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Opsins as a new class of evolutionarily conserved taste receptors: Diversity Supplement for Pre-Doctoral Training

视蛋白作为一类新的进化保守味觉受体：博士前培训的多样性补充

基本信息

批准号：
10750801
负责人：
CRAIG MONTELL
金额：
$ 6.05万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA BARBARA
依托单位国家：
美国
项目类别：
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/10750801
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 Training 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 pre-doctoral receptor response sugar tool transcriptome transcriptome sequencing vector

项目摘要

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.

抽象的视蛋白是经典的 G 蛋白偶联受体，一个多世纪以来一直被认为具有功能专门用作光探测器。然而，利用果蝇，我们发现了多种与光无关的作用用于视蛋白，包括检测苦味剂。这一发现提出了令人兴奋的问题，我们建议地址在这里。视蛋白只对苦味化合物有反应，还是它们在味觉上发挥更广泛的作用感觉？特别有趣的是，视蛋白在其他动物（包括蚊子）中充当味觉受体吗？埃及伊蚊（Ae.）？该载体传播导致登革热、黄热病和其他疾病的病毒每年困扰数百万人。雄性和雌性伊蚊都利用味觉来识别花蜜并避免有毒物质化学物质，而女性则使用多种感官来识别需要血粉的人。然而，需要的受体蚊子的味道很大程度上是未知的。另一个有趣的问题是视蛋白是否起到味觉的作用哺乳动物体内的受体。为了解决这些问题，我们建议采用一系列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、和最先进的分子遗传学。我们还将利用我们在果蝇味觉、视蛋白和我们最近在AE中进行分子遗传学和行为的经验。埃及伊蚊和小鼠。目标1将定义果蝇视蛋白在氨基酸味觉中的作用，在果蝇中，氨基酸味觉是一种知之甚少的味觉模式。如果果蝇的饮食缺乏氨基酸，对氨基酸的吸引力就会增加。我们概述了实验定义视蛋白对氨基酸吸引力动态变化的贡献。我们还将测试建议视蛋白直接被氨基酸激活。目标 2 将建立在我们两个 Ae 的初步数据之上。埃及伊视蛋白在主要味觉器官中表达，是感知类黄酮所必需的。的目标这个目的是揭示这些视蛋白在体外和体内的化学特异性，确定它们是否起作用独立或一起，并识别这些视蛋白在其中发挥作用的味觉受体神经元。目标 3 将描述视蛋白在哺乳动物味觉中的作用。这一目标得到了初步数据的支持，显示多种小鼠视蛋白在小鼠舌头中表达，并在体外被苦味化合物激活，包括没有已知受体的类黄酮。此外，我们还提供了初步数据视蛋白突变体对这种黄酮类化合物的反应受到损害。我们建议检验该视蛋白的假设在感知黄酮类化合物方面具有广泛的功能，识别表达这种和其他黄酮类化合物的味觉受体细胞 “味觉视蛋白”，并对突变体进行行为测试，以确定每种视蛋白的要求用于检测苦味化合物。该项目的结果将视蛋白确立为一种新的味觉类别从昆虫到哺乳动物都保守的受体。我们建议将视蛋白定义为第一个苦味受体蚊子提供了利用这些信息来操纵蚊子行为的潜力，从而减少它们的数量，从而减少虫媒疾病。