The Wiggle Index: An Open Source Bioassay to Assess Sub-Lethal Insecticide Response in Drosophila melanogaster.
The Wiggle Index: An Open Source Bioassay to Assess Sub-Lethal Insecticide Response in Drosophila melanogaster.
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DOI：
10.1371/journal.pone.0145051
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Batterham P
中科院分区：
文献类型：
--
作者：
Denecke S;Nowell CJ;Fournier-Level A;Perry T;Batterham P
测量死亡率的毒理学测定通常用于描述杀虫剂的反应，但亚致死暴露于杀虫剂可以选择抗性，并产生额外的生物信息，描述杀虫剂影响昆虫的方式。在这里，我们提出了摆动指数（WI），高通量的方法来量化杀虫剂的反应，通过测量减少运动在亚致死暴露在醋蝇果蝇幼虫。将敏感野生型菌株暴露于杀虫剂氯虫苯甲酰胺、吡虫啉、多杀菌素和伊维菌素。每种杀虫剂降低幼虫的运动，但响应时间和配置文件不同的杀虫剂。两套目标位点突变体先前确定的死亡率研究的基础上，吡虫啉或多杀菌素抗性表型进行了测试。在每种情况下，抗性突变体的反应明显低于对照。WI还能够在不存在主要多杀菌素靶位点的情况下检测多杀菌素应答。在死亡率测定中未检测到这种反应，这表明多杀菌素与许多其他杀虫剂一样，可能具有影响行为的次要靶标。通过在消化组织或中枢神经系统中过表达吡虫啉代谢Cyp6g1基因，证实了WI检测杀虫剂代谢变化的能力。这里提出的数据验证WI作为一种廉价的，通用的，亚致死的测定，可以补充从死亡率测定获得的信息，扩大我们的理解的遗传基础的杀虫剂反应在D。黑腹菌
Toxicological assays measuring mortality are routinely used to describe insecticide response, but sub-lethal exposures to insecticides can select for resistance and yield additional biological information describing the ways in which an insecticide impacts the insect. Here we present the Wiggle Index (WI), a high-throughput method to quantify insecticide response by measuring the reduction in motility during sub-lethal exposures in larvae of the vinegar fly Drosophila melanogaster. A susceptible wild type strain was exposed to the insecticides chlorantraniliprole, imidacloprid, spinosad, and ivermectin. Each insecticide reduced larval motility, but response times and profiles differed among insecticides. Two sets of target site mutants previously identified in mortality studies on the basis of imidacloprid or spinosad resistance phenotypes were tested. In each case the resistant mutant responded significantly less than the control. The WI was also able to detect a spinosad response in the absence of the primary spinosad target site. This response was not detected in mortality assays suggesting that spinosad, like many other insecticides, may have secondary targets affecting behaviour. The ability of the WI to detect changes in insecticide metabolism was confirmed by overexpressing the imidacloprid metabolizing Cyp6g1 gene in digestive tissues or the central nervous system. The data presented here validate the WI as an inexpensive, generic, sub-lethal assay that can complement information gained from mortality assays, extending our understanding of the genetic basis of insecticide response in D. melanogaster.