Astrocyte Control of Toxin-Mediated Neuron Death: Role of the Gamma-Glutamyl Path

星形胶质细胞控制毒素介导的神经元死亡：γ-谷氨酰路径的作用

• 批准号: 7281387
• 负责人: GEORGE I HENDERSON
• 金额: $ 28.81万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-10 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7281387
• 关键词: Address; Aging; Alzheimer' s Disease; Animals; Antioxidants; Apoptosis; Apoptotic; Area; Astrocytes; Biological Models; Brain; Cell Culture System; Cell Death; Cell membrane; Cells; Cerebral cortex; Cessation of life; Clinical; Coculture Techniques; Commit; Cysteine; Cystine; Data; Development; Disease; Elements; Ethanol; Event; Exopeptidase; Exposure to; Functional disorder; Gamma-glutamyl transferase; Glutamates; Glutathione; Glycine; Herbicides; Homeostasis; Human; Image; In Vitro; Injury; Intervention; Lead; Life; Mediating; Microscopy; Mitochondria; Multiple Sclerosis; Nerve Degeneration; Neurodegenerative Disorders; Neuroglia; Neurons; Neurotoxins; Organic Anion Transporters; Oxidative Stress; P-Glycoproteins; Paraquat; Parkinson Disease; Pathway interactions; Pattern; Pesticides; Primary Lateral Sclerosis; Principal Investigator; Process; Proteins; Publishing; Reactive Oxygen Species; Regulation; Regulatory Pathway; Research; Research Personnel; Rodent Model; Role; Rotenone; Source; Stress; Stroke; System; Testing; Therapeutic Intervention; Time; Toxic Environmental Substances; Toxic effect; Toxicology; Toxin; Transgenic Mice; Triad Acrylic Resin; Work; alanine aminopeptidase; alcohol response; cell injury; cysteinylglycine; environmental stressor; gamma glutamyl path; in vivo Model; multi-photon; neuron component; neuroprotection; neurotoxic; neurotoxicity; nonhuman primate; novel; prevent; programs; protein expression; research study; response; stressor; toxin mediated neuron death; two-photon

DESCRIPTION (provided by principal investigator): Background Toxin-mediated oxidative stress (OS) is a trigger that ultimately commits neurons to apoptotic death. The event can be associated with decreased cell glutathione (GSH) and cell death is mitigated by augmenting neuron GSH. Astrocytes protect neurons from toxin-related OS and subsequent apoptosis via the ?-glutamyl cycle, which maintains neuron GSH homeostasis. An understanding of this neuroprotective pathway, especially its regulatory components and species comparisons, could ultimately lead to novel clinical interventions for devastating neurodegenerative disorders as well as establish accurate model systems. Hypothesis: We hypothesize that within the astrocyte-neuron axis, there is a highly effective, regulated pathway consisting of components which can enhance neuron GSH homeostasis in response to EtOH (E) and Parkinson's Disease-producing environmental toxins, thereby minimizing death of neurons. However, there are exposure patterns to these environmental cross-stressors which damage components of the pathways, thereby impairing its neuroprotective capacity. Specific Aim 1: Aim one will utilize cultures of neurons and astrocytes, alone and in co-culture, to address the direct impact of rotenone, paraquat, and E on astrocyte and neuron components of the ?-glutamyl cycle. Experimental parameters will be effects of the environmental toxins, alone or in combination, on inward transport of key precursors of GSH, GSH enzymatic synthesis, and components of this machinery at which neuroprotection occurs. Specific Aim 2: Aim two will address regulation of three essential components of the neuroprotective pathway. To be determined will be basic mechanisms underlying regulatory and damaging responses to the toxins. These will be regulation of GSH efflux via multidrug resistance protein(s) (Mrp), and two plasma membrane ectopeptidases, ?-glutamyl transpeptidase (GGT) and aminopeptidase N (ApN). Specific Aim 3: Aim 3 will use an in vivo model to extend the in vitro findings to the intact animal. Experiments will utilize two-photon excitation microscopy to determine toxin effects specifically in cortical astrocytes and neurons in the living brain. These will be time-lapse determinations of expressions of GGT and ApN, GSH content and apoptosis-related events during toxin exposure. They will utilize transgenic mice deficient in Mrpl, Mrp4, or GGT. General Description: The proposal addresses a new system by which glial cells protect neurons from the toxic effects of three environmental toxins. These compounds can cause neurons to die as a result of oxidative damage and cells called astrocytes prevent this by maintaining neuron antioxidants. We will determine how the toxins damage it (to prevent this from happening), and how this system is normally controlled.

描述（由首席调查员提供）： 背景毒素介导的氧化应激（OS）是最终将神经元变成凋亡死亡的触发因素。该事件可以与细胞谷胱甘肽（GSH）减少有关，并且通过增加神经元GSH来减轻细胞死亡。星形胶质细胞通过？-谷氨基循环保护神经元免受毒素相关的OS和随后的细胞凋亡，该周期维持神经元GSH稳态。对这种神经保护途径的理解，尤其是其调节成分和物种比较，最终可能导致新的临床干预措施，以造成破坏性神经退行性疾病以及建立准确的模型系统。假设：我们假设在星形胶质细胞神经元轴内，有一种高效，调节的途径，该途径由组件组成，可以增强神经元GSH稳态，以响应ETOH（E）和帕金森氏病产生的环境毒素，从而使神经元死亡最小。但是，这些环境交叉压力会损害途径的成分，从而损害其神经保护能力。特定目的1：目标一个人将单独和在共培养中利用神经元和星形胶质细胞的培养物来解决烤面包酮，帕拉夸酸和E对？ - 谷氨酰基循环的星形胶质细胞和神经元成分的直接影响。实验参数将是环境毒素单独或组合的影响，对GSH的关键前体，GSH酶合成的内在转运以及发生神经保护剂的这种机械的成分。特定目标2：目标两个将解决对神经保护途径的三个基本组成部分的调节。待确定将是对毒素的调节和破坏性反应的基本机制。这些将是通过多药耐药蛋白（S）（MRP）和两个质膜to肽酶的调节GSH外排调节？ - 谷氨酰基转肽酶（GGT）和氨基肽酶N（APN）。特定目标3：AIM 3将使用体内模型将体外发现扩展到完整的动物。实验将利用两光子激发显微镜来确定活体大脑中皮质星形胶质细胞和神经元中的毒素作用。这些将是毒素暴露期间GGT和APN，GSH含量和与凋亡相关的事件的表达的延时确定。他们将利用缺乏MRPL，MRP4或GGT的转基因小鼠。一般描述：该提案解决了一个新系统，通过该系统，神经胶质细胞可保护神经元免受三种环境毒素的毒性作用。这些化合物会导致神经元因氧化损伤而死亡，而称为星形胶质细胞的细胞通过维持神经元抗氧化剂来阻止这种损伤。我们将确定毒素如何损害毒素（以防止这种情况发生），以及该系统通常如何控制。