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: 6076334
• 负责人: JOHN I GLENDINNING
• 金额: $ 15.83万
• 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 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6076334
• 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.

人类患有多种味觉障碍，这些障碍损害了他们获取必需营养素、调节消化酶分泌以及享受生活中简单乐趣之一——吃饭的能力。 由于针对这些疾病的有效疗法很少，即将对小鼠基因组进行的全面分析为帮助填补这一空白提供了巨大的希望。 由于缺乏高通量筛选方案，味觉遗传学家识别小鼠味觉敏感性基因改变的能力受到阻碍。 结果，大多数味觉遗传学家被迫使用通量极低且解释性有限的程序，称为两瓶偏好测试。 在过去的十年中，味觉心理物理学家开发了一种评估大鼠味觉敏感性的高通量程序，称为短暂访问味觉测试。 在此过程中，动物在短暂（例如 10 秒）试验期间获得单一味觉刺激，并用自动味觉计测量舔反应。 使用该设备，人们可以在 30 分钟的测试过程中以随机顺序呈现多种味觉刺激（例如，不同浓度的蔗糖），得出强大的浓度反应函数，并相对确信舔舐反应受到口感觉控制，并且不会被味觉刺激的食后效应所混淆。 我们已成功应用这种方法来研究大鼠的味觉功能，我们在此建议将这种可靠、易于使用且高通量的程序应用于小鼠。 为此，将开发二次筛选来测试野生型和突变型 C57BL/6 小鼠。 由于该筛选将使用各种味觉刺激物（奎宁、氯化钠和蔗糖）的广泛浓度进行，因此它将能够检测任一方向味觉敏感性的总体和细微变化。 作为通过二次筛选的开发获得的知识的基础，将为初级筛选选择临界浓度，以提供相对快速的味觉敏感性评估。通过主屏幕识别的异常值可以通过辅助屏幕进行确认。 将建立两个屏幕的规范数据库并向科学界提供。 开发的测定程序将由经验有限的单个实验室技术人员应用。