Neural and molecular rules of mosquito olfactory rhythms

项目摘要 蚊子传播许多人类病原体，如登革热病毒和恶性疟原虫， 每年造成至少一百万人死亡。除了杀虫剂抗药性上升， 随着其他因素的影响，越来越明显的是，目前控制蚊子数量的策略正在被 蚊子高度的行为和生理可塑性所赋予的适应力令人困惑。在 特别是，生物节律对疾病传播至关重要，因为它们允许蚊子活跃， 在一天中主人有空的时候对主人的暗示做出反应。然而，尽管有明确的流行病学 相关性，我们对时间生物学和生物学之间相互作用的机制知之甚少。 蚊子的嗅觉行为为了解决这一关键的知识差距，我们制定了一个综合和 多学科的方法，将提供创新的关键步骤，阐明神经和分子 引导蚊子嗅觉节奏的感觉处理规则。我们建议在这里使用这种策略， 分析日常节律、嗅觉检测和 处理和嗅觉介导的行为在埃及伊蚊。 我们将首先确定嗅觉敏感性和嗅觉编码的节奏如何在大脑皮层的水平上变化。 触角叶对寄主相关挥发物的行为反应的节律性有贡献。第二个目标，我们 将进一步分析生物钟如何调节蚊子的大脑和触角转录组， 通过使用CRISPR/Cas9敲除clock基因表达，产生第一系蚊子。 最后，我们将结合联合收割机行为学方法和转录组学分析（单细胞RNA测序）， 负责蚊子行为与宿主可用性同步的细胞类型。第三个目标将 进一步提供了蚊子脑活动的节律调节的大规模表征，并定义了 细胞调节状态的节奏。 完成后，这些目标将阐明嗅觉的神经和分子过程， 行为每天都在变化。埃及蚊子。不仅拟议的工作预计将提供新的机制 深入了解蚊子的生物节律，并产生第一个时钟敲除蚊子系，但它也 具有很大的潜力，可以揭示新的目标和破坏蚊子与宿主相互作用的方法， 一种综合的病媒管理战略，可以抵消蚊子的行为可塑性。

PROJECT SUMMARY Mosquitoes transmit numerous human pathogens, such as dengue virus and Plasmodium falciparum, collectively responsible for at least one million deaths each year. In addition to rising insecticide resistance and other factors, it has become increasingly evident that current strategies to control mosquito populations are being confounded by the resilience conferred by mosquitoes’ high levels of behavioral and physiological plasticity. In particular, biological rhythms are crucial to disease transmission as they allow mosquitoes to be active and responsive to host cues at times of the day when hosts are available. However, despite clear epidemiological relevance, we know very little about the mechanisms underlying the interaction between the chronobiology and the olfactory behavior of mosquitoes. To address this key knowledge gap, we have developed an integrative and multidisciplinary approach that will provide innovative critical steps towards elucidating the neural and molecular rules of sensory processing that guide mosquitoes’ olfactory rhythms. We propose here to use this strategy for analyzing the mechanistic underpinnings of the interaction between daily rhythms, olfactory detection and processing, and olfactory-mediated behaviors in Aedes aegypti mosquitoes. We will first determine how rhythms in olfactory sensitivity and in olfactory encoding at the level of the antennal lobe contribute to rhythms in the behavioral responses to host-related volatiles. In a second aim, we will further analyze how circadian clocks modulate the brain and antennal transcriptome of mosquitoes, and generate the first line of arrhythmic mosquitoes, by using CRISPR/Cas9 to knockout clock gene expression. Finally, we will combine behavioral methods and transcriptomic analysis (single-cell RNA sequencing) to identify the cell-types responsible for the synchronization of mosquito behavior with host-availability. This third aim will further provide a large-scale characterization of the rhythmic regulation of mosquito brain activity, and define rhythms in cell regulatory states. Upon completion, these aims will shed light on the neural and molecular processes by which olfactory behaviors vary daily in Ae. aegypti mosquitoes. Not only the proposed work is expected to offer new mechanistic insights into the biological rhythms of mosquitoes and generate a first clock knockout mosquito line, but it also bears strong potential for revealing new targets and methods for disrupting mosquito-host interactions and inform an integrated vector management strategy that can counteract mosquito behavioral plasticity.