CORE--HISTOLOGY/NEUROPATHOLOGY

核心--组织学/神经病理学

• 批准号: 6234542
• 负责人: Neil W. Kowall
• 金额: $ 16.44万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6234542
• 关键词: Alzheimer's disease; DNA damage; Parkinson's disease; amyotrophic lateral sclerosis; animal tissue; biomedical facility; brain disorder diagnosis; carbonyl group; electron microscopy; fluorescence microscopy; histochemistry /cytochemistry; histopathology; human tissue; immunocytochemistry; in situ hybridization; light microscopy; lipid peroxides; northern blottings; oxidative stress; patient /disease registry; peroxidation; postmortem; superoxide dismutase; tissue /cell culture

The Histology/Neuropathology Core of the program project is located at the Bedford Veterans Affairs Medical Center. The specific aims of the core are: 1) to fully characterize the anatomical pathology of animal lesions produced in projects 1 and 3 by defining the spatial distribution of lesions and the extent of oxidative injury affecting histochemically defined neuronal subsets;. 2) to determine the light and ultrastructural changes produced by manipulation of superoxide dismutase (SOD) expression in cell culture systems (project 2), 3) to perform detailed postmortem analysis on human brains studied in the center for the conformation of diagnosis, the detection of specific patterns of oxidative injury in affected neuronal classes and the detection of clinical variants associated with specific genetic abnormalities. Postmortem specimens will be studied with the same methods used in experimental animals to determine the validity of SOD manipulation as animal models for neurodegenerative diseases and aging. Specific methods include three dimensional serial reconstruction of lesions and quantitation with computerized image analysis, definition of localized tissue injury with glial fibrillary acidic protein staining, detection of DNA oxidation in situ with the terminal transferase method and specific antibodies to 8-OH deoxyguanosine; detection of protein oxidation using thiocarbohydrazide binding assay and specific antibodies to nitrotyrosine; detection of lipid peroxidation with the thiobarbituric acid method in animals and the hydroxynaphthoic acid hydrazide/Fast Blue B method in postmortem tissue; SOD immunocytochemistry and enzyme histochemistry; and detection of thiol specific antioxidant protein. These determinations will permit us to accurately define lesions, the extent and nature of oxidative injury and the effects of experimental therapies such as those proposed in project 3 (tetanus-SOD fusion protein). Postmortem human tissue will be obtained from the Bedford VA, Massachusetts General Hospital (MGH) and Johns Hopkins University. In the past year, 56 autopsies, including 20 Alzheimer's disease (AD), 9 Pick's disease, 5 Lewy body disease, 1 amyotrophic lateral sclerosis (ALS) and 9 controls have been performed at Bedford - an autopsy rate of over 80%. Additional ALS specimens will be obtained from affiliated clinics at MGH and Hopkins. A range of both fixed and deep-frozen tissue samples are acquired and stored using uniform acquisition procedures to ensure accuracy and reproducibility and fulfill the specific requirements of affiliated investigators. A comprehensive database is maintained that includes quantitative morphometric, histochemical, and biochemical analyses to facilitate clinical-pathological correlative studies of neurochemical, histological, and neuropsychiatric aspects of neurodegenerative diseases including AD, Parkinson's disease, Huntington's disease, and ALS. The comprehensive facilities available to perform detailed quantitative histology and anatomically define injury in a consistent manner will provide a unique resource.

该计划项目的组织学/神经病理学核心位于 贝德福德退伍军人事务医疗中心。 该计划的具体目标 核心是：1）全面表征动物的解剖病理学 通过定义空间分布在项目 1 和 3 中产生的损伤 损伤的程度和氧化损伤的程度影响组织化学 定义的神经元子集；。 2) 光和超微结构的测定 操纵超氧化物歧化酶 (SOD) 表达产生的变化 在细胞培养系统（项目 2）、3）中进行详细的尸检 对中心研究的人脑的构象进行分析 诊断，检测氧化损伤的特定模式 受影响的神经元类别和临床变异的检测 与特定的基因异常有关。 尸检标本 将使用与实验动物相同的方法进行研究 确定 SOD 操作作为动物模型的有效性 神经退行性疾病和衰老。 具体方法包括三种 病变的维度连续重建和定量 计算机图像分析，局部组织损伤的定义 胶质纤维酸性蛋白染色，检测DNA氧化 原位末端转移酶法和 8-OH 特异性抗体 脱氧鸟苷；使用硫代碳酰肼检测蛋白质氧化 硝基酪氨酸的结合测定和特异性抗体；检测 硫代巴比妥酸法对动物体内脂质过氧化的影响 死后组织中的羟基萘甲酰肼/固蓝B法； SOD免疫细胞化学和酶组织化学；和硫醇的检测 特定的抗氧化蛋白。 这些决定将使我们能够 准确定义病变、氧化损伤的程度和性质， 实验疗法的效果，例如项目中提出的疗法 3（破伤风-SOD 融合蛋白）。 将获得死后人体组织 来自弗吉尼亚州贝德福德、马萨诸塞州总医院 (MGH) 和约翰斯 霍普金斯大学。 去年，进行了 56 次尸检，其中 20 次 阿尔茨海默病 (AD), 9 皮克氏病, 5 路易体病, 1 肌萎缩侧索硬化症 (ALS) 和 9 例对照已进行 贝德福德 - 尸检率超过 80%。 额外的 ALS 样本将 可从麻省总医院 (MGH) 和霍普金斯大学 (Hopkins) 的附属诊所获得。 两者皆有的范围 使用固定和深度冷冻的组织样本获取和存储 统一的采集程序以确保准确性和再现性 满足附属调查人员的具体要求。一个 维护综合数据库，其中包括定量 形态测定、组织化学和生化分析，以促进 神经化学、组织学、 和神经退行性疾病的神经精神方面，包括 AD， 帕金森病、亨廷顿病和肌萎缩侧索硬化症。 全面的 可用于进行详细的定量组织学分析和 以一致的方式在解剖学上定义损伤将提供独特的方法 资源。