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Hormonal regulation of microRNA expression dynamics in the gut of the yellow fever mosquito Aedes aegypti

机译:黄热病蚊子埃及伊蚊肠道中microRNA表达动态的激素调控

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The yellow fever mosquito Aedes aegypti is an obligatory blood feeder and a major arboviral disease vector, evoking severe public health concerns worldwide. In adult female mosquitoes, the gut is critical for blood digestion and pathogen entry. We aimed for a systematic exploration of microRNA expression dynamics in the gut during the gonadotrophic cycle. Small RNA libraries were constructed from female mosquito gut tissues at five time points. Unsupervised hierarchical clustering revealed three expression clusters (early, mid and late) peaking at sequential time points - 24, 48 and 72 h posteclosion. Differentially expressed miRNAs were identified at 24 h post-blood meal (PBM). Depletions of Methoprene-tolerant Met; the juvenile hormone (JH) receptor and Ecdysone receptor EcR; the receptor to 20-hydroxyecdysone (20E) were performed using dsRNA to these genes to investigate impacts on microRNA expressions. Our results suggest that Met-mediated signalling downregulates miRNA expression from the early cluster and upregulates that from the late cluster. EcR signalling either up- or downregulated miRNA levels at 24 h PBM, indicating a differential effect of this receptor in miRNA gene expression. Furthermore, miR-281, which is the most abundant miRNA in the gut tissue, is induced and repressed by Met- and EcR-mediated signalling, respectively. Systematic depletion using synthetic antagomir and phenotype examinations indicate that miR-281 is obligatory for the normal progression of blood digestion, ovarian development and reproduction. Collectively, this study unveils expression dynamics of microRNAs in the female gut tissue during the gonadotrophic cycle and demonstrates that they are affected by JH and 20E signalling.
机译:黄热病蚊子埃及伊蚊是强制性的血液喂养者和主要的虫媒病毒病媒介,在世界范围内引起严重的公共卫生问题。在成年雌性蚊子中,肠道对血液消化和病原体进入至关重要。我们旨在系统地探索促性腺激素周期中肠道中的microRNA表达动态。在五个时间点由雌性蚊子肠道组织构建小型RNA文库。无监督分层聚类显示3个表达簇(早期、中期和晚期)在闭合后24、48和72 h的连续时间点达到峰值。在血餐后 24 小时 (PBM) 鉴定差异表达的 miRNA。使用对这些基因的 dsRNA 进行甲氧耐受 [Met;保幼激素 (JH) 受体] 和 Ecdysone 受体 [EcR;20-羟基蜕皮激素 (20E)] 的消耗,以研究对 microRNA 表达的影响。我们的结果表明,Met介导的信号转导下调了早期簇的miRNA表达,并上调了晚期簇的miRNA表达。EcR 信号传导在 24 小时 PBM 时上调或下调 miRNA 水平,表明该受体在 miRNA 基因表达中的差异作用。此外,miR-281 是肠道组织中最丰富的 miRNA,分别由 MetR 和 EcR 介导的信号传导诱导和抑制。使用合成拮抗剂和表型检查的系统耗竭表明,miR-281 对于血液消化、卵巢发育和繁殖的正常进展是必需的。总的来说,这项研究揭示了促性腺激素周期中女性肠道组织中 microRNA 的表达动力学,并证明它们受到 JH 和 20E 信号传导的影响。

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