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.

人类患有各种味觉障碍，这些疾病损害了他们获取必需营养素、调节消化酶分泌和享受生活中简单乐趣之一--吃的能力。 由于对这些疾病几乎没有有效的治疗方法，即将进行的小鼠基因组全面分析为帮助填补这一空白提供了巨大的希望。 味觉遗传学家鉴定小鼠味觉敏感性中基于遗传的改变的能力由于缺乏高通量筛选方案而受到阻碍。 因此，大多数味觉遗传学家被迫使用一种极低通量和解释有限的程序，称为2瓶偏好测试。 在过去的十年里，味觉心理学家已经开发出一种高通量的程序来评估大鼠的味觉敏感性，称为味觉接近测试。 在该程序中，在短暂的过程中向动物提供单一味觉刺激（例如，10秒）试验，并且用自动味觉计测量舔体反应。 使用该装置，可以呈现几种味觉刺激（例如，不同浓度的蔗糖），得到一个强有力的浓度-反应函数，并且相对确信舔体反应是在口腔感觉控制下的，并且不会被味觉刺激的摄食后效应所混淆。 我们已经成功地应用这种方法来研究大鼠的味觉功能，我们建议在这里适应这种可靠的，易于使用的，高通量的程序，小鼠。 为此，将开发用于测试野生型和突变型C57 BL/6小鼠的二次筛选。 因为这个屏幕将与每一个味道刺激（奎宁，NaCl和蔗糖）的浓度的扩展阵列进行，它将能够检测在任何方向的味觉敏感性的总体和微妙的变化。 在通过开发二级筛选获得的知识的基础上，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。通过初步筛选确定的离群值可进行二次筛选以进行确认。 将为这两种筛查建立一个规范性数据库，并提供给科学界。 开发的测定方法将适用于经验有限的实验室技术人员。