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Transcriptional plasticity of different ABC transporter genes from Tribolium castaneum contributes to diflubenzuron resistance.
Insect Biochemistry and Molecular Biology ( IF 3.2 ) Pub Date : 2019-11-15 , DOI: 10.1016/j.ibmb.2019.103282 Janin Rösner 1 , Hans Merzendorfer 1
Insect Biochemistry and Molecular Biology ( IF 3.2 ) Pub Date : 2019-11-15 , DOI: 10.1016/j.ibmb.2019.103282 Janin Rösner 1 , Hans Merzendorfer 1
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The development of insecticide resistance challenges the sustainability of pest control and several studies have shown that ABC transporters contribute to this process. ABC transporters are known to transport a large range of chemically diverse molecules across cellular membranes, and therefore the identification of ABC transporters involved in insecticide resistance is difficult. Here, we describe a comprehensive strategy for the identification of whole sets of ABC transporters involved in insecticide resistance using the pest beetle, Tribolium castaneum (Tc) as a model. We analyzed the expression of ABCA to ABCC genes in different tissues and developmental stages using larvae that were sensitive or resistant to diflubenzuron (DFB). The mRNA levels of several ABC genes expressed in excretory or metabolic tissues such as midgut, Malpighian tubules or fat body were markedly upregulated in response to DFB. Next, we monitored mortality in the presence of the ABC inhibitor verapamil, and found that it causes sensitization to DFB. We furthermore established a competitive assay for the elimination of DFB, based on Texas Red (TR) fluorescence. We monitored TR elimination in larvae that were treated with DFB or different ABC inhibitors, and combinations of them. TR elimination was decreased significantly in the presence of DFB, verapamil and the ABCC inhibitor MK-571. The effect was synergized when DFB and verapamil were both present suggesting that the transport of TR and DFB involves overlapping sets of ABC transporters. Finally, we silenced the expression of DFB-responding ABC genes by RNA interference and then followed the survival rates after DFB exposure. Mortality increased particularly when specific ABCA and ABCC genes were silenced. Taken together, we were able to show that different ABC transporters expressed in metabolic and excretory tissues contribute to the elimination of DFB. Up- or down-regulation of gene expression occurs within a few days already at very low DFB concentrations. These results suggests that transcriptional plasticity of several ABC genes allows adaptation of the efflux capacity in different tissues to eliminate insecticides and/or their metabolites.
中文翻译：
Tribolium castaneum的不同ABC转运蛋白基因的转录可塑性有助于地氟苯隆抗性。
抗杀虫剂的发展对害虫控制的可持续性提出了挑战，一些研究表明，ABC转运蛋白在这一过程中发挥了作用。已知ABC转运蛋白可跨细胞膜转运大量化学多样性的分子，因此很难鉴定与杀虫剂抗性有关的ABC转运蛋白。在这里，我们描述了一种综合策略，用于使用害虫甲虫Tribolium castaneum（Tc）作为模型来鉴定参与抗药性的整套ABC转运蛋白。我们使用对地氟苯隆（DFB）敏感或耐药的幼虫，分析了ABCA在ABCC基因在不同组织和发育阶段中的表达。在排泄或代谢组织（如中肠）中表达的几种ABC基因的mRNA水平 响应DFB，马尔皮基小管或脂肪体显着上调。接下来，我们在存在ABC抑制剂维拉帕米的情况下监测死亡率，并发现其引起DFB致敏。我们还基于德克萨斯红（TR）荧光建立了消除DFB的竞争性检测方法。我们监测了用DFB或不同ABC抑制剂以及它们的组合治疗的幼虫的TR消除。在DFB，维拉帕米和ABCC抑制剂MK-571的存在下，TR消除显着降低。当同时存在DFB和维拉帕米时，效果是协同的，这表明TR和DFB的转运涉及ABC转运蛋白的重叠集合。最后，我们通过RNA干扰沉默了DFB响应的ABC基因的表达，然后追踪了DFB暴露后的存活率。当特定的ABCA和ABCC基因沉默时，死亡率增加。综上所述，我们能够证明在代谢和排泄组织中表达的不同ABC转运蛋白有助于DFB的消除。在很低的DFB浓度下，几天内就已经发生了基因表达的上调或下调。这些结果表明，几种ABC基因的转录可塑性允许适应不同组织中的外排能力，以消除杀虫剂和/或其代谢产物。
更新日期：2019-11-15
中文翻译：
Tribolium castaneum的不同ABC转运蛋白基因的转录可塑性有助于地氟苯隆抗性。
抗杀虫剂的发展对害虫控制的可持续性提出了挑战，一些研究表明，ABC转运蛋白在这一过程中发挥了作用。已知ABC转运蛋白可跨细胞膜转运大量化学多样性的分子，因此很难鉴定与杀虫剂抗性有关的ABC转运蛋白。在这里，我们描述了一种综合策略，用于使用害虫甲虫Tribolium castaneum（Tc）作为模型来鉴定参与抗药性的整套ABC转运蛋白。我们使用对地氟苯隆（DFB）敏感或耐药的幼虫，分析了ABCA在ABCC基因在不同组织和发育阶段中的表达。在排泄或代谢组织（如中肠）中表达的几种ABC基因的mRNA水平 响应DFB，马尔皮基小管或脂肪体显着上调。接下来，我们在存在ABC抑制剂维拉帕米的情况下监测死亡率，并发现其引起DFB致敏。我们还基于德克萨斯红（TR）荧光建立了消除DFB的竞争性检测方法。我们监测了用DFB或不同ABC抑制剂以及它们的组合治疗的幼虫的TR消除。在DFB，维拉帕米和ABCC抑制剂MK-571的存在下，TR消除显着降低。当同时存在DFB和维拉帕米时，效果是协同的，这表明TR和DFB的转运涉及ABC转运蛋白的重叠集合。最后，我们通过RNA干扰沉默了DFB响应的ABC基因的表达，然后追踪了DFB暴露后的存活率。当特定的ABCA和ABCC基因沉默时，死亡率增加。综上所述，我们能够证明在代谢和排泄组织中表达的不同ABC转运蛋白有助于DFB的消除。在很低的DFB浓度下，几天内就已经发生了基因表达的上调或下调。这些结果表明，几种ABC基因的转录可塑性允许适应不同组织中的外排能力，以消除杀虫剂和/或其代谢产物。
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