Mechanisms of visual transduction and light-driven behaviors in Aedes aegypti

摘要 这个项目的目标是通过研究视觉的分子基础，从根本上开辟新的领域。 日间蚊媒伊蚊Aedes（Ae.）埃及人。这种蚊子是一种主要的疾病媒介， 导致登革热、黄热病、寨卡病毒和其他影响数千万人的疾病的病毒 每年.由于气候变化和旅行，这种入侵物种正在向新的地点扩散，包括 在美国的多个地区。因此，控制蚊媒疾病的创新策略是 迫切需要。女性Ae。埃及人探测并整合多种感官线索来定位人类宿主。一旦 它们能在10-15米的距离内感知到人类呼吸发出的二氧化碳气味， 对潜在宿主的关注大大增加。然后，他们依赖视觉信息沿着与其他人类- 衍生线索，如有机嗅觉刺激，以家庭在人。尽管做出了重要贡献， 愿景在提升Ae.埃及寻找人类，一直没有全面的方法， 应用分子遗传学来确定视觉和气味刺激视觉引导宿主的潜在机制 景点拟议研究的目标是解决这一差距。目标1是确定 TRP和TRPL通道在伊蚊视觉。我们最近使用CRISPR/Cas9来产生突变， 破坏Trp和Trpl基因。我们建议检查这些通道的光响应中的作用， 感光细胞我们还概述了实验，以调查这些渠道在几个潜在的作用 视觉和光驱动的行为，包括二氧化碳刺激的视觉引导的目标吸引。目标2的目标是 为了验证这两个磷脂酶C基因在Ae.埃及（挪威国家石油管理局和 PLC 21 C）在视觉传导和多种视觉和光驱动行为中具有不同的作用，包括 昼夜节律我们还将讨论突变破坏norpA和plc 21 C组合的影响， 突变破坏了隐花色素，隐花色素在昆虫大脑中编码一种光敏蛋白。AE. 埃及人最活跃，主要在日出后和日落前咬人。因此了解 在这种生物体中，昼夜节律的调节是重要的，并且视网膜蛋白对此的贡献 行为代表了我们对Ae生物学理解的另一个主要差距。埃及人。目标3侧重于 测试Gaq在光响应中的偶像角色。除了作为效应器的经典作用外， 为了研究视紫红质的蛋白质，我们将测试Gaq直接调节TRP和TRPL通道的想法。到 为了实现我们的目标，我们建议采用多学科的方法，包括电生理学， 分子遗传学、生物化学、细胞生物学和多种行为测定。我们建议，这些 研究将为设计创新战略提供概念框架， 蚊子有效地定位宿主和传播疾病。

Abstract The goal of this project is to break fundamentally new ground by investigating the molecular basis of vision in the daytime mosquito vector, Aedes (Ae.) aegypti. This mosquito is a major disease vector, which transmits the viruses that cause dengue, yellow fever, Zika and other diseases that affect many tens of millions of people each year. Due to climate change and travel, this invasive species is spreading to new locations, including multiple areas in the United States. Therefore, innovative strategies to control mosquito borne disease are urgently needed. Female Ae. aegypti detect and integrate multiple sensory cues to locate human hosts. Once they sense CO2 odor plumes emanating from human breath at distances of up to 10-15 meters, their visual attention to potential hosts is greatly increased. They then rely on visual information along with other human- derived cues, such as organic olfactory stimuli, to home in on people. Despite the important contribution of vision in promoting the ability of Ae. aegypti to find humans, there has been no comprehensive approach to apply molecular genetics to define the mechanisms underlying vision and odor-stimulated vision-guided host attraction. The objective of the proposed research is to address this gap. Aim 1 is to define the roles of the TRP and TRPL channels in Aedes vision. We have recently used CRISPR/Cas9 to generate mutations that disrupt the trp and trpl genes. We propose to examine the roles of these channels for the light response in photoreceptor cells. We also outline experiments to investigate potential roles for these channels in several vision- and light-driven behaviors, including CO2-stimulated vision-guided target attraction. The goal of aim 2 is to test the idea that the two phospholipase C genes expressed in the eyes of Ae. aegypti (NORPA and PLC21C) have distinct roles in visual transduction and in multiple vision- and light-driven behaviors, including circadian rhythms. We will also address the impact of mutations disrupting norpA and plc21C in combination with mutations that disrupt cryptochrome, which encodes a light sensitive protein in the insect brain. Ae. aegypti are most active and bite primarily after sunrise and before sunset. Therefore, understanding the regulation of circadian rhythms in this organism is important, and the contributions of retinal proteins to this behavior represents another major gap in our understanding of the biology of Ae. aegypti. Aim 3 focuses on testing an iconoclastic role for the Gaq in the light response. In addition to its classical role as an effector protein for rhodopsin, we will test the idea that Gaq directly regulates the TRP and TRPL channels. To accomplish our goals, we propose to employ a multidisciplinary approach, including electrophysiology, molecular genetics, biochemistry, cell biology and a wide diversity of behavioral assays. We propose that these studies will provide the conceptual framework for devising innovative strategies to limit the ability of these mosquitoes from efficiently locating hosts and spreading disease.