Shi Xuekai, Li Shuai, Yang Lin, Liu Xiaojian, Merzendorfer Hans, Zhu Kun Yan, Zhang Jianzhen
Institute of Applied Biology, Shanxi University, Taiyuan, China.
College of Life Science, Shanxi University, Taiyuan, China.
Insect Sci. 2022 Dec;29(6):1601-1611. doi: 10.1111/1744-7917.13030. Epub 2022 Apr 5.
Clathrin heavy chain (Chc) is a constituent of clathrin-coated vesicles and serves important functions in endocytosis and intracellular membrane trafficking but appears to have physiological roles also at the organismal level. Most of what we know about Chc functions originates from studies performed in fungal or vertebrate cells. However, the physiological functions of Chc in insects remain poorly understood. Here, we identified a Chc ortholog from a Locusta migratoria transcriptome database. RT-qPCR revealed that LmChc was constitutively expressed in fifth-instar nymphs. In this developmental stage, LmChc showed the highest expression in the ovary followed by hemolymph, testis, hindgut, midgut, and foregut. In isolated hemocytes, we detected the Chc protein in patches at the plasma membrane. To examine the role of LmChc in L. migratoria during development, RNA interference was performed by injecting dsRNA into the early fifth-instar nymphs. Silencing of LmChc caused a lethal phenotype with molting defect from nymph to adult. In addition, silencing of LmChc resulted in abnormal development of the ovaries, the size of which was significantly smaller than that in controls. Taken together, our results suggest that LmChc is a vital gene in L. migratoria that plays an important role in growth, development, and reproduction. LmChc may be used as an efficient RNAi target gene for developing dsRNA-based biological insecticides to manage insect pests.
网格蛋白重链（Chc）是网格蛋白包被小泡的一个组成部分，在胞吞作用和细胞内膜运输中发挥重要功能，但在生物体水平似乎也具有生理作用。我们对Chc功能的了解大多源于在真菌或脊椎动物细胞中进行的研究。然而，Chc在昆虫中的生理功能仍知之甚少。在这里，我们从飞蝗转录组数据库中鉴定出一个Chc直系同源基因。RT-qPCR显示，LmChc在五龄若虫中组成性表达。在这个发育阶段，LmChc在卵巢中的表达最高，其次是血淋巴、睾丸、后肠、中肠和前肠。在分离的血细胞中，我们在质膜上的斑块中检测到了Chc蛋白。为了研究LmChc在飞蝗发育过程中的作用，通过向五龄初期若虫注射dsRNA进行RNA干扰。LmChc的沉默导致了从若虫到成虫蜕皮缺陷的致死表型。此外，LmChc的沉默导致卵巢发育异常，其大小明显小于对照组。综上所述，我们的结果表明，LmChc是飞蝗中的一个重要基因，在生长、发育和繁殖中发挥重要作用。LmChc可作为一个有效的RNAi靶基因，用于开发基于dsRNA的生物杀虫剂来防治害虫。
