Mutations in NlInR1 affect normal growth and lifespan in the brown planthopper Nilaparvata lugens
Mutations in NlInR1 affect normal growth and lifespan in the brown planthopper Nilaparvata lugens
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NlInR1 突变影响褐飞虱 Nilaparvata lugens 的正常生长和寿命
DOI：
10.1016/j.ibmb.2019.103246
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发表时间：
2019
影响因子：
3.8
通讯作者：
Zhang Wenqing
中科院分区：
文献类型：
--
作者：
Zhao Yu;Huang Gang;Zhang Wenqing
褐飞虱（Nilaparvata lugens）含有两个胰岛素受体同源物，命名为NlInR 1和NlInR 2。NlInR 1与昆虫和脊椎动物中典型的胰岛素受体具有惊人的同源性，含有一个配体激活的胞内酪氨酸激酶催化结构域。在此，我们报告了一种优化的CRISPR/Cas9系统，用于诱导BPH中NlInRl基因座的突变，该系统由通过Nlvasa启动子在种系中特异性表达的Cas9质粒和在U6启动子控制下的通用sgRNA表达质粒组成。我们系统地评估了注射混合物组合物的效率，并证明了Cas9/sgRNA靶向必需基因的适当组合。此外，我们发现，纯合突变体的theNlInR 1基因是早期胚胎致死，而杂合突变体生长缓慢，表现出严重的体重和翅膀的大小减少，寿命比野生型更长。有趣的是，当靶向theNlInR 1基因座的不同重要结构域时，突变表型的严重程度是不同的。突变表型的严重程度与脊椎动物中胰岛素/胰岛素样生长因子（IGF）信号通路缺陷的严重程度相似，表明NlInR 1在调节发育和寿命中具有保守的功能。NlInR 1基因在BPH中的表达谱表明，它调控着许多细胞过程，包括胰岛素抵抗、光传导、代谢、内吞、寿命、生物合成和蛋白质加工等。我们的研究结果也为了解昆虫翅多型化中胰岛素信号传导的精确分子机制铺平了道路。
The brown planthopper (BPH)Nilaparvata lugenscontains two insulin receptor homologues, designatedNlInR1 andNlInR2.NlInR1 is strikingly homologous to the typical InR in insects and vertebrates, containing a ligand-activated intracellular tyrosine kinase catalytic domain. Herein, we report an optimized CRISPR/Cas9 system to induce mutations in theNlInR1 locus in BPH,consisting of a Cas9 plasmid that is specifically expressed in the germline via theNlvasa promoter and versatile sgRNA expression plasmids under the control of the U6 promoter. We systematically evaluated the efficiency of injection mix compositions and demonstrated an appropriate combination of Cas9/sgRNA to target essential genes. Furthermore, we showed that homozygous mutants for theNlInR1 gene are early embryonic lethal, whereas heterozygous mutants grow more slowly, exhibit a severe reduction in body weight and wing size and live longer than the wild type. Interestingly, the severity of the mutant phenotype was different when targeting distinct important domains of theNlInR1 locus. The severity of the mutant phenotype is similar to that of insulin/insulin-like growth factor (IGF) signaling pathway deficiencies in vertebrates, suggesting a conserved function ofNlInR1 in the regulation of development and longevity. Global expression profiling suggests thatNlInR1 regulates many cellular processes in BPH, including insulin resistance, phototransduction, metabolism, endocytosis, longevity, biosynthesis and protein processing. Our results also pave the way for understanding the precise molecular mechanism of insulin signaling in wing polyphenism in insects.