Zhang Yu, Dong Xiaoqian, Hou Lie, Cao Zhengfeng, Zhu Guoqiang, Vongsangnak Wanwipa, Xu Qi, Chen Guohong
Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education, Yangzhou University, Yangzhou, China.
Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, China.
Front Vet Sci. 2021 Jun 16;8:692501. doi: 10.3389/fvets.2021.692501. eCollection 2021.
serovar Enteritidis (. Enteritidis) is a pathogen that can colonize the preovulatory follicles of poultry, thereby causing both reduced egg production and an elevated risk of foodborne salmonellosis in humans. Although a few studies have revealed . Enteritidis preferentially invades the granulosa cell layer within these follicles, it can readily persist and proliferate through mechanisms that are not well-understood. In this study, we characterized competing endogenous RNA (ceRNA) regulatory networks within duck granulosa cells following time-course of . Enteritidis challenge. The 8108 long non-coding RNAs (lncRNAs), 1545 circular RNAs (circRNAs), 542 microRNAs (miRNAs), and 4137 mRNAs (fold change ≥2; < 0.01) were differentially expressed during S. Enteritidis challenge. Also, eight mRNAs, eight lncRNAs and five circRNAs were selected and the consistent expression trend was found between qRT-PCR detection and RNA-seq. Moreover, the target genes of these differentially expressed ncRNAs (including lncRNAs, circRNAs and miRNAs) were predicted, and significantly enriched in the innate immune response and steroidogenesis pathways. Then, the colocalization and coexpression analyses were conducted to investigate relationships between ncRNAs and mRNAs. The 16 differentially expressed miRNAs targeting 60 differentially expressed mRNAs were identified in granulosa cells at 3 and 6 h post-infection (hpi) and enriched in the MAPK, GnRH, cytokine-cytokine receptor interaction, Toll-like receptor, endocytosis, and oxidative phosphorylation signaling pathways. Additionally, underlying lncRNA-miRNA-mRNA and circRNA-miRNA-mRNA ceRNA networks were then constructed to further understand their interaction during . Enteritidis infection. Lnc_012227 and novel_circ_0004892 were identified as ceRNAs, which could compete with miR-let-7g-5p and thereby indirectly modulating expression to control . Enteritidis infection. Together, our data thus identified promising candidate ncRNAs responsible for regulating . Enteritidis infection in the preovulatory follicles of ducks, offering new insights regarding the ovarian transmission of this pathogen.
肠炎血清型沙门氏菌（Salmonella Enteritidis，S. Enteritidis）是一种能够定殖于家禽排卵前卵泡的病原体，从而导致产蛋量下降以及人类食源性沙门氏菌病风险升高。尽管一些研究表明，肠炎血清型沙门氏菌优先侵入这些卵泡内的颗粒细胞层，但它能够通过一些尚未完全了解的机制轻易地持续存在并增殖。在本研究中，我们对肠炎血清型沙门氏菌攻击后的鸭颗粒细胞内竞争性内源RNA（ceRNA）调控网络进行了表征。在肠炎血清型沙门氏菌攻击期间，8108条长链非编码RNA（lncRNA）、1545条环状RNA（circRNA）、542条微小RNA（miRNA）和4137条信使RNA（mRNA）（倍数变化≥2；P<0.01）差异表达。此外，选择了8条mRNA、8条lncRNA和5条circRNA，qRT-PCR检测与RNA测序之间发现了一致的表达趋势。此外，预测了这些差异表达的非编码RNA（包括lncRNA、circRNA和miRNA）的靶基因，并在先天免疫应答和类固醇生成途径中显著富集。然后，进行共定位和共表达分析以研究非编码RNA与mRNA之间的关系。在感染后3小时和6小时（hpi）的颗粒细胞中鉴定出16条差异表达的miRNA靶向60条差异表达的mRNA，并在丝裂原活化蛋白激酶（MAPK）、促性腺激素释放激素（GnRH）、细胞因子-细胞因子受体相互作用、Toll样受体、内吞作用和氧化磷酸化信号通路中富集。此外，构建了潜在的lncRNA-miRNA-mRNA和circRNA-miRNA-mRNA ceRNA网络，以进一步了解它们在肠炎血清型沙门氏菌感染期间的相互作用。Lnc_012227和novel_circ_0004892被鉴定为ceRNA，它们可以与miR-let-7g-5p竞争，从而间接调节表达以控制肠炎血清型沙门氏菌感染。总之，我们的数据确定了有望调控鸭排卵前卵泡中肠炎血清型沙门氏菌感染的候选非编码RNA，为该病原体的卵巢传播提供了新的见解。
