Sun Hongjuan, Zhou Zunchun, Dong Ying, Yang Aifu, Jiang Jingwei, Chen Zhong, Guan Xiaoyan, Wang Bai, Gao Shan, Jiang Bei
Liaoning Key Lab of Marine Fishery Molecular Biology, Liaoning Ocean and Fisheries Science Research Institute, Dalian, Liaoning 116023, PR China.
Liaoning Key Lab of Marine Fishery Molecular Biology, Liaoning Ocean and Fisheries Science Research Institute, Dalian, Liaoning 116023, PR China.
Fish Shellfish Immunol. 2016 Feb;49:205-12. doi: 10.1016/j.fsi.2015.12.036. Epub 2015 Dec 24.
MicroRNAs (miRNAs) are small non-coding RNAs that are involved in many biological processes. To investigate the miRNAs related to skin ulceration syndrome (SUS) of Apostichopus japonicus, small RNA libraries of body wall, intestine, respiratory tree and coelomocytes from healthy and diseased A. japonicus were sequenced on Illumina Hiseq 2000 platform. A total of 247 conserved and 10 novel miRNAs were identified across all libraries. After pair-wise comparisons, 215 miRNAs in body wall, 36 in intestine, 2 in respiratory tree and 38 in coelomocytes showed significant expression differences. Further analyses were conducted on some tissue-specific differentially expressed miRNAs: miR-8 and miR-486-5p in body wall, miR-200-3p, let-7-5p and miR-125 in intestine, miR-278a-3p and bantam in respiratory, miR-10a and miR-184 in coelomocytes. Notably, these miRNAs in some species were reported to function in various physiological or pathological processes associated with immune regulations. Using stem-loop quantitative real time PCR, six representative miRNAs in four tissues were selected to validate the sequencing results. The Pearson's correlation coefficient (R) of the six miRNAs ranged from 0.777 to 0.948, which confirmed the consistency and accuracy between these two approaches. This study provides comprehensive expression and regulation patterns of functional miRNAs in different tissues and gives insights into the tissue-specific immune response mechanisms in SUS-infected A. japonicus.
微小RNA（miRNA）是参与许多生物学过程的小非编码RNA。为了研究与刺参皮肤溃疡综合征（SUS）相关的miRNA，在Illumina Hiseq 2000平台上对健康和患病刺参的体壁、肠道、呼吸树和体腔细胞的小RNA文库进行了测序。在所有文库中总共鉴定出247个保守miRNA和10个新的miRNA。经过两两比较，体壁中有215个miRNA、肠道中有36个、呼吸树中有2个、体腔细胞中有38个miRNA表现出显著的表达差异。对一些组织特异性差异表达的miRNA进行了进一步分析：体壁中的miR-8和miR-486-5p、肠道中的miR-200-3p、let-7-5p和miR-125、呼吸中的miR-278a-3p和bantam、体腔细胞中的miR-10a和miR-184。值得注意的是，据报道这些miRNA在某些物种中在与免疫调节相关的各种生理或病理过程中发挥作用。使用茎环定量实时PCR，选择四个组织中的六个代表性miRNA来验证测序结果。这六个miRNA的皮尔逊相关系数（R）在0.777至0.948之间，证实了这两种方法之间的一致性和准确性。本研究提供了不同组织中功能性miRNA的全面表达和调控模式，并深入了解了SUS感染的刺参中的组织特异性免疫反应机制。
