Ligand selectivity of vomeronasal receptors

描述（由申请人提供）：犁鼻器官中表达的三种受体家族（v1rs， V2Rs和fprs）的配体选择性在很大程度上是未知的。为了加深我们对犁鼻受体与其配体之间相互作用的理解，我们建议建立系统来识别V2Rs和fpr的活性配体。此外，我们建议确定受体蛋白的哪些结构域决定配体选择性。为了验证特异性V2R或FPR犁鼻受体被一组特定配体激活的假设，提出了三个特定目的：表征V2Rs的配体选择性。建立V2Rs的异源表达体系。我们将使用该系统来识别V2R的活性配体，并研究V2R家族成员对配体选择性的结构基础。特异性目的2：表征VNO-FPRs的配体选择性。我们将建立VNO-FPRs的异源表达系统。我们将使用该系统来识别VNO-FPRs的活性配体。特异性目的3：利用VNO感觉神经元表征VNO受体的配体选择性。我们将建立一个系统来测量解离型犁鼻神经元的反应，然后测量相同细胞中的mRNA表达。我们将使用这种方法来鉴定在给定配体激活的细胞中表达的受体。提出的实验进一步我们的长期目标，这是了解犁鼻受体的配体识别的机制基础。这些研究与健康的关系在于，更好地了解犁鼻受体的配体结合特性，将有助于深入了解g蛋白偶联受体（一类经常被药物靶向的受体）识别配体的一般特征。这项工作将提高我们对G蛋白偶联犁鼻受体如何识别和区分特定配体的理解。这些基本知识将转化为对G蛋白偶联受体的更深入了解，G蛋白偶联受体是心脏、精神和癌症疾病状态的主要药物靶点。对配体和受体之间相互作用的理解提供了开发广泛的健康相关产品所需的关键信息。

DESCRIPTION (provided by applicant): The ligand selectivity of the three families of receptors expressed in the vomeronasal organ-V1Rs, V2Rs and FPRs-is largely unknown. To deepen our understanding of the interactions between the vomeronasal receptors and their ligands, we propose to establish systems to identify active ligands for the V2Rs and the FPRs. In addition, we propose to identify which domains of the receptor proteins determine ligand selectivity. To test the hypothesis that specific V2R or FPR vomeronasal receptors are activated by specific set of ligands, three Specific Aims are proposed: Specific Aim 1. Characterize the ligand selectivity of V2Rs. We will establish a heterologous expression system for the V2Rs. We will use this system to identify active ligands of the V2Rs and examine the structural basis of ligand selectivity by V2R family members. Specific Aim 2: Characterize the ligand selectivity of VNO-FPRs. We will establish a heterologous expression system for the VNO-FPRs. We will use this system to identify the active ligands of VNO-FPRs. Specific Aim 3: Characterize the ligand selectivity of VNO receptors using VNO sensory neurons. We will establish a system for measuring the response of dissociated vomeronasal neurons using calcium imaging and then measuring the mRNA expression in the same cells. We will use this method to identify the receptors expressed in the cells activated by a given ligand. The proposed experiments further our long term goal, which is to understand the mechanistic basis for ligand recognition by the vomeronasal receptors. The health relatedness of these studies is that a better understanding of the ligand binding attributes of vomeronasal receptors will offer insights into the general features of ligand recognition by G-protein coupled receptors, a class of receptor that is frequently targeted by drugs. This work will improve our understanding of how G protein-coupled vomeronasal receptors recognize and discriminate specific ligands. This basic knowledge will translate to greater understanding of G protein-coupled receptors, which are major pharmaceutical targets for cardiac, psychiatric, and cancer disease states. An understanding of the interactions between the ligand and receptor provides crucial information needed to develop a wide range of health-related products.