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）：大量创伤性脑损伤（TBI）研究集中在机械和化学损伤（例如钙稳态紊乱）对轴突的病理意义。然而轴突损伤的确切生化机制仍不清楚。 TBI中白质丢失和脱髓鞘的研究还不够充分，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已经记录了脑损伤后各种蛋白酶（例如钙蛋白酶、半胱天冬酶和基质金属蛋白酶）的过度表达和激活。在这项拟议的研究中，我们假设轴突和髓磷脂中的结构蛋白子集在 TBI 后对蛋白水解的脆弱性不同。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，导致持久的轴突损伤和脱髓鞘。这项研究代表了对轴突和 mvelin 结构蛋白的蛋白水解降解机制的首次系统研究。我们的初步数据已经鉴定出几种易被蛋白水解的轴突富集蛋白（神经丝-H和-L、微管相关蛋白tau、轴突相关淀粉样前体蛋白[APR]和细胞骨架蛋白α11-和（βII-血影蛋白）重要的是，我们还鉴定出至少两种在 TBI 后被蛋白水解的髓磷脂蛋白（髓磷脂碱性蛋白）。 [MBP] 和髓磷脂少突胶质细胞特异性蛋白 [MOSP]）将通过最先进的蛋白质组学技术来鉴定易于蛋白水解的蛋白质底物中独特的体内切割位点。特异性”抗体，将用于免疫组织化学检查这些蛋白水解产物的精确亚细胞分布。相同的抗体工具将用于配置酶联免疫测定(ELISA) 来量化脑组织和脑脊液 (CSF) 中的这些轴突和髓磷脂蛋白水解产物，后一种方法如果成功，将为量化 TBI 相关的轴突和髓磷脂损伤提供一种新颖的微创方法。这些在理解 TBI 中轴突和髓磷脂病理学基础的蛋白水解机制方面的进展将极大地加速保护轴突和髓磷脂结构和功能完整性的疗法的开发。