Opsins as a new class of evolutionarily conserved taste receptors

视蛋白作为一类新的进化上保守的味觉受体

• 批准号: 10657704
• 负责人: CRAIG MONTELL
• 金额: $ 44.12万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA BARBARA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10657704
• 关键词: Address; Aedes; Amino Acids; Animals; Behavior; Behavioral Assay; Biological Assay; Blood; CRISPR/Cas technology; Catechin; Cellular biology; Chemicals; Clustered Regularly Interspaced Short Palindromic Repeats; Culicidae; Data; Dengue; Detection; Development; Diet; Disease; Disease Vectors; Drosophila genus; Electrophysiology (science); Esthesia; Evolution; Family; Female; Flavonoids; Fruit; G-Protein-Coupled Receptors; Genetic; Goals; Grapefruit; Health; Homologous Gene; Human; Image; Impairment; In Vitro; Insecta; Ligand Binding; Ligands; Light; Mammals; Modality; Modeling; Molecular Genetics; Motivation; Mus; Mutagenesis; Mutant Strains Mice; Mutation; Neurons; Nutrient; Nutritional; Opsin; Organ; Partner in relationship; Persons; Photoreceptors; Plants; Poison; Population; Production; Property; Proteins; Receptor Activation; Receptor Cell; Rhodopsin; Role; Specificity; Taste Buds; Taste Perception; Testing; Tongue; Transgenic Organisms; Vertebrate Photoreceptors; Virus; Visual System; Vitamin A; Work; Yellow Fever; ZIKA; antagonist; behavior test; chromophore; defined contribution; detector; dietary; egg; experience; experimental study; fascinate; feeding; fly; genetic manipulation; human disease; improved; in vitro Assay; in vivo; insect disease vector; interest; male; member; mutant; receptor; response; sugar; tool; transcriptome; transcriptome sequencing; vector

Abstract Opsins are classical G-protein coupled receptors, which for more than a century have been thought to function exclusively as light detectors. However, using Drosophila, we have uncovered multiple, light-independent roles for opsins including detection of bitter tastants. This discovery raises exciting questions, which we propose to address here. Do opsins only respond to bitter compounds, or do they function more broadly in taste sensation? Of particular interest, do opsins serve as taste receptors in other animals, including the mosquito, Aedes (Ae.) aegypti? This vector spreads viruses that cause dengue, yellow fever, and other diseases that afflict millions of people each year. Both male and female Aedes use taste to identify nectar and avoid toxic chemicals, while females use many senses to identify people for blood meals. However, the receptors required for mosquito taste are largely unknown. Another fascinating question is whether opsins serve as taste receptors in mammals. To address these questions, we propose to employ an extensive suite of approaches including electrophysiology, Ca2+ imaging, behavioral assays, cell biology, an in vitro receptor activation assay, and state-of-the art molecular genetics. We will also bring to bear our expertise in Drosophila taste, opsins, and our recent experience performing molecular genetics and behavior in Ae. aegypti and in the mouse. Aim 1 will define the role of a Drosophila opsin in amino acid taste, which in flies is a poorly understood taste modality. Attraction to amino acids increases if the flies are kept on an amino acid deficient diet. We outline experiments to define the contribution of the opsin to this dynamic change in amino acid appeal. We will also test the proposal that the opsin is activated directly by amino acids. Aim 2 will build on our preliminary data that two Ae. aegypti opsins are expressed in the major taste organ, and are required for sensing a flavonoid. The goals of this aim are to reveal the chemical specificity of these opsins in vitro and in vivo, determine whether they act independently or together, and identify the gustatory receptor neurons in which these opsins function. Aim 3 will characterize the roles of opsins in mammalian taste. This aim is supported by preliminary data showing that multiple mouse opsins are expressed in the mouse tongue, and are activated in vitro by bitter compounds, including a flavonoid for which there is no known receptor. In addition, we present preliminary data that the response to this flavonoid is impaired in the opsin mutant. We propose to test the hypothesis that this opsin functions broadly in sensing flavonoids, identify the taste receptor cells that express this and the other “gustatory opsins,” and perform behavioral tests with mutants to characterize the requirements for each opsin for sensing bitter compounds. The results from this project would establish opsins as a new class of taste receptor conserved from insects to mammals. We propose that defining opsins as the first bitter receptors in mosquitoes offers the potential to harness this information to manipulate mosquito behavior and thereby reduce their populations and thus insect-borne disease.

抽象的 OPSIN是经典的G蛋白耦合受体，在一个多世纪以来，它可以发挥作用 仅作为光检测器。但是，使用果蝇，我们发现了多个独立的角色 对于术语，包括检测苦味。这一发现提出了令人兴奋的问题，我们建议 地址在这里。 Opsins仅对苦味化合物反应，或者它们在口味上的功能更广泛 感觉？特别感兴趣的是，Opsins在包括蚊子在内的其他动物中充当味觉受体 埃及埃及？该矢量传播了引起风扇，黄热病和其他疾病的病毒 每年折磨数百万的人。雄性和雌性艾德斯都使用味道来识别花蜜并避免有毒 化学物质，而女性则使用许多感觉来识别血液餐的人。但是，接收者需要 因为蚊子的味道在很大程度上是未知的。另一个有趣的问题是Opsins是否充当口味 哺乳动物的受体。为了解决这些问题，我们建议采用广泛的方法 包括电生理学，CA2+成像，行为测定，细胞生物学，体外受体激活测定法， 和最先进的分子遗传学。我们还将带来我们在果蝇的口味，opsin和Opsins和 我们最近在AE中执行分子遗传学和行为的经验。埃及和鼠标。目标1意志 定义果蝇蛋白在氨基酸味道中的作用，果蝇在苍蝇中是一种鲜为人知的味道形态。 如果将果蝇保留在氨基酸缺乏饮食中，则对氨基酸的吸引力会增加。我们概述了实验 为了定义Opsin对氨基酸外观这种动态变化的贡献。我们还将测试 提议OPSIN被氨基酸直接激活。 AIM 2将基于我们两个AE的初步数据。 埃及Opsins在主要味道器官中表达，是感测类黄酮所必需的。目标 此目的是揭示这些在体外和体内这些蛋白酶的化学特异性，确定它们是否作用 独立或一起识别这些Opsins起作用的味觉受体神经元。目标3 将表征Opsin在哺乳动物味道中的作用。该目标由显示的初步数据支持 多种小鼠蛋白在小鼠的舌头中表达，并被苦的化合物在体外激活， 包括没有已知受体的类黄酮。此外，我们介绍了初步数据 对这种类黄酮的反应在Opsin突变体中受损。我们建议检验这种opsin的假设 在传感类黄酮中广泛发挥作用，识别表达这一点的味觉受体细胞 “味道蛋白”，并用突变体进行行为测试，以表征每种opsin的需求 用于感知苦涩的化合物。该项目的结果将建立Opsins作为新的品味类 从昆虫到哺乳动物的受体。我们建议将Opsins定义为第一个苦涩的接收者 蚊子提供了利用这些信息来操纵蚊子行为的潜力 减少其人群，从而减少昆虫传播疾病。