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

N-乙酰乙醇胺(NAE)是一种有效的、生物活性的脂质信号物质，在哺乳动物生理中具有不同的作用。我们建议研究植物来源的NAE作为现有神经保护治疗的潜在替代品和补充，用于治疗包括阿尔茨海默病在内的神经疾病。我们的初步数据表明，NAE调节细胞内钙通道的功能，从而调节细胞内钙信号。NAE是天然存在的化合物，因此对这些化合物有适当的代谢和清除过程，以减少与NAE相关的毒性的可能性。这一应用的中心假设是NAE对神经元起到保护作用。NAE介导的神经保护机制将作为神经毒性损伤和神经变性的模型在神经细胞系、原代神经元培养和活体模型中从分子水平进行分析和评估。特别是，将评估NAE在防止细胞死亡和引发神经保护相关信号通路方面的作用。 其具体目的是：1)分析NAE对细胞内钙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介导的反应对神经元细胞内钙信号的作用； 3)确定NAE在神经细胞系中作为神经毒性损伤和神经变性模型的神经保护作用；4)确定NAE在原代培养的海马神经元中诱导的神经保护信号转导通路作为神经毒性损伤和神经变性的模型；5)确定NAE对卒中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、细胞内钙浓度的光学成像、细胞内信号变化的分析和神经保护分析。这项研究的总体目标是为开发阿尔茨海默病神经变性的新的替代或补充治疗方法提供必要的基础。

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.