Single Cell Analysis of Salt Transduction

描述（由申请人提供）：盐/水平衡对哺乳动物生理至关重要。过量食用盐与健康问题有关，尤其是高血压。盐味替代品或增强剂的开发一直是无效的，部分原因是对盐味受体和其他信号转导成分的不完全了解。虽然上皮钠通道已被证明参与了阿米洛利敏感盐味转导，但阿米洛利不敏感（AI）盐味转导机制仍有待阐明。本研究的目标是通过识别人工智能盐转导成分来提高我们对盐味道的理解。我们将对味蕾细胞进行功能表征，并分别鉴定和单独收集AI盐敏感型II苦味受体细胞和AI盐III型味蕾细胞，以及盐不敏感型II和III型对照细胞（Aim 1）。我们将使用深度测序扩增和询问这些单细胞的转录组，并鉴定在盐反应型AI II和III细胞中优先表达的基因。我们将对这些差异表达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（目标2）。这些候选基因在AI盐味中的作用将在我们的研究中进一步研究

DESCRIPTION (provided by applicant): 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 cels salt-unresponsive type II and III cells, respectively (Aim 1). We will amplify and interrogate thes 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.