In Vivo Imaging Alzheimers Disease Pathology with 2-Photon/Lifetime Microscopy

使用 2 光子/终身显微镜对阿尔茨海默病病理学进行体内成像

• 批准号: 9150491
• 负责人: Mohammad Abbas Yaseen
• 金额: $ 24.9万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9150491
• 关键词: ATP Synthesis Pathway; Adult; Age; Aging; Alzheimer' s Disease; Alzheimer' s disease model; Animal Model; Astrocytes; Binding; Boa; Brain; Calcium; Cell Respiration; Cerebrovascular system; Cerebrum; Complex; Computer software; Consumption; Coupled; Custom; Detection; Development; Disease; Disease Progression; Elderly; Electrons; Endoplasmic Reticulum; Energy Metabolism; Engineering; Ensure; Enzymes; Exposure to; Family; Fluorescence; Formulation; Glucose; Glycolysis; Health; Homeostasis; Human Pathology; Image; Imaging Device; Incidence; Investigation; Knowledge; Laboratories; Measurement; Measures; Mentors; Metabolic; Metabolic Marker; Metabolic Pathway; Metabolism; Methods; Microscope; Microscopic; Microscopy; Mitochondria; Modeling; Monitor; Mus; NADH; Nature; Nerve Degeneration; Neurodegenerative Disorders; Neurofibrillary Tangles; Neurons; Neurosciences; Nicotinamide adenine dinucleotide; Optics; Oxidative Phosphorylation; Oxidative Stress; Oxygen; Oxygen Consumption; Partial Pressure; Pathogenesis; Pathology; Pd-porphyrin; Phosphorescent Assays; Physiological Processes; Population; Populations at Risk; Preclinical Drug Evaluation; Prevention strategy; Preventive therapy; Process; Reaction; Reagent; Regulation; Research; Research Personnel; Resolution; Respiration; Signal Transduction; Solid; Source; Staging; Synaptic Transmission; Systems Biology; Techniques; Technology; Training; Transgenic Mice; Writing; age group; amyloid peptide; brain cell; brain tissue; effective therapy; human disease; imaging system; in vivo; in vivo imaging; insight; instrumentation; meetings; mitochondrial dysfunction; mitochondrial metabolism; mouse model; normal aging; novel; novel diagnostics; novel therapeutics; optical imaging; peptide A; success; temporal measurement; therapy development; tool; two-photon

DESCRIPTION (provided by applicant): This project aims to characterize the changes in calcium regulation and cerebral metabolism associated with Alzheimer's Disease (AD) using advanced microscopy. AD has a devastating impact on over 20 million people and their families worldwide, and the incidence rate is expected to double every 20 years. Understanding the mechanisms of AD-related deficits in calcium homeostasis and mitochondrial metabolism is important for developing new diagnostic criterion and effective preventative therapies. Optical microscopy permits non- destructive, measurement of metabolic markers with high spatial and temporal resolution in animal models of human pathologies. Two-photon / lifetime microscopy has recently proven useful for measuring absolute calcium concentration ([Ca]) and measuring the reduced form of the endogenous electron carrier nicotinamide adenine dinucleotide (NADH), reportedly allowing the distinction of NADH species involved in glycolysis from NADH species involved in oxidative metabolism, with unprecedented resolution. Through the use of novel dendritic Pd-porphyrin phosphors, cerebral oxygen partial pressure (pO2) in brain tissue and vasculature can also be measured with high resolution using two-photon microscopy. The central focus of this project is to use two-photon / lifetime microscopy to measure cerebral NADH, [Ca], and pO2 in vivo in an AD mouse model at different disease stages. Characterizing the relationship between these metabolic markers as the disease progresses in vivo will provide detailed insight into the complex metabolic alterations involved in AD pathogenesis. The specific aims of this project are: 1. Develop and validate methods and instrumentation for near-simultaneous 2P measurement of pO2, and either [Ca] or NADH species. A two-photon imaging system will be upgraded and validated to feature two excitation sources, four detection channels, and custom-written control software, enabling non-destructive measurement of cerebral metabolic indicators in vivo. 2. Apply the microscope to identify cerebral NADH species and validate their association with specific metabolic processes in healthy mice Fluorescence lifetime imaging enables resolution between specific enzyme-bound formulations of NADH. The specific nature of these enzyme bound formulations will be comprehensively evaluated and characterized. 3. Apply the microscope to characterize cerebral metabolism in the exposed cortices of transgenic mice modeling AD pathogenesis The technology and knowledge obtained from aims 1 and 2 will be utilized to evaluate and compare cerebral metabolism in vivo in APPswe:PS1dE9 transgenic mice representing distinct stages of AD progression. This project will yield insight into the mechanisms of AD pathogenesis with unprecedented detail, and it will facilitate the development of new therapeutic techniques widely applicable to the growing population of at-risk aging citizens.

描述（由申请人提供）：该项目旨在表征使用高级显微镜使用与阿尔茨海默氏病（AD）相关的钙调节和大脑代谢的变化。广告对全球超过2000万人及其家庭有毁灭性的影响，预计发病率将每20年翻一番。了解钙稳态和线粒体代谢中与广告相关的缺陷的机制对于制定新的诊断标准和有效的预防疗法很重要。光学显微镜允许在人体病理动物模型中对具有高空间和时间分辨率的代谢标记的非破坏性测量。两光子 /生命周期显微镜最近已证明可用于测量绝对钙浓度（[CA]）并测量内源性电子载体烟酰胺尼核苷酸（NADH）的降低形式，据报道允许NADH涉及的NADH涉及的NADH涉及的NADH涉及的NADH涉及的NADH涉及的NADH涉及的NADH涉及的NADH涉及的氧化度含量，氧化不良的氧化不良。通过使用新型的树突状PD-卟啉磷酸盐，也可以使用两光子显微镜通过高分辨率来测量脑组织和脉管系统中的脑氧部分压（PO2）。该项目的主要重点是使用两光子 /终生显微镜在不同疾病阶段的AD小鼠模型中测量体内脑NADH，[CA]和PO2。表征这些代谢标志物在体内进展的过程中的关系将提供详细的见解，以了解AD发病机理所涉及的复杂代谢改变。该项目的具体目的是：1。开发和验证近似PO2的2p测量的方法和仪器，以及[CA]或NADH物种。将升级并验证了两个光子成像系统，以具有两个激发源，四个检测通道和定制的控制控制软件，从而实现了体内大脑代谢指标的无损测量。 2。应用显微镜识别脑NADH物种，并验证其与健康小鼠荧光寿命成像中的特定代谢过程的关联，可以在NADH的特定酶结合配方之间进行分辨率。这些酶结合的制剂的具体性质将得到全面评估和表征。 3。应用显微镜在转基因小鼠的裸露皮质中表征脑代谢，以建模AD发病机理，将利用AIM 1和2获得的技术和知识来评估和比较Appswe中体内的脑代谢：PS1DE9转源小鼠代表不同ad ad togressions的不同阶段。该项目将以前所未有的细节来洞悉AD发病机理的机制，并有助于开发新的治疗技术，广泛适用于不断增长的高危衰老公民人口。