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.

这一核心为项目编制补助金的所有四个项目提供所需的专门服务。这些都是最好的协调，以提高时间和成本的效率，促进科学的协同作用，并最大限度地利用专用的高度专业化的设施。该项目是一项多学科研究，研究AD中内体、自噬和溶酶体异常的体内和体外模型，这些模型被认为对AD发病机制至关重要。在这些模型中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），必须在严格控制的条件下进行维护 以便对各项目的信息进行可靠的比较。具体目标是：1.以协调的方式采集、储存和有效分配来自AD、FAD、21三体和非痴呆和非AD神经系统对照病例的充分表征的组织（脑和成纤维细胞），用于所有四个项目的研究。2.对细胞和小鼠模型以及人脑进行电子显微镜和免疫金电子显微镜检查。3.为了进行Ab 40和Ab 42的ELISA测量以定量内源性鼠Ab和人Ab， AD脑、小鼠模型脑、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APP代谢物上的Ab肽和肽裂解位点。4.根据需要提供专门的组织学程序和形态学技术，以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 08-10年的PPG，为选定的动物模型提供体内功能性MRI分析，以纵向表征病理学变化。核心服务于所有项目，对项目的成功至关重要。