High-Speed Assays for Neuromessengers Involved in Stroke

脑缺血期间释放的谷氨酸有毒水平与中风造成的损害有关。不幸的是，调节谷氨酸作用的药物已被证明无效或 造成不可接受的副作用。目前的研究正在向调查一些 参与谷氨酸信号转导的神经信使包括锌、D-丝氨酸和一氧化氮。 由于我们对它们在体内的功能了解有限，基于这些神经信使的中风治疗方法的开发一直受到阻碍。目前还没有方法来测量 体内锌、D-丝氨酸和一氧化氮在秒尺度上的动态变化。 我们建议设计在线毛细管电泳微渗析分析锌，D-丝氨酸 以及每10秒进行一次测量的一氧化氮，比现有技术提高了120倍。这些检测将允许对体内锌、D-丝氨酸和一氧化氮的作用进行详细的药理学研究。组织(火蜥蜴视网膜)和单细胞模型(神经元和GILA)也将被研究，以比较不同程度的复杂性。 将测定锌、D-丝氨酸和一氧化氮在缺血和缺氧时的动态变化。 第一次再灌流。据推测，有毒浓度的锌和硝酸盐 在缺血过程中会释放出氧化物。了解大脑在化学过程中发生了什么 考虑到中风造成的大部分损害被认为是 化学物质的来源。 我们将使用高时间分辨率的谷氨酸、锌、D-丝氨酸和硝酸盐的分析 氧化物，以测试几种拟议的预防性中风治疗的效果。班级 将接受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)。

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).