ROLE INTRACELLULAR Ca2+ CONCENTRATION IN DEVELOPMENT OF ALZHEIMER'S DISEASE

N-酰基乙醇胺（NAE）是一种有效的生物活性脂质信号物质，在哺乳动物生理学中具有多种作用。我们建议研究植物源性NAE作为潜在的替代品和补充剂，以目前现有的神经保护治疗神经系统疾病，包括阿尔茨海默病。我们的初步数据表明，NAE调节细胞内钙通道的功能，允许调节细胞内钙信号。NAE是天然存在的化合物，因此这些化合物存在适当的代谢和清除过程，降低了NAE相关毒性的可能性。该应用的中心假设是NAE对神经元发挥保护作用。NAE介导的神经保护机制将在神经元细胞系、原代神经元培养物和体内神经毒性损伤和神经变性模型中在分子水平上进行分析和评价。特别是，将评估NAE的作用，以确定其预防细胞死亡和引发神经保护相关信号通路的能力。 具体目标是：1）分析NAE对细胞内钙离子通道的生物物理和药理学特性的调节作用，2）鉴定和测量NAE介导的神经元细胞内钙信号反应的贡献; 3)以神经毒性损伤和神经退行性变为模型，确定NAE在神经元细胞系中的神经保护作用; 4）以神经毒性损伤和神经退行性变为模型，鉴定海马的原代神经元培养物中NAE引起的神经保护信号转导途径。5)确定NAE对中风动物模型缺血性损伤的影响。实验将结合生物化学、电生理学、细胞内Ca 2+浓度的光学成像、细胞内信号传导变化的分析和神经保护测定。本研究的总体目标是为开发新的替代或补充治疗阿尔茨海默病神经退行性变提供必要的基础。

N-Acylethanolamines (NAEs) are potent, bioactive lipid signaling substances with diverse roles in mammalian physiology. We propose to investigate plant-derived NAEs functioning both as potential alternatives and supplements to currently existing neuroprotecting treatments for neurological disorders including Alzheimer's disease. Our preliminary data indicate that NAEs regulate the function of intracellular calcium channels allowing modulation of intracellular calcium signaling. NAEs are naturally occurring compounds, and therefore there are in place metabolic and clearance processes for these compounds lessening the likelihood of NAE-associated toxicities. The central hypothesis of this application is that NAEs exert protective effects on neurons. The mechanisms of neuroprotection mediated by NAEs will be analyzed and evaluated at the molecular level, in neuronal cell lines, primary neuronal cultures and in vivo as models of neurotoxic insults and neurodegeneration. In particular, the effect of NAEs will be evaluated for their ability to prevent cell death and elicit neuroprotection-related signaling pathways. The specific aims are: 1) Analyze the modulation of biophysical and pharmacological characteristics of intracellular calcium channels by NAEs; 2) to identify and measure the contribution of NAE mediated responses to intracellular Ca 2+ signaling of neurons; 3) to determine the neuroprotective effects of NAEs in neuronal cell lines as models of neurotoxic insults and neurodegeneration; 4) to identify the neuroprotective signal transduction pathways elicited by NAEs in primary neuronal cultures of the hippocampus as models of neurotoxic insults and neurodegeneration.; 5) to determine the effects of NAEs on ischemic damage in an animal model of stroke. Experiments will use a combination of biochemistry, electrophysiology, optical imaging of intracellular Ca 2+ concentrations, analyses of changes in intracellular signaling and neuroprotection assays. The overall goal of this study is to provide the necessary foundation for the development of novel alternative or supplemental treatments for neurodegeneration in Alzheimer's disease.