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（由适用提供）：已经确定了嗅觉感觉神经元（OSN）对气味检测和翻译至关重要的许多分子，并且已经确定了它们在这些过程中定义的基本作用。但是，我们对嗅觉翻译过程的理解仍然存在很大的差距。例如，在许多神经元中发现的信号转导级联反应在由接收器，效应酶，通道，脚手架元件和其他分子组成的信号传导复合物中起作用。 These "signalplexes" may enhance both the efficiency and specificity of signaling by increasing the local concentration of signaling elements (e.g., receptors, enzymes and solid messengers such as cAMP or Ca2+), restricting proteins to functionally important cellular domains (e.g., dendritic spines or dendritic cilia), orby regulatory access of "modulatory" proteins (e.g., receptor激酶，� -arrestin）。但是，几乎没有 知道嗅觉转移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移级联的关键组成部分来定义蛋白质 - 蛋白质相互作用，以解决嗅觉生物学的这一重要领域。我们的研究将集中于鉴定直接与两个关键嗅觉转移分子相互作用的蛋白质：嗅觉标记蛋白（OMP）和典型的异常受体（OR）。尽管OMP和ORS均已证明与其他OSN蛋白（在OMP和受体运输蛋白（RTP）家族成员的情况下，在ORS的情况下）相互作用（在ORS的情况下），但功能研究表明，两种蛋白质都存在其他伴侣。例如，OMP会影响OSN中的CAMP动力学和Ca2+动力学，而天然ORS则假设需要其他OSN特异性的共受体或伴侣来有效地靶向质膜。现在，蛋白质组学的最新进展为验证先前实施的OMP和或相互作用的人提供了独特的机会，并确定与这些关键转移分子相关的新型蛋白质。在此提案中，P.I.S将利用我们在嗅觉转移和最先进的蛋白质组学方法方面的完整专业知识，以完成两个平行的特定目的，专注于鉴定两个“诱饵”的蛋白质相互作用：opt（aim 1）和七体反应性或i7（aim 2）。我们将使用表达不同水平的任何一种诱饵的小鼠的稳定同位素标记，然后对来自天然嗅觉上皮细胞的相互作用复合物进行免疫沉淀，并随后进行液态色谱量表质谱，以定量分离出具有高灵敏度的特定相互作用。这些研究将导致我们对气味检测和转移的理解，并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的降低或损失可能对健康和生活质量造成严重影响，包括食欲失调，含糖，脂肪或咸味食物的过量消耗，避免食物或保持适当卫生的能力，以及抑郁症和社交戒断的增加的能力下降。此外，正常的嗅觉功能会导致功能失调的过度消耗，并导致相关疾病的发展，包括肥胖，糖尿病，高血压和心血管疾病。由于正常和异常的嗅觉功能都可以导致威胁生命的疾病并降低生活质量，因此我们必须更好地理解这种批判意义。这种理解可能会导致新的干预措施，包括加工食品的变化，这将增强人类健康。