title: Comparison of research methods for functional characterization of insect olfactory receptors.
abstract: 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.
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Wang
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Kang
authors.initials: K
authors.lastname: He
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Guirong
authors.initials: G
authors.lastname: Wang
mesh_terms: D000483:Alleles; D000818:Animals; D030801:Animals, Genetically Modified / Q000502:physiology; D001522:Behavior, Animal; D004330:Drosophila / Q000502:physiology; D004566:Electrodes; D005260:Female; D007915:Lepidoptera / Q000502:physiology; D008297:Male; D009154:Mutation; D009812:Odorants; D018034:Olfactory Receptor Neurons / Q000502:physiology*; D010641:Phenotype; D010675:Pheromones; D018035:Receptors, Odorant / Q000502:physiology*; D043563:Receptors, Pheromone / Q000502:physiology*; D015203:Reproducibility of Results; D012724:Sex Attractants / Q000502:physiology*
chemical_list: D010675:Pheromones; D018035:Receptors, Odorant; D043563:Receptors, Pheromone; D012724:Sex Attractants