BIOCHEMISTRY OF THE INTERVERTEBRAL DISC

椎间盘的生物化学

• 批准号: 3157733
• 负责人: David R Eyre
• 金额: $ 17.73万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-01-01 至 1993-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3157733
• 关键词: aging; backache; cartilage disorder; chondrocytes; collagen; cow; crosslink; extracellular matrix; fibrous protein; high performance liquid chromatography; histopathology; human tissue; intervertebral disk; laboratory rabbit; molecular pathology; osteoarthritis; polymerization; protein biosynthesis; protein structure function; tissue /cell culture

The different molecular types of collagen in cartilage and intervertebral disc are the focus of the investigation. The long- term objective is a basic understanding of how the various kinds of collagen molecule are polymerized in the matrix, to what degree they are designed to interact specifically with each other and with other matrix macromolecules and self-assemble into a functioning fibrillar network. Of particular interest is the manner of covalent intermolecular cross-linking in collagens and the potential for heterotypic polymerization. Aims include defining the sites of covalent interaction between types IX and II collagen molecules, and between types V and I and types XI and II collagens in intervertebral disc and other skeletal tissues. The distribution, function and changes with age in type VI collagen of the human disc will be examined. A new collagenous protein of annulus fibrosus will be characterized and its function explored. The latest techniques in protein chemistry will be the primary approach, including extensive use of HPLC peptide separations, gas- phase automated amino acid sequencing and specialized methods to detect and quantify the unique collagen cross-linking amino acids. Rates of synthesis of the various collagens will be determined in tissue explants incubated in vitro. As the work progresses, an effort will also be made to study the matrix expression of isolated disc cells in primary high-density monolayer cultures. All human lumbar discs show progressive degenerative changes by 35 yr of age, which many experts believe is a significant physical factor behind the low back pain syndrome. The cardinal features of these changes are splits, fissures, dislocations, water loss and other signs of local damage to the organized collagen framework of the nucleus pulposus and annulus fibrosus. The basic studies in this proposal are aimed at an improved knowledge at the molecular level of how this complex collagen framework is assembled, how cells maintain it in the mature tissue and what happens to it in degeneration.

软骨中胶原蛋白的不同分子类型， 椎间盘是研究的重点。 很长的- 术语目标是对各种 胶原分子在基质中聚合到什么程度 它们被设计成专门相互作用， 其他基质大分子并自组装成功能性的 纤维状网络 特别感兴趣的是 胶原蛋白中的共价分子间交联， 异型聚合的可能性。 目标包括定义 IX型和II型胶原之间共价相互作用的位点 分子之间，以及V型和I型与XI型和II型胶原之间 在椎间盘和其它骨骼组织中。 的 人脐静脉血中VI型胶原的分布、功能及随年龄的变化 将检查人类椎间盘。 一种新的胶原蛋白， 纤维环将被表征并探索其功能。 蛋白质化学的最新技术将是主要的 方法，包括广泛使用的HPLC肽分离，气体- 阶段自动氨基酸测序和专门的方法， 检测和量化独特的胶原蛋白交联氨基酸。 各种胶原蛋白的合成速率将在 体外培养的组织外植体。 随着工作的进展， 还将努力研究分离的基质表达， 原代高密度单层培养的椎间盘细胞。 所有人类腰椎间盘在35岁时都表现出进行性退行性变化 年龄，许多专家认为这是一个重要的身体 下背痛综合症背后的因素。 主要特征 这些变化是分裂，裂缝，错位，水分流失， 其他迹象表明，局部损害的组织胶原蛋白框架， 髓核和纤维环。 基础研究 该建议旨在提高分子水平， 这个复杂的胶原蛋白框架是如何组装的， 细胞将其维持在成熟的组织中， 退化