HIGH-THROUGHPUT SCEENS FOR ASSESSING TASTE SENSITIVITY

用于评估味觉敏感性的高通量屏幕

• 批准号: 6379551
• 负责人: JOHN I GLENDINNING
• 金额: $ 17.08万
• 依托单位: BARNARD COLLEGE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-27 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6379551
• 关键词: behavior test; behavioral /social science research tag; computer program /software; gene mutation; gene targeting; genetic screening; genetic strain; genetically modified animals; inbreeding; information systems; laboratory mouse; nutrition related tag; oral behavior; phenotype; psychophysiology; quinine; site directed mutagenesis; sodium chloride; stimulus /response; sucrose; taste; taste disorders; taste threshold; water drinking behavior

Humans suffer from a variety of taste disorders that impair their ability to acquire essential nutrients, regulate secretion of digestive enzymes, and enjoy one of life's simple pleasures-- eating. As there are few effective therapies for these disorders, the impending comprehensive analysis of the mouse genome offers great promise for helping fill this void. The ability of taste geneticists to identify genetically-based alterations in murine taste sensitivity has been impeded by the absence of a high-throughput screening protocol. As a result, most taste geneticists are forced to use an extremely low- throughput and interpretively limited procedure called 2-bottle preference testing. Over the last decade, taste psychophysicists have developed a high-throughput procedure for assessing taste sensitivity in rats, called brief-access taste testing. In this procedure, animals are provided with access to a single taste stimulus during brief (e.g., 10 sec) trials, and licking responses are measured with an automated gustometer. Using this device, one can present several taste stimuli (e.g., different concentrations of sucrose) during a 30-min test session in a randomized order, derive a robust concentration-response function, and be relatively confident that the licking responses are under orosensory control and not confounded by postingestive effects of the taste stimulus. We have successfully applied this methodology to study taste function in rats, and we propose here to adapt this reliable, easy-to-use, and high-throughput procedure to mice. To this end, a secondary screen will be developed for testing wild-type and mutant C57BL/6 mice. Because this screen will be conducted with an expansive array of concentrations of each taste stimulus (quinine, NaCl, and sucrose), it will be able to detect gross and subtle alterations in taste sensitivity in either direction. One the basis of the knowledge gained through the development of the secondary screen, critical concentrations will be chosen for a primary screen to provide a relatively quick assessment of taste sensitivity. Outliers identified with the primary screen can be subjected to the secondary screen for confirmation. A normative data-base for both screens will be established and made available to the scientific community. The assay procedures developed will be applicable by a single laboratory technician with limited experience.

人类患有多种味道障碍，这些味道障碍会损害其获得必需营养素，调节消化酶的分泌以及享受生活中一种简单的乐趣之一 - 饮食的能力。 由于对于这些疾病的有效疗法很少，因此对小鼠基因组的全面分析为帮助填补这一空隙提供了巨大的希望。 没有高通量筛查方案，味觉遗传学家鉴定基于遗传学的改变的能力受到阻碍。 结果，大多数味觉遗传学家被迫使用极低的吞吐量和解释性限制的程序，称为2瓶偏好测试。 在过去的十年中，味觉心理物理学家开发了一种高通量程序，用于评估大鼠的味觉敏感性，称为简短访问味觉测试。 在此过程中，在简短试验（例如10秒）试验中，为动物提供了单个味道刺激的机会，并通过自动阵风测量舔反应。 使用该设备，可以在30分钟的测试过程中以随机顺序呈现几种味道刺激（例如，不同浓度的蔗糖），得出强大的浓度 - 响应功能，并相对确信舔反应在口感控制下控制，并且不会因味觉刺激的后效果而混淆。 我们已经成功地应用了这种方法来研究大鼠的味觉功能，并在此提议将这种可靠，易于使用和高通量程序调整到小鼠中。 为此，将开发一个辅助屏幕，用于测试野生型和突变体C57BL/6小鼠。 因为该屏幕将以每种口味刺激（奎宁，NACL和蔗糖）的浓度浓度进行进行进行，因此它将能够检测到任何方向上的味觉敏感性的毛刺和微妙的变化。 通过二级屏幕的开发获得的知识的基础之一，将选择临界浓度作为主要屏幕，以提供相对快速的味觉敏感性评估。用主屏幕确定的离群值可以接受辅助屏幕以进行确认。 两个屏幕的规范性数据库将被建立并提供给科学界。 开发的测定程序将由具有有限经验的单个实验室技术人员适用。