription level was consistent with the RNA-sequencing results.In conclusion,this study is the first to systematically analyze the differential expression profiles of circRNA in protoscolex,cyst wall and adult Echinococcus granulosus,the potential regulatory network of circRNA-miRNA-mRNA of differentially expressed circRNA was predicted,the high expression and specific expression circRNA molecules in protoscolex,cyst wall and adult worms were also screened,this study lays a foundation for further research on the role of circRNA in the growth and development of Echinococcus granulosus.
作者
王正荣
马勋
张艳艳
孙艳
孟季蒙
薄新文
WANG Zhengrong;MA Xun;ZHANG Yanyan;SUN Yan;MENG Jimeng;BO Xinwen(State Key Labaratory of Sheep Genetic Improvement and Healthy Production,Shihezi 832000,China;Institute of Animal Husbandry and Veterinary,Xinjiang Academy of Agricultural and Reclamation Sciences,Shihezi 832000,China;College of Animal Science and Technology,Shehezi University,Shihezi 832000,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2023年第8期3474-3489,共16页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金(31860701)
新疆生产建设兵团国际科技合作(2021BC008)
省部共建绵羊遗传改良与健康养殖国家重点实验室重大专项(2021ZD02)
兵团农业科技创新工程专项(NCG202213)。
