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）：大多数脊椎动物都对五种基本口味反应：甜，苦，鲜味，酸和咸，每种都提供了有关摄入食物的营养含量和安全性的独特信息。五种味道质量中的每一个都通过不同的味觉细胞的不同子集检测到，这些细胞表达不同的受体和信号分子。尽管在理解介导苦，甜，鲜味和咸味的分子和途径方面取得了巨大进展，但对酸味的味道相对较少。检测到酸味的细胞已被证明是通过TRP离子通道PKD2L1的表达来定义的，但是PKD2L1基因本身对于酸味和对酸味的感觉反应构成的离子机制是可分配的。在最近的工作中，使用小鼠，其中黄色荧光蛋白（YFP）由PKD2L1的启动子驱动，我们表明酸味细胞表达了一种新型的质子电导，这有助于对酸的反应。当前建议的具体目标是确定编码该质子电导的基因。为此，我们提出了两个具体目标。在AIM 1中，我们将使用深层测序对YFP标记的酸性细胞进行转录分析，我们将确定AE富含PKD2L1细胞的转录本，并可能编码新型的跨膜蛋白。在AIM 2下，我们将在异源细胞类型中表达顶级候选物，并通过贴片夹记录它们是否生成质子通道。该质子通道的识别将代表理解味觉系统的重要步骤，并可能有助于定义一类新的离子通道。味道是人类和其他生物体调节其摄入行为的重要方法，并且鉴定味觉信号的关键分子成分可以直接影响人类健康和福祉。