Research Institute of Applied Biology, Shanxi University, Taiyuan, Shanxi 030006, China; College of Life Science, Shanxi University, Taiyuan, Shanxi 030006, China; Guangdong Provincial Key Laboratory of Insect Development Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, China.
Guangdong Provincial Key Laboratory of Insect Development Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, China.
Insect Biochem Mol Biol. 2019 Oct;113:103210. doi: 10.1016/j.ibmb.2019.103210. Epub 2019 Aug 15.
Insect cytochrome P450 monooxygenases (CYPs) play essential roles in both xenobiotic metabolism and developmental processes. However, the exact physiological function of many CYP genes remains largely unknown. Screening the expression of the CYP genes from the CYP2 and mitochondrial CYP clans of Drosophila melanogaster revealed that Cyp303a1 is highly expressed in the pupal stage. Knockdown of CYP303A1 transcripts by RNAi using the Gal4/UAS system with a ubiquitous driver (tubulin-Gal4) in Drosophila or by dsRNA injection in the last nymph stage of Locusta migratoria resulted in severe defects in eclosion and lethality during and after adult emergence. In Drosophila, tissue-specific RNAi of Cyp303a1 with a wing-specific driver (MS1096-Gal4) revealed that Cyp303a1 was essential for wing extension. Stage-specific RNAi of Cyp303a1 using Gal80 for thermal-dependent-suppression found that the expression of Cyp303a1 at the middle pupal stage was absolutely required. Meanwhile, Cyp303a1 mutants exhibited more than 80% lethality at the late embryonic development stages. Embryonic lethality of the Cyp303a1 mutants was fully rescued by the ubiquitous overexpression of exogenous Cyp303a1. Taken together, we conclude that Cyp303a1 is indispensable for embryonic development and adult eclosion in D. melanogaster, the latter role being conserved over 400 million years of insect evolution.
昆虫细胞色素 P450 单加氧酶 (CYPs) 在异生物质代谢和发育过程中都起着重要作用。然而，许多 CYP 基因的确切生理功能仍知之甚少。筛选黑腹果蝇 CYP2 和线粒体 CYP 族的 CYP 基因的表达情况显示，Cyp303a1 在蛹期表达量较高。使用 Gal4/UAS 系统和 ubiquitous 驱动（tubulin-Gal4）在果蝇中进行 RNAi 敲低 Cyp303A1 转录本，或在 locusta migratoria 的最后一个若虫期注射 dsRNA，导致羽化严重缺陷，并在成虫出现时和之后死亡。在果蝇中，使用 wing-specific 驱动（MS1096-Gal4）进行 Cyp303a1 的组织特异性 RNAi 显示，Cyp303a1 对于翅膀伸展是必需的。使用 Gal80 进行热依赖性抑制的 Cyp303a1 阶段特异性 RNAi 发现，Cyp303a1 在中期蛹期的表达是绝对必需的。同时，Cyp303a1 突变体在胚胎发育后期阶段的致死率超过 80%。Cyp303a1 突变体的胚胎致死率通过 ubiquitous 过表达外源 Cyp303a1 得到完全挽救。总之，我们得出结论，Cyp303a1 对于黑腹果蝇的胚胎发育和成虫羽化是不可或缺的，后者的作用在昆虫进化的 4 亿多年中是保守的。
