ROLE OF CYCLIC ADP-RIBOSE IN CALCIUM DISPOSITION IN NG108-15 CELLS

环状 ADP-核糖在 NG108-15 细胞钙沉积中的作用

• 批准号: 6237945
• 负责人: TIMOTHY Francis WALSETH
• 金额: $ 8.62万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-03-10 至 1998-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6237945
• 关键词: ADP ribosylation; adenosine diphosphate; caffeine; calcium flux; high performance liquid chromatography; homeostasis; hybrid cells; immunologic assay /test; inositol phosphates; microsomes; neurons; phosphorus oxygen lyase; purinergic receptor; radioimmunoassay; radiotracer; ribose; tissue /cell culture

The overall objective of the proposed research is to define the role of cyclic ADP-ribose (cADPR) in the regulation of calcium homeostasis in NG108 cells, a neuronal cell line. cADPR is a naturally occurring nucleotide found to be a potent mediator of calcium release from intracellular stores. The specific aims of the proposal are: (1) to characterize intracellular calcium pools in NG 108 cells, (2) to characterize the cADPR synthetic and degradative enzymes in NG108 cells, (3) to characterize the cADPR binding protein(s) in NG108 cells, and (4) to examine the regulation of the endogenous level of cADPR in NG108 cells. The intracellular pools of calcium in NG108 cells will be probed using detergent permeabilized cells or purified microsomes and characterized in terms of the ability of cADPR, caffeine and inositol trisphosphate to release stored calcium. The characterization of the proteins that synthesize, degrade and bind cADPR will be achieved through studies on their subcellular localization, regulation, and purification. The regulation of the intracellular concentration of cADPR will be examined by measuring cADPR under a variety of conditions known to raise intracellular calcium. The regulation of the intracellular concentration of calcium is a critical process in all cells . In neuronal cells, changes in calcium play an important role in a number of neuronal processes including neurotransmitter release, changes in the cytoskeleton, gene expression and a number of metabolic events. Recent evidence suggests that cADPR may be the endogenous modulator of calcium-induced calcium release and act through a mechanism similar to caffeine. Caffeine is a powerful stimulant of the central nervous system, and probably because of this action, is one of the most widely used drugs in the world. The mechanism by which caffeine stimulates the central nervous system is not completely understood. Since cADPR appears to be the endogenous modulator of calcium release from caffeine-sensitive stores, a systematic study of cADPR function and metabolism in neuronal cells should provide useful information on stimulation of the central nervous system.

拟议研究的总体目的是定义 在调节钙稳态中的环状ADP-核糖（CADPR） NG108细胞，神经元细胞系。 CADPR是自然发生的 核苷酸发现是钙释放的有效介质 细胞内存储。 该提案的具体目的是：（1） 表征Ng 108细胞中的细胞内钙池，（2）至 表征NG108细胞中的CADPR合成和降解酶， （3）表征NG108细胞中的CADPR结合蛋白（S）和（4） 检查NG108中CADPR的内源性水平的调节 细胞。 NG108细胞中钙的细胞内池将被探测 使用清洁剂通透性细胞或纯化的微粒体和 以CADPR，咖啡因和肌醇的能力为特征 三磷酸盐以释放储存的钙。 表征 合成，降解和结合CADPR的蛋白质将通过 研究其亚细胞定位，调节和纯化。 CADPR细胞内浓度的调节将是 通过在已知升高的各种条件下测量CADPR检查 细胞内钙。 细胞内浓度的调节 钙的关键过程是所有细胞的关键过程。在神经元细胞中， 钙的变化在许多神经元中起重要作用 包括神经递质释放的过程，变化 细胞骨架，基因表达和许多代谢事件。 最近的 有证据表明CADPR可能是内源调节剂 钙诱导的钙释放并通过类似的机制起作用 咖啡因。 咖啡因是中枢神经的有力兴奋剂 系统，可能是由于这种动作，是最广泛的 在世界上使用了药物。 咖啡因刺激的机制 中枢神经系统尚未完全理解。 由于CADPR出现 成为从咖啡因敏感的钙释放的内源调节剂 商店，对神经元中CADPR功能和代谢的系统研究 细胞应提供有关中央刺激的有用信息 神经系统。