The Role of Odorant Binding Proteins in Anopheline Mosquito Odor Perception

这里提出的工作将调查气味识别的特异性在 冈比亚按蚊及其与特定气味刺激的相关性 可量化的生理和行为反应。一项由多名调查人员组成的努力将是 将整合各相关领域的互补专业知识，以 彻底了解化学传感系统和 冈比亚按蚊的化学感官行为。 气味识别的特异性很可能在这两种气味结合蛋白中编码 (OBP)和气味受体(ORs)。我们会进行产权证本地化研究和 用免疫细胞化学和原位杂交技术在触角切片上检测ORS 研究气味感受器神经元的最佳调谐到某个特定的假设 气味可以通过OBP和OBP的匹配组合的共同表达来实现 明确的嗅觉感受器中的ORS。因为OBP代表了第一批蛋白质 嗅觉系统会遇到并结合气味分子，在体外表达OBP 用于高通量筛选分析以鉴定天然配体并推断结合 已知的和新确定的配体的能力。结合计算机建模研究 突变方法将定义结合口袋并预测修饰的配体 具有改进的性能。所有体外研究的体内验证将通过以下方式获得 电生理和行为测试使用表达A基因的雌性蚊子 OBP的正常补充或由于以下原因而导致特定OBP表达不足 RNAi介导的基因敲除。 从这种多学科方法中获得的知识将导致对 化学物质会干扰雌性蚊子寻找寄主的行为，因此 减少人类宿主和寄生虫媒介之间的接触。

Work proposed here will investigate the specificity of odorant recognition in the malaria mosquito Anopheles gambiae and correlate specific odor stimuli with quantifiable physiological and behavioral responses. A multi-investigator effort will be undertaken that will integrate complementary expertise in various relevant fields toward the common goal of thoroughly understanding the chemosensory system and chemosensory-mediated behavior of Anopheles gambiae. Odor recognition specificity is very likely encoded in both odorant binding proteins (OBPs) and odorant receptors (ORs). We will carry out localization studies of OBPs and ORs by immunocytochemical and in situ hybridization assays on antennal sections to investigate the hypothesis that optimal tuning of odorant receptor neurons to a certain odor may be achieved by the co-expression of a matching combination of OBPs and ORs in defined olfactory sensilla. Because OBPs represent the first proteins of the olfactory system to encounter and bind odor molecules, in vitro expressed OBPs will be used in high-throughput screening assays to identify natural ligands and deduce binding capacities for known and newly identified ligands. Computer modeling studies combined with mutagenesis approaches will define the binding pocket and predict modified ligands with improved properties. In vivo validation of all in vitro studies will be obtained through electrophysiological and behavioral assays employing female mosquitoes that express a normal complement of OBPs or are deficient in specific OBP expression as a result of RNAi-mediated knockdown. Knowledge gained from this multi-disciplinary approach will lead to the identification of chemicals, which can interfere with host seeking behavior by female mosquitoes, thus reducing the incidence of contact between the human host and the parasite vector.