二化螟GABA受体的组成及其药理学特性研究

项目名称： 二化螟GABA受体的组成及其药理学特性研究

项目编号： No.31501672

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 赵春青

作者单位： 南京农业大学

项目金额： 21万元

中文摘要： 针对GABA受体作为农药的重要靶标却在农业昆虫中研究浅显的现状，围绕近几年以GABA受体为靶标的新化合物（fluralaner、BPB等）与传统农药（氟虫腈、阿维菌素等）无交互抗性和作用靶位点不明确这一科学问题。本研究拟：1.选用水稻害虫二化螟为研究对象，克隆其GABA受体RDL、GRD和LCCH3亚基，比对其与哺乳动物亚基的差异，检测其表达谱，揭示受体的时空分布规律；2.采用亚基的胞内环区域片段（R1、G1和L1）为抗原制备多克隆抗体，通过免疫共沉淀技术明确受体的亚基组成；3.利用双电极电压钳技术测定表达GABA受体的爪蟾卵母细胞对多种杀虫剂的电生理反应，明确GABA受体的药理学特性。预期结果可明确二化螟GABA受体的天然组成及其药理学特性，揭示其分子结构和与农药的作用机制，为不同亚基上靶位点的寻找及以GABA受体为靶标的新农药研发和应用提供理论依据和技术支持。

中文关键词： 有机合成杀虫剂；作用机理；分子靶标；离子通道；选择性

英文摘要： GABA receptor is an important insecticide target but without deep study in the agricultural insect. In recent years, the new compounds (fluralaner and BPB etc.) target to the GABA receptor as well as the conventional insecticides (fipronil, avermectin, etc.) but without cross-resistance were synthesized, however, the binding sites are not clear. To explain the phenomenon, our project aims to: First, clone the GABA receptor subunits, RDL, GRD and LCCH3, from the rice stem borer, compare the difference of three subunits to those from mammalian and detect spatial and temporal distribution expression profile of three subunits; Second, utilize the intracellular loop region (R1, G1 and L1) of subunits as antigen to obtain polyclonal antibodies, and examine the subunits composition of GABA receptor by co-immunoprecipitation method; Third, examine the electrophysiological responses of Xenopus oocytes with GABA receptor to various insecticides using two-electrode voltage clamp technique to identify the GABA receptor pharmacological properties. The completion of this present project will define the natural composition and pharmacological properties of GABA receptors in the rice stem borer and reveal its molecular structure and mechanism of insecticides action for finding the target site from different subunits and providing theoretical basis and technical support for the development and application of new insecticides, which function on the GABA receptor.

英文关键词： synthetic insecticide;functional mechanism;molecular target;ion channel;selectivity