Rengaraj Deivendran, Truong Anh Duc, Ban Jihye, Lillehoj Hyun S, Hong Yeong Ho
Department of Animal Science and Technology, Chung-Ang University, Anseong 17546, Korea.
Animal Biosciences and Biotechnology Laboratory, Agricultural Research Services, United States Department of Agriculture, Beltsville, MD 20705, USA.
Asian-Australas J Anim Sci. 2017 Jul;30(7):1037-1047. doi: 10.5713/ajas.16.0685. Epub 2017 Jan 13.
Despite an increasing number of investigations into the pathophysiology of necrotic enteritis (NE) disease, etiology of NE-associated diseases, and gene expression profiling of NE-affected tissues, the microRNA (miRNA) profiles of NE-affected poultry have been poorly studied. The aim of this study was to induce NE disease in the genetically disparate Fayoumi chicken lines, and to perform non-coding RNA sequencing in the intestinal mucosal layer.
NE disease was induced in the Fayoumi chicken lines (M5.1 and M15.2), and non-coding RNA sequencing was performed in the intestinal mucosal layer of both NE-affected and uninfected chickens to examine the differential expression of miRNAs. Next, quantitative real-time polymerase chain reaction (real-time qPCR) was performed to further examine four miRNAs that showed the highest fold differences. Finally, bioinformatics analyses were performed to examine the four miRNAs target genes involvement in the signaling pathways, and to examine their interaction.
According to non-coding RNA sequencing, total 50 upregulated miRNAs and 26 downregulated miRNAs were detected in the NE-induced M5.1 chickens. While 32 upregulated miRNAs and 11 downregulated miRNAs were detected in the NE-induced M15.2 chickens. Results of real-time qPCR analysis on the four miRNAs (gga-miR-9-5p, gga-miR-20b-5p, gga-miR-196-5p, and gga-let-7d) were mostly correlated with the results of RNAseq. Overall, gga-miR-20b-5p was significantly downregulated in the NE-induced M5.1 chickens and this was associated with the upregulation of its top-ranking target gene, mitogen-activated protein kinase, kinase 2. Further bioinformatics analyses revealed that 45 of the gene targets of gga-miR-20b-5p were involved in signal transduction and immune system-related pathways, and 35 of these targets were predicted to interact with each other.
Our study is a novel report of miRNA expression in Fayoumi chickens, and could be very useful in understanding the role of differentially expressed miRNAs in a NE disease model.
尽管对坏死性肠炎（NE）疾病的病理生理学、NE相关疾病的病因以及NE感染组织的基因表达谱进行了越来越多的研究，但对NE感染家禽的微小RNA（miRNA）谱的研究却很少。本研究的目的是在遗传上不同的法尤米鸡品系中诱导NE疾病，并在肠黏膜层进行非编码RNA测序。
在法尤米鸡品系（M5.1和M15.2）中诱导NE疾病，并对NE感染鸡和未感染鸡的肠黏膜层进行非编码RNA测序，以检测miRNA的差异表达。接下来，进行定量实时聚合酶链反应（实时qPCR）以进一步检测四个差异倍数最高的miRNA。最后，进行生物信息学分析，以检测这四个miRNA的靶基因参与的信号通路，并检测它们之间的相互作用。
根据非编码RNA测序，在NE诱导的M5.1鸡中检测到总共50个上调的miRNA和26个下调的miRNA。而在NE诱导的M15.2鸡中检测到32个上调的miRNA和11个下调的miRNA。对四个miRNA（gga-miR-9-5p、gga-miR-20b-5p、gga-miR-196-5p和gga-let-7d）的实时qPCR分析结果大多与RNAseq结果相关。总体而言,gga-miR-20b-5p在NE诱导的M5.1鸡中显著下调，这与其排名靠前的靶基因丝裂原活化蛋白激酶激酶2的上调有关。进一步的生物信息学分析表明，gga-miR-20b-5p的45个基因靶标参与信号转导和免疫系统相关通路，其中35个靶标预计会相互作用。
我们的研究是关于法尤米鸡中miRNA表达的新报告，对于理解差异表达的miRNA在NE疾病模型中的作用可能非常有用。
