β淀粉样蛋白调节囊泡谷氨酸转运体功能的研究

项目名称： β淀粉样蛋白调节囊泡谷氨酸转运体功能的研究

项目编号： No.81473191

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 程肖蕊

作者单位： 中国人民解放军军事科学院军事医学研究院

项目金额： 80万元

中文摘要： 阿尔茨海默病(AD)发病机制不明,临床尚无有效治疗药物。大量研究表明,淀粉样蛋白(Aβ)与谷氨酸能神经传递通路多个环节相互作用在AD发病机制中发挥关键作用。但代谢产生Aβ的APP及Aβ与谷氨酸释放的控制源头囊泡谷氨酸转运体(VGLUT1)的相互作用并不清楚。为了研究APP/Aβ是否与VGLUT1存在相互作用,本课题首先观察APP与VGLUT1在脑组织、神经细胞、突触囊泡的共定位情况,其次采用计算机模拟、免疫共沉淀及生物膜干涉技术观察APP/Aβ与VGLUT1是否存在相互作用,接着基于神经元和纯化的突触囊泡观察Aβ对VGLUT1转运功能的调节作用,最后以Aβ可能和VGLUT1的变构激活子Cl-竞争结合进而调节VGLUT1活性为假说来揭示Aβ调节VGLUT1功能的具体机制。本课题将揭示Aβ是否调节以及调节VGLUT1功能的具体机制,可为AD发病机制的深入阐明和防治药物靶点的发现奠定基础。

中文关键词： 阿尔茨海默病；突触可塑性；药物靶标；学习记忆；神经保护

英文摘要： Alzheimer's disease (AD) is an irreversible, progressive brain disease and the most common cause of dementia among older people. The drugs now available to treat AD provide for some people symptomatic benefit, but there are no approved disease-modifying therapies. The AD etiology remains unknown or incomplete understanding now. Accumulated evidence indicates there are interactions between glutamatergic neurotransmission and APP metabolic process. However, a direct link between VGLUT and APP, the key molecular in the amyloid cascade hypothesis and glutamate excitotoxicity hypothesis respectively on AD etiology, has yet to be established. In order to investigate the direct link between VGLUT and APP/Aβ, the probability of colocalization between APP and VGLUT1 in normal physiological state will be observed in the normal adult mouse brain, neurons, astrocytes, microglia, and synaptic vesicles isolated in the present study. Then, the interactions between APP/A β and VGLUT1 will be tested using computer simulation, immunoprecipitation and biolayer interferometry technology. Furthermore, the regulation of A β on the VGLUT1 transporting glutamate into synaptic vesicle will be investigated based on purified synaptic vesicle and primary cultured neuron. At last, the mechanism underlying Aβ regulating VGLUT1 function will be disclosed based on the hypothesis that Aβ regulates VGLUT1 transporting glutamate by competing with Cl- at the site of allosteric regulation. Therefore, This study aims to reveal the interaction between VGLUT and APP/Aβ and clarify the specific mechanisms of A β regulating VGLUT1 function. It is important for the pathogenesis of AD and discovery of AD therapeutic target.

英文关键词： Alzheimer's disease;Synaptic plasticity;drug target;learning and memory;neuroprotection