GROWTH PLATE STUDIES IN THE CHONDRODYSTROPHIES

软骨营养不良的生长板研究

• 批准号: 3319061
• 负责人: WILLIAM A HORTON
• 金额: $ 10.16万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-08-01 至 1990-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3319061
• 关键词: autoradiography; bone development disorder; cartilage development; cartilage disorder; cartilage metabolism; cell differentiation; cell growth regulation; chondrocytes; chondrodystrophy; chondroitin; chondroitin sulfates; collagen; connective tissue disorder; connective tissue metabolism; electron microscopy; extracellular matrix; fibronectins; gel electrophoresis; high performance liquid chromatography; histochemistry /cytochemistry; hyaluronate; image processing; immunochemistry; in situ hybridization; keratan sulfate; microscopy; monoclonal antibody; normal ossification; nucleic acid probes; organ culture; procollagen; protein biosynthesis; proteoglycan; radiotracer; scintillation counter; tritium

The chondrodystrophies (CDX) are a heterogeneous group of disorders of endochondral bone growth that are characterized clinically by shortening and deformities of the limbs and trunk. The delineation of the pathogenetic mechanisms responsible for the disorders would be highly desirable because not only would it lead to definitive therapy for the CDX, but it would provide valuable insights into the causes of skeletal deformities in general as well as normal bone growth. Unfortunately, the understanding of the abnormalities in the CDX has been hampered by a variety of problems primarily related to obtaining and investigating the affected tissue, the skeletal growth plate. Addressing these problems, the proposed investigator has developed an approach tailored to studying these disorders. It involves the use of highly specific histochemistry to localize matrix constituents in plastic embedded tissues combined with biochemical analysis of extremely small samples of cartilage. Preliminary studies have demonstrated its potential value as a means to probe the growth plate as a dynamic structure. It is now proposed to expand this approach to systematically and comprehensively examine endochondral ossification in growth plate tissue from normal control and CDX patients. The tissue will be studied by light and electron microscopic immune and lectin histochemistry, autoradiography, and in situ hybridization. Monoclonal antibodies to matrix components: types I-VI, IX (G), X (M), and 1Alpha, 2Alpha, 3Alpha collagens; chondroitin-4- and -6- sulfate, keratan sulfate and hyaluronic acid binding region substructures of cartilage proteoglycan; link protein; fibronectin; and cDNA probes to specific skeletal procollagen mRNAs will be employed. To biochemically confirm the microscopic observations, microsamples of the cartilages corresponding to different stages of chondrocyte proliferation and differentiation will be studied in organ culture by HPLC and SDS-PAGE methods to determine the presence of and relative proportions of isotopically labeled major and minor collagens and glycosaminoglycans in newly synthesized matrix. These studies should permit an accurate representation of the events of normal endochondral ossification to be constructed and defects therein to be identified.

软骨造成的（CDX）是一组异质性疾病 内侧软骨骨生长通过缩短在临床上表征 和四肢和躯干的畸形。 描述 导致疾病的致病机制将是高度的 理想 但这将为骨骼原因提供宝贵的见解 通常，畸形以及正常的骨骼生长。 不幸的是， 对CDX异常的理解已受到A的阻碍 主要与获得和研究有关的各种问题 影响组织，骨骼生长板。 解决这些问题， 拟议的研究者已经开发了一种针对研究的方法 这些疾病。 它涉及使用高度特定的组织化学 将基质成分定位在塑料嵌入式组织中 非常小的软骨样品的生化分析。 初步的 研究表明，其潜在价值作为探测的手段 生长板作为动态结构。 现在建议扩展这一点 系统和全面检查内软骨的方法 正常对照和CDX患者的生长板组织的骨化。 该组织将通过光和电子显微镜免疫进行研究， 凝集素组织化学，放射自显影和原位杂交。 矩阵组件的单克隆抗体：类型I-VI，IX（G），X（M）和 1alpha，2alpha，3alpha胶原蛋白；软骨素-4-和-6-硫酸盐，Keratan 软骨的硫酸盐和透明质酸结合区域 蛋白聚糖；链接蛋白；纤连蛋白；和cDNA探针到特定 将使用骨骼胶原mRNA。 在生化上确认 微观观察，软骨的微观样本对应于 软骨细胞增殖和分化的不同阶段将是 通过HPLC和SDS-PAGE方法在器官培养中进行了研究，以确定 同位素标记的主要和相对比例的主要比例 新合成基质中的次要胶原蛋白和糖胺聚糖。 这些 研究应允许准确表示正常事件 内软骨骨化要构造并在其中缺陷 确定。