RNAi is an essential technology for studying gene function in eukaryotes, and is also considered to be a potential strategy for pest control. However, the mechanism behind the cellular uptake of dsRNA in aphids, a group of important agricultural sucking pests, remains unknown. Here, using the pea aphid Acyrthosiphon pisum as model for aphids, we identified two core genes of clathrin-dependent endocytosis (CDE), Apchc and Apvha16. We confirmed that expression of Apchc, Apvha16 and RNAi core component genes (ApAgo2, ApDcr2 and ApR2d2) were simultaneously induced at 12 h after feeding dsRNA. By using an RNAi-of-RNAi approach, we demonstrated that suppression of Apchc and Apvha16 transcripts by RNAi significantly impaired RNAi efficiency of selected reporter genes (RGs), including ApGNBP1, Apmts and Aphb, suggesting the involvement of CDE in cellular dsRNA uptake in aphids. Further confirmation was also provided using two inhibitors, chlorpromazine (CPZ) and bafilomycin A1 (BafA1). Administration of CPZ and of BafA1 both led to an impaired silencing efficiency of the RGs in the pea aphid. Finally, these RNAi-of-RNAi results were reconfirmed in the peach aphid Myzus persicae. Taking these findings together, we conclude that CDE is involved in cellular dsRNA uptake in aphids.

RNAi 是研究真核生物基因功能的一项重要技术，也被认为是一种潜在的害虫防治策略。然而，蚜虫（一组重要的农业吸吮害虫）中 dsRNA 的细胞摄取背后的机制仍然未知。在这里，我们使用豌豆蚜虫Acyrthosiphon pisum作为蚜虫模型，鉴定了网格蛋白依赖性内吞作用 (CDE)、Apchc和Apvha16 的两个核心基因。我们证实Apchc、Apvha16和 RNAi 核心成分基因（ApAgo2、ApDcr2和ApR2d2) 在喂食 dsRNA 后 12 小时同时诱​​导。通过使用RNAi-的-RNAi的一个方法中，我们表明，抑制Apchc和Apvha16通过RNAi选定的显著的RNAi效率受损转录报道基因（RG中），包括ApGNBP1，Apmts和Aphb ，提示CDE在细胞的dsRNA摄取的参与蚜虫。还使用两种抑制剂氯丙嗪 (CPZ) 和巴弗洛霉素 A1 (BafA1) 进行了进一步确认。氯丙嗪BafA1和行政两种导致的受损沉默效率的RG在豌豆蚜虫。最后，这些 RNAi-of-RNAi 结果在桃蚜中再次得到证实桃蚜。综合这些发现，我们得出结论，CDE 参与了蚜虫的细胞 dsRNA 摄取。