Molecular Genesis and Organization of Olfactory Transduction Components and Cilia

嗅觉传导成分和纤毛的分子起源和组织

• 批准号: 8629885
• 负责人: RANDALL R REED
• 金额: $ 34.43万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8629885
• 关键词: Action Potentials; Address; Adenylate Cyclase; Afferent Neurons; Animals; Behavior; Brain; C-terminal; Cell Culture Techniques; Cell physiology; Cells; Cellular Structures; Chemicals; Cilia; Communication; Complex; Cultured Cells; Cyclic AMP; Cyclic Nucleotides; Data; Defect; Detection; Development; Disease; Environment; Enzymes; Family; Functional disorder; G Protein-Coupled Receptor Genes; GTP-Binding Proteins; Genetic; Golf; Grant; Health; Hereditary Disease; Human; Human Genetics; Imagery; In Vitro; Individual; Kidney; Laboratories; Length; Ligands; Link; Mammalian Cell; Measurement; Mediating; Metabolism; Modeling; Molecular; Molecular Genetics; Mus; Neuroglia; Neurons; Neurosecretory Systems; Neurotransmitters; Odorant Receptors; Odors; Olfactory Receptor Neurons; Organelles; Organism; Pathway interactions; Physiological; Play; Process; Property; Protein Isoforms; Proteins; Proteomics; Regulation; Reporter; Role; Sensory; Sensory Process; Signal Transduction; Smell Perception; Source; Specificity; Stimulus; Structure; System; Tissues; Validation; ciliopathy; genetic manipulation; in vivo; insight; mutant; neuronal circuitry; novel; polypeptide; public health relevance; research study; sensory system

DESCRIPTION (provided by applicant): The mammalian olfactory system achieves remarkable sensitivity through novel genetic mechanisms, cellular specializations, and subsequent processing of sensory information by neuronal circuitry. The earliest steps in the transduction of odorant information into signals that are propagated to the brain occur in olfactory cilia. The dozen long immotile cilia of each olfactory neuron are the only part of the sensory cell exposed to the outside environment and the sources of odorant stimuli and their existence is essential for efficient odor detection. The olfactory neuron has developed efficient but poorly understood mechanisms for enriching the components of olfactory transduction (odorant receptor (OR), G protein (Golf), adenylyl cyclase (AC3) and olfactory cyclic nucleotide channel) in cilia. The abundant expression of the single polypeptide comprising the AC3 enzyme in all olfactory neurons makes it particularly amenable to elucidate the pathways and mechanisms responsible for cilia localization and enrichment. In this grant we will use molecular and genetic approaches to investigate the hypothesis that a specialized but broadly utilized mechanism and pathway is responsible for AC3 localization to cilia and that this localization is critical for cilia dynamics and function. In specific aim 1, we target the cilia localization signa of AC3 in vivo by a conditional genetic disruption approach and determine the consequences of re- localization of AC3 into different cellular compartments. In specific aim 2, we will utilize a robut expression system to identify components of the AC3 translocation/enrichment pathway leading to cilia localization and examine the dynamics of AC3 in cilia and role of AC3 localization in modulation of cilia length. Together, these experiments will expand our understanding of how an important class of proteins are enriched in cilia and the importance of their selective localizatio to this organelle. Cilia are present in essentially all terminally differentiated mammalian cells ad critical for the development of the organism. The pathways and mechanisms underlying adenylyl cyclase localization will have important consequences for olfaction, sensory communication and broadly for human health and understanding disease. The abundance of AC3 in olfactory neurons makes it particularly amenable for detailed studies. The AC3 enzyme is broadly expressed and highly enriched in many neuronal cilia of the brain and kidney where it likely mediates neuroendocrine/neurotransmitter signaling and its subcellular localization in these cells appear to be mediated by similar mechanisms. Modulation of this pathway should perturb complex behaviors and metabolic processes. Additionally, it is likely that the cilia localization pathway utilized by AC3 is shared by other proteins whose presence in cilia is critical for cellular function. Elucidation of these pathways will provide insight into cellular processes that have widespread consequences for human health.

描述（由申请人提供）：哺乳动物嗅觉系统通过新的遗传机制、细胞特化和神经元回路对感觉信息的后续处理实现了非凡的灵敏度。气味信息转导成信号并传播到大脑的最早步骤发生在嗅觉纤毛中。每个嗅觉神经元的12根长而不动的纤毛是感觉细胞中唯一暴露于外界环境和气味刺激来源的部分，它们的存在对有效的气味检测至关重要。嗅觉神经元在纤毛中丰富嗅觉转导成分（气味受体（OR）、G蛋白（Golf）、腺苷酸环化酶（AC3）和嗅觉环核苷酸通道）方面已经发展出高效但尚不清楚的机制。包含AC3酶的单多肽在所有嗅觉神经元中的丰富表达使得它特别适合于阐明纤毛定位和富集的途径和机制。在这项资助中，我们将使用分子和遗传学方法来研究AC3定位到纤毛的假设，即一种专门但广泛使用的机制和途径负责AC3定位到纤毛，这种定位对纤毛的动力学和功能至关重要。在具体目标1中，我们通过条件遗传破坏方法靶向体内AC3的纤毛定位信号，并确定AC3重新定位到不同细胞室的后果。在具体的目标2中，我们将利用一个健壮的表达系统来识别导致纤毛定位的AC3易位/富集途径的组成部分，并研究AC3在纤毛中的动态以及AC3定位在纤毛长度调节中的作用。总之，这些实验将扩大我们对一类重要蛋白质如何在纤毛中富集的理解，以及它们对纤毛细胞器选择性定位的重要性。纤毛基本上存在于所有终末分化的哺乳动物细胞中，对生物体的发育至关重要。腺苷酸环化酶定位的途径和机制将对嗅觉、感觉交流以及广泛的人类健康和疾病理解产生重要影响。嗅觉神经元中丰富的AC3使其特别适合进行详细的研究。AC3酶在脑和肾的许多神经元纤毛中广泛表达和高度富集，可能介导神经内分泌/神经递质信号传导，其在这些细胞中的亚细胞定位似乎通过类似的机制介导。这一途径的调节会扰乱复杂的行为和代谢过程。此外，AC3利用的纤毛定位途径可能与其他蛋白质共享，这些蛋白质在纤毛中的存在对细胞功能至关重要。阐明这些途径将有助于深入了解对人类健康产生广泛影响的细胞过程。