Modulation of Olfactory Bulb Neuron Current Properties

嗅球神经元电流特性的调节

• 批准号: 6580705
• 负责人: DEBRA Ann FADOOL
• 金额: $ 29.85万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6580705
• 关键词: SDS polyacrylamide gel electrophoresis; biological signal transduction; electrophysiology; electrostimulus; enzyme activity; enzyme linked immunosorbent assay; genetically modified animals; immunocytochemistry; immunoprecipitation; insulin receptor; laboratory mouse; membrane channels; neural plasticity; neurochemistry; neuroregulation; neurotrophic factors; olfactory lobe; phosphorylation; potassium channel; protein protein interaction; protein tyrosine kinase; site directed mutagenesis; tissue /cell culture; voltage /patch clamp; western blottings

DESCRIPTION (provided by applicant): The designed research is a multidisciplinary analysis of the modulation of potassium currents in granule and mitral cells of the olfactory bulb. The broad, long-term objective of this research is to elucidate how neurotrophins and growth factors can utilize ion channels as substrates for phosphorylation to give rise to short-term and long-term plastic changes in synaptic efficacy or to aid in the establishment of neural circuits in the olfactory bulb. Understanding the general principles governing these transduction cascades and the involvement of ion channels will provide information of how protein kinases and protein phosphatases contribute to the onset or severity of specific neuronal diseases, such as Alzheimer's, or how uncontrolled signaling of these enzymes leads to deregulated cell proliferation and diseases such as cancer and diabetes. Because of the unique trophic and regenerative capacity of neurons in the olfactory system, continual expression of neuromodulators could alter patterns of electrical excitability in addition to their well-studied roles in growth and differentiation. The specific aims of this proposal are to characterize using patch-clamp electrophysiology how receptor-linked tyrosine phosphorylation signaling in the olfactory bulb is altered by sensory experience, patterned electrical stimulation, and trophic factor infusion. By utilizing the cloned, olfactory bulb potassium channel Kv1.3 as a parallel model, combined biochemical measurement of kinase-induced tyrosine phosphorylation, co-immunoprecipitation, and molecular mutagenesis will elucidate the mechanistic details of how ion channels form molecular scaffolds with kinases and adaptor proteins through discrete protein-protein interactions at SH2, SH3, PDZ, and PTB domains. Gene-targeted deletions in Kv1.3 channel, insulin receptor kinase, and TrkB kinase will provide mechanistic details for the role for tyrosine phosphorylation signaling in olfaction and for neuromodulation in the CNS in general, as defined by loss of function experiments (behavioral, biochemical, electrophysiological) using knock-out mice strains. The proposal will provide new important information regarding the integration of signaling molecules by construction of protein-protein interactions with ion channels. Modulation of ion channel function would thus be dependent upon the repertoire of signaling proteins expressed in a given neuron, a background that could change with sensory experience or electrical patterning.

描述（由申请人提供）：设计的研究是对嗅球颗粒和二尖瓣细胞中钾电流调节的多学科分析。这项研究的广泛长期目标是阐明神经营养蛋白和生长因子如何利用离子通道作为磷酸化的底物，从而导致突触功效的短期和长期塑性变化，或者有助于在嗅球中建立神经回路。了解管理这些转导级联的一般原理以及离子通道的参与将提供有关蛋白激酶和蛋白质磷酸酶如何促进特定神经元疾病的发作或严重程度的信息，例如阿尔茨海默氏症的发作或严重性，或者这些酶的信号导致这些酶的信号如何导致细胞腐蚀和疾病的疾病和疾病癌症和疾病。由于神经元在嗅觉系统中具有独特的营养和再生能力，神经调节剂的持续表达还可以改变电兴奋性的模式，除了它们在生长和分化中的良好作用。该提案的具体目的是使用贴片钳电生理学表征嗅球中受体连接的酪氨酸磷酸化信号传导如何通过感觉体验，图案化的电刺激和营养因子输注来改变。通过利用克隆的嗅球钾通道KV1.3作为平行模型，结合了激酶诱导的酪氨酸磷酸化，共产生沉淀和分子诱变的生物化学测量，将阐明与kIn蛋白相互作用的蛋白质的机械细节，并将SH2，SH3，PDZ和PTB域。 KV1.3通道，胰岛素受体激酶和TRKB激酶中的基因靶向缺失将为嗅觉磷酸化信号传导在嗅觉和CN中的神经调节中的作用提供机械详细信息，这是通过功能实验的丧失（使用BioChemical，BioChemical，biofiencational of Beachemical，biofiencational of Becogical of功能，使用了二元，生物化学，电力学）。该提案将通过构建蛋白质 - 蛋白质相互作用与离子通道来提供有关信号分子整合的新重要信息。因此，离子通道功能的调节将取决于在给定神经元中表达的信号蛋白的曲目，这种背景可以随感觉体验或电气模式而变化。