Opsins and TRP channels controlling sensation and behavior in Aedes aeygpti

摘要 埃及伊蚊传播登革热、寨卡病毒、黄热病等疾病， 每年百万人。这些蚊子依靠多种敏锐的感官来定位人类宿主的血液 食物，以及寻找交配的同类。目前，我们只有一个初步的了解， 控制这些关键行为的受体。这项研究的目的是解决这一差距。的 这个建议的统一主题是测试视蛋白和TRP通道是两个关键类别的想法。 在Ae中的感觉和控制行为中具有广泛作用的信号蛋白。埃及人。视紫红质是 G蛋白偶联受体（GPCR）。我们最近发现视蛋白是多模态的 感官受体，挑战了100年来的教条，他们只检测光。为了寻找人类，雌性艾人。 埃及人整合来自不同刺激的信息，包括CO2，视觉线索，有机分子， 来自皮肤的对流热量。我们发现了另一个线索。目标1建立在我们的初步数据，即Ae。aegypti 使用红外（IR）辐射作为额外的宿主刺激。我们概述了实验，以揭示视蛋白的作用， 和TRPA1通道的红外探测。我们建议鉴定表达视蛋白的红外敏感神经元 和TRPA1，并测试模型来解释视蛋白在IR感觉中的作用。为了达到这个目标，我们设计了一个 监测红外吸引力的高效测定和克服困难的新分子遗传学方法 结合多种基因元素。Aim 2利用了我们在另一个TRP中创建的突变 （TRPV-A），使男性和女性耳聋。我们将测试听觉和TRPV-A在蜂群中的作用 形成，交配，和寻找人类。目标2也将建立在雄性交配所需的观察基础上 通过TRPV-A介导的听证会，制定一项战略，以克服限制TRPV-A疗效的主要障碍， 昆虫不育技术（SIT）。SIT是抑制Ae的有希望的策略。埃及人。它包括淹没一个当地的 人口与不育男性，然后使女性不育后交配。使用SIT的一个障碍是 野生型雄性胜过不育雄性。我们建议，操纵TRPV-A- 表达听觉神经元提高不育雄性交配成功率，从而增加SIT的功效。 抑制Ae。埃及人。目的3涉及识别驱避剂的感觉受体。我们建议测试 认为视蛋白是驱虫剂的高敏感受体。如果得到 在所提出的实验中，这将证明视蛋白包括一类新的嗅觉受体。到 为了实现我们的目标，我们制定了一套广泛的最先进的方法。这些 包括新的分子遗传学工具，一套行为分析，原始视频跟踪软件，以及体内 电生理记录。总之，本项目将揭示视蛋白和TRP通道在细胞内的作用。 使蚊子能够感知人类和同种配偶。从这项工作中获得的见解 令人兴奋的潜力，导致创新的战略，以控制Ae。减少昆虫传播的疾病。

Abstract The mosquito Aedes aegypti spreads diseases such as dengue, Zika, yellow fever, and others that afflict >100 million people each year. These mosquitoes rely on multiple keen senses to locate human hosts for blood meals, and for finding conspecifics for mating. Currently, we have only a rudimentary understanding of the receptors that control these critical behaviors. The goal of the proposed research is to address this gap. The unifying theme of this proposal is to test the idea that opsins and TRP channels are two key classes of signaling proteins that have broad roles in sensation and in controlling behavior in Ae. aegypti. Rhodopsins are the founding G-protein coupled receptors (GPCRs). We recently discovered that opsins are multi-modal sensory receptors, challenging 100 years of dogma that they detect only light. To find humans, female Ae. aegypti integrate information from diverse stimuli, including CO2, visual cues, organic molecules, and convection heat from skin. We discovered another cue. Aim 1 builds on our preliminary data that Ae. aegypti use infrared (IR) radiation as an additional host stimulus. We outline experiments to reveal the roles of opsins and the TRPA1 channel in IR detection. We propose to identify the IR-sensing neurons that express the opsins and TRPA1, and to test a model to explain the role of opsins in IR sensation. To pursue this aim, we devised a highly effective assay for monitoring IR attraction and a new molecular genetic approach to bypass difficulties in combining multiple genetic elements. Aim 2 takes advantage of a mutation that we created in another TRP (TRPV-A), which renders males and females deaf. We will test the roles of hearing and TRPV-A in swarm formation, in mating, and in finding humans. Aim 2 will also build on the observations that male mating requires audition mediated by TRPV-A to devise a strategy to overcome a major impediment limiting the efficacy of the sterile insect technique (SIT). SIT is a promising strategy to suppress Ae. aegypti. It involves inundating a local population with sterile males, which then render females sterile upon mating. An obstacle to using SIT is that wild-type males outcompete sterile males. We propose that manipulation of the activity of the TRPV-A- expressing auditory neurons elevates sterile male mating success, and will thereby increase the efficacy of SIT in suppressing Ae. aegypti. Aim 3 concerns identifying the sensory receptors for repellents. We propose to test the idea that an opsin functions as a highly sensitive receptor for insect repellents. If confirmed by the proposed experiments, this would demonstrate that opsins comprise a new class of olfactory receptor. To accomplish our goals, we have developed an extensive repertoire of state-of-the-art approaches. These include new molecular genetic tools, a suite of behavioral assays, original video tracking software, and in vivo electrophysiological recordings. In summary, this project will reveal the roles of opsins and TRP channels in allowing mosquitoes to sense humans and conspecific mates. The insights gleaned from this work have exciting potential to lead to innovative strategies to control Ae. aegypti and reduce insect-borne disease.