Ren Yipeng, Chen Juhong, Wang Yuan, Fu Siying, Bu Wenjun, Xue Huaijun
Institute of Entomology, College of Life Sciences, Nankai University, Tianjin, China.
Front Physiol. 2023 Aug 17;14:1244190. doi: 10.3389/fphys.2023.1244190. eCollection 2023.
Long noncoding RNAs (lncRNAs) play significant roles in the regulation of mRNA expression or in shaping the competing endogenous RNA (ceRNA) network by targeting miRNA. The insect gut is one of the most important tissues due to direct contact with external pathogens and functions in the immune defense against pathogen infection through the innate immune system and symbionts, but there are limited observations on the role of the lncRNA-involved ceRNA network of the Toll/Imd pathway and correlation analysis between this network and bacterial microbiota in the gut. In this research, we constructed and sequenced six RNA sequencing libraries using normal and antibiotic-reared samples, generating a total of 17,193 lncRNAs and 26,361 mRNAs from massive clean data by quality control and bioinformatic analysis. Furthermore, a set of 8,539 differentially expressed lncRNAs (DELs) and 13,263 differentially expressed mRNAs (DEMs), of which related to various immune signaling pathways, such as the Toll/Imd, JAK/STAT, NF-κB, and PI3K-Akt signaling pathways, were obtained between the two experimental groups in . In addition, numerous GO and KEGG enrichment analyses were used to annotate the DELs and their target genes. Moreover, six family members and nineteen signal genes from the Toll/Imd signaling pathway were identified and characterized using online tools, and phylogenetic analyses of the above genes proved their classification. Next, a lncRNA-miRNA-mRNA network of the Toll/Imd pathway was built, and it contained different numbers of DEMs in this pathway and related DELs based on prediction and annotation. In addition, qRT-PCR validation and sequencing data were conducted to show the expression patterns of the above DELs and DEMs related to the Toll/Imd signaling pathway. Finally, the correlated investigations between DELs or DEMs of the Toll/Imd signaling pathway and most changes in the gut bacterial microbiota revealed significantly positive or negative relationships between them. The present findings provide essential evidence for innate immune ceRNAs in the beetle gut and uncover new potential relationships between innate immune pathways and the gut bacterial microbiota in insects.
长链非编码RNA（lncRNAs）在调节mRNA表达或通过靶向miRNA构建竞争性内源RNA（ceRNA）网络中发挥着重要作用。昆虫肠道是最重要的组织之一，因为它直接接触外部病原体，并通过先天免疫系统和共生体在抵御病原体感染的免疫防御中发挥作用，但关于Toll/Imd途径中lncRNA参与的ceRNA网络的作用以及该网络与肠道细菌微生物群之间的相关性分析的观察结果有限。在本研究中，我们使用正常和抗生素饲养的样本构建并测序了六个RNA测序文库，通过质量控制和生物信息学分析从大量清洁数据中总共获得了17,193个lncRNAs和26,361个mRNAs。此外，在两个实验组之间获得了一组8,539个差异表达的lncRNAs（DELs）和13,263个差异表达的mRNAs（DEMs），其中它们与各种免疫信号通路相关，如Toll/Imd、JAK/STAT、NF-κB和PI3K-Akt信号通路。此外，使用了大量的GO和KEGG富集分析来注释DELs及其靶基因。此外，使用在线工具鉴定并表征了来自Toll/Imd信号通路的六个家族成员和十九个信号基因，并且对上述基因的系统发育分析证明了它们的分类。接下来，构建了Toll/Imd途径的lncRNA-miRNA-mRNA网络，并且基于预测和注释，该网络在该途径中包含不同数量的DEMs和相关的DELs。此外，进行了qRT-PCR验证和测序数据以显示上述与Toll/Imd信号通路相关的DELs和DEMs的表达模式。最后，Toll/Imd信号通路的DELs或DEMs与肠道细菌微生物群的大多数变化之间的相关性研究揭示了它们之间显著的正相关或负相关关系。本研究结果为甲虫肠道中的先天免疫ceRNAs提供了重要证据，并揭示了昆虫先天免疫途径与肠道细菌微生物群之间的新潜在关系。
