Xu Yang, Du Jiajia, Zhang Kewei, Li Jinze, Zou Feifei, Li Xixi, Meng Yufen, Chen Ying, Tao Li, Zhao Fengming, Ma Lei, Shen Bo, Zhou Dan, Sun Yan, Yan Guiyun, Zhu Changliang
Department of Pathogen Biology, Nanjing Medical University, Nanjing 211100, China.
School of Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China.
J Agric Food Chem. 2024 Dec 11;72(49):27150-27162. doi: 10.1021/acs.jafc.4c05708. Epub 2024 Nov 27.
Mosquitoes within the complex play a crucial role in human disease transmission. Insecticides, especially pyrethroids, are used to control these vectors. Mosquito legs are the main entry point and barrier for insecticides to gain their neuronal targets. However, the resistance mechanism in mosquito legs is unclear. Herein, we employed transcriptomic analyses and isobaric tags for relative and absolute quantitation techniques to investigate the resistance mechanism, focusing on legs. We discovered 2346 differentially expressed genes (DEGs) between deltamethrin-resistant (DR) and deltamethrin-sensitive (DS) mosquito legs, including 41 cytochrome P450 genes. In the same comparison, we identified 228 differentially expressed proteins (DEPs), including six cytochrome P450 proteins. Combined transcriptome and proteome analysis revealed only two upregulated P450 genes, and . The main clusters of DEGs and DEPs were associated with metabolic processes, such as cytochrome P450-mediated metabolism of drugs and xenobiotics. Transcription analysis revealed high and expression in the DR strain at 72 h posteclosion compared with that in the DS strain, particularly in the legs. Mosquitoes knocked down for were more sensitive to deltamethrin than the controls. knockdown reduced the expression of several chlorinated hydrocarbon (CHC)-related genes, which altered the cuticle thickness and structure. Conversely, knockdown increased CHC gene expression without altering cuticle thickness and structure. P450 activity analysis demonstrated that CYP325G4 and CYP6AA9 contributed to metabolic resistance in the midgut and legs. This study identified CYP325G4 as a novel mosquito deltamethrin resistance factor, being involved in both metabolic and cuticular resistance mechanisms. The previously identified CYP6AA9 was investigated for its involvement in metabolic resistance and potential cuticular resistance in mosquito legs. These findings enhance our comprehension of resistance mechanisms, identifying P450s as promising targets for the future management of mosquito vector resistance, and laying a theoretical groundwork for mosquito resistance management.
该复合体中的蚊子在人类疾病传播中起着至关重要的作用。杀虫剂，尤其是拟除虫菊酯，被用于控制这些病媒。蚊子的腿部是杀虫剂到达其神经元靶点的主要切入点和屏障。然而，蚊子腿部的抗性机制尚不清楚。在此，我们采用转录组分析和相对与绝对定量等压标签技术来研究抗性机制，重点关注腿部。我们发现溴氰菊酯抗性（DR）和溴氰菊酯敏感（DS）蚊子腿部之间有2346个差异表达基因（DEG），其中包括41个细胞色素P450基因。在同一比较中，我们鉴定出228个差异表达蛋白（DEP），其中包括6个细胞色素P450蛋白。转录组和蛋白质组联合分析仅揭示了两个上调的P450基因，即 和 。差异表达基因和差异表达蛋白的主要聚类与代谢过程相关，如细胞色素P450介导的药物和异源生物代谢。转录分析显示，与DS品系相比，DR品系在羽化后72小时时 和 的表达较高，尤其是在腿部。敲低 的蚊子对溴氰菊酯比对照更敏感。敲低 会降低几个与氯代烃（CHC）相关基因的表达，从而改变表皮厚度和结构。相反，敲低 会增加CHC基因表达，而不会改变表皮厚度和结构。P450活性分析表明，CYP325G4和CYP6AA9在中肠和腿部的代谢抗性中起作用。本研究确定CYP325G4是一种新的蚊子溴氰菊酯抗性因子，参与代谢和表皮抗性机制。对先前鉴定的CYP6AA9在蚊子腿部的代谢抗性和潜在表皮抗性中的作用进行了研究。这些发现增强了我们对抗性机制的理解，确定细胞色素P450是未来蚊子病媒抗性管理的有希望的靶点，并为蚊子抗性管理奠定了理论基础。
