Molecular and functional characterization of olfactory pathways in the arbovirus vector mosquito Aedes aegypti

虫媒病毒载体蚊子埃及伊蚊嗅觉通路的分子和功能特征

• 批准号: 10638710
• 负责人: Carolyn Sarah McBride
• 金额: $ 40.5万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-02 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10638710
• 关键词: Acceleration; Aedes; Afferent Neurons; Arboviruses; Area; Axon; Behavior; Brain; CRISPR/Cas technology; Cell Nucleus; Chemicals; Culicidae; Data; Detection; Disease Vectors; Drosophila genus; Equation; Female; Flowers; Gene Library; Genetic; High-Throughput Nucleotide Sequencing; Human; Human Bites; Image; Individual; Label; Ligands; Lobe; Logic; Maps; Maxilla; Molecular; Mosquito Control; Mosquito-borne infectious disease; Neurons; Odors; Olfactory Pathways; Oviposition; Pattern; Peripheral; Persons; Population; Property; Reagent; Research; Resolution; Resources; Site; Smell Perception; Sorting; Technology; Terminator Codon; Volatilization; Work; cell type; design; experimental study; flexibility; genome editing; in vivo; neural; olfactory receptor; olfactory sensory neurons; pathogen; rational design; receptor; response; single nucleus RNA-sequencing; tool; transcription factor; transcriptome; vector control; vector mosquito; vinegar fly

Project Summary Abstract Mosquitoes infect hundreds of millions of people with deadly pathogens every year. Since mosquitoes identify humans and other important resources primarily via their sense of smell, the disruption of mosquito olfactory systems has long been recognized as a potential strategy for controlling these pathogens. For example, repellants that scramble or block the detection of odors may be used to push mosquitoes away from humans and the areas where we live and work. Conversely, irresistibly attractive blends of volatile chemicals may be deployed to pull mosquitoes into lethal traps. Despite some advances in this area over the past decade, progress has been limited by the fact that the olfactory systems of our most important vector mosquitoes remain largely uncharacterized. We know that mosquitoes detect odors using tens to hundreds of ligand-specific olfactory receptors expressed in approximately 60 different types of olfactory sensory neurons (OSNs) found on their antennae and maxillary palps. But we don’t yet know exactly which neurons mosquitoes rely on for detecting humans, flowers, and oviposition sites, nor which receptors are expressed in those neurons. Moreover, exciting preliminary data from our lab and others indicates that the 1-to-1 matching between receptors and sensory neurons observed in Drosophila vinegar flies does not apply in mosquitoes. Instead, mosquito sensory neurons appear to express multiple, ligand-specific receptors. This means that the tuning of the neurons that drive behavior cannot be equated to the tuning of individual receptors and thus helps to explain why previous receptor-focused studies have largely failed to unlock the logic of mosquito host attraction. Here, we propose to characterize the molecular and functional properties of all major OSN cell types on the antennae of the arbovirus vector mosquito Aedes aegypti. In Aim 1, we will conduct single- nucleus RNA sequencing of antennal neurons to identify putative OSN cell types and the receptors expressed therein. In Aim 2, we will use CRISPR/Cas9 genome editing to generate OSN type-specific expression drivers that can be used to match OSN types to their target glomeruli in the antennal lobe of the brain. In Aim 3, we will use in vivo antennal lobe imaging to characterize the tuning of a subset of OSN types to a panel of 200-300 biologically relevant odorants and natural blends. Taken together we expect to generate a receptor-neuron-glomerulus map for this important disease vector and a library of genetic tools with which to manipulate it—facilitating the identification of the neurons that drive behavior and opening the door to the efficient and rational design of chemical repellants and attractants for use in vector control.

项目摘要 蚊子每年用致命的病原体感染数亿人。因为蚊子 识别人类和其他重要资源，主要是通过他们的嗅觉， 蚊子的嗅觉系统长期以来被认为是控制这些疾病的潜在策略 病原体例如，可以使用扰乱或阻挡气味检测的驱避剂来推动气味。 蚊子远离人类和我们生活和工作的地方。相反，有吸引力 挥发性化学品的混合物可用于将蚊子引入致命陷阱。尽管有一些 尽管在过去十年中在这一领域取得了进展，但进展受到以下事实的限制： 我们最重要的病媒蚊子的系统在很大程度上仍然没有特征。我们知道 蚊子利用几十到几百个在细胞中表达的配体特异性嗅觉受体来检测气味。 在它们的触角上发现了大约60种不同类型的嗅觉感觉神经元（OSN）， 下颚须但我们还不知道蚊子依靠哪些神经元来探测人类， 花和产卵地点，也没有受体在这些神经元中表达。此外，令人兴奋的 来自我们实验室和其他实验室的初步数据表明，受体和受体之间的1对1匹配， 在果蝇中观察到的感觉神经元并不适用于蚊子。相反，蚊子 感觉神经元似乎表达多种配体特异性受体。这意味着， 驱动行为的神经元不能等同于个体受体的调谐，因此有助于 解释为什么以前以受体为重点的研究在很大程度上未能解开蚊子宿主的逻辑 景点在这里，我们建议表征所有主要OSN细胞的分子和功能特性， 虫媒病毒载体埃及伊蚊触角上的类型。在目标1中，我们将进行单- 触角神经元的核RNA测序以鉴定推定的OSN细胞类型和受体 在其中表达。在目标2中，我们将使用CRISPR/Cas9基因组编辑来生成OSN类型特异性的 表达驱动程序，可用于匹配OSN类型，以其目标肾小球在触角叶， 大脑在目标3中，我们将使用在体触角叶成像来表征调谐的一个子集， OSN对200-300种生物相关气味剂和天然混合物进行分类。总的来说， 我希望能为这种重要的疾病载体产生一个受体-神经元-肾小球图，并建立一个 基因工具来操纵它-促进识别驱动行为的神经元 并为有效合理地设计化学驱避剂和引诱剂打开了大门， 病媒控制