A new method to measure taste bud function

测量味蕾功能的新方法

• 批准号: 7085009
• 负责人: STEPHEN D. ROPER
• 金额: $ 10万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-15 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7085009
• 关键词: acetylcholine; adenosine triphosphate; animal tissue; biological signal transduction; calcium flux; calcium indicator; cell cell interaction; cholecystokinin; gap junctions; glutamates; intracellular transport; method development; neurotransmitter transport; norepinephrine; receptor expression; sensory mechanism; serotonin; taste; taste buds; tissue /cell culture; vasoactive intestinal peptide

DESCRIPTION (provided by applicant): The long-term objectives of this proposal are twofold: (1) to identify neurotransmitters released from 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 identify the neurotransmitter(s) released from 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. The specific aims include investigating whether taste stimulation elicits release of serotonin, norepinephrine, acetylcholine, glutamate, ATP, CCK, and VIP from taste buds. Further, we aim to investigate whether such release is mediated by Ca2+ influx, intracellular Ca2* release, or via transporter mediated mechanisms. We also will study whether gap junctions mediate signal transfer between taste cells during taste stimulation. The project will illuminate information processing in peripheral taste organs and may solve the conundrum of if/how signals generated in one cell (receptor cells) are transmitted to other cells (output cells) in the taste bud. The results may resolve the dilemma that taste cells appear to express a limited set of taste receptors (i.e. bitter, or sweet GPCRs, but not both) yet some taste cells and many sensory afferent nerve fibers respond to multiple taste stimuli: signals generated in dedicated receptor cells are transmitted to other (output) cells and integrated within processing units prior to the final output from the taste bud.

描述（由申请人提供）：该提案的长期目标有两个：（1）识别味蕾刺激期间味蕾细胞释放的神经递质；（2）研究味蕾内的信号处理。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。我们开发了一种新的方法来识别小鼠味蕾在味觉刺激过程中释放的神经递质。我们的方法使用稳定表达候选神经递质高亲和力受体的中国仓鼠卵巢 (CHO) 细胞，并负载钙敏感染料 Fura 2。具体目的包括研究味觉刺激是否会引发血清素、去甲肾上腺素、乙酰胆碱、谷氨酸、ATP 的释放、CCK、VIP来自味蕾。此外，我们的目的是研究这种释放是否是由 Ca2+ 流入、细胞内 Ca2* 释放介导的，还是通过转运蛋白介导的机制介导的。我们还将研究在味觉刺激过程中间隙连接是否介导味觉细胞之间的信号传递。该项目将阐明外周味觉器官的信息处理，并可能解决一个细胞（受体细胞）产生的信号是否/如何传输到味蕾中其他细胞（输出细胞）的难题。结果可能会解决味觉细胞似乎表达一组有限的味觉受体（即苦味或甜味 GPCR，但不是两者）的困境，但一些味觉细胞和许多感觉传入神经纤维对多种味觉刺激做出反应：在专用味觉刺激中产生的信号受体细胞被传输到其他（输出）细胞，并在味蕾最终输出之前整合到处理单元中。

项目成果

ATP release through connexin hemichannels and gap junction transfer of second messengers propagate Ca2+ signals across the inner ear.

通过连接蛋白半通道释放 ATP 和第二信使的间隙连接转移，将 Ca2 信号传播到内耳。

• 发表时间: 2008-12-02
• 影响因子: 11.1
• 作者: Anselmi, Fabio;Hernandez, Victor H;Crispino, Giulia;Seydel, Anke;Ortolano, Saida;Roper, Stephen D;Kessaris, Nicoletta;Richardson, William;Rickheit, Gesa;Filippov, Mikhail A;Monyer, Hannah;Mammano, Fabio
• 通讯作者: Mammano, Fabio