GROWTH PLATE STUDIES IN THE CHONDRODYSTROPHIES

软骨营养不良的生长板研究

• 批准号: 3319058
• 负责人: WILLIAM A HORTON
• 金额: $ 0.5万
• 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/3319058
• 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.

软骨营养不良 (CDX) 是一组异质性疾病 软骨内骨生长的临床特征是缩短 以及四肢和躯干的畸形。 的划定 导致这些疾病的发病机制将高度 之所以令人向往，是因为它不仅能带来 CDX 的确定性治疗， 但它将为骨骼疾病的成因提供有价值的见解 一般畸形以及正常的骨骼生长。 不幸的是， 对 CDX 异常的理解受到以下因素的阻碍： 主要与获取和调查相关的各种问题 受影响的组织，骨骼生长板。 针对这些问题， 拟议的研究者开发了一种适合研究的方法 这些疾病。 它涉及使用高度特异性的组织化学来 结合塑料嵌入组织中的基质成分定位 对极小软骨样本进行生化分析。 初步的 研究已经证明了其作为探索问题的手段的潜在价值 生长板作为动态结构。 现在建议扩大这一范围 系统、全面地检查软骨内的方法 正常对照和 CDX 患者的生长板组织中的骨化。 该组织将通过光和电子显微镜免疫和 凝集素组织化学、放射自显影和原位杂交。 针对基质成分的单克隆抗体：I-VI、IX (G)、X (M) 型和 1Alpha、2Alpha、3Alpha 胶原蛋白； 4-和-6-硫酸软骨素、角质素 软骨的硫酸盐和透明质酸结合区域亚结构 蛋白多糖；连接蛋白；纤连蛋白；和特定的 cDNA 探针 将使用骨骼原胶原mRNA。 生化证实 显微镜观察，软骨的微样本对应于 软骨细胞增殖和分化的不同阶段 通过 HPLC 和 SDS-PAGE 方法对器官培养进行研究，以确定 同位素标记的主要和相对比例的存在和相对比例 新合成的基质中含有少量胶原蛋白和糖胺聚糖。 这些 研究应该能够准确地描述正常情况下发生的事件 软骨内骨化的构建及其缺陷 确定。