AGONISTS/ANTAGONISTS FOR CHARACTERIZING CHEMORECEPTORS

用于表征化学受体的激动剂/拮抗剂

基本信息

批准号：
2125450
负责人：
BRUCE P BRYANT
金额：
$ 17.41万
依托单位：
MONELL CHEMICAL SENSES CENTER
依托单位国家：
美国
项目类别：
财政年份：
1985
资助国家：
美国
起止时间：
1985-09-16 至 1994-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2125450
关键词：
alternatives to animals in research aminoacid biological signal transduction catfish chemical binding chemoreceptors electrophysiology fresh water environment laboratory mouse monoclonal antibody neurochemistry neurophysiology olfactions receptor stimulus /response taste tissue /cell culture

项目摘要

The senses of taste and olfaction are intimately involved in food selection, identification digestion and metabolism, as well as in reproduction (olfaction) and the general awareness and appreciation of the environment. Loss or alteration in function of these senses can lead to a nutritional compromises and to a decrease in the quality of life. The overall goal of our work has been the understanding of these senses at the molecular level, so that therapies for deficiencies of these sense may be approached from knowledgeable and rational bases. The current research proposal emphasizes production of antagonists and agonist to qustatory and olfactory receptors, and the use of these in characterizing the receptor binding proteins. The techniques of biochemistry, neurophysiology, synthetic organic chemistry and cell culture are central to the integrated collaborative approaches used to address the aims of this work. Our experimental model, the catfish, I. punctatus, possesses unique and identifiable receptors for various chemostimualtrou amino acids. This model has proved invalualboej to understanding the fundamental mechanisms of taste and olfaction. The specific aims of this proposal seek to (1) identify novel agonist and antagonists to taste and olfactory receptors form three major molecular classes - amino acid analogy (including photoaffinity probes), monoclonal antibodies and lectins; (2) characterize these agonist. antagonists using neurophysiological and biochemical binding assays, defining their specificity, mode of action and inhibitory kinetic; and (3) utilize these parve in characterizing events in reception and transduction and in labeling and identifying receptor proteins after electrophoresis. These aims will be accomplished through a series of experimental procedures that emphasize multidisiplinary approach. Specificity, mode of action and type of inhibition are studied using both stimulatory effectiveness and cross-adaptation procedures in a neurophysiological assay and competitive binding in biochemical assays. The resultant characterized probes and competitive binding in biochemical assays. The resultant characterized probe are then used to define the receptors as to subcalsand selectivity. Future studies will emphasize use of these proves in purifying and localizing receptors proteins in situ-upon completion of this study, we will have produced the chemical probes necessity for receptor identification, purification and documentation, and have achieved a more complete understanding of the receptor mechanisms in taste and olfaction.

味觉和嗅觉与食物密切相关选择、识别、消化和代谢，以及繁殖（嗅觉）和一般意识和欣赏环境的。这些感官功能丧失或改变可能会导致营养不良并导致生活质量。我们工作的总体目标是在分子水平上理解这些感觉，以便针对这些感觉缺陷的治疗方法可以从知识和理性的基础。目前的研究计划强调 qustatory 的拮抗剂和激动剂的产生嗅觉受体，以及它们在表征受体结合蛋白。生物化学技术，神经生理学、合成有机化学和细胞培养用于解决问题的综合协作方法的核心这项工作的目的。我们的实验模型，鲶鱼，I。 punctatus，拥有独特且可识别的各种受体化学刺激氨基酸。事实证明这个模型是无价的了解味觉和嗅觉的基本机制。该提案的具体目标旨在 (1) 确定新颖的味觉和嗅觉受体的激动剂和拮抗剂形成三主要分子类别 - 氨基酸类比（包括光亲和探针）、单克隆抗体和凝集素； (2) 表征这些激动剂。利用神经生理学的拮抗剂和生化结合测定，定义其特异性、模式作用和抑制动力学； (3)利用这些parve 表征接收、转导和标记中的事件并在电泳后鉴定受体蛋白。这些目标将通过一系列实验来实现强调多学科方法的程序。特异性，使用两者研究作用方式和抑制类型刺激有效性和交叉适应程序神经生理学测定和生化竞争性结合化验。由此产生的表征探针和竞争性结合在生化分析中。然后得到的特征探针是用于定义受体的下层和选择性。未来研究将强调使用这些证据来纯化和完成此操作后将受体蛋白定位在原位研究，我们将生产出必要的化学探针受体鉴定、纯化和记录，并具有对受体机制有了更全面的了解在味觉和嗅觉上。