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

该提案的长期目标是双重的：（1）识别从老鼠释放的神经递质味觉刺激期间的小鼠味蕾细胞和（2）研究信号处理味蕾。这两者都是化学感觉接收领域的重大未解决问题。迄今谷氨酸，去甲肾上腺素，乙酰胆碱，ATP和肽。我们已经开发了一种新颖的方法在味觉刺激过程中，牙齿从大鼠和小鼠味蕾中释放出神经递质（S）。我们的方法论使用中国仓鼠卵巢（CHO）细胞稳定地表达高亲和力受体候选神经递质并用钙敏感的染料加载fura 2。我们将使用这些细胞作为生物传感器可检测散发味的芽和味道细胞中诱人的神经递质释放。具体目的包括调查味觉刺激是否引起5-羟色胺的释放，去甲肾上腺素，乙酰胆碱，谷氨酸，ATP，CCK和VIP，从哀悼和老鼠味蕾中取出 Vallate，Foliate，Paletal和Fungiorper Taste Fields。此外，我们旨在调查此次发布是否是由Ca2+流入，细胞内Ca2+释放或通过转运蛋白介导的机制介导。最后，我们建议通过旁分泌物质（例如ATP）研究味蕾内的细胞间通信和谷氨酸，以及信息如何从感觉受体细胞传输到突触输出细胞以及到主要感觉传入终端。该项目将在外围口味器官中照亮信息处理，并可能解决难题关于在一个细胞中产生的信号（受体细胞）如何传输到其他细胞中的信号（突触输出单元）味蕾。结果可能可以解决味觉细胞似乎仅表达指定的一组的困境味觉受体（即苦或甜或鲜味GPCR）。然而，一些味道细胞和许多感官传入神经纤维应对多种味道刺激。该项目将开始提高我们对口味的理解信号在周围进行了完善，也许与视网膜中的视觉信号相媲美