Astrocytic Calcineurin and Connexin43 Gap Junctions in Alzheimer's Disease

描述（由申请人提供）：作为影响超过520万人的第六大死亡原因，阿尔茨海默病（AD）正在成为一场健康危机。令人失望的是，以前的临床试验在寻找这种毁灭性疾病的治疗方法方面收效甚微。因此，探索新的和替代的分子靶标是至关重要的。拟议的项目将测试总体假设，即钙调神经磷酸酶（CN）和连接蛋白43（Cx43）的相互作用破坏了AD进展过程中星形胶质细胞的间隙连接（GJ）偶联，导致神经功能的有害变化。本研究将采用多学科的方法，使用一系列AD模型系统，包括人类海马标本、APP/PS1转基因小鼠和大鼠原代星形胶质细胞培养物，来研究这一假说。除了优秀的模型系统外，该项目还将使用各种尖端技术，包括荧光共振能量转移（FRET），腺相关病毒（AAV）介导的基因传递和切片电生理学。主要的假设将通过三个独立的具体目标进行检验，这些目标如下：使用诸如FRET和免疫共沉淀（Co-IPs）的技术来检验在AD进展期间CN与Cx43相互作用的假设;使用诸如染料偶联和Western印迹的技术来检验β淀粉样蛋白（A-β）以CN依赖的方式破坏间隙连接（GJ）偶联的假设;使用认知测量如主动回避范式和技术如染料偶联、蛋白质印迹和切片电生理学来检验CN/Cx43相互作用破坏AD小鼠模型中的GJ偶联和神经功能的假设。该项目不仅提供神经科学研究方面的上级培训，还将为AD的潜在新治疗靶点提供极其有价值的信息。

DESCRIPTION (provided by applicant): As the sixth leading cause of death that affects over 5.2 million people, Alzheimer's disease (AD) is becoming a health crisis. To much disappointment, previous clinical trials have shown little success in finding a treatment for this devastating disease. Thus, it is critical that new and alternative molecular targets be explored. The proposed project will test the overarching hypothesis that calcineurin (CN) and connexin43 (Cx43) interactions disrupt gap junction (GJ) coupling in astrocytes during the progression of AD leading to detrimental changes in neurologic function. This project will use a multi-disciplinary approach to investigate the hypothesis using an array of AD model systems, including human hippocampal specimens, APP/PS1 transgenic mice, and rat primary astrocyte cultures. In addition to the excellent model systems, this project will use a variety of cutting-edge techniques, including fluorescence resonance energy transfer (FRET), adeno-associated virus (AAV) - mediated gene delivery, and slice electrophysiology. The overarching hypothesis will be examined through three independent specific aims which are as follows: to test the hypothesis that CN interacts with Cx43 during the progression of AD using techniques such as FRET and co-immunoprecipitations (Co-IPs); to test the hypothesis that beta amyloid (A-beta) disrupts gap junction (GJ) coupling in a CN-dependent manner using techniques such as dye coupling and Western blot; to test the hypothesis that CN/Cx43 interactions disrupt GJ coupling and neurologic function in a mouse model of AD using cognitive measures such as the active avoidance paradigm and techniques such as dye coupling, Western blot, and slice electrophysiology. This project not only provides superior training in neuroscience research, but also will provide extremely valuable information into a potential new therapeutic target for AD.