Gustatory coding in brainstem nuclei using a novel c-Fos-tau-LacZ mouse model

使用新型 c-Fos-tau-LacZ 小鼠模型进行脑干核中的味觉编码

• 批准号: 8332526
• 负责人: Jennifer Stratford
• 金额: $ 5.39万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8332526
• 关键词: Animals; Area; Behavioral; Brain; Brain Stem; Cell Nucleus; Cells; Cellular Morphology; Characteristics; Code; Cytoplasm; Data; Detection; Development; Eating; Esthesia; Etiology; FOS Protein; FOS gene; Fingers; Food; Food Preferences; Genetic; Health; Hour; Imagery; Immediate-Early Genes; Individual; Individual Differences; Knock-out; Knockout Mice; LacZ Genes; Link; Maps; Measures; Medial; Mediating; Messenger RNA; Methods; Modality; Morphology; Mus; Nerve; Neural Pathways; Neurons; Nuclear; Nucleus solitarius; Nutrient; Obesity; Overweight; Patients; Pattern; Peripheral Nerves; Personal Communication; Play; Population; Role; Satiation; Savory; Sensory; Sodium Glutamate; Spatial Distribution; Staining method; Stains; Stimulus; Sucrose; System; Taste Perception; Techniques; Time; Transgenic Mice; beta-Galactosidase; driving force; hedonic; immunoreactivity; insight; mRNA Expression; mouse model; novel; obesity treatment; preference; receptor; regional difference; relating to nervous system; response; sensory system; tau Proteins; tau expression

Project Summary Obesity is a major health issue with approximately two thirds of the U.S. population considered overweight and a third of those classified as obese. Overconsumption of high calorie foods contributes significantly to the development of obesity. Thus, identification of factors that influence food choice is important for the development of effective obesity treatments. In this regard, the sense of taste has a direct impact on food preference. Therefore, the neural pathways involved in how the brain represents taste can contribute to our understanding of the etiology of obesity. One of the most effective and widely-used methods to determine the neural substrate that underlies sensation of taste is visualization of the spatial patterns of expression of the activity-dependent marker, c-Fos. Recently, the Fos-tau-LacZ transgenic mouse line was developed in which activation of the c-Fos gene results in expression of tau-linked Beta-galactosidase (Beta-gal) in the cytoplasm of activated cells; thereby enabling the examination of the morphology of c-Fos positive cells. Furthermore, the perdurance of Beta-gal in these transgenic mice is much longer than c-Fos protein itself; Beta-gal can be seen up to 24 hours post stimulation in the brainstem, whereas c-Fos protein lasts 1-3 hours and c-Fos mRNA peaks 30 min. post stimulation. This difference in timing between Beta-gal and mRNA purdurance enables visualization of c-Fos expression in response to two stimuli, separated temporally, within the same animal (e.g. Beta-gal) expression and c-Fos mRNA expression). Accordingly, I can assess whether individual cells respond to multiple taste stimuli. The current proposal will use Fos-tau-LacZ transgenic mice to quantify morphological and spatial differences in cellular activation (e.g. Beta-gal expression and c-Fos mRNA) in brainstem taste nuclei in response to stimuli of different modalities (sweet, ¿umami¿/savory and bitter) and different taste hedonics (appetitive vs. aversive). Moreover, the proposal will also assess changes in this central taste representation following loss of sweet- and umami- specific sensory input via deletion of the T1R3 receptor- an obligatory subunit for the detection of these qualities. The data obtained from this proposal will provide valuable insight into the spatial and morphological organization of taste representation within the nucleus of the solitary tract.

项目概要 肥胖是一个主要的健康问题，大约三分之二的美国人口被认为超重，其中三分之一被归类为肥胖。过度食用高热量食物会显着导致肥胖的发生。因此，确定影响食物选择的因素对于开发有效的肥胖治疗方法非常重要。就此而言，味觉对食物偏好有直接影响。因此，大脑如何代表味觉所涉及的神经通路有助于我们理解肥胖的病因。确定味觉基础神经底物的最有效和最广泛使用的方法之一是可视化活动依赖性标记物 c-Fos 表达的空间模式。 最近，开发了Fos-tau-LacZ转基因小鼠系，其中c-Fos基因的激活导致tau连接的β-半乳糖苷酶（Beta-gal）在激活细胞的细胞质中表达；从而能够检查c-Fos阳性细胞的形态。此外，Beta-gal 在这些转基因小鼠中的持久性比 c-Fos 蛋白本身长得多；脑干刺激后 24 小时内可以看到 Beta-gal，而 c-Fos 蛋白可持续 1-3 小时，c-Fos mRNA 在 30 分钟达到峰值。刺激后。 Beta-gal 和 mRNA 表达之间的这种时间差异使得能够在同一动物内对 c-Fos 表达响应于时间上分开的两种刺激进行可视化（例如 Beta-gal 表达和 c-Fos mRNA 表达）。因此，我可以评估单个细胞是否对多种味觉刺激做出反应。 目前的提案将使用 Fos-tau-LacZ 转基因小鼠来量化脑干味觉核中细胞激活（例如 Beta-gal 表达和 c-Fos mRNA）的形态和空间差异，以响应不同形式（甜味、“鲜味”/咸味和苦味）和不同味觉享乐（食欲与厌恶）的刺激。此外，该提案还将评估通过删除 T1R3 受体（检测这些品质的必需亚基）而失去甜味和鲜味特异性感官输入后，这一中心味觉表征的变化。从该提议中获得的数据将为了解孤束核内味觉表征的空间和形态组织提供有价值的见解。