Calcineurin and inflammatory signaling processes in aging and Alzheimer's Disease

衰老和阿尔茨海默病中的钙调神经磷酸酶和炎症信号传导过程

基本信息

批准号：
8825991
负责人：
Christopher Mark Norris
金额：
$ 29.04万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8825991
关键词：
APP-PS1 Age Aging Alzheimer&apos s Disease Alzheimer&apos s disease model Animals Astrocytes Automobile Driving Avoidance Learning Behavior Behavioral Biochemical Process Biological Assay Biological Markers Brain Brain Diseases Calcineurin Cells Ceramics Cognitive Data Dependence Dependovirus Deposition Development Disease Electrodes Electrophysiology (science)Enzyme-Linked Immunosorbent Assay Enzymes Funding Gene Delivery Glial Fibrillary Acidic Protein Glutamate Receptor Glutamate Transporter Glutamates Hippocampus (Brain)Human Hypertrophy Impaired cognition Implant In Vitro Inflammatory Ipsilateral Label Measurement Measures Methods Microelectrodes Microglia Mus NFAT Pathway Nerve Degeneration Nervous System Physiology Neurodegenerative Disorders Neurologic Dysfunctions Outcome Measure Pathologic Pathology Pathway interactions Phenotype Process Production Protein phosphatase Proteins Reaction Regulation Resolution Rodent Model Role Signal Transduction Slice Staging Synapses Synaptic plasticity Techniques Testing Therapeutic Time Tissues Transgenic Mice Transgenic Organisms Western Blotting Wild Type Mouse adeno-associated viral vector aging brain amyloid pathology astrogliosis base beta-site APP cleaving enzyme 1 cytokine functional outcomes glutamatergic signaling improved in vivo inhibitor/antagonist innovation interest mouse model multi-electrode arrays neuroinflammation neuropathology neuroprotection novel strategies prevent promoter protein expression relating to nervous system research study response signal processing synaptic function transcription factor treatment strategy vector

项目摘要

DESCRIPTION (provided by applicant): The experiments in this project use innovative gene delivery techniques and a multipronged approach to assess the fundamental role of astrocyte activation in neurologic function in an intact mouse model of Alzheimer's disease (AD). Studies use adeno-associated virus vectors (AAV) bearing the astrocyte-specific promoter Gfa2 to target the protein phosphatase calcineurin (CN) and NFAT transcription factors in astrocytes of wild-type and APP/PS1 mice. AAV-Gfa2 vectors are bilaterally delivered to the hippocampus at different ages/disease stages and mice are assessed on different AD biomarkers. In Aim 1, cognitive status is assessed using the active avoidance task, while synaptic function is evaluated using hippocampal slice electrophysiology and Western blot measures of synaptic proteins. In Aim 2, hippocampal glutamate regulation is investigated using ceramic enzyme-based microelectrode arrays and measures of glutamate transporter levels. In Aim 3, levels of glial activation and neuroinflammation are determined with immunohistochemical (IHC) analyses and assessment of cytokine levels using Multiplex ELISAs. In Aim 4, IHC is used to determine the extent of A? deposition, while ELISAs are used to quantify levels of A?40 and A?42 in soluble and insoluble hippocampal tissue fractions, and Westerns used to assess BACE protein expression. AAV-Gfa2 vectors encode either potent inhibitors or activators of CN/NFAT signaling and therefore will determine the necessity and sufficiency of this astrocytic pathway in driving and/or maintaining neurologic dysfunction in AD mice. These studies provide a highly novel approach to the study of activated astrocytes and could have a major impact on the development of treatment strategies for AD and other neurodegenerative conditions.

描述（由申请人提供）：该项目中的实验使用创新的基因输送技术和一种多重方法来评估星形胶质细胞激活在神经系统疾病（AD）完整小鼠模型中的神经系统功能中的基本作用。研究使用带有星形胶质细胞特异性启动子GFA2的腺相关病毒载体（AAV）靶向蛋白质磷酸酶钙调蛋白（CN）和NFAT转录因子的野生型和APP/PS1小鼠的NFAT转录因子。 AAV-GFA2载体在不同年龄/疾病阶段将双侧交付到海马，并且在不同的AD生物标志物上评估小鼠。在AIM 1中，使用主动回避任务评估认知状态，而突触功能则使用海马切片电生理学和突触蛋白的蛋白质印迹测量评估。在AIM 2中，使用基于陶瓷酶的微电极阵列和谷氨酸转运蛋白水平的测量来研究海马谷氨酸调节。在AIM 3中，使用多重ELISAS进行了免疫组织化学（IHC）分析，并评估细胞因子水平，确定神经胶质激活和神经炎症水平。在AIM 4中，IHC用于确定A的程度？沉积，虽然ELISA用于量化可溶性和不溶性海马组织中的A 40和A？42的水平，而西方则用于评估BACE蛋白表达。 AAV-GFA2矢量编码了CN/NFAT信号的有效抑制剂或活化剂，因此将确定该星形胶质细胞途径在AD小鼠中驾驶和/或维持神经功能障碍时的必要性和充分性。这些研究提供了一种高度新颖的方法来研究活化的星形胶质细胞，并可能对AD和其他神经退行性疾病的治疗策略的发展产生重大影响。