J Med Entomol. 2014 May;51(3):605-15. doi: 10.1603/me13228.
A field strain of Aedes aegypti (L.) was collected from Puerto Rico in October 2008. Based on LD50 values by topical application, the Puerto Rico strain was 73-fold resistant to permethrin compared with a susceptible Orlando strain. In the presence of piperonyl butoxide, the resistance of Puerto Rico strain of Ae. aegypti was reduced to 15-fold, suggesting that cytochrome P450-mediated detoxification is involved in the resistance of the Puerto Rico strain to permethrin. To determine the cytochrome P450s that might play a role in the resistance to permethrin, the transcriptional levels of 164 cytochrome P450 genes in the Puerto Rico strain were compared with that in the Orlando strain. Of the 164 cytochrome P450s, 33 were significantly (P < 0.05) up-regulated, including cytochrome P450s in families four, six, and nine. Multiple studies have investigated the functionality of family six and nine cytochrome P450s, therefore, we focused on the up-regulated family 4 cytochrome P450s. To determine whether up-regulation of the four cytochrome P450s had any functional role in permethrin resistance, transgenic Drosophila melanogaster Meigen lines overexpressing the four family 4 P450 genes were generated, and their ability to survive exposure to permethrin was evaluated. When exposed to 5 microg per vial permethrin, transgenic D. melanogaster expressing CYP4D24, CYP4H29, CYP4J15v1, and CYP4H33 had a survival rate of 60.0 +/- 6.7, 29.0 +/- 4.4, 64.4 +/- 9.7, and 11.0 +/- 4.4%, respectively. However, none of the control flies survived the permethrin exposure at the same concentration. Similarly, none of the transgenic D. melanogaster expressing CYP4J15v1 or CYP4H33 ?5 survived when they were exposed to permethrin at 10 microg per vial. However, transgenic D. melanogaster expressing CYP4D24 and CYP4H29 had a survival rate of 37.8 +/- 4.4 and 2.2 +/- 2.2%, respectively. Taken together, our results suggest that CYP4D24 might play an important role in cytochrome P450-mediated resistance to permethrin.
2008 年 10 月，从波多黎各采集到埃及伊蚊（L.）的田间株系。基于经皮应用的 LD50 值，与敏感的奥兰多株系相比，波多黎各株系对氯菊酯的抗性高 73 倍。在胡椒基丁醚存在的情况下，波多黎各埃及伊蚊的抗性降低至 15 倍，表明细胞色素 P450 介导的解毒作用参与了波多黎各株系对氯菊酯的抗性。为了确定可能在对氯菊酯的抗性中起作用的细胞色素 P450，比较了波多黎各株系和奥兰多株系中 164 个细胞色素 P450 基因的转录水平。在 164 个细胞色素 P450 中，33 个（P < 0.05）显著上调，包括家族四、六和九的细胞色素 P450。多项研究调查了家族六和九的细胞色素 P450 的功能，因此，我们专注于上调的家族四细胞色素 P450。为了确定四个细胞色素 P450 的上调是否对氯菊酯的抗性具有任何功能作用，生成了过表达四个家族 4 P450 基因的转基因黑腹果蝇 Meigen 系，并评估了它们对暴露于氯菊酯的存活能力。当暴露于 5μg/管氯菊酯时，表达 CYP4D24、CYP4H29、CYP4J15v1 和 CYP4H33 的转基因黑腹果蝇的存活率分别为 60.0±6.7%、29.0±4.4%、64.4±9.7%和 11.0±4.4%。然而，在相同浓度下，没有一只对照果蝇在氯菊酯暴露下存活。同样，当暴露于 10μg/管氯菊酯时，表达 CYP4J15v1 或 CYP4H33 的转基因黑腹果蝇无一存活。然而，表达 CYP4D24 和 CYP4H29 的转基因黑腹果蝇的存活率分别为 37.8±4.4%和 2.2±2.2%。总之，我们的结果表明 CYP4D24 可能在细胞色素 P450 介导的氯菊酯抗性中起重要作用。
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