A new method to measure taste bud function

测量味蕾功能的新方法

基本信息

批准号：
7577508
负责人：
STEPHEN D. ROPER
金额：
$ 31.16万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-04-01 至 2011-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7577508
关键词：
Acetylcholine Address Adrenergic Agents Affinity Bathing Biosensor Calcium Cell Communication Cell secretion Cells Chemical Synapse Chinese Hamster Ovary Cell Cholecystokinin Dependence Dependency Dyes Engineering Extracellular Fluid Fiber Fungiform Papilla Fura-2 Gap Junctions Glutamates Grant Image In Situ Hybridization Logic Measures Mediating Membrane Methodology Methods Monitor Mus Muscarinics Nature Nerve Fibers Neurotransmitters Norepinephrine Organ Output Palate Paracrine Communication Peptides Peripheral Rattus Receptor Cell Research Research Personnel Retina Sensory Sensory Receptors Serotonin Signal Transduction Specific qualifier value Stimulus Synapses Synaptic Transmission Taste Bud Cell Taste Buds Taste Perception Testing United States National Institutes of Health Visual adrenergic afferent nerve computerized data processing extracellular information processing intercellular communication interstitial meetings neurotransmitter release novel paracrine presynaptic programs promoter receptor research study response

项目摘要

The long-term objectives of this proposal are twofold: (1) to identify neurotransmitters released from rat and mouse taste bud cells during gustatory stimulation and (2) to investigate signal processing within the taste bud. These both are significant unanswered questions in the field of chemosensory reception. To date, there is only indirect evidence for candidate neurotransmitters in taste cells, including serotonin, glutamate, norepinephrine, acetylcholine, ATP, and peptides. We have developed a novel methodology to dentify the neurotransmitter(s) released from rat and mouse taste buds during gustatory stimulation. Our methodology uses Chinese Hamster Ovary (CHO) cells stably expressing high affinity receptors for candidate neurotransmitters and loaded with the calcium-sensitive dye, Fura 2. We will use these cells as biosensors to detect tastant-evoked release of neurotransmitters from isolated tasted buds and taste cells. The specific aims include investigating whether taste stimulation elicits release of serotonin, norepinephrine, acetylcholine, glutamate, ATP, CCK, and VIP from mourse and rat taste buds removed from vallate, foliate, palatal, and fungiform taste fields. Further, we aim to investigate whether such release is mediated by Ca2+ influx, intracellular Ca2+ release, or via transporter-mediated mechanisms. Finally, we propose to investigate cell-to-cell communication within taste buds via paracrine substances such as ATP and glutamate, and how information is transmitted from sensory receptor cells to synaptic output cells and on to primary sensory afferent terminals. The project will illuminate information processing in peripheral taste organs and may solve the conundrum of how signals generated in one cell (receptor cells) are transmitted to other cells (synaptic output cells) in the taste bud. The results may resolve the dilemma that taste cells appear to express only a specified set of taste receptors (i.e., bitter, or sweet, or umami GPCRs). Yet, some taste cells and many sensory afferent nerve fibers respond to multiple taste stimuli. The project will begin to raise our understanding of how taste signals are refined in the periphery, perhaps comparable to how visual signals are processed in the retina

该提案的长期目标有两个：（1）鉴定大鼠释放的神经递质和小鼠味蕾细胞在味觉刺激过程中的作用，以及（2）研究味觉刺激过程中的信号处理味蕾。这些都是化学感应接收领域尚未解答的重要问题。迄今为止，只有间接证据表明味觉细胞中存在候选神经递质，包括血清素、谷氨酸、去甲肾上腺素、乙酰胆碱、ATP 和肽。我们开发了一种新颖的方法确定大鼠和小鼠味蕾在味觉刺激过程中释放的神经递质。我们的该方法使用稳定表达高亲和力受体的中国仓鼠卵巢（CHO）细胞候选神经递质并负载钙敏感染料 Fura 2。我们将使用这些细胞作为生物传感器检测促味剂诱发的神经递质从分离的味蕾和味觉细胞的释放。具体目标包括调查味觉刺激是否会引起血清素的释放，去甲肾上腺素、乙酰胆碱、谷氨酸、ATP、CCK 和 VIP，取自小鼠和大鼠味蕾腭味区、叶味区、腭味区和菌状味觉区。此外，我们的目的是调查此类发布是否是由 Ca2+ 流入、细胞内 Ca2+ 释放或通过转运介导的机制介导。最后，我们提议研究味蕾内通过旁分泌物质（如 ATP）进行的细胞间通讯和谷氨酸，以及信息如何从感觉受体细胞传递到突触输出细胞等至初级感觉传入终端。该项目将阐明外周味觉器官的信息处理，并可能解决这个难题一个细胞（受体细胞）产生的信号如何传输到其他细胞（突触输出细胞）味蕾。这些结果可能会解决味觉细胞似乎只表达一组特定的味道的困境。味觉受体（即苦味、甜味或鲜味 GPCR）。然而，一些味觉细胞和许多感觉传入细胞神经纤维对多种味觉刺激做出反应。该项目将开始提高我们对味道的理解信号在外围被细化，也许可以与视觉信号在视网膜中的处理方式相媲美