犬新孢子虫miRNAs的鉴定与分析  

Identification and Analysis of miRNAs of Neospora caninum LI Jingyu,CHEN Jinming,ZHANG Mingyi,ZHAO Shanshan,TAO Deliang,

作　　者：李京宇 陈金铭 张明一 赵姗姗 陶德良 宋军科[1] 杨新 樊莹莹 赵光辉[1] SONG Junke;YANG Xin;FAN Yingying;ZHAO Guanghui(College of Veterinary Medicine,Northwest A&F University,Yangling 712100,China)

机构地区：[1]西北农林科技大学动物医学院,杨凌712100

出　　处：《畜牧兽医学报》2024年第7期3085-3093,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(32373040)。

摘　　要：为鉴定犬新孢子虫(Neospora caninum)特异表达的miRNAs,并研究其在宿主细胞中的表达情况,本研究收集N.caninum速殖子感染的山羊子宫内膜上皮细胞(caprine endometrial epithelial cells,EECs)24和48 h的样本,用Illumina高通量测序平台和生物信息学分析犬新孢子虫特异表达的miRNAs及其体外表达情况,并研究了一个上调的miRNA novel3_mature对犬新孢子虫在山羊EECs增殖的影响。结果发现,共鉴定出3404个犬新孢子虫特异表达的miRNAs(155个新预测miRNAs),其中77个(47个上调和30个下调)miRNAs显著差异表达;随机选取7个显著差异表达的miRNAs进行实时荧光定量PCR分析,结果与高通量测序结果一致;预测分析发现61个miRNAs的3978个靶基因,功能富集分析发现这些靶基因可能主要参与MAPK信号通路、突触导向、黏着斑、cMAP信号通路、钙信号通路等;转染novel3_mature的模拟物显著促进犬新孢子虫在山羊EECs中的增殖。犬新孢子虫存在多个特异表达的miRNAs,其在虫体增殖中发挥重要作用,研究成果为深入理解犬新孢子虫胞内生存提供了新的思路。To identify specific expressed miRNAs of Neospora caninum and to investigate their expression in host cells,the samples of caprine endometrial epithelial cells(EECs)infected with tachyzoites of N.caninum at 24 and 48 h were collected.Specific expressed miRNAs of N.caninum and their in vitro expression were analyzed by using Illumina highthroughput sequencing technology and bioinformatics analysis.The effect of an up-regulated miRNA,novel3_mature,on the propagation of N.caninum in caprine EECs was also studied.The results showed that a total of 3404 specific expressed miRNAs(155 novel predicted miRNAs)of N.caninum were identified.Of them,77(47 up-and 30 down-regulated)miRNAs were significantly differentially expressed.The consistent results were found with highthroughput sequencing by using quantitative real-time PCR analysis of seven significantly differentially expressed miRNAs randomly selected.Predictive analysis found 3978 target genes of 61 miRNAs.Functional enrichment analysis showed that these target genes were possible involved in MAPK signaling pathway,axon guidance,focal adhesion,cAMP signaling pathway,calcium signaling pathway.Transfection of novel3_mature mimics significantly promoted the propagation of N.caninum in caprine EECs.Many specific expressed miRNAs were encoded by N.caninum and they played important roles in the propagation of N.caninum,providing a novel way to understand the intracellular survival of N.caninum.

关 键 词：犬新孢子虫 山羊子宫内膜上皮细胞 MIRNAS novel3_mature 增殖 

分 类 号：S852.723[农业科学—基础兽医学]