NEUROPATHOLOGY CORE

神经病理学核心

基本信息

批准号：
7894580
负责人：
Stephen J. DeArmond
金额：
$ 14.36万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7894580
关键词：
ATP phosphohydrolase Aldehydes Amyloid Animals Antibodies Antigens Apoptotic Arts Autopsy Axon Binding Brain Cells Computer Assisted Computers Data Databases Diagnostic Digestion Disease Doctor of Medicine Electron Microscopy Endopeptidase K Etiology Event FVB Mouse Filament Fixatives Formalin Formic Acids Freezing Frozen Sections Glutaral Golgi Apparatus Graph Hamsters Histocytochemistry Histological Techniques Image Analysis Immobilon P Immunohistochemistry In Situ In Situ Nick-End Labeling Label Luxol Fast Blue MBS Measurement Membrane Microscopic Molecular Molecular Analysis Mus Muscle Myelin Nerve Degeneration Neurites Neurons Organ Oxyuranus scutellatus toxin 2 Paper Paraffin Embedding Pathogenesis Peptide Hydrolases Peripheral Nerves Phenotype Photography Physical Chemistry Plastic Embedding Plastics Play Population PrP PrPSc Proteins Principal Investigator Prion Diseases Prions Procedures Process Program Research Project Grants Proteins Publications Pyroxylin Recombinants Reporting Research Personnel Resistance Rodent Role Science Scrapie Serial Passage Services Silver Staining Skeletal Muscle Spinal Cord Staining method Stains Synapses Synaptophysin System Techniques Testing Thick Time Tissues Tolonium chloride Transgenic Organisms Trees Trichrome stain method Waxes Wild Type Mouse Work Yeasts abnormal PrP age-dependent CNS degeneration animal tissue araldite base cryostat density digital guanidinium interest light microscopy morphometry neurochemistry neuronal cell body neuropathology paraform programs recombinant PrP technique development three dimensional structure yeast prion

项目摘要

The Neuropathology Core (NP Core) under the direction of Dr. Stephen DeArmond will perform a service function, a hypothesis-generating function, and a hypothesis-testing function in molecular pathogenesis studies of prion diseases. The NP Core has contributed directly to approximately 100 publications on prion diseases. This NP Core will provide a full range of qualitative and quantitative neuropathology expertise including neurohistology for light microscopy, immunohistochemistry, and electron microscopy. Quantitative unbiased stereology and morphometrics are available as requested by the Projects. A full range of digital gross and microscopic photography and graphing are provided to investigators for notebooks and publications. Animals are autopsied, dissected, fixed or snap-frozen as needed, embedded in wax or plastic as needed, sectioned, and stained. For previous programs, the NP Core has pioneered development of techniques to identify PrPc and protease-resistant PrPSc in tissue sections. The NP Core maintains a bank of embedded and frozen animal tissues and a database with almost 10,000 animal entries in it to retrieve sections or blocks of tissues. The first Project proposes to generate 5 to 10 new recombinant PrP's folded into amyloid filaments per year. The NP Core must determine whether or not each construct causes a prion disease in transgenic (Tg) and wild-type mice and whether each serially transmits a similar disease to other Tg and wild-type mice. Those that do will have fulfilled the basic criteria of a mammalian prion. In addition, the NP Core must determine whether the neuropathological phenotype caused by a synthetic prion is sufficiently different to designate it a new prion strain. The fourth Project will test whether neurodegeneration caused by natural and synthetic prion strains follow the same stereotypical sequence of events during the second serial passage in Tg and wild-type mice. Neurohistological and immunohistochemical stains and histoblot analysis for protease-resistant PrPSc at 6 time-points during the course of disease will be processed in the NP Core. Animals will also be prepared and stained for Golgi silver impregnation-and synaptophysin immunohistochemistry. In addition, animals will be perfused with glutaraldehyde and paraformaldehyde, embedded in plastic, and prepared for routine electron microscopy. The NP Core will also help develop PrPSc immunogold histochemistry for electron microscopy.

在斯蒂芬·戴蒙德（Stephen Dearmond）博士的指导下，神经病理学核心（NP核心）将执行服务功能，假设生成功能以及对prion疾病的分子发病机理研究的假设检测功能。 NP Core直接为大约100个有关Prion疾病的出版物做出了贡献。该NP核心将提供全范围的定性和定量神经病理学专业知识，包括光学显微镜，免疫组织化学和电子显微镜的神经组织学。根据项目的要求，可以使用定量无偏的立体论和形态计量学。向调查人员提供了全方位的数字总和微观摄影和图形，以获取笔记本和出版物。根据需要对动物进行尸检，解剖，固定或折断，根据需要，切片和染色嵌入蜡或塑料中。对于以前的程序，NP核心已经开发了技术在组织切片中识别PRPC和耐蛋白酶的PRPSC的发展。 NP核心保持了一堆嵌入和冷冻的动物组织和一个数据库，其中有近10,000个动物条目，以检索组织或组织块。第一个项目建议每年产生5至10个新的重组PRP折叠成淀粉样蛋白丝。 NP核心必须确定每个构建体是否引起转基因（TG）和野生型小鼠的病毒疾病，以及每个构造是否串行传播与其他TG和野生型小鼠相似的疾病。那些这样的人将满足哺乳动物王室的基本标准。此外，NP核心必须确定由合成prion引起的神经病理表型是否足够不同，可以将其指定为新的prion菌株。第四个项目将测试由自然和合成prion菌株引起的神经变性是否遵循在TG和野生型小鼠中第二个串行通道期间，事件的刻板印象序列。 NP核心将处理疾病过程中6个时间点的抗蛋白酶耐药性PRPSC的神经组织学和免疫组织化学染色和组体印迹分析。动物还将用于高尔基银浸渍和突触素免疫组织化学的动物。此外，动物将用戊二醛和多聚甲醛灌注，并嵌入塑料中，并准备常规电子显微镜。 NP核心还将有助于开发电子电子的PRPSC免疫元组织化学显微镜。