Laboratory of Tropical Veterinary Medicine and Vector Biology, School of Life and Health Sciences, Hainan University, Haikou, Hainan, China.
Hainan One Health Key Laboratory, Hainan University, Haikou, Hainan, China.
Insect Mol Biol. 2024 Dec;33(6):678-686. doi: 10.1111/imb.12930. Epub 2024 May 31.
Arylalkylamine N-acetyltransferase (aaNAT) is a crucial enzyme that catalyses the transfer of acetyl groups from acetyl coenzyme A to arylalkylamines and arylamines. Evolutionary studies have identified a distinct class of aaNATs specific to mosquitoes, yet their functions remain elusive. This study focuses on Ae-aaNAT7, a mosquito-unique gene in Aedes aegypti (Diptera:Culicidae), to explore its functionality. Temporal and spatial expression analysis of Ae-aaNAT7 mRNA revealed high expression during embryonic development and in first-instar larvae, with notable expression in the limbs of adult mosquitoes based on tissue expression profiling. By further employing CRISPR/Cas9 technology for loss-of-function studies, our investigation revealed a reduction in the area of white spotting in the limbs of Ae-aaNAT7 mutant adult mosquitoes. Further investigation revealed a significant decrease in the fecundity and hatchability of the mutants. Dissection of the ovaries from Ae-aaNAT7 heterozygous mutants showed a noticeable reduction in the oocyte area compared with wild type. Dissection of the exochorion of the eggs from Ae-aaNAT7 homozygous mutants consistently revealed a striking absence of mature embryos. In addition, RNA interference experiments targeting Ae-aaNAT7 in males resulted in a reduction in fecundity, but no effect on hatchability was observed. These collective insights underscore the substantial impact of Ae-aaNAT7 on reproduction and its pivotal contribution to adult limb pigmentation in Ae. aegypti. These revelations offer insights pivotal for the strategic design of future insecticide targets.
芳基烷基胺 N-乙酰基转移酶(aaNAT)是一种关键酶,可催化乙酰辅酶 A 中的乙酰基转移到芳基烷基胺和芳胺上。进化研究已经确定了一类特定于蚊子的独特的 aaNATs,但它们的功能仍然难以捉摸。本研究集中于埃及伊蚊(双翅目:蚊科)中蚊子特有的基因 Ae-aaNAT7,以探索其功能。Ae-aaNAT7 mRNA 的时空表达分析显示,在胚胎发育和第一龄幼虫期表达量较高,在成年蚊子的肢体组织表达谱中表达显著。通过进一步采用 CRISPR/Cas9 技术进行功能丧失研究,我们的研究发现 Ae-aaNAT7 突变体成年蚊子的肢体上的白色斑点面积减少。进一步的研究发现突变体的繁殖力和孵化率显著降低。从 Ae-aaNAT7 杂合突变体的卵巢解剖中发现,与野生型相比,卵母细胞的面积明显减小。从 Ae-aaNAT7 纯合突变体的卵壳外膜解剖中一致发现成熟胚胎明显缺失。此外,针对 Ae-aaNAT7 的 RNA 干扰实验在雄性中导致繁殖力降低,但孵化率没有观察到影响。这些综合的见解强调了 Ae-aaNAT7 对繁殖的重大影响及其对埃及伊蚊成年肢体色素沉着的关键贡献。这些发现为未来杀虫剂靶标的战略设计提供了重要的见解。