ANALYTICAL CORE

分析核心

• 批准号: 6920484
• 负责人: RALPH A. NIXON
• 金额: $ 22.89万
• 依托单位: NATHAN S. KLINE INSTITUTE FOR PSYCH RES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6920484
• 关键词: Alzheimer' s disease; Downs syndrome; Niemann Pick disease; antibody; autophagy; biomedical facility; cell line; cellular pathology; confocal scanning microscopy; electron microscopy; endocytosis; enzyme linked immunosorbent assay; fluorescence microscopy; histology; immunoelectron microscopy; immunologic substance development /preparation; liquid chromatography mass spectrometry; longitudinal animal study; lysosomes; magnetic resonance imaging; molecular pathology; morphometry; neuropathology; sample collection; tissue resource /registry; training; video microscopy

This core provides specialized services required for all four projects of the PPG. These are best coordinated by a core to increase efficiencies of time and cost, promote scientific synergies, and maximally utilize dedicated highly specialized facilities. This program grant is a multidisciplinary investigation of in vivo and in vitro models of endosomal, autophagic, and lysosomal abnormalities in AD considered critical to AD pathogenesis. Neuropathological assessment and biochemical quantification of AD pathologies in these models is a major goal of the core. Other core functions are the maintenance of critical reagents (antibodies and cell lines) that are used in every project and must be maintained under rigorously controlled conditions to allow reliable comparisons of information across projects. The specific aims are: 1. To acquire, store and efficiently distribute well-characterized tissues (brain and fibroblasts) from cases of AD, FAD, Trisomy 21 and non-demented and non-AD neurological controls in a coordinated manner for studies in all four projects. 2. To perform electron microscopy and immunogold electron microscopy on cell and mouse models and human brain. 3. To perform ELISA measurements of Ab40 and Ab42 to quantify endogenous murine Ab and human Ab in AD brain, brains of mouse models, medium and lysates of fibroblasts and other cell models, and, when appropriate, to perform liquid chromatography-mass spectroscopy to identify Ab peptides and peptide cleavage sites on APP metabolites. 4. To provide specialized histological procedures and morphometric techniques as needed to characterize neurodegeneration and other AD pathologies as well as provide training on laser confocal microscopy and video-fluorescent microscopy. 5. To maintain and distribute polyclonal and monoclonal antibodies and generate additional polyclonal antibodies as necessary. 6. In years 08-10 of the PPG, to provide in vivo functional MRI analysis for selected animal models to characterize alterations in pathology longitudinally. The core serves all projects and is essential to their success.

该核心提供 PPG 所有四个项目所需的专业服务。这些最好由核心协调，以提高时间和成本效率，促进科学协同作用，并最大限度地利用专用的高度专业化设施。该项目资助是对 AD 内体、自噬和溶酶体异常的体内和体外模型进行多学科研究，这些异常被认为对 AD 发病机制至关重要。这些模型中 AD 病理的神经病理学评估和生化定量是核心的主要目标。其他核心功能是维护每个项目中使用的关键试剂（抗体和细胞系），并且必须在严格控制的条件下进行维护 允许跨项目的信息进行可靠的比较。具体目标是： 1. 以协调的方式从 AD、FAD、21 三体症和非痴呆和非 AD 神经系统对照病例中获取、存储和有效分配特征明确的组织（大脑和成纤维细胞），用于所有研究四个项目。 2. 对细胞、小鼠模型和人脑进行电镜和免疫金电镜观察。 3. 对 Ab40 和 Ab42 进行 ELISA 测量，以量化内源性鼠抗体和人抗体 AD 大脑、小鼠模型大脑、成纤维细胞和其他细胞模型的培养基和裂解物中的 Ab，并在适当时进行液相色谱-质谱分析以鉴定 Ab 肽和 APP 代谢物上的肽裂解位点。 4. 根据需要提供专门的组织学程序和形态测量技术来表征神经变性和其他 AD 病理，并提供激光共聚焦显微镜和视频荧光显微镜的培训。 5. 维护和分发多克隆和单克隆抗体，并根据需要生成额外的多克隆抗体。 6. 在 PPG 的 08-10 年，为选定的动物模型提供体内功能 MRI 分析，以纵向表征病理学的变化。核心服务于所有项目，对于项目的成功至关重要。