Biochemical Basis of Axonal & Myelin Injury In Head Trauma

轴突的生化基础

基本信息

批准号：
7234283
负责人：
KEVIN Ka Wang WANG
金额：
$ 13.61万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-05-01 至 2007-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7234283
关键词：
11-aminoundecanoic acid 2&apos ,3&apos -Cyclic-Nucleotide Phosphodiesterases Affect Affinity Amines Amyloid beta-Protein Precursor Animals Antibodies Antibody Specificity Antigens Arts Axon Binding Biochemical Biological Assay Biological Preservation Brain Brain Injuries Brain region C-terminal Calcium Calpain Carbon Carrier Proteins Caspase Cathepsins Cathepsins B Cell Adhesion Molecules Cell membrane Cerebrospinal Fluid Chemical Injury Chemicals Classification Coomassie blue Corpus Callosum Cortical Contusions Coupled Craniocerebral Trauma Craniotomy Cyclic Nucleotides Cysteine Cytoskeletal Proteins Cytoskeleton Data Databases Demyelinations Development Digestion Endopeptidases Ensure Enzyme Immunoassay Enzymes Event Future Gelatinase B Hippocampus (Brain)Homeostasis Hour Immunoassay Immunoblotting Immunoprecipitation In Vitro Injury Integral Membrane Protein Invasive Ischemic Brain Injury Keyhole Limpet Hemocyanin Knowledge Label Laboratories Length Link Mass Spectrum Analysis Matrix Metalloproteinases Mechanics Mediating Membrane Membrane Proteins Metalloproteases Methods Microglia Microtubules Myelin Myelin Associated Glycoprotein Myelin Basic Proteins Myelin Proteins Myelin Sheath N-terminal Nature Neurofilament-H Neurons Oligodendroglia Operative Surgical Procedures Oryctolagus cuniculus Parents Pathologic Pathology Pattern Peptide Hydrolases Peptides Plant Resins Precipitation Protease Inhibitor Proteins Proteolipids Proteolysis Proteolytic Processing Proteomics Rattus Relative (related person)Reporting Research Residual state Risk Running Sampling Sepharose Shapes Site Source Spectrin Staining method Stains Standards of Weights and Measures Structural Protein Structure System Techniques Technology Time Traumatic Brain Injury Tubulin base brain tissue carboxyl group caspase-3 caspase-6 caspase-7 cell type day design gel electrophoresis in vivo interest myelin degeneration neurofilament novel polyclonal antibody response to injury scale up tau Proteins therapy development tool white matter

项目摘要

DESCRIPTION (provided by applicant): A considerable amount of traumatic brain injury (TBI) research has focused on the pathologic significance of mechanical and chemical insults (e.g. disturbed calcium homeostasis) to the axons. Yet the precise biochemical mechanisms of axonal injury remain unclear. White matter loss and demyelination have been insufficiently studied in TBI, and we have little knowledge of the biochemical mechanisms of myelin damage. In the past, our laboratories and others have documented over-expression and activation of various proteases (e.g. calpains, caspases, and matrix metalloproteases) following brain injury. In this proposed research, we hypothesize that a subset of structural proteins in axons and myelin are differentially vulnerable to proteolysis after TBI. Inevitably, such proteolysis would result in degradation that significantly compromises the structural and functional integrity of axons and myelin sheaths leading to long-lasting axonal damage and demyelination. The proposed research represents the first systematic examination of the mechanisms of proteolvtic degradation of structural proteins in both axons and mvelin. Our preliminary data have already identified several axonally-enriched proteins that are vulnerable to proteolysis (neurofilament- H, and -L, microtubule-associated protein tau, axolemma-associated amyloid precursor protein [APR] and cytoskeletal proteins alphall- and (betaII-spectrin). Importantly, we have also identified at least two myelin proteins that are proteolysed after TBI (myelin basic protein [MBP] and myelin oligodendrocyte specific protein [MOSP]). Unique in vivo cleavage sites in proteolysis-prone protein substrates will be identified with state-of-the- art proteomic techniques. This knowledge will then allow us to generate novel "fragment-specific" antibodies, which will be used to immunohistochemically examine the precise subcellular distribution of these proteolytic products. The same antibody tools will be used to configure enzyme-linked immunoassays (ELISAs) to quantify these axonal and myelin proteolytic products in both brain tissue and cerebrospinal fluid (CSF). The latter method, if successful, will provide a novel and minimally-invasive way of quantifying TBI-associated axonal and myelin damage. These advances in understanding the proteolvtic mechanisms underlying axonal and myelin pathology in TBI will greatly accelerate development of therapies preserving the structural and functional integrity of axons and myelin.

描述（由申请人提供）：相当多的创伤性脑损伤（TBI）研究重点是机械和化学损伤（例如，脑稳态受干扰）的病理意义。然而，轴突损伤的精确生化机制尚不清楚。在TBI中，对白质丧失和脱髓鞘的研究不足，我们对髓磷脂损伤的生化机制知之甚少。过去，我们的实验室和其他实验室记录了脑损伤后各种蛋白酶（例如Calpains，caspase和基质金属蛋白酶）的过表达和激活。在这项拟议的研究中，我们假设轴突和髓磷脂中的一部分结构蛋白在TBI后很容易受到蛋白水解的影响。不可避免地，这种蛋白水解会导致降解，从而显着损害轴突和髓鞘的结构和功能完整性，从而导致持久的轴突损伤和脱髓鞘。拟议的研究代表了轴突和Mvelin中结构蛋白蛋白质降解机制的首次系统检查。 Our preliminary data have already identified several axonally-enriched proteins that are vulnerable to proteolysis (neurofilament- H, and -L, microtubule-associated protein tau, axolemma-associated amyloid precursor protein [APR] and cytoskeletal proteins alphall- and (betaII-spectrin). Importantly, we have also identified at least two myelin在TBI（髓磷脂碱性蛋白[MBP）和髓磷脂少突胶质细胞特异性蛋白[MOSP]中，蛋白质的蛋白质在蛋白质分解的位点中，蛋白质蛋白质蛋白质的蛋白质将允许使用该蛋白质的protecieniqu'免疫组织化学研究这些蛋白水解产品的精确亚细胞分布将使用相同的抗体工具来配置酶 - 链接的免疫测定（ELISA），以量化这些轴突和髓磷脂蛋白质的蛋白质水解产物。 TBI相关的轴突和髓磷脂损伤。这些进步在TBI中理解轴突和髓磷脂病理学的蛋白旋转机制将极大地加速疗法的发展，从而保留轴突和髓磷脂的结构和功能完整性。