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: 6176690
• 负责人: JOHN I GLENDINNING
• 金额: $ 16.45万
• 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/6176690
• 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.

人类患有各种各样的味觉障碍，损害了他们获取必需营养、调节消化酶分泌的能力，以及享受生活中最简单的乐趣之一——吃。由于这些疾病几乎没有有效的治疗方法，即将进行的对小鼠基因组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变化的能力受到阻碍。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，味觉心理物理学家开发了一种高通量的方法来评估大鼠的味觉敏感性，称为短暂接触味觉测试。在这个过程中，在短暂的（例如10秒）试验中，给动物提供单一味觉刺激，并用自动味觉仪测量舔舐反应。使用该装置，可以在30分钟的测试过程中以随机顺序呈现几种味觉刺激（例如，不同浓度的蔗糖），得出一个强大的浓度-反应函数，并且相对确信舔舐反应是在口腔感官控制下的，不会被味觉刺激的进食后效应所混淆。我们已经成功地将这种方法应用于大鼠的味觉功能研究，我们建议将这种可靠、易于使用和高通量的方法应用于小鼠。为此，将开发用于检测野生型和突变型C57BL/6小鼠的二次筛选。因为这种筛选将通过各种浓度的味觉刺激（奎宁、氯化钠和蔗糖）来进行，它将能够检测到味觉敏感性在任何方向上的明显和细微的变化。在通过二级筛选获得的知识的基础上，将为一级筛选选择临界浓度，以提供相对快速的味觉敏感性评估。通过主屏幕识别出的异常值可以通过次屏幕进行确认。将为这两种筛选建立一个规范的数据库，并提供给科学界。开发的分析程序将由经验有限的单个实验室技术人员适用。