State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
Sci Rep. 2016 Sep 16;6:32806. doi: 10.1038/srep32806.
Insect olfactory receptors (ORs) in the peripheral olfactory system play an important role detecting elements of information from the environment. At present, various approaches are used for deorphanizing of ORs in insect. In this study, we compared methods for functional analysis of ORs in vitro and in vivo taking the candidate pheromone receptor OR13 of Helicoverpa assulta (HassOR13) as the object of our experiments. We found that the natural system was more sensitive than those utilizing transgenic Drosophila. The two-electrode voltage-clamp recording is more suitable for functional screening of large numbers of ORs, while the in vivo transgenic Drosophila system could prove more accurate to further validate the function of a specific OR. We also found that, among the different solvents used to dissolve pheromones and odorants, hexane offered good reproducibility and high sensitivity. Finally, the function of ORs was indirectly confirmed in transgenic Drosophila, showing that odor-activation of ORs-expressing olfactory receptor neurons (ORNs) can mediate behavioral choices. In summary, our results compare advantages and drawbacks of different approaches, thus helping in the choice of the method most suitable, in each specific situation, for deorphanizing insect ORs.
昆虫嗅觉受体（ORs）在周围嗅觉系统中起着重要作用，可检测环境中的信息元素。目前，人们采用多种方法对昆虫 ORs 进行功能分析。本研究以甜菜夜蛾（ Helicoverpa assulta ）候选信息素受体 OR13（ HassOR13 ）为实验对象，比较了体外和体内功能分析 ORs 的方法。我们发现，天然系统比利用转基因果蝇的方法更灵敏。双电极电压钳记录更适合于大量 ORs 的功能筛选，而体内转基因果蝇系统则更能准确地验证特定 OR 的功能。我们还发现，在溶解信息素和气味剂的不同溶剂中，正己烷的重现性和灵敏度都较好。最后，在转基因果蝇中间接证实了 ORs 的功能，表明表达 OR 的嗅觉感觉神经元（ORNs）的气味激活可以介导行为选择。综上所述，我们的结果比较了不同方法的优缺点，有助于在每种特定情况下选择最适合的方法对昆虫 ORs 进行功能分析。
