Feng Xinyu, Wu Jiatong, Zhou Shuisen, Wang Jingwen, Hu Wei
National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Key Laboratory of Parasite and Vector Biology, National Health and Family Planning Commission, WHO Collaborating Center for Tropical Diseases, National Center for International Research On Tropical Diseases, Shanghai, 200025 People's Republic of China.
2Joint Research Laboratory of Genetics and Ecology on Parasites-hosts Interaction, National Institute of Parasitic Diseases-Fudan University, Shanghai, 200025 People's Republic of China.
Cell Biosci. 2018 Apr 12;8:29. doi: 10.1186/s13578-018-0227-1. eCollection 2018.
microRNAs (miRNAs) are one kind of small non-coding RNAs widely distributed in insects. Many studies have shown that miRNAs play critical roles in development, differentiation, apoptosis, and innate immunity. However, there are a few reports describing miRNAs in , the most common, and one of the dominant malaria mosquito in China. Here, we investigated the global miRNA expression profile across four different developmental stages including embryo, larval, pupal, and adult stages using Illumina Hiseq 2500 sequencing.
In total, 164 miRNAs were obtained out of 107.46 million raw sequencing reads. 99 of them identified as known miRNAs, and the remaining 65 miRNAs were considered as novel. By analyzing the read counts of miRNAs in all developmental stages, 95 miRNAs showed stage-specific expression (q < 0.01 and |log2 (fold change)| > 1) in consecutive stages, indicating that these miRNAs may be involved in critical physiological activity during development. Sixteen miRNAs were identified to be commonly dysregulated throughout four developmental stages. Many miRNAs showed stage-specific expression, such as asi-miR-2943 was exclusively expressed in the embryo stage, and asi-miR-1891 could not be detected in larval stage. The expression of six selected differentially expressed miRNAs identified by qRT-PCR were consistent with our sequencing results. Furthermore, 5296 and 1902 target genes were identified for the dysregulated 68 known and 27 novel miRNAs respectively by combining miRanda and RNAhybrid prediction. GO annotation and KEGG pathway analysis for the predicted genes of dysregulated miRNAs revealed that they might be involved in a broad range of biological processes related with the development, such as membrane, organic substance transport and several key pathways including protein processing in endoplasmic reticulum, propanoate metabolism and folate biosynthesis. Thirty-two key miRNAs were identified by microRNA-gene network analysis.
The present study represents the first global characterization of miRNAs in its four developmental stages. The presence and differential expression of miRNAs imply that such miRNAs may play critical roles in life cycle. A better understanding of the functions of these miRNAs will have great implication for the effective control of vector population and therefore interrupting malaria transmission.
微小RNA（miRNA）是一类广泛分布于昆虫中的小型非编码RNA。许多研究表明，miRNA在发育、分化、凋亡和先天免疫中发挥着关键作用。然而，关于中国最常见且占主导地位的疟疾传播媒介之一——中华按蚊（Anopheles sinensis）的miRNA报道较少。在此，我们使用Illumina Hiseq 2500测序技术研究了中华按蚊胚胎、幼虫、蛹和成虫四个不同发育阶段的全局miRNA表达谱。
在1.0746亿条原始测序读数中，共获得164个miRNA。其中99个被鉴定为已知miRNA，其余65个miRNA被视为新发现的。通过分析所有发育阶段中miRNA的读数计数，95个miRNA在连续阶段表现出阶段特异性表达（q < 0.01且|log2（倍数变化）| > 1），表明这些miRNA可能参与发育过程中的关键生理活动。16个miRNA在四个发育阶段均表现出普遍失调。许多miRNA呈现阶段特异性表达，例如asi-miR-2943仅在胚胎阶段表达，而asi-miR-1891在幼虫阶段未被检测到。通过qRT-PCR鉴定的六个差异表达miRNA的表达与我们的测序结果一致。此外，通过结合miRanda和RNAhybrid预测，分别为68个失调的已知miRNA和27个新发现的miRNA鉴定了5296个和1902个靶基因。对失调miRNA的预测基因进行GO注释和KEGG通路分析表明，它们可能参与与发育相关的广泛生物学过程，如膜、有机物质运输以及包括内质网中的蛋白质加工、丙酸代谢和叶酸生物合成在内的几个关键通路。通过miRNA-基因网络分析鉴定了32个关键miRNA。
本研究首次对中华按蚊四个发育阶段的miRNA进行了全局表征。中华按蚊miRNA的存在和差异表达意味着此类miRNA可能在其生命周期中发挥关键作用。更好地理解这些miRNA的功能对于有效控制媒介种群从而阻断疟疾传播具有重要意义。
