Single Cell Analysis of Salt Transduction

项目摘要 盐/水平衡对哺乳动物的生理至关重要。盐的过度消费与 健康问题，特别是高血压。已经开发了盐味替代品或增强剂 无效的部分原因是对受体和其他信号转导的不完全了解 负责盐味的成分。而上皮性钠通道已经被证明是 参与阿米洛利敏感的盐味转导，阿米洛利不敏感(AI)的盐味 其转导机制尚待阐明。这项拟议的研究的目标是推进我们的 通过鉴定AI盐转导成分来理解盐味。我们将在功能上 鉴定味蕾细胞，鉴定并个别收集AI盐响应型苦味受体 细胞和AI盐III型味蕾细胞以及对照细胞对盐不响应的III型和III型细胞， 分别(目标1)。我们将用Depth扩增和询问这些单细胞的转录产物 测序，并确定在盐响应的AIII和III型中优先表达的基因 细胞。我们将对这些差异表达的基因进行深入的生物信息学分析以确定 候选AI盐受体基因(AIM 2)。这些候选基因在人工智能盐味中的作用将是 在我们的后续研究中进一步调查。这些研究的结果可以填补一个重要的空白 了解食盐的味觉和知觉，从而得出降低食盐的有效方法 消费。

Project Summary Salt/water balance is crucial to mammalian physiology. Overconsumption of salt has been linked with health problems, particularly hypertension. Development of salt taste substitutes or enhancers has been ineffective partly due to the incomplete knowledge of the receptors and other signal transduction components responsible for salt taste. While an epithelial sodium channel has been shown to be involved in the amiloride-sensitive salt taste transduction, the amiloride-insensitive (AI) salt taste transduction mechanism remains to be elucidated. The goal of this proposed research is to advance our understanding of salt taste by identifying the AI salt transduction components. We will functionally characterize taste bud cells and identify and individually collect AI salt-responsive type II bitter receptor cells and AI salt type III taste bud cells as well as control cells salt-unresponsive type II and III cells, respectively (Aim 1). We will amplify and interrogate these single cells' transcriptomes using deep sequencing, and identify genes that are preferentially expressed in the salt-responsive AI type II and III cells. We will perform in-depth bioinformatic analysis on these differentially expressed genes to identify candidate AI salt receptor genes (Aim 2). The role of these candidate genes in the AI salt taste will be further investigated in our follow-up studies. Results from these studies can fill an important gap in understanding salt taste sensation and perception, thus leading to effective means to reduce salt consumption.