Defects of Neurotransmission in Ischemia and Reperfusion

缺血和再灌注中神经传递的缺陷

• 批准号: 7350929
• 负责人: Alexander A Mongin
• 金额: $ 17.28万
• 依托单位: ALBANY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-05 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7350929
• 关键词: 7-nitroindazole; Acetylcysteine; Affect; Antioxidants; Arts; Attention; Biological Preservation; Blood flow; Brain; Cardiovascular system; Cell Death; Cell physiology; Cessation of life; Chemistry; Chimeric Proteins; Communication; Cysteine; Data; Defect; Disruption; Docking; Environment; Equipment; Ethylmaleimide; Foundations; Freezing; Functional disorder; Genes; Glutamate Receptor; Hour; Impairment; In Vitro; Interdisciplinary Study; Interruption; Intervention; Ischemia; Ischemic Penumbra; Knowledge; Laboratories; Lesion; Link; Literature; Measurement; Measures; Medical; Metabolism; Methods; Modeling; Modification; Molecular; Molecular Conformation; Monitor; Motor Cortex; N-ethylmaleimide-sensitive protein; Neurons; Neuropharmacology; Neurosciences; Nitric Oxide; Nitric Oxide Synthase; Nitrosation; Pathologic Processes; Pathology; Patients; Phosphorylation; Physiological reperfusion; Post-Translational Protein Processing; Preparation; Proteins; Proteomics; Rattus; Reactive Nitrogen Species; Reagent; Recording of previous events; Reducing Agents; Rehabilitation therapy; Reperfusion Therapy; Reporting; Research; Research Personnel; Running; SNAP receptor; Science; Stroke; Sulfhydryl Compounds; Synapses; Synapsins; Synaptic Transmission; Synaptic Vesicles; Synaptosomes; Tamoxifen; Testing; Therapeutic; Tissues; Transient Ischemic Attack; Vesicle; Work; base; brain tissue; college; day; expectation; experience; in vivo; inhibitor/antagonist; knockout animal; neurotransmission; neurotransmitter release; oxidation; postsynaptic; presynaptic; prevent; programs; protein function; response; size; tissue preparation

Stroke is a pathologicalreduction in blood flow which causes irreversible damage of brain tissue, long-term disruptions of brain functions, and frequently patient death. The major focus of ischemia research is on the mechanisms of cell death and the search for pharmacological approaches to salvage vulnerable neural cells. Substantially less attention is paid to long-term impairment of neural cell function that is not associatedwith cell death. Existing literature suggest, nonetheless, that even relatively mild ischemia causes long-lasting suppression of synaptic communication. For unknown reasons, only presynaptic release is affected, while postsynaptic responses remain preserved. In the current study we hypothesize that suppression of synaptic transmission occurs due to nitrosation of cysteine residues in N-ethylmaleimide sensitive fusion protein (NSF) by nitric oxide (NO) and NO-related reactive nitrogen species. The NSFprotein is a trimeric ATPasewhich is crucial for continuous synaptic vesicle docking/fusion. Modification of critical cysteine residues in the NSF by N-ethylmaleimide causes irreversible inhibition of vesicle docking and fusion. Similarly, NO-dependent modification of NSF thiols may cause long-term inhibition of vesicular neurotransmitter release. To test our hypothesis we propose the following specific aims. (1) We will test for the increased modification of thiols in the ischemic tissue up to 24 hours after ischemia and will further demonstrate thiol modification in the NSFprotein immunoprecipitated from the ischemic brain. (2) Using synaptosomal preparation we will examine whether nitric oxide and related reactive nitrogen species modify NSFand via this mechanism inhibit vesicular neurotransmitter release. (3) We will further explore reversibility of NSFnitrosation and defects in neurotransmitter release in vitro and in vivo by applying nitric oxide synthase inihibitors and thiol reducing agents. This project will investigate the molecular mechanisms of poorly understood long-term disruption of synaptic transmission in ischemia, and will provide a foundation for an R01 project developing additional therapeutic approaches for patient treatment and rehabilitation after stroke and transient ischemic attacks.

中风是一种病理性的血流减少，导致脑组织长期不可逆的损伤， 大脑功能的破坏，以及患者的死亡。缺血研究的主要焦点是 细胞死亡的机制和寻找药理学方法来挽救脆弱的神经细胞。 对与神经细胞功能损伤无关的神经细胞功能的长期损害的关注少得多。 细胞死亡然而，现有的文献表明，即使是相对轻微的缺血也会导致长期的缺血性脑损伤。 抑制突触通讯由于未知的原因，只有突触前释放受到影响，而 突触后反应保持不变。在目前的研究中，我们假设， 突触传递的发生是由于N-乙基马来酰亚胺敏感的 融合蛋白（NSF）通过一氧化氮（NO）和NO相关的活性氮物种。NSF蛋白是 一种三聚体ATP酶，对连续突触囊泡对接/融合至关重要。修改关键 NSF中的半胱氨酸残基通过N-乙基马来酰亚胺引起囊泡对接的不可逆抑制， 核聚变类似地，NSF巯基的NO依赖性修饰可能导致囊泡性细胞凋亡的长期抑制。 神经递质释放为了验证我们的假设，我们提出了以下具体目标。(1)我们将测试 缺血后24小时内缺血组织中硫醇的修饰增加， 进一步证明了从缺血性脑损伤免疫沉淀的NSF蛋白中的巯基修饰， 个脑袋(2)使用突触体制备，我们将检查是否一氧化氮和相关的反应性 氮物质修饰NSF并通过该机制抑制囊泡神经递质释放。（三） 我们将进一步探讨NSF亚硝化的可逆性和体外神经递质释放的缺陷 以及通过应用一氧化氮合酶抑制剂和硫醇还原剂在体内进行。该项目将 研究突触传递长期中断的分子机制， 缺血，并将为R 01项目开发其他治疗方法提供基础， 中风和短暂性脑缺血发作后的患者治疗和康复。

项目成果

Long-lasting inhibition of presynaptic metabolism and neurotransmitter release by protein S-nitrosylation.

通过蛋白质 S-亚硝基化作用持久抑制突触前代谢和神经递质释放。

• DOI: 10.1016/j.freeradbiomed.2010.05.032
• 发表时间: 2010
• 期刊: Free radical biology & medicine
• 影响因子: 7.4
• 作者: Rudkouskaya,Alena;Sim,Vasiliy;Shah,AabhaA;Feustel,PaulJ;Jourd'heuil,David;Mongin,AlexanderA
• 通讯作者: Mongin,AlexanderA

Nitric oxide may contribute to the long-term impairment of synaptic transmission after transient ischemia.

一氧化氮可能导致短暂性缺血后突触传递的长期损害。

• DOI: 10.1161/01.str.0000033074.40202.8e
• 发表时间: 2002
• 期刊: Stroke
• 影响因子: 8.3
• 作者: Mongin,AA