Liao Xin, Yang Liu, Zhang Qilin, Chen Junyuan
School of Life Sciences, Nanjing University, China; Beihai Marine Station of Evo-devo Institute, Nanjing University, China.
Department of Biomedical Research Center, The First People's Hospital of Kunming, China.
Dev Comp Immunol. 2017 May;70:39-44. doi: 10.1016/j.dci.2017.01.007. Epub 2017 Jan 6.
Recently, amphioxus has served as a model for studying the origin and evolution of vertebrate immunity. However, little is known about how microRNAs (miRNAs) are involved in the immune defense in amphioxus. In this article, we identified the amphioxus miRNAs in the acute-phase response to lipopolysaccharide (LPS). We determined the time point for the peak of immune response in amphioxus after LPS challenge by evaluating the expression of Branchiostoma belcheri toll-like receptor 1, NF-κb (c-Rel), and big defensin which react with pathogen-associated molecular patterns(PAMPs). Then we chose 12 h as the point to perform miRNA microarray analysis to select the differentially expressed miRNAs. Furthermore, we used quantitative real-time PCR to detect the expression patterns of selected amphioxus miRNAs under effective LPS challenge during the time course. The microarray data revealed that the miRNA expression file was significantly changed after LPS stimulation. The changes of the 10 most upregulated and 7 most downregualted miRNAs in gills of the amphioxus following challenge with LPS revealed a temporal induction kinetic. Our current study will provide valuable information to take an insight into molecular mechanism of innate immune and the evolution of the miRNA family.
最近，文昌鱼已成为研究脊椎动物免疫起源和进化的模型。然而，关于微小RNA（miRNA）如何参与文昌鱼的免疫防御却知之甚少。在本文中，我们鉴定了文昌鱼在对脂多糖（LPS）急性期反应中的miRNA。我们通过评估与病原体相关分子模式(PAMPs)反应的文昌鱼贝氏文昌鱼Toll样受体1、NF-κB（c-Rel）和大防御素的表达，确定了LPS攻击后文昌鱼免疫反应峰值的时间点。然后我们选择12小时作为时间点进行miRNA微阵列分析，以筛选差异表达的miRNA。此外，我们使用定量实时PCR检测在有效LPS攻击期间选定的文昌鱼miRNA在时间进程中的表达模式。微阵列数据显示，LPS刺激后miRNA表达谱发生了显著变化。LPS攻击后文昌鱼鳃中10个上调最多和7个下调最多的miRNA的变化呈现出时间诱导动力学。我们目前的研究将为深入了解先天免疫的分子机制和miRNA家族的进化提供有价值的信息。
