Neuron-glia interactions in regulation of activity-dependent signaling pathways

神经元-胶质细胞相互作用调节活性依赖性信号通路

• 批准号: 8337833
• 负责人: MARTA MARGETA
• 金额: $ 33.8万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8337833
• 关键词: Address; Affect; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animals; Antioxidants; Astrocytes; Attenuated; Biochemical; Biological; Biology; Blood - brain barrier anatomy; Brain; Cells; Chemosensitization; Coculture Techniques; Complex; Data; Development; Disease; Drug Design; Drug Metabolic Detoxication; Environment; Environmental Risk Factor; Enzymes; Feedback; Fostering; Foundations; Functional disorder; Future; Genetic; Glutamates; Goals; Hippocampus (Brain); In Vitro; Individual; Injury; Knock-out; Knowledge; Laboratories; Learning; Mediating; Mediator of activation protein; Metabolic; Mission; Mitochondria; Molecular; Molecular Target; N-Methyl-D-Aspartate Receptors; Nerve Degeneration; Neurodegenerative Disorders; Neuroglia; Neurons; Oxidative Stress; Parkinson Disease; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Phase; Physiology; Play; Potassium Channel; Predisposition; Prevention; Process; Public Health; Reactive Oxygen Species; Receptor Signaling; Regulation; Research; Resistance; Role; Second Messenger Systems; Shapes; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Molecule; Slice; Stroke; Synapses; Synaptic Transmission; Synaptic plasticity; Testing; Toxin; Transgenic Mice; United States National Institutes of Health; Work; base; cell type; cellular targeting; density; excitotoxicity; improved; innovation; mouse model; nervous system disorder; neuroprotection; neurotoxicity; protein aggregation; response; second messenger; synaptic function

DESCRIPTION (provided by applicant): Oxidative stress plays an important role in the pathogenesis of neurodegenerative diseases and stroke. However, low levels of reactive oxygen species (ROS) function as second messengers in many neuronal signal transduction pathways, which could thus be affected either by the disease process itself or by activation of endogenous antioxidant responses. The long term goal of this research is to define how endogenous antioxidant signaling regulates synaptic transmission and thus shapes vulnerability of synaptic networks to oxidative stress. The specific objective of this application is to elucidate molecular mechanisms underlying synaptic function of Nrf2, the transcriptional regulator of inducible antioxidant response that is a key determinant of neuronal susceptibility to injury. Our central hypothesis is that astrocytes respond to neuronal activity by enhancing current flow through ROS-sensitive glutamatergic NMDA receptors (NMDARs, the key mediators of both synaptic plasticity and glutamate excitotoxicity), while simultaneously activating Nrf2 pathway to protect neurons from ROS-induced damage and neurotoxicity. This hypothesis was formulated on the basis of the strong preliminary data obtained in our laboratory and will be tested by pursuing four specific aims. First, we will elucidate the molecular mechanism that underlies activity-mediated induction of Nrf2 pathway in neuron-astrocyte co-cultures. Second, we will establish whether neuroprotection induced by synaptic activity is enhanced when neurons are co-cultured with astrocytes. Third, we will determine how glial cells increase neuronal NMDAR current density in the mixed neuron-glia environment. Fourth, we will dissect the neuron-glia signal transduction cascade that underlies Nrf2-mediated regulation of NMDAR signaling and determine the effect of Nrf2 pathway activation on circuit plasticity. These aims will be accomplished through a combination of molecular, biochemical, electrophysiological, cell biological, and toxicological approaches whose feasibility in our hands has been established through the preliminary data; to dissect the roles of individual cell types, we will use primary neuronal, glial, and mixed hippocampal cultures, as well as neuron-glia co-cultures of defined cellular composition. The overall approach takes the field in a new direction by focusing on the role of neuron- glia interactions in the regulation and function of Nrf2 signaling in the brain, an aspect of Nrf2 biology that has not yet been investigated. Completion of the proposed research is expected to advance our understanding of ROS signaling and Nrf2 physiology in the brain; ultimately, such knowledge will enable development of pharmacologic treatments capable of harnessing neuroprotective power of endogenous antioxidants without negatively affecting neuronal activity and synaptic signaling.

描述（由申请人提供）：氧化应激在神经退行性疾病和中风的发病机制中起着重要作用。然而，低水平的活性氧（ROS）在许多神经元信号转导途径中充当第二信使，因此可能受到疾病过程本身或内源性抗氧化反应激活的影响。这项研究的长期目标是确定内源性抗氧化信号如何调节突触传递，从而塑造突触网络对氧化应激的脆弱性。本申请的具体目的是阐明 Nrf2 突触功能的分子机制，Nrf2 是诱导性抗氧化反应的转录调节因子，是神经元损伤易感性的关键决定因素。我们的中心假设是，星形胶质细胞通过增强流经 ROS 敏感谷氨酸能 NMDA 受体（NMDAR，突触可塑性和谷氨酸兴奋性毒性的关键介质）的电流来响应神经元活动，同时激活 Nrf2 通路以保护神经元免受 ROS 诱导的损伤和神经毒性。这一假设是根据我们实验室获得的强有力的初步数据制定的，并将通过追求四个具体目标进行测试。首先，我们将阐明神经元-星形胶质细胞共培养物中 Nrf2 通路活性介导诱导的分子机制。其次，我们将确定当神经元与星形胶质细胞共培养时，突触活动诱导的神经保护是否会增强。第三，我们将确定神经胶质细胞如何在混合神经元-神经胶质环境中增加神经元 NMDAR 电流密度。第四，我们将剖析 Nrf2 介导的 NMDAR 信号传导调节背后的神经元-胶质细胞信号转导级联，并确定 Nrf2 通路激活对电路可塑性的影响。这些目标将通过分子、生物化学、电生理学、细胞生物学和毒理学方法的结合来实现，这些方法在我们手中的可行性已通过初步数据确定；为了剖析单个细胞类型的作用，我们将使用原代神经元、神经胶质和混合海马培养物，以及具有特定细胞组成的神经元-神经胶质细胞共培养物。总体方法通过关注神经胶质细胞相互作用在大脑中 Nrf2 信号传导的调节和功能中的作用（Nrf2 生物学的一个尚未被研究的方面），将该领域引向了一个新的方向。拟议研究的完成预计将增进我们对大脑中 ROS 信号传导和 Nrf2 生理学的理解；最终，这些知识将使药物治疗的开发能够利用内源性抗氧化剂的神经保护能力，而不会对神经元活动和突触信号传导产生负面影响。