PHYSIOLOGY OF ISOLATED HUMAN TASTE RECEPTOR CELLS

分离的人类味觉受体细胞的生理学

基本信息

批准号：
6125009
负责人：
JOSEPH G BRAND
金额：
$ 7.29万
依托单位：
MONELL CHEMICAL SENSES CENTER
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-12-01 至 2000-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6125009
关键词：
calcium flux human tissue membrane channels membrane potentials method development neurophysiology taste buds tissue /cell culture voltage /patch clamp voltage gated channel

项目摘要

Little is known of the transduction mechanisms used by human taste cells. It is the purpose of this application to enhance techniques for taste cell isolation from fungiform papillae of human volunteers and to then investigate the physiological characteristics of these cells. Successful completion of this feasibility study will open new important routes for studying normal human taste response and give impetus to determining the course of human chemosensory disease. The three aims of this study are: to develop protocols to maximize viability of isolated human taste cells; to characterize stimulus-regulated changes in intracellular calcium ion activities and membrane potential in these cells; and to perform loose patch analyses on these cells to identify stimulus-regulated currents in these isolated cells. Procedures developed to date by the investigators on this proposal for isolated human taste cells have yet to be optimized, resulting in unacceptable variations among preparations. In order to allow characterization studies to proceed efficiently, isolation protocols need to be better defined. Experimentation with enzyme type/amount, trituration, media and holding conditions will be performed. Imaging studies of intracellular calcium and membrane potential using the dyes Fura 2 & di-8-ANEPPS will permit basic characterization studies to be performed. These will emphasize characterization of ion channels and normal responses of taste cells to pharmacological challenges. Responses to primarily sweet and bitter stimuli will be evaluated. Loose patch studies will be used to characterize individual cells as to taste modality and chemical specificity. Human taste cells have not been the subject of study for physiological experiments. The unique specificities of human taste can be used to test individual cells. Volunteers can be taste tested prior to have fungiform papillae removed to determine their response to particular stimuli. This research is of high impact since it deals directly with the receptor cells of humans, thereby strengthening the likelihood that therapy regimens can be developed to treat chemosensory disorders. Successful completion of this study will allow correlations to be made between psychophysical measures of taste and biophysical characteristics of the receptor cells. A more complete understanding of peripheral versus central processes of taste will be a likely outcome.

人们对人类味觉细胞使用的转导机制知之甚少。本申请的目的是增强从人类志愿者的菌状乳头中分离味觉细胞的技术，然后研究这些细胞的生理特征。这项可行性研究的成功完成将为研究人类正常味觉反应开辟新的重要途径，并为确定人类化学感应疾病的病程提供动力。这项研究的三个目标是：开发方案以最大限度地提高分离的人类味觉细胞的活力；表征这些细胞中细胞内钙离子活性和膜电位的刺激调节变化；并对这些细胞进行松散斑块分析，以确定这些分离细胞中的刺激调节电流。迄今为止，研究人员针对分离的人类味觉细胞的提议开发的程序尚未优化，导致制剂之间存在不可接受的差异。为了使表征研究有效进行，需要更好地定义分离方案。将进行酶类型/量、研磨、介质和保存条件的实验。使用染料 Fura 2 和 di-8-ANEPPS 对细胞内钙和膜电位进行成像研究将允许进行基本表征研究。这些将强调离子通道的特征和味觉细胞对药理学挑战的正常反应。将评估对主要甜味和苦味刺激的反应。松散斑块研究将用于表征单个细胞的味觉形态和化学特异性。人类味觉细胞尚未成为生理实验的研究对象。人类味觉的独特性可用于测试单个细胞。在去除菌状乳头之前，可以对志愿者进行味觉测试，以确定他们对特定刺激的反应。这项研究具有很高的影响力，因为它直接涉及人类的受体细胞，从而增强了开发治疗化学感应疾病的治疗方案的可能性。这项研究的成功完成将能够在味觉的心理物理测量与受体细胞的生物物理特征之间建立关联。可能的结果是更全面地了解味觉的外围过程和中枢过程。