Mechanisms of odor detection and transduction

气味检测和传导机制

• 批准号: 8400573
• 负责人: Steven D Munger
• 金额: $ 23.03万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 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 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8400573
• 关键词: Address; Afferent Neurons; Aging; Amino Acids; Antibodies; Area; Arrestins; Binding; Biological; Biology; Brain; Calmodulin; Cardiovascular Diseases; Cell membrane; Cells; Cilia; Co-Immunoprecipitations; Complex; Consumption; Cyclic AMP; Dendritic Spines; Desire for food; Detection; Development; Diabetes Mellitus; Disease; Drosophila genus; Elements; Enzymes; Family member; Food; Food Processing; Gene Targeting; Goals; Health; Human; Hygiene; Hypertension; Immune Sera; Immunohistochemistry; Immunoprecipitation; In Situ Hybridization; In Vitro; Incidence; Injury; Intervention; Kinetics; Label; Lead; Life; Liquid Chromatography; Macromolecular Complexes; Mass Spectrum Analysis; Membrane; Membrane Lipids; Mental Depression; Molecular; Molecular Chaperones; Mus; NGFRAP1 gene; Neurons; Obesity; Odorant Receptors; Odors; Olfactory Epithelium; Peripheral; Phenotype; Phosphotransferases; Procedures; Process; Protein Family; Proteins; Proteomics; Quality of life; Research; Role; Scaffolding Protein; Signal Transduction; Specificity; Stable Isotope Labeling; Stimulus; System; Translating; Validation; Withdrawal; Yang; adenylyl cyclase III; base; biological systems; cell type; gel electrophoresis; human RIPK1 protein; in vivo; insight; interdisciplinary approach; novel; novel strategies; olfactory marker protein; programs; protein protein interaction; receptor; relating to nervous system; response; scaffold; social; tandem mass spectrometry; trafficking

DESCRIPTION (provided by applicant): Many of the molecules that are critical for the detection and transduction of odors by olfactory sensory neurons (OSNs) have been identified, and their basic roles in these processes defined. However, significant gaps in our understanding of the olfactory transduction process remain. For example, signal transduction cascades found in many neurons function within signaling complexes composed of receptors, effector enzymes, channels, scaffolding elements and other molecules. These "signalplexes" may enhance both the efficiency and specificity of signaling by increasing the local concentration of signaling elements (e.g., receptors, enzymes and soluble messengers such as cAMP or Ca2+), restricting proteins to functionally important cellular domains (e.g., dendritic spines or dendritic cilia), orby regulating access of "modulatory" proteins (e.g., receptor kinases, ¿-arrestin). However, little is known about how olfactory transduction proteins interact to impact olfactory function. Our proposed studies will address this important yet understudied area of olfactory biology by using a cutting-edge, multidisciplinary approach to define protein-protein interactions for key components of the olfactory transduction cascade. Our studies will focus on identifying proteins that directly interact with two key olfactory transduction molecules: olfactory marker protein (OMP) and canonical odorant receptors (OR). While both OMP and the ORs have been shown to interact with other OSN proteins (Bex proteins in the case of OMP and receptor trafficking protein (RTP) family members in the case of ORs), functional studies suggests that other partners exist for both proteins. For example, OMP influences cAMP kinetics and Ca2+ dynamics in OSNs, while native ORs are hypothesized to require additional OSN-specific co-receptors or chaperones to efficiently target the plasma membrane. Recent advances in proteomics now offer a unique opportunity to both validate previously implicated OMP and OR interactors as well as to identify novel proteins that associate with these key transduction molecules. In this proposal, the P.I.s will take advantage of our complementary expertise in olfactory transduction and state-of-the-art proteomics approaches to complete two parallel Specific Aims focused on identifying protein interactors for two "baits:" OMP (Aim 1) and the heptanal-responsive OR I7 (Aim 2). We will use stable isotope labeling of mice expressing different levels of the either bait, followed by immunoprecipitation of interacting complexes from native olfactory epithelium and subsequent liquid chromatography-tandem mass spectroscopy to quantitatively isolate specific interactors with high sensitivity. These studies will result in significant advances in our understanding of odor detection and transduction, and will establish an important new approach for characterizing the interactions of rare proteins in nearly any biological system. PUBLIC HEALTH RELEVANCE: The reduction or loss of olfactory function due to injury, disease or aging can have serious consequences for health and quality of life, including appetite dysregulation, excessive consumption of sugary, fatty or salty foods, a decreased ability to avoid spoiled foods or to maintain proper hygiene, and an increased incidence of depression and social withdrawal. Furthermore, normal olfactory function can contribute to dysfunctional overconsumption and lead to the development of associated diseases, including obesity, diabetes, hypertension and cardiovascular disease. As both normal and abnormal olfactory function can contribute to life-threatening disease and reduce quality of life, it is imperative tht we better understand this critical sense; such understanding could lead to new interventions, including changes in processed foods, that would enhance human health.

描述（由申请人提供）：许多对于嗅觉感觉神经元（OSN）检测和转导气味至关重要的分子已经被识别，并且它们在这些过程中的基本作用也被定义。然而，我们对气味的理解存在重大差距。例如，在许多神经元中发现的信号转导级联在由受体、效应酶、通道、支架元件和其他分子组成的信号复合物中发挥作用。可以通过增加信号元件（例如受体、酶和可溶性信使，如 cAMP 或 Ca2+）的局部浓度、将蛋白质限制在功能重要的细胞域（例如树突棘或树突纤毛）来增强信号传导的效率和特异性，或通过调节“调节”蛋白（例如受体激酶、¿-arrestin）的访问然而，很少有。 了解嗅觉转导蛋白如何相互作用以影响嗅觉功能，我们提出的研究将通过使用尖端的多学科方法来定义嗅觉转导级联关键组成部分的蛋白质-蛋白质相互作用，从而解决嗅觉生物学中这一重要但尚未充分研究的领域。我们的研究将集中于识别与两个关键嗅觉转导分子直接相互作用的蛋白质：嗅觉标记蛋白（OMP）和典型气味受体（OR）。 OR 已被证明与其他 OSN 蛋白（OMP 中的 Bex 蛋白和 OR 中的受体运输蛋白 (RTP) 家族成员）相互作用，功能研究表明这两种蛋白都存在其他伙伴。 OMP 影响 OSN 中的 cAMP 动力学和 Ca2+ 动力学，而天然 OR 则需要额外的 OSN 特异性共受体或伴侣来有效靶向质膜。蛋白质组学的最新进展现在提供了独特的机会。两者均验证先前涉及的 OMP 和 OR 相互作用因子，并鉴定与这些关键转导分子相关的新型蛋白质。在本提案中，PI 将利用我们在嗅觉转导和最先进的蛋白质组学方法方面的互补专业知识来进行研究。完成两个平行的具体目标，重点是识别两个“诱饵”的蛋白质相互作用物：OMP（目标 1）和庚醛响应 OR I7（目标 2）。使用表达不同水平的任一诱饵的小鼠的稳定同位素标记，然后对天然嗅上皮的相互作用复合物进行免疫沉淀，以及随后的液相色谱-串联质谱法以高灵敏度定量分离特定的相互作用物。这些研究将导致我们的重大进展。了解气味检测和转导，并将建立一种重要的新方法来表征几乎所有生物系统中稀有蛋白质的相互作用。 公共卫生相关性：由于受伤、疾病或衰老而导致的嗅觉功能减弱或丧失可能会对健康和生活质量产生严重后果，包括食欲失调、过量食用含糖、脂肪或咸味食物、避免变质食物的能力下降此外，正常的嗅觉功能可能会导致过度消费，并导致相关疾病的发生，包括肥胖、糖尿病、高血压和心血管疾病。正常和异常的嗅觉功能可能导致危及生命的疾病并降低生活质量，我们必须更好地理解这种关键感觉；这种理解可能会导致新的干预措施，包括改变加工食品，从而增强人类健康。