MINERAL-MATRIX RELATIONS IN CALCIFYING TISSUES

钙化组织中矿物质与基质的关系

• 批准号: 2413975
• 负责人: WILLIAM J LANDIS
• 金额: $ 35.7万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-05-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2413975
• 关键词: animal tissue; atomic force microscopy; calcification; collagen; computer simulation; cytoskeleton; decorin; extracellular matrix proteins; infrared spectrometry; integrins; intermolecular interaction; minerals; molecular dynamics; normal ossification; osteocalcin; osteopontin; tissue /cell culture; transmission electron microscopy; vesicle /vacuole

DESCRIPTION: (Adapted from investigator's abstract) This proposal intends to obtain new information concerning vertebrate skeletal calcification in bone, cartilage, and tendon. Normal chicks, mice, rats, and turkeys; bone and cartilage cultures; and an abnormal osteogenesis imperfecta mouse mutant will be used as models. Studies in these examples will examine certain structural and chemical interactions in their respective extracellular matrices, intracellular matrices, and intra-and extracellular interfaces. Work will be done systematically, defining protein secretory pathways, cell matrix adhesion relations, and mineral-matrix associations. Ultrastructural, biochemical, immunochemical, and biophysical methods will be applied to extend the applicants' previous result describing the presence, nature, location, and interaction of organic matrix and mineral components in vertebrate calcification. The specific hypothesis to be examined is: the organic matrix of vertebrate calcifying tissues critically influences the deposition of mineral in terms of crystal size, shape, location, orientation, and distribution. Specific Aims, to address structure- function relations at atomic, molecular, and macromolecular levels of hierarchy, are to: (1) identify the presence and location of collagenous and non-collagenous proteins involved in mineralization, as well as selected integrins and cytoskeletal elements at the cell-matrix interface, utilizing conventional and high voltage electron microscopy, three-dimensional (3D) image reconstruction and video techniques, and 2D and 3D immunochemistry; (2) determine the sites of mineral deposition by similar means and also electron diffraction, x-ray probe microanalysis, and novel Fourier transform infrared spectroscopy; (3) characterize interaction between extracellular matrix proteins and mineral through correlation of the methods above and by atomic force microscopy and computer modeling (molecular dynamics); and (4) assess the effects in bone cell culture on matrix-mineral interaction following modulation of protein transport with the inhibitor, monensin. It is suggested that outcome data will contribute to knowledge of structure and function of cells, the cell-matrix interface, and extracellular matrices and events fundamental to vertebrate calcification.

描述：（改编自研究者的摘要）该提案 旨在获得有关脊椎动物骨骼的新信息 骨、软骨和肌腱钙化。 正常的小鸡、小鼠、大鼠、 和火鸡；骨和软骨培养；和异常的成骨作用 不完美小鼠突变体将用作模型。 研究这些 示例将检查某些结构和化学相互作用 它们各自的细胞外基质、细胞内基质和 细胞内和细胞外界面。 工作将系统地进行， 定义蛋白质分泌途径、细胞基质粘附关系，以及 矿物基质协会。 超微结构、生化、 将应用免疫化学和生物物理方法来扩展 申请人之前的结果描述了存在、性质、位置， 以及脊椎动物有机基质和矿物质成分的相互作用 钙化。 要检验的具体假设是：有机 脊椎动物钙化组织基质严重影响 矿物沉积的晶体尺寸、形状、位置、 方向、分布。 具体目标，解决结构- 原子、分子和大分子水平上的函数关系 层次结构的目的是：（1）识别胶原蛋白的存在和位置 和参与矿化的非胶原蛋白，以及 细胞基质中选定的整合素和细胞骨架元件 接口，利用传统和高压电子显微镜， 三维 (3D) 图像重建和视频技术，以及 2D 和 3D 免疫化学； (2) 确定矿物沉积部位 类似的方法还有电子衍射、X 射线探针微量分析、 和新颖的傅里叶变换红外光谱； (3) 表征 细胞外基质蛋白和矿物质之间的相互作用 上述方法与原子力显微镜的相关性和 计算机建模（分子动力学）； (4) 评估影响 骨细胞培养对基质-矿物质相互作用的调节 与抑制剂莫能菌素一起运输蛋白质。 建议 结果数据将有助于了解结构和功能 细胞、细胞-基质界面、细胞外基质和事件 脊椎动物钙化的基础。