Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing 210023, China.
Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing 210023, China; College of Biology and Food Engineering, Anyang Institute of Technology, Anyang 455000, China.
Gene. 2022 Oct 30;842:146794. doi: 10.1016/j.gene.2022.146794. Epub 2022 Aug 8.
Choline acetyltransferase (ChAT) and vesicular acetylcholine transporter (VAChT) are essential enzymes for synthesizing and transporting acetylcholine (ACh). But their functions in metamorphosis, reproduction, and the insecticide susceptibility were poorly understood in the insects. To address these issues, we identified the orthologues of chat and vacht in Tribolium castaneum. Spatiotemporal expression profiling showed Chat has the highest expression at the early adult stage, while vacht shows peak expression at the early larval stage. Both of them were highly expressed at the head of late adult. RNA interference (RNAi) of chat and vacht both led to a decrease in ACh content at the late larval stage. It is observed that chat knockdown severely affected larval development and pupal eclosion, but vacht RNAi only disrupted pupal eclosion. Further, parental RNAi of chat or vacht led to 35 % or 30 % reduction in fecundity, respectively, and knockdown of them completely inhibited egg hatchability. Further analysis has confirmed that both the reduction in fecundity and hatchability caused through the maternal specificity in T. castaneum. Moreover, the transcript levels of chat and vacht were elevated after carbofuran or dichlorvos treatment. Reduction of chat or vacht decreased the resistance to carbofuran and dichlorvos. This study provides the evidence for chat and vacht not only involved in development and reproduction of insects but also could as the potential targets of insecticides.
胆碱乙酰转移酶（ChAT）和囊泡乙酰胆碱转运体（VAChT）是合成和转运乙酰胆碱（ACh）的必需酶。然而，它们在昆虫变态、生殖和杀虫剂敏感性中的功能仍不清楚。为了解决这些问题，我们在赤拟谷盗中鉴定了 chat 和 vacht 的同源物。时空表达谱分析表明，Chat 在成虫早期表达水平最高，而 vacht 在幼虫早期表达水平最高。它们在成虫晚期头部的表达水平都很高。Chat 和 vacht 的 RNA 干扰（RNAi）都导致幼虫后期 ACh 含量降低。观察到 Chat 敲低严重影响幼虫发育和蛹羽化，但 vacht RNAi 仅破坏蛹羽化。此外，chat 或 vacht 的亲代 RNAi 分别导致生育力降低 35%或 30%，而敲低则完全抑制卵的孵化能力。进一步分析证实，赤拟谷盗中的生育力和孵化能力降低都是通过母体特异性引起的。此外，carbofuran 或敌敌畏处理后 chat 和 vacht 的转录水平升高。Chat 或 vacht 的减少降低了对 carbofuran 和敌敌畏的抗性。这项研究为 chat 和 vacht 不仅参与昆虫的发育和生殖提供了证据，而且它们也可能成为杀虫剂的潜在靶标。
