Identification of mosquito brain neurons that drive blood-feeding behavior

项目总结 蚊子通过定位和叮咬脊椎动物的能力向人类传播大量病原体。 目前预防这些疾病的战略面临各种限制，需要采取新的方法。一 可能是抑制蚊子大脑中引导对人类产生吸引力的行为的神经元 并以血液为食，但这些神经元还没有被识别出来，因为这样做的工具和知识 不存在。这份提案描述了我的计划，即使用基于遗传和测序的方法来识别 供血行为所需的神经元。我会利用蚊子需要 多种人类来源的线索，特别是热量和二氧化碳，会吞噬血液。利用基因表达的变化 在单个细胞中，我将识别在这两种刺激下特别表现出不同反应的神经元。至 启用对感兴趣的特定细胞类型的基因靶向，我们将创建一份增强子元素的图谱，该图谱 利用配对的单细胞RNA和ATAC-SEQ数据，对蚊子大脑中的每一种细胞类型具有特异性。最后， 我们将使用增强器驱动程序构建来针对关键神经元进行血液喂养，并将它们与 基因效应器，以测试神经元是否对血液喂养行为是必要的和充分的。这 提案结合了我在计算基因组学、转基因蚊子和行为方面的专业知识，以实现 目标是识别供血的神经回路中关键节点的神经元。这些人的身份 驱动血液供应的神经元将有助于理解它们是如何被激活的，从而使 设计更吸引人的捕蚊器。这些神经元如果失活，将防止蚊子叮咬 因此，它们也可以成为阻止蚊子传播疾病传播的潜在目标。

PROJECT SUMMARY Mosquitoes transmit numerous pathogens to humans by virtue of their ability to locate and bite vertebrates. Current strategies to prevent these diseases face various limitations and new approaches are needed. One possibility would be to inhibit the neurons in the mosquito brain that direct the behavior of attraction to humans and feeding on blood, but such neurons have not been identified because the tools and knowledge to do so do not exist. This proposal describes my plan to use genetic and sequencing-based approaches to identify the neurons that are required for blood-feeding behavior. I will take advantage of the fact that mosquitoes require multiple human-derived cues, particularly heat and CO2, to engorge on blood. Using gene expression changes in single cells, I will identify neurons that show altered responses specifically in the presence of both stimuli. To enable the genetic targeting of specific cell types of interest, we will create an atlas of enhancer elements that are specific to each cell type in the mosquito brain, using paired single cell RNA and ATAC-seq data. Finally, we will target key neurons for blood feeding using an enhancer driver constructs and combine them with genetic effectors to test whether the neurons are necessary and sufficient for blood feeding behavior. This proposal combines my expertise in computational genomics, transgenic mosquitoes, and behavior to achieve the goal of identifying neurons at a key node in the neural circuits for blood feeding. The identification of these neurons that drive blood feeding would be useful for understanding how they are activated and thus allow the design of more attractive mosquito traps. These neurons, if inactivated, would prevent mosquitoes from biting humans so they also could serve as a potential target to disrupt the spread of mosquito borne illness.