Odorant Receptor Expression and Sensitivity to Odorants

气味受体表达和对气味剂的敏感性

• 批准号: 8759723
• 负责人: Timothy S McClintock
• 金额: $ 13.92万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8759723
• 关键词: Afferent Neurons; Agonist; Alleles; Anosmia; Axon; ChIP-seq; Chemical Structure; Chemicals; Color Visions; Data; Data Analyses; Detection; Development; Dimensions; Discrimination; Ear; Event; Future; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Variation; Genomics; Human; Life; Measures; Mentors; Methods; Modification; Molecular Biology; Molecular Biology Techniques; Mus; Nature; Neurons; Odorant Receptors; Odors; Olfactory Epithelium; Outcome; Pattern; Pilot Projects; Receptor Activation; Receptor Gene; Regulation; Research; Research Personnel; Research Project Grants; Resources; Role; Signal Transduction; Solutions; Stimulus; System; Techniques; Technology; Testing; Training; Work; base; chromatin modification; detector; experience; flexibility; homeodomain; hyposmia; improved; in vivo; member; neural patterning; next generation sequencing; olfactory bulb; programs; public health relevance; receptor; receptor expression; receptor sensitivity; research study; response; tool; transcription factor; transcriptome sequencing

DESCRIPTION (provided by applicant): The application proposes to allow an investigator experienced in the molecular biology of the olfactory epithelium to cross the gap from his expertise in microarray-based genomics to more computationally challenging next generation sequencing approaches. The plan includes: (1) training in R programming to allow the investigator take advantage of flexible analysis methods for ChIP-seq and RNA-seq techniques, (2) mentored pilot experiments in both of these approaches, and (3) mentored analyses of the data. The pilot studies take advantage of a unique resource that allows purification of olfactory neurons expressing subsets of odorant receptors to work toward testing two fundamental hypotheses about odorant receptors. (1) Active chromatin modifications (which can only be detected by enrichment for neurons expressing an odorant receptor) are critical for the tightly regulated expression of odorant receptors. Active chromatin modifications will be measured by ChIP-seq. The exquisitely specific control of odorant receptor expression is perhaps the greatest remaining mystery about the function of olfactory sensory neurons. (2) Odorants activate sets of odorant receptors that may overlap but must have at least one distinct member if the odorants can be discriminated. Enrichment for olfactory sensory neurons marked by expression of GFP from an activity-dependent gene locus will allow RNA-seq methods to identify sets of odorant receptors activated by specific odorants. Understanding the fundamental question of how hundreds of odorant receptors allow the detection and discrimination of thousands of odorant chemicals should help explain some causes of hyposmia and anosmia; and should allow the future development of odorant agonists and antagonists that could be used to improve the quality of human life.

描述(由申请人提供)：该申请建议允许在嗅觉上皮分子生物学方面有经验的研究人员跨越他在基于微阵列的基因组学专业知识到更具计算挑战性的下一代测序方法之间的差距。该计划包括：(1)R编程培训，使研究人员能够利用芯片序列和RNA-序列技术的灵活分析方法，(2)这两种方法的指导性试点实验，以及(3)数据的指导性分析。试点研究利用一种独特的资源，允许纯化表达气味受体亚群的嗅觉神经元，以测试关于气味受体的两个基本假说。(1)主动的染色质修饰(只有通过丰富表达气味受体的神经元才能检测到)对于气味受体的严格调控表达至关重要。活性染色质修饰将通过CHIP-SEQ进行测量。气味受体表达的精细特异控制可能是关于嗅觉感觉神经元功能的最大谜团。(2)气味物质激活成套的气味感受器，这些气味感受器可能重叠，但必须至少有一个不同的成员，才能区分气味。通过从活性依赖基因位点表达GFP来标记嗅觉感觉神经元的丰富，将使RNA-SEQ方法能够识别由特定气味激活的成套气味受体。了解数以百计的气味受体如何允许检测和辨别数千种气味化学物质这一根本问题，应该有助于解释嗅觉不足和嗅觉障碍的一些原因；并应该允许未来开发可用于改善人类生活质量的气味激动剂和拮抗剂。