Liberation of Intracellular Zinc and Neuronal Cell Death

发明描述（由申请人提供）已知囊泡Zn 2+在其从突触前释放位点移位到注定死亡的突触后细胞的细胞质中之后对神经元有毒。除了这种胞外来源的Zn 2+，我们已经观察到，这种金属可以从细胞内的金属结合蛋白的硫醇氧化剂释放。我们已经进一步表明，胞内释放的Zn 2+是一个强大的神经元凋亡刺激。在这里，使用分子，细胞，和整个动物的方法，我们建议评估是否释放细胞内锌+代表氧化应激后神经元细胞死亡的一个共同特征。 该提案的具体目标是：1。确定过氧亚硝酸盐和一氧化氮是否触发神经毒性级联反应，导致细胞内锌在体外释放。2.鉴定与体外细胞内锌释放相关的细胞死亡途径的分子组分。3在轴突切断诱导的神经元细胞死亡的体内模型中评估细胞内锌的释放是否触发细胞凋亡。在这些目标中描述的实验将测试细胞内Zn 2+的释放代表神经元损伤中细胞死亡的重要且普遍存在的触发的假设。这项研究计划的长期目标是提供额外的治疗靶点，以预防或最大限度地减少与氧化损伤相关的大量神经系统疾病中的神经元细胞死亡。

DESCRIPTION (provided by the applicant) Vesicular Zn2+ is known to be toxic to neurons following its translocation from presynaptic release sites into the cytoplasm of postsynaptic cells that are destined to die. In addition to this extracellular source of Zn2+, we have observed that this metal can be released from intracellular metal binding proteins by thiol axidants. We have shown further that intracellularly liberated Zn2+ is a powerful apoptotic stimulus in neurons. Here, using molecular, cellular, and whole animal approaches, we propose to evaluate whether the release of intracellular Zn2+ represents a common feature in neuronal cell death following oxidative stress. The Specific Aims of this proposal are: 1. To determine whether peroxynitrite and nitric oxide trigger a neurotoxic cascade that results from the intracellular release of zinc in vitro. 2. To identify molecular components of the cell death pathway that are associated with the intracellular release of zinc in vitro. 3 To evaluate whether the intracellular release of zinc triggers apoptosis in an axotomy-induced in vivo model of neuronal cell death. Experiments described within these aims will test the hypothesis that the liberation of intracellular Zn2+ represents an important and ubiquitous trigger for cell death in neuronal injury. The long-term goal of this research program is to provide additional therapeutic targets to prevent or minimize neuronal cell death in the large number of neurological disorders associated with oxidative injury.