High-Speed Assays for Neuromessengers Involved in Stroke

中风神经信使的高速检测

• 批准号: 7062505
• 负责人: MICHAEL T BOWSER
• 金额: $ 23.47万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-15 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7062505
• 关键词: antioxidants; biological signal transduction; capillary electrophoresis; glutamates; histology; ischemia; laboratory rat; microdialysis; neurochemistry; nitric oxide; pharmacokinetics; reperfusion; serine; stroke; technology /technique development; tissue /cell culture; zinc

Toxic levels of glutamate released during ischemia have been implicated in the damage caused by stroke. Unfortunately, drugs that mediate the effect of glutamate have proven ineffective or cause unacceptable side effects. Current research is moving towards investigating a number of neuromessengers involved in glutamate signaling including Zn2+, D-serine and nitric oxide. Developing treatments for stroke based on these neuromessengers has been hampered by our limited understanding of their function in vivo. Currently there are no methods for measuring the dynamics of Zn2+, D-serine and nitric oxide in vivo on a time scale of seconds. We propose designing online capillary electrophoresis-microdialysis assays for Zn2+, D-serine and nitric oxide capable of making measurements every 10 seconds, a 120-fold improvement over existing techniques. These assays will allow a detailed pharmacological study of the function Zn2+, D-serine and nitric oxide play in vivo. Tissue (salamander retina) and single cell models (neuron and gila) will also be studied to compare different levels of complexity. The dynamics of Zn2+, D-serine and nitric oxide will be measured during ischemia and reperfusion for the first time. It is hypothesized that toxic concentrations of Zn2+ and nitric oxide are released during ischemia. Understanding what happens in the brain chemically during ischemia is cdtical considering that most of the damage caused by stroke is thought to be chemical in origin. We will use the high temporal resolution assays for glutamate, Zn2+, D-serine and nitric oxide to test the effect of several proposed preventative stroke treatments. Classes of compounds that will be tested include blood thinners (aspirin, t-PA), anaerobic energy sources (creatine, arginine), antioxidants (vitamins C and E) and metal chelators (EDTA).

缺血期间释放的毒性谷氨酸水平与中风引起的损害有关。不幸的是，介导谷氨酸作用的药物已被证明是无效的， 会产生不可接受的副作用目前的研究正朝着调查一些 参与谷氨酸信号传导的神经信使包括Zn 2+、D-丝氨酸和一氧化氮。 基于这些神经信使的中风治疗方法的开发受到了我们对其体内功能的有限理解的阻碍。目前还没有测量 以秒为时间尺度的体内Zn 2+、D-丝氨酸和一氧化氮的动力学。 我们建议设计在线毛细管电泳-微透析分析法， 一氧化氮每10秒测量一次，比现有技术提高了120倍。这些测定将允许详细的药理学研究的功能Zn 2+，D-丝氨酸和一氧化氮在体内发挥。组织（蝾螈视网膜）和单细胞模型（神经元和神经胶质细胞）也将进行研究，以比较不同水平的复杂性。 在缺血期间测量Zn 2+、D-丝氨酸和一氧化氮的动力学， 第一次再灌注。据推测，有毒浓度的锌2+和硝酸盐 氧化物在局部缺血期间释放。了解大脑在化学上发生了什么， 考虑到大多数由中风引起的损伤被认为是 化学起源 我们将使用高时间分辨率分析谷氨酸，锌2+，D-丝氨酸和硝酸盐 氧化物来测试几种预防性中风治疗方法的效果。类 将被测试的化合物包括血液稀释剂（阿司匹林，t-PA），厌氧能源 （肌酸、精氨酸）、抗氧化剂（维生素C和E）和金属螯合剂（EDTA）。