Equipment request to increase scientific rigor and reproducibility

提高科学严谨性和可重复性的设备要求

• 批准号: 10060295
• 负责人: Erin R Hascup
• 金额: $ 24.81万
• 依托单位: SOUTHERN ILLINOIS UNIVERSITY SCH OF MED
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-15 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10060295
• 关键词: APP-PS1; Acute; Address; Affect; Age; Age-Months; Aging; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Amyloid beta-42; Amyloid beta-Protein; Animals; Axon; Behavioral; Brain; Chronic; Cognition; Cognitive; Dementia; Disease; Disease Progression; Early identification; Electrochemistry; Equipment; Evaluation; Event; Functional disorder; Glutamate Transporter; Glutamates; Hippocampus (Brain); Human; Immunohistochemistry; Impaired cognition; Impairment; Intervention; Kinetics; Knock-in; Knock-in Mouse; Knowledge; Lead; Learning; Long-Term Depression; Measurement; Measures; Memory; Memory Loss; Memory impairment; Messenger RNA; Metabolic; Mus; Nerve Degeneration; Neurobiology; Neuroglia; Oxidative Stress; Pathologic; Pathway interactions; Patients; Pharmacology; Phase; Play; Preventive Intervention; Protein Analysis; Protein Isoforms; Proteins; Reproducibility; Research; Resolution; Role; Senile Plaques; Site; Stimulus; Symptoms; Synapses; System; Techniques; Testing; Therapeutic; Time; Tissues; Transgenic Organisms; abeta accumulation; alpha Bungarotoxin; awake; biomarker identification; cognitive testing; cohort; early detection biomarkers; excitotoxicity; experimental study; extracellular; hyperphosphorylated tau; improved; in vivo; insight; memory recall; mild cognitive impairment; millisecond; mitochondrial dysfunction; morris water maze; mouse model; neuroinflammation; neuropathology; neuroregulation; neurotoxic; neurotransmission; new therapeutic target; novel; object recognition; optimal treatments; postsynaptic; pre-clinical; presynaptic; presynaptic neurons; tau Proteins; therapeutic target; uptake; vesicular glutamate transporter 1

Project Summary/Abstract In spite of the evidence supporting the involvement of the glutamatergic system in Alzheimer’s disease (AD), research has focused on indirect measures of glutamate (Glu) involvement, mainly through increased downstream pathways related to excitotoxicity, without addressing possible changes in extracellular Glu occurring during disease progression. Knowledge of basal and phasic extracellular Glu levels and clearance kinetics would allow us to establish an early biomarker, better determine and explore novel therapeutic targets, and establish the opportune treatment window that has the potential to alter AD progression. Until recently, research measuring extracellular Glu levels has been limited because few techniques are capable of in vivo analysis within small sub-regions of the brain. We have developed a MEA that, when combined with electrochemistry, has the ability to measure micromolar changes in basal and phasic extracellular Glu with high temporal (msec) and spatial (micron) resolution during acute and chronic recordings. We plan to use this technique to address our central hypothesis that alterations in extracellular Glu in awake animals occur prior to cognitive decline and neuropathology associated with AD, and that Aβ accumulation with age potentiates these changes resulting in the cognitive decline typical in AD. This hypothesis will be evaluated using a novel knock-in mouse model of AD, APPNL-F/NL-F mice, and APP/PS1 mice, and their respective controls at 2-4, 8-10, and 18-20 months of age. At these ages, one cohort of mice will undergo cognitive evaluation using the Morris water maze followed by awake stimulus (KCl)-evoked Glu recordings in the CA1 region of the hippocampus. These studies will help us determine if basal Glu and Glu release (presynaptic) and uptake (glia and postsynaptic) kinetics are altered in AD mice and if so, how and when these alterations occur over the continuum of cognitive and pathophysiological decline. Next, to determine if alterations in Glu neurotransmission is behaviorally detrimental, we will examine a second group of mice at the same ages and hippocampal sub-region during a memory related task, the spontaneous alternation y maze. This will allow us to determine the impact of aging and AD progression on formation and recall of memories in the form of phasic Glu measurements. Taken together, we anticipate that these studies will give us valuable insight into the role of Glu as an early biomarker, a mechanism for disease progression, a site for potential novel therapeutic targets, and optimal intervention timeframes for AD.

项目概要/摘要 尽管有证据支持谷氨酸能系统参与阿尔茨海默病 疾病（AD），研究重点是谷氨酸（Glu）参与的间接测量，主要是 通过增加与兴奋性毒性相关的下游途径，而不解决可能的问题 疾病进展期间细胞外谷氨酸发生变化。基础知识和 阶段性细胞外 Glu 水平和清除动力学将使我们能够建立早期 生物标志物，更好地确定和探索新的治疗靶点，并建立合适的治疗靶点 有可能改变 AD 进展的治疗窗口。直到最近，研究 测量细胞外谷氨酸水平受到限制，因为很少有技术能够 大脑小分区内的体内分析。我们开发了一种 MEA，当 与电化学相结合，能够测量基础和微摩尔变化 急性期阶段性细胞外 Glu 具有高时间（毫秒）和空间（微米）分辨率 和慢性录音。我们计划使用这种技术来解决我们的中心假设： 清醒动物细胞外谷氨酸的改变发生在认知能力下降之前 与 AD 相关的神经病理学，以及 Aβ 随着年龄的增长而增强这些 导致 AD 中典型的认知能力下降的变化。该假设将使用以下方法进行评估 AD、APPNL-F/NL-F 小鼠和 APP/PS1 小鼠的新型敲入小鼠模型，以及它们各自的 控制2-4个月、8-10个月和18-20个月龄。在这些年龄，一组小鼠将经历 使用 Morris 水迷宫进行认知评估，然后使用清醒刺激 (KCl) 诱发 Glu 海马 CA1 区的记录。这些研究将帮助我们确定基础是否 AD 中 Glu 和 Glu 释放（突触前）和摄取（神经胶质细胞和突触后）动力学发生改变 小鼠，如果是这样，这些改变是如何以及何时在认知和认知的连续体中发生的 病理生理衰退。接下来，确定 Glu 神经传递的改变是否是 行为上有害，我们将检查第二组相同年龄的小鼠 在执行记忆相关任务时，海马亚区域会出现自发交替迷宫。这 将使我们能够确定衰老和 AD 进展对大脑形成和记忆的影响 以阶段性 Glu 测量形式的记忆。综合起来，我们预计这些 研究将为我们提供有价值的见解，让我们了解 Glu 作为早期生物标志物的作用，这是一种机制 疾病进展、潜在新治疗靶点和最佳干预 AD 的时间表。