School of Chemistry, ARC Centre of Excellence for Free Radical Chemistry and Biotechnology, and Bio21 Molecular Science and Biotechnology Institute, University of Melbourne , Melbourne, Victoria 3010, Australia.
Anal Chem. 2014 Apr 1;86(7):3525-32. doi: 10.1021/ac404188g. Epub 2014 Mar 11.
Insecticide resistance is one of the most prevalent examples of anthropogenic genetic change, yet our understanding of metabolic-based resistance remains limited by the analytical challenges associated with rapidly tracking the in vivo metabolites of insecticides at nonlethal doses. Here, using twin ion mass spectrometry analysis of the extracts of whole Drosophila larvae and excreta, we show that (i) eight metabolites of the neonicotinoid insecticide, imidacloprid, can be detected when formed by susceptible larval genotypes and (ii) the specific overtranscription of a single gene product, Cyp6g1, associated with the metabolic resistance to neonicotinoids, results in a significant increase in the formation of three imidacloprid metabolites that are formed in C-H bond activation reactions; that is, Cyp6g1 is directly linked to the enhanced metabolism of imidacloprid in vivo. These results establish a rapid and sensitive method for dissecting the metabolic machinery of insects by directly linking single gene products to insecticide metabolism.
杀虫剂抗性是最常见的人为遗传改变的例子之一，但我们对基于代谢的抗性的理解仍然受到与非致死剂量下快速跟踪杀虫剂体内代谢物相关的分析挑战的限制。在这里，我们使用双离子质谱分析整个果蝇幼虫和排泄物的提取物，表明：（i）当易感幼虫基因型形成时，可以检测到新烟碱类杀虫剂吡虫啉的八种代谢物；（ii）与新烟碱类杀虫剂代谢抗性相关的单个基因产物 Cyp6g1 的特异性过度转录，导致与 C-H 键活化反应形成的三种吡虫啉代谢物的形成显著增加；也就是说，Cyp6g1 与体内吡虫啉代谢的增强直接相关。这些结果通过将单个基因产物直接与杀虫剂代谢联系起来，建立了一种快速灵敏的方法来剖析昆虫的代谢机制。
