Neural and molecular rules of mosquito olfactory rhythms

项目总结 蚊子传播许多人类病原体，如登革热病毒和恶性疟原虫， 集体每年至少造成一百万人死亡。除了杀虫剂抗药性的上升和 其他因素，越来越明显的是，目前控制蚊子数量的战略正在 被蚊子高水平的行为和生理可塑性赋予的韧性所迷惑。在……里面 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和 在一天中主机可用时响应主机提示。然而，尽管有明确的流行病学 相关性，我们对时间生物学和时间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，我们开发了一种综合和 多学科方法，将提供创新的关键步骤，以阐明神经和分子 指导蚊子嗅觉节律的感官处理规则。我们在此建议使用此策略来 分析日常节律、嗅觉检测和脑电活动相互作用的机制基础 埃及伊蚊的加工和嗅觉调节行为。 我们将首先确定在嗅觉敏感度和嗅觉编码水平上的节奏 触角叶在对寄主相关挥发物的行为反应中起到节律作用。在第二个目标中，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，以及 通过使用CRISPR/Cas9敲除时钟基因表达，产生第一个心律失常蚊子。 最后，我们将结合行为方法和转录分析(单细胞RNA测序)来识别 负责蚊子行为与宿主可获得性同步的细胞类型。这第三个目标将 进一步提供蚊子大脑活动节律性调节的大规模表征，并定义 细胞调节状态的节律。 一旦完成，这些目标将阐明嗅觉的神经和分子过程 在Ae，人们的行为每天都不同。埃及伊蚊。不仅拟议的工作有望提供新的机制 洞察蚊子的生物节律并产生第一个时钟敲除蚊子品系，但它也 具有揭示破坏蚊子-宿主相互作用的新目标和方法并提供信息的强大潜力 一种可以抵消蚊子行为可塑性的综合媒介管理策略。

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.