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（由适用提供）：突触体内稳态是神经元的受保护机制的员工，以平衡全球神经元活动的变化。在过去的十年中，在该领域对谷氨酸能突触进行了激烈的研究，但是，对于合成稳态的骨稳态是否也是由其他兴奋受体（例如烟碱乙酰胆碱受体（NACHR））介导的几乎一无所知。实际上，在重要的病理状况（例如阿尔茨海默氏病（AD）和尼古丁依赖性）中已经实施了胆碱能突触稳态。在此R21应用中，我们的目标是验证一种新的系统，用于研究胆碱能突触体内稳态，并在尚未探索的重要领域中脱颖而出。果蝇中枢神经系统提供了研究胆碱能突触稳态的理想模型，因为NACHRS是果蝇中心神经元的主要兴奋受体。由于哺乳动物α7NACHR是NACHR的最丰富和宽度之一，并且与阿尔茨海默氏病，添加尼古丁，尼古丁诱发的癫痫发作和精神分裂症有关，因此我们将重点放在果蝇α7之类的受体（Dα7）上。我们建议使用原发性神经元培养物与培养的大脑制剂并行，我们建议操纵神经元活性，并测定突触强度的变化以及NACHR定位和分布的变化。我们还将通过检查对由Dα7NACHR介导的已知行为输出的影响（巨型纤维介导的逃逸行为），测试胆碱能突触稳态的物理相关性。这种新型系统的开发涉及原代培养物的结合，前体脑的制备和可量化的行为输出，将是该领域独有的，并使我们能够回答有关胆碱能突触稳态的期待已久的问题。