COLLAGEN IN OSTEOARTHRITIC CARTILAGE

骨关节炎软骨中的胶原蛋白

• 批准号: 3154901
• 负责人: Paul David Benya
• 金额: $ 16.36万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-03-15 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3154901
• 关键词: actins; cartilage disorder; chondrocytes; collagen; crosslink; cytoskeleton; electron microscopy; gene expression; glycosylation; immunofluorescence technique; laboratory mouse; laboratory rabbit; microfilaments; monoclonal antibody; nucleoproteins; osteoarthritis; phosphorylation; protein kinase C; protein structure; retinoate; tyrosine analog

It has been shown that the phenotypic expression of rabbit articular chondrocytes may be modulated either by continued subculture or by treatment of primary chondrocytes with retinoic acid, as monitored by changes in the collagen phenotype. Although the two modulated phenotypes differ, both are reversed by treatment with the microfilament modifying drug dihydrocytochalasin B, suggesting a common mechanism for the reexpression of the chondrocyte phenotype, we will investigate the possibility that modifications in microfilament structure are physically transmit:ed into the nucleus, by way of cytoskeletal-nuclear matrix interactions, and result in changes in gene expression, and the possibility that modification may generate chemical signals mediated by altered membrane association and phosphorylation by protein kinase C or tyrosine kinases. Multiple processes and molecular associations will be tested for involvement in the microfilament mechanism. Alterations in the compartmentalization of cellular proteins will be investigated using high resolution 2-D protein mapping techniques. Monoclonal antibodies to cytoskeletal proteins will be used in their identification and quantitation. The cellular distribution and architectural patterns of cytoskeletal/nuclear-matrix component will also be investigated using indirect immunofluorescence and transmission electron microscopy. Crosslinking analysis will be used to identify both actin- and DNA-associated proteins, both to establish the presence or absence of actin within the nucleus and to study changes in proteins that are likely to be involved in the mechanism of reexpression. Additional studies will test the role of poly-ADP ribosylation, DNA supercoiling, and the requirement for a serum-derived factor in the reexpression mechanism. The proposed studies are part of a long term strategy to understand and manipulate the pathogenesis of osteoarthritis. Developing an understanding of the mechanisms of phenotypic change, using this unique model system, will greatly enhance the probability of future clinical intervention at natural control points.

已经表明兔子的表型表达 关节软骨细胞可以通过继续调节 亚文化或通过视黄氨酸的原发软骨细胞的治疗 通过胶原蛋白表型的变化监测的酸。 虽然 两种调制表型不同，两者都通过处理逆转 带有微丝修饰药物二氢细胞藻素B， 提出了重新表达的共同机制 软骨细胞表型，我们将调查 微丝结构的修饰是物理传递的： 通过细胞骨架核基质进入核 相互作用，并导致基因表达的变化，以及 修改可能会产生化学信号的可能性 通过改变的膜关联和磷酸化介导 蛋白激酶C或酪氨酸激酶。 将测试多个过程和分子关联 参与微丝机制。 改变 将研究细胞蛋白的分室化 使用高分辨率的2-D蛋白映射技术。 单克隆 将使用细胞骨架蛋白的抗体 识别和定量。 细胞分布和 细胞骨架/核矩阵组件的建筑模式 还将使用间接免疫荧光和 透射电子显微镜。 交联分析将是 用于鉴定肌动蛋白和DNA相关蛋白，均为 在细胞核内建立或不存在肌动蛋白 研究可能参与的蛋白质变化 重新表达的机制。 其他研究将测试角色 多-ADP核糖基化，DNA超螺旋和对 重新表达机制中的血清衍生因子。 拟议的研究是理解长期策略的一部分 并操纵骨关节炎的发病机理。 开发一个 理解表型变化的机制，使用此 独特的模型系统将大大提高未来的可能性 自然控制点的临床干预。