Development of a Novel System to Study Cholinergic Synaptic Homeostasis

开发研究胆碱能突触稳态的新系统

• 批准号: 8823476
• 负责人: SUSAN L TSUNODA
• 金额: $ 18.16万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8823476
• 关键词: Alzheimer' s Disease; Alzheimer' s disease model; Area; Behavior; Behavioral; Biological Assay; Brain; Confocal Microscopy; Curare; Development; Disease; Distal; Drosophila genus; Event; Fiber; Frequencies; Genetic; Glutamate Receptor; Glutamates; Goals; Health; Homeostasis; Immunoelectron Microscopy; Kinetics; Learning; Mediating; Memory; Mental Depression; Modeling; Nerve Degeneration; Neurons; Nicotine; Nicotine Dependence; Nicotinic Receptors; Output; Pathway interactions; Patients; Physiological; Physiological Processes; Preparation; Process; Role; Schizophrenia; Seizures; Signal Transduction; Specificity; Staging; Study models; Synapses; Synaptic Transmission; System; Techniques; Testing; cholinergic; cognitive function; desensitization; high risk; in vivo; insight; neurotransmission; novel; receptor; relating to nervous system; response; success

DESCRIPTION (provided by applicant): Synaptic homeostasis is a protective mechanism employed by neurons to counterbalance changes in global neural activity. The last decade has seen intensive study in this field for glutamatergic synapses, however, almost nothing is known about whether synaptic homeostasis is also mediated by other excitatory receptors, such as nicotinic acetylcholine receptors (nAChRs). Indeed, cholinergic synaptic homeostasis has been implicated in important pathological conditions, such as Alzheimer's disease (AD) and nicotine dependence. In this R21 application, our goal is to validate a new system for studying cholinergic synaptic homeostasis and break new ground in an important area that has not yet been explored. The Drosophila CNS provides an ideal model for studying cholinergic synaptic homeostasis since nAChRs are the major excitatory receptor in Drosophila central neurons. Since mammalian α7 nAChRs are among the most abundant and widespread of nAChRs, and have been implicated in Alzheimer's disease, nicotine addiction, nicotine-induced seizures, and schizophrenia, we focus on the Drosophila α7 like receptor (Dα7) to validate our model. Using primary neuronal cultures in parallel with a cultured brain preparation, we propose to manipulate neural activity, and assay for changes in synaptic strength and changes in nAChR localization and distribution. We will also test the physiological relevance of cholinergic synaptic homeostasis by examining effects on a known behavioral output mediated by Dα7 nAChRs, the giant fiber mediated escape behavior. The development of this novel system, which involves the combination of primary cultures, an ex-vivo brain preparation, and a quantifiable behavioral output, will be unique to the field and allow us to answer long-awaited questions about cholinergic synaptic homeostasis.

描述（由申请人提供）：突触稳态是神经元用来平衡整体神经活动变化的保护机制。过去十年，谷氨酸能突触在这一领域进行了深入研究，然而，对于突触稳态是否也由其他兴奋性受体（例如烟碱乙酰胆碱受体（nAChR）介导）几乎一无所知。事实上，胆碱能突触稳态与重要的病理状况有关，例如阿尔茨海默病（AD）和尼古丁依赖。在此 R21 应用中，我们的目标是验证用于研究胆碱能突触稳态的新系统，并在尚未探索的重要领域开辟新领域。果蝇中枢神经系统为研究胆碱能突触稳态提供了理想的模型，因为 nAChR 是果蝇中枢神经元的主要兴奋性受体。由于哺乳动物 α7 nAChR 是最丰富和广泛的 nAChR 之一，并且与阿尔茨海默病、尼古丁成瘾、尼古丁诱发的癫痫发作和精神分裂症有关，因此我们重点关注果蝇 α7 样受体 (Dα7) 来验证我们的模型。我们建议使用原代神经元培养物与培养的大脑制剂并行操作神经活动，并测定突触强度的变化以及 nAChR 定位和分布的变化。我们还将通过检查 Dα7 nAChR（巨纤维介导的逃避行为）介导的已知行为输出的影响来测试胆碱能突触稳态的生理相关性。这种新颖系统的开发涉及原代培养物、离体大脑准备和可量化的行为输出的结合，这对于该领域来说是独一无二的，并使我们能够回答期待已久的有关胆碱能突触稳态的问题。