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Ocular Albinism Type 1 Regulates Deltamethrin Tolerance in Lymantria dispar and Drosophila melanogaster

机译:眼白化病1型可调节Disman Lymantria dispar和Drosophila melanogaster中的溴氰菊酯耐受性。

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摘要

The ocular albinism type 1 (OA1), a pigment cell-specific integral membrane glycoprotein, is a member of the G-protein-coupled receptor (GPCR) superfamily that binds to heterotrimeric G proteins in mammalian cells. We aimed to characterize the physiological functions an insect OA1 from Lymantria dispar (LdOA1) employs in the regulation of insecticide tolerance. In the present study, we investigated the roles of LdOA1 in response to deltamethrin exposure in both L. dispar and Drosophila melanogaster. LdOA1 was expressed at the lowest level during the 4th instar stage, while LdOA1 was significantly upregulated in the 5th instar and male stages. Knockdown of LdOA1 by injecting dsRNA of LdOA1 into gypsy moth larvae caused a 4.80-fold higher mortality than in control larvae microinjected with dsRNA of GFP under deltamethrin stress. Nine out of 11 L. dispar CYP genes were significantly downregulated under deltamethrin stress in LdOA1 silenced larvae as compared to control larvae. Moreover, the LdOA1 gene was successfully overexpressed in D. melanogaster using transgenic technique. The deltamethrin contact assay showed that the LdOA1 overexpression in flies significantly enhanced the tolerance to deltamethrin compared to the control flies. Furthermore, the downstream Drosophila CYP genes were upregulated in the LdOA1 overexpression flies, suggesting LdOA1 may play a master switch role in P450-mediated metabolic detoxification. This study is the first report of an insect OA1 gene regulating insecticide tolerance and potentially playing a role in the regulation of downstream cytochrome P450 expression. These results contribute to the future development of novel insecticides targeting insect GPCRs.
机译:眼白化病1型(OA1)是色素细胞特异性整合膜糖蛋白,是G蛋白偶联受体(GPCR)超家族的成员,该家族与哺乳动物细胞中的异三聚体G蛋白结合。我们旨在表征来自Lymantria dispar(LdOA1)的昆虫OA1在调节杀虫剂耐受性方面的生理功能。在当前的研究中,我们调查了LdOA1响应L.dispar和果蝇果蝇中溴氰菊酯暴露的作用。 LdOA1在第4龄期的最低水平表达,而LdOA1在第5龄期和雄性期显着上调。通过在吉普赛蛾幼虫中注射LdOA1的dsRNA来抑制LdOA1的死亡率是在溴氰菊酯胁迫下微注射GFP dsRNA的对照幼虫的死亡率的4.80倍。与对照幼虫相比,LdOA1沉默幼虫中溴氰菊酯胁迫下11个dispar CYP基因中有9个显着下调。此外,使用转基因技术成功地在黑腹果蝇中过表达了LdOA1基因。溴氰菊酯接触试验表明,与对照果蝇相比,果蝇中LdOA1过表达显着增强了对溴氰菊酯的耐受性。此外,在 LdOA1 过表达果蝇中下游果蝇CYP基因上调,提示 LdOA1 可能在P450介导的代谢排毒中起主要转换作用。这项研究是昆虫OA1基因调节杀虫剂耐受性并可能在下游细胞色素P450表达调节中发挥作用的第一个报道。这些结果有助于针对昆虫GPCR的新型杀虫剂的未来发展。

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