Transcriptome profiling of sour taste cells

酸味细胞的转录组分析

• 批准号: 8495312
• 负责人: EMILY R. LIMAN
• 金额: $ 27.68万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8495312
• 关键词: Acids; Action Potentials; Apical; Biological Assay; Candidate Disease Gene; Cell membrane; Cells; Complementary DNA; Data; Detection; Feeding behaviors; Food; Gene Expression Profile; Generations; Genes; Genetic Transcription; Goals; Health; Human; In Situ Hybridization; Integral Membrane Protein; Ion Channel; Ions; Label; Length; Libraries; Measures; Mediating; Membrane; Messenger RNA; Modeling; Molecular; Molecular Profiling; Monitor; Mouse Protein; Mus; Nutritional; Organism; Pathway interactions; Personal Satisfaction; Proteins; Protons; Receptor Signaling; Safety; Sensory; Signal Transduction; Signaling Molecule; Specificity; Stimulus; Structure of thyroid parafollicular cell; System; Taste Perception; Testing; Tissues; Transcript; Transgenic Mice; Work; base; cDNA Library; cell type; deep sequencing; interest; next generation sequencing; novel; patch clamp; promoter; protein expression; public health relevance; receptor; research study; response; sensor; sweet taste perception; transcriptome sequencing

DESCRIPTION (provided by applicant): Most vertebrate species are responsive to five basic tastes: sweet, bitter, umami, sour and salty, each of which provides unique information on the nutritional content and safety of ingested food. Each of the five taste qualities is detected by a distinct subset of taste cells, which express distinct receptors and signaling molecules. While great progress has been made in understanding the molecules and pathways that mediate bitter, sweet, umami and salty tastes, relatively little is known about sour taste. The cells that detect sour taste have been shown to be defined by expression of the TRP ion channel PKD2L1, but the Pkd2l1 gene itself is dispensable for sour taste and the ionic mechanisms that underlie sensory responses to sour remain poorly understood. In recent work, using a mouse in which yellow fluorescent protein (YFP) was driven by the promoter of Pkd2l1, we showed that sour taste cells express a novel proton conductance which contributes to the response to acids. The specific goal of the current proposal is to identify the gene that encodes this proton conductance. To do this, we propose two specific aims. In aim 1, we will use deep sequencing to perform transcription profiling of YFP-tagged sour cells and we will identify transcripts that ae enriched in Pkd2l1 cells and are likely to encode novel transmembrane proteins. Under Aim 2, we will express the top candidates in a heterologous cell type and determine with patch clamp recording whether they generate a proton channel. The identification of this proton channel will represent an important step in understanding the taste system, and may help define a new class of ion channels. Taste is an essential way in which humans and other organisms regulate their ingestive behavior and the identification of key molecular components of taste signaling can therefore have a direct impact on human health and well-being.

描述(由申请人提供)：大多数脊椎动物物种对五种基本口味做出反应：甜、苦、鲜味、酸和咸，每一种都提供关于所摄取食物的营养含量和安全性的独特信息。五种口味中的每一种都是由不同的味觉细胞来检测的，这些味觉细胞表达不同的受体和信号分子。虽然在了解调节苦味、甜味、鲜味和咸味的分子和途径方面取得了很大进展，但对酸味的了解相对较少。检测酸味的细胞已被证明是由Trp离子通道PKD2L1的表达定义的，但PKD2L1基因本身对于酸味来说是必不可少的，而且对酸味感觉反应的离子机制仍然知之甚少。在最近的工作中，我们用一只由PKD2L1启动子驱动黄色荧光蛋白(YFP)的小鼠，证明了酸味细胞表达一种新的质子电导，这种电导有助于对酸的反应。目前这项提议的具体目标是识别编码这种质子电导的基因。为了做到这一点，我们提出了两个具体目标。在目标1中，我们将使用深度测序来对YFP标记的酸性细胞进行转录谱分析，并鉴定在PKD2L1细胞中丰富的、可能编码新的跨膜蛋白的转录本。在目标2下，我们将在异源细胞类型中表达排名靠前的候选细胞，并通过膜片钳记录来确定它们是否产生质子通道。这种质子通道的鉴定将代表着理解味觉系统的重要一步，并可能有助于定义一类新的离子通道。味觉是人类和其他生物调节其摄食行为的重要方式，因此识别味觉信号的关键分子成分可以直接影响人类的健康和福祉。