Wang Fangfang, Gong Haiyan, Zhang Houshuang, Zhou Yongzhi, Cao Jie, Zhou Jinlin
Key Laboratory of Animal Parasitology of Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China.
Key Laboratory of Animal Parasitology of Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, China.
PLoS One. 2015 Oct 2;10(10):e0139241. doi: 10.1371/journal.pone.0139241. eCollection 2015.
Lipopolysaccharide (LPS) stimulates the innate immune response in arthropods. In tick vectors, LPS activates expression of immune genes, including those for antibacterial peptides. miRNAs are 21-24 nt non-coding small RNAs that regulate target mRNAs at the post-transcriptional level. However, our understanding of tick innate immunity is limited to a few cellular immune reactions and some characterized immune molecules. Moreover, there is little information on the regulation of the immune system in ticks by miRNA. Therefore, this study aimed to analyze the differential expression of miRNAs in male and female ticks after LPS injection. LPS was injected into male and female Rhipicephalus haemaphysaloides ticks to stimulate immune response, with phosphate buffered saline (PBS)-injected ticks as negative controls. miRNAs from each group were sequenced and analyzed. In the PBS- and LPS-injected female ticks, 11.46 and 12.82 million reads of 18-30 nt were obtained respectively. There were 13.92 and 15.29 million reads of 18-30 nt obtained in the PBS- and LPS-injected male ticks, respectively. Expression of miRNAs in male ticks was greater than that in female ticks. There were 955 and 984 conserved miRNA families in the PBS- and LPS-injected female ticks, respectively, and correspondingly 1684 and 1552 conserved miRNA families in male ticks. Nine novel miRNAs were detected as common miRNAs in two or more tested samples. There were 37 known miRNAs up-regulated >10-fold and 33 down-regulated >10-fold in LPS-injected female ticks; and correspondingly 52 and 59 miRNAs in male ticks. Differential expression of miRNAs in PBS- and LPS-injected samples supports their involvement in the regulation of innate immunity. These data provide an important resource for more detailed functional analysis of miRNAs in this species.
脂多糖（LPS）刺激节肢动物的先天免疫反应。在蜱虫媒介中，LPS激活免疫基因的表达，包括抗菌肽的基因。微小RNA（miRNA）是21 - 24个核苷酸的非编码小RNA，在转录后水平调控靶mRNA。然而，我们对蜱虫先天免疫的理解仅限于少数细胞免疫反应和一些已表征的免疫分子。此外，关于miRNA对蜱虫免疫系统调节的信息很少。因此，本研究旨在分析LPS注射后雄性和雌性蜱虫中miRNA的差异表达。将LPS注射到血红扇头蜱的雄性和雌性蜱虫中以刺激免疫反应，以注射磷酸盐缓冲盐水（PBS）的蜱虫作为阴性对照。对每组的miRNA进行测序和分析。在注射PBS和LPS的雌性蜱虫中，分别获得了1146万和1282万个18 - 30个核苷酸的读数。在注射PBS和LPS的雄性蜱虫中，分别获得了13...92万和1529万个18 - 30个核苷酸的读数。雄性蜱虫中miRNA的表达高于雌性蜱虫。在注射PBS和LPS的雌性蜱虫中，分别有955个和984个保守miRNA家族，在雄性蜱虫中相应地有1684个和1552个保守miRNA家族。在两个或更多测试样本中检测到9个新的miRNA作为常见miRNA。在注射LPS的雌性蜱虫中有37个已知miRNA上调超过10倍，33个下调超过10倍；在雄性蜱虫中相应地有52个和59个miRNA。注射PBS和LPS样本中miRNA的差异表达支持它们参与先天免疫的调节。这些数据为该物种中miRNA更详细的功能分析提供了重要资源。 （注：原文中“13.92”处表述不完整，可能影响理解，这里按原文呈现翻译）
