Lv Yuan, Li Xixi, Zhang Hongbo, Zou Feifei, Shen Bo
Department of Nuclear Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjing, China.
Department of Pathogen Biology, Nanjing University of Chinese Medicine, Nanjing, China.
Comp Biochem Physiol B Biochem Mol Biol. 2022 Aug-Sep;261:110750. doi: 10.1016/j.cbpb.2022.110750. Epub 2022 May 2.
The excessive and improper application of insecticides has caused the evolution of resistance in many mosquito populations, including Culex pipiens pallens (L.). Deltamethrin, a representative pyrethroid insecticide, is the most widely used synthetic insecticide in mosquito-borne control field. Comprehensively identifying genes and regulators associated with deltamethrin resistance and elucidating the manner in which they regulate this process is critical for effective control of mosquitoes. CircRNAs are the upstream regulatory factors of miRNAs and mRNAs, which play a role via the competitive endogenous RNA mechanism. In this study, we used high-throughput circRNA sequencing to identify circRNAs that were expressed differently in deltamethrin-susceptible strain (DS strain) and -resistant strain (DR strain) mosquitoes [NCBI Sequence Read Archive (SRA) database accession number: PRJNA714543]. We detected a total of 12,816 significantly differentially expressed circRNAs (DE-circRNAs). Among them, 6769 circRNAs were up-regulated and 6047 circRNAs were down-regulated in the DR strain compared to DS strain. Among the DE-circRNAs, we further screened that supercont3.352:252102|253283 was significantly over-expressed in the DR strain through qPCR multiple verification (P < 0.05).We used the divergent primer to amplify the rolling circle product and obtained the full-length sequence of supercont3.352:252102|253283 (GeneBank accession number: MW729338). Through software comparison and bioinformatics analysis, we predicted that supercont3.352:252102|253283 might participate in deltamethrin resistance by sponging cpp-miR-1671 and blocking its inhibition on CYP4G15.We further found that the expression of cpp-miR-1671 was significantly lower in DR strain (P < 0.01), while the expression of CYP4G15 was significantly higher in DR strain (P < 0.05).Taken together, the present study provided the most comprehensive circRNA expression profile of mosquitoes, and suggested that supercont3.352:252102|253283 might participate in deltamethrin resistance through the supercont3.352:252102|253283/cpp-miR-1671/CYP4G15 pathway.
杀虫剂的过度和不当使用已导致许多蚊虫种群产生抗药性，包括淡色库蚊（Culex pipiens pallens (L.)）。溴氰菊酯作为一种代表性的拟除虫菊酯类杀虫剂，是蚊媒控制领域使用最广泛的合成杀虫剂。全面鉴定与溴氰菊酯抗性相关的基因和调控因子，并阐明它们调控这一过程的方式，对于有效控制蚊虫至关重要。环状RNA是微小RNA和信使RNA的上游调控因子，通过竞争性内源RNA机制发挥作用。在本研究中，我们使用高通量环状RNA测序来鉴定在溴氰菊酯敏感品系（DS品系）和抗性品系（DR品系）蚊虫中表达差异的环状RNA[美国国立生物技术信息中心序列读取存档（SRA）数据库登录号：PRJNA714543]。我们共检测到12816个显著差异表达的环状RNA（DE-circRNAs）。其中，与DS品系相比，DR品系中有6769个环状RNA上调，6047个环状RNA下调。在这些DE-circRNAs中，我们通过qPCR多次验证进一步筛选出supercont3.352:252102|253283在DR品系中显著过表达（P < 0.05）。我们使用发散引物扩增滚环产物，获得了supercont3.352:252102|253283的全长序列（基因银行登录号：MW729338）。通过软件比对和生物信息学分析，我们预测supercont3.352:252102|253283可能通过吸附cpp-miR-1671并阻断其对CYP4G15的抑制作用来参与溴氰菊酯抗性。我们进一步发现，cpp-miR-1671在DR品系中的表达显著降低（P < 0.01），而CYP4G15在DR品系中的表达显著升高（P < 0.05）。综上所述，本研究提供了最全面的蚊虫环状RNA表达谱，并表明supercont3.352:252102|253283可能通过supercont3.352:252102|253283/cpp-miR-1671/CYP4G15途径参与溴氰菊酯抗性。
