Regulation of P450-mediated permethrin resistance in Culex quinquefasciatus by the GPCR/Gαs/AC/cAMP/PKA signaling cascade.
Regulation of P450-mediated permethrin resistance in Culex quinquefasciatus by the GPCR/Gαs/AC/cAMP/PKA signaling cascade.
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DOI：
10.1016/j.bbrep.2017.08.010
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发表时间：
2017-12
影响因子：
2.7
通讯作者：
Liu N
中科院分区：
文献类型：
--
作者：
Li T;Liu N
本研究探讨了G蛋白偶联受体胞内信号在致倦库蚊P450介导的抗药性形成中的作用，重点探讨了G蛋白偶联受体及其下游效应蛋白GαS和腺苷环化酶在P450介导的致倦库蚊抗药性中的重要作用。我们的RNA干扰功能研究表明，GαS基因的敲除导致G-α信号通路下游效应因子ACs和PKA的表达减少以及抗性P450基因的表达，而ACs的敲除则导致PKAs和抗性P450基因的表达减少。敲除G-α、S或ACs均可增加蚊虫对二氯苯菊酯的敏感性。这些结果有助于我们进一步了解蚊虫抗药性中通过gpr/Gα、S/AC/cAMP-PKA信号通路调控抗性P450基因的分子基础。对两个抗药性品系的GPCRs、GαS、ACEs、PKA和P450 s的时空动态分析表明，GPCRs、GαS、Acs和PKA在两个抗药性品系的脑组织中表达最高，表明这些基因在信号转导和调控中发挥了作用。抗性P450基因主要在脑、中肠和马氏管(MTS)中表达，提示它们在中枢神经系统中起关键作用，对解毒具有重要意义。对该基因表达的时间动力学分析表明，该基因在蚊子发育过程中具有不同的表达谱，这表明它们在蚊子生命阶段对杀虫剂暴露的最初功能重要性。致倦库蚊抗药性P450基因的表达受G-α、Gs-α亚单位蛋白(G-GPCr)、腺苷环化酶(Acs)和蛋白激酶A(PKA)的调节。GPCR4、Gα、S、ACs和PKA在蚊子的脑中高度表达，与它们在信号转导和调节中的作用相对应。GPCR4、Gα、S、ACEs、PKA和P450在蚊虫不同的生活史中均有表达，揭示了它们在蚊虫生活史中对杀虫剂暴露的重要作用。
This study explores the role of G-protein-coupled receptor-intracellular signaling in the development of P450-mediated insecticide resistance in mosquitoes, Culex quinquefasciatus, focusing on the essential function of the GPCRs and their downstream effectors of Gs alpha subunit protein (Gαs) and adenylyl cyclase (ACs) in P450-mediated insecticide resistance of Culex mosquitoes. Our RNAi-mediated functional study showed that knockdown of Gαs caused the decreased expression of the downstream effectors of ACs and PKAs in the GPCR signaling pathway and resistance P450 genes, whereas knockdown of ACs decreased the expression of PKAs and resistance P450 genes. Knockdown of either Gαs or ACs resulted in an increased susceptibility of mosquitoes to permethrin. These results add significantly to our understanding of the molecular basis of resistance P450 gene regulation through GPCR/Gαs/AC/cAMP-PKA signaling pathways in the insecticide resistance of mosquitoes. The temporal and spatial dynamic analyses of GPCRs, Gαs, ACs, PKAs, and P450s in two insecticide resistant mosquito strains revealed that all the GPCR signaling pathway components tested, namely GPCRs, Gαs, ACs and PKAs, were most highly expressed in the brain for both resistant strains, suggesting the role played by these genes in signaling transduction and regulation. The resistance P450 genes were mainly expressed in the brain, midgut and malpighian tubules (MTs), suggesting their critical function in the central nervous system and importance for detoxification. The temporal dynamics analysis for the gene expression showed a diverse expression profile during mosquito development, indicating their initially functional importance in response to exposure to insecticides during their life stages. GPCR, Gs alpha subunit protein (Gαs), adenylyl cyclase (ACs), and protein kinase A (PKAs) regulates resistance P450 gene expression and the development of insecticide resistance in mosquitoes, Culex quinquefasciatus. GPCR, Gαs, ACs, and PKAs, are highly expressed in the brain of mosquitoes, corresponding their role in signaling transduction, and regulation. GPCR, Gαs, ACs, PKAs, and P450s are expressed in the different life stages of mosquitoes, revealing their functional importance in response to exposure to insecticides during mosquito life stages.