Shandong Institute of Parasitic Diseases, Shandong First Medical University & Shandong Academy of Medical Sciences, Jining, Shandong, 272033, P.R. China.
Collaborative Innovation Center for the Origin and Control of Emerging Infectious Diseases, Taishan Medical University, Taian, Shandong, 271000, P.R. China.
Sci Rep. 2019 Apr 24;9(1):6485. doi: 10.1038/s41598-019-42961-w.
To reveal overwintering dormancy (diapause) mechanisms of Culex pipiens pallens (L.), global protein expression differences at three separate time points represent nondiapause, diapause preparation and overwintering diapause phases of Cx. pipiens pallens were compared using iTRAQ. Cx. pipiens pallens females accumulate more lipid droplets during diapause preparation and overwintering diapause maintenance than during the nondiapause phase. A total of 1030 proteins were identified, among which 1020 were quantified and compared. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes (KEGG), Domain and Clusters of Orthologous Groups (COG) analyses revealed key groups of proteins, pathways and domains differentially regulated during diapause preparation and overwintering diapause maintenance phases in this mosquito, including major shifts in energy production and conversion, fatty acid metabolism, the citrate (TCA) cycle, and the cytoskeletal reorganization pathway. Our results provide novel insight into the molecular bases of diapause in mosquitoes and corroborate previously reported diapause-associated features in invertebrates. More interestingly, the phototransduction pathway exists in Cx. pipiens pallens, in particular, actin, rather than other proteins, appears to have substantial role in diapause regulation. In addition, the differential changes in calmodulin protein expression in each stage implicate its important regulatory role of the Cx. pipiens pallens biological clock. Finally, 24 proteins were selected for verification of differential expression using a parallel reaction monitoring strategy. The findings of this study provide a unique opportunity to explore the molecular modifications underlying diapause in mosquitoes and might therefore enable the future design and development of novel genetic tools for improving management strategies in mosquitoes.
为了揭示淡色库蚊（Culex pipiens pallens）的越冬休眠（滞育）机制，使用 iTRAQ 比较了淡色库蚊非滞育、滞育准备和越冬滞育三个不同时间点的全球蛋白质表达差异。与非滞育期相比，淡色库蚊雌蚊在滞育准备和越冬滞育维持期间积累了更多的脂滴。共鉴定出 1030 种蛋白质，其中 1020 种进行了定量比较。基因本体论、京都基因与基因组百科全书（KEGG）、结构域和直系同源群（COG）分析揭示了在这种蚊子的滞育准备和越冬滞育维持阶段差异调节的关键蛋白质组、途径和结构域，包括能量产生和转化、脂肪酸代谢、柠檬酸（TCA）循环和细胞骨架重组途径的重大转变。我们的研究结果为蚊子滞育的分子基础提供了新的见解，并证实了先前在无脊椎动物中报道的与滞育相关的特征。更有趣的是，光转导途径存在于淡色库蚊中，特别是肌动蛋白，而不是其他蛋白质，似乎在滞育调节中起着重要作用。此外，在每个阶段钙调蛋白蛋白表达的差异变化表明其在淡色库蚊生物钟的重要调节作用。最后，选择了 24 种蛋白质使用平行反应监测策略验证差异表达。本研究的结果为探索蚊子滞育的分子修饰提供了一个独特的机会，因此可能为未来设计和开发改善蚊子管理策略的新型遗传工具提供了可能。
