Defining the Connectivity and Dynamics of Peripheral Sour Taste Circuits

定义外围酸味电路的连接性和动态

• 批准号: 10465014
• 负责人: Shannon Marie Landon
• 金额: $ 3.71万
• 依托单位: UNIVERSITY OF TEXAS SAN ANTONIO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-28 至 2025-01-27
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10465014
• 关键词: Brain; Calcium; Cell Differentiation process; Cells; Circumvallate Papilla; Complex; Consummatory Behavior; Data; Development; Engineering; Fiber; Food; Functional disorder; Fungiform Papilla; Ganglia; Goals; Grant; Half-Life; Hand; Health; Homeostasis; Hour; Image; Individual; Label; Larynx; Lead; Light; Location; Longevity; Maintenance; Malnutrition; Maps; Methods; Microscopy; Modeling; Molecular; Mus; Neurons; Nutrient; Peripheral; Population; Process; Proteins; Quality of life; Receptor Cell; SHH gene; Sampling; Signal Transduction; Site; Stimulus; Structure of geniculate ganglion; Synapses; Synaptic Cleft; Synaptic Membranes; System; Taste Buds; Taste Disorders; Taste Perception; Techniques; Technology; Testing; Time; Tissues; Toxin; Transgenic Mice; Variant; Viral; Work; cohort; density; design; experimental study; in vivo; inhibitor; innovation; nerve supply; neurogenetics; novel; precursor cell; prevent; promoter; reconstitution; sensory system; taste system; tool; two photon microscopy

Abstract The sense of taste is a complex and integrative chemosensory system that guides consummatory behavior. Taste receptor cells (TRCs) in the taste bud are continuously renewed and must reconnect with peripheral gustatory neurons (GNs) to relay taste signals to the brain. The turnover and re-establishment of peripheral taste synapses is vital to maintain this complex sensory system. Studies have demonstrated the molecular mechanisms governing the differentiation, maturation and turnover of TRCs as well as the identification of molecularly and functionally separate populations of geniculate ganglion neurons. However, the synaptic connectivity and dynamics between TRCs and GNs remains unknown due to the lack of technologies to examine them. To overcome this, I have developed GFP Reconstitution Across Synaptic Partners (GRASP) to visualize synaptic contacts in the taste bud. GRASP consists of pre- and post-GRASP split GFP molecules expressed, directed, and anchored into the synaptic membranes of pre- and post-synaptic cells, respectively. Because expression of the GRASP components can be controlled genetically, we can use it to assess the connectivity of specific populations. Moreover, GRASP produces bright fluorescent signals in native tissues which grants the unique ability to study synapse dynamics in time-lapse studies. In preliminary work, I have created transgenic mice that express the cyan variant of the pre-GRASP protein under the PKD2L1 promoter, a marker for sour TRCs. Additionally, I have engineered Flex-switch post-GRASP AAVs which enables us to express the post-synaptic GRASP in GNs. With this novel technique at hand, my goal is to genetically identify potential GN-TRC synaptic partners and to characterize synaptic dynamics during taste cell turnover, using sour TRCs as a model. To achieve these goals, this proposal will investigate two aims, 1) Identify synaptic partners for sour TRCs in the fungiform, circumvallate, and laryngeal taste buds, and 2) Determine the longevity and dynamics of sour TRC synapses over time. This proposal is designed to use GRASP, a novel and innovative technique, to elucidate synaptic connectivity and dynamics in the peripheral taste system, which will have far-reaching impact in the fields of taste development, connectivity, and taste disorders.

抽象的 味道感是一种指导完整行为的复杂而综合的化学感应系统。品尝 味蕾中的受体细胞（TRC）连续更新，必须与外围味道重新连接 神经元（GNS）将味道信号传递给大脑。周围口味突触的营业额和重新建立 维护这个复杂的感觉系统至关重要。研究表明了分子机制 管理TRC的分化，成熟和营业额以及分子和分子的识别 在功能上分开的晶神经神经元的种群。但是，突触连通性和 由于缺乏检查它们的技术，因此TRC和GN之间的动态仍然未知。到 克服这一点，我已经开发了跨突触伙伴（grasp）的GFP重建，以可视化突触 味蕾中的联系。掌握由表达，定向的格拉斯和后分裂后的GFP分子组成，并且 分别固定在前和突触后细胞的突触膜中。因为表达 可以通过遗传来控制掌握组件，我们可以使用它来评估特定人群的连通性。 此外，GRASP在天然组织中产生明亮的荧光信号，该信号具有独特的研究能力 延时研究中的突触动态。在初步工作中，我创建了表达的转基因小鼠 pkd2l1启动子下的Pre-Grasp蛋白的青色变体，酸TRC的标记。另外，我还有 工程性的弹性开关后的格子aavs使我们能够在GNS中表达突触后的掌握。和 这项手头的新技术是，我的目标是从基因上识别潜在的GN-TRC突触伙伴和 使用Sour TRC作为模型来表征味觉细胞更新过程中突触动力学的表征。为了实现这些目标， 该提案将调查两个目标，1）确定真菌状酸trc的突触伙伴， 环绕和喉味芽，以及2）确定酸TRC突触的寿命和动态 时间。该建议旨在使用Grasp（一种新颖而创新的技术）来阐明突触 外围口味系统中的连通性和动态，这将在领域具有深远的影响 品味发展，连通性和味觉障碍。