PROTEOGLYCANS AND OSTEOPONTIN IN BONE FORMATION

骨形成中的蛋白聚糖和骨桥蛋白

• 批准号: 3072204
• 负责人: CHARLES W PRINCE
• 金额: $ 5.44万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3072204
• 关键词: autoradiography; bone development; calcium binding protein; collagen; elastin; extracellular matrix; glycoproteins; hormone regulation /control mechanism; hydroxyapatites; molecular biology; monoclonal antibody; mucopolysaccharides; normal ossification; organ culture; osteoblasts; parathyroid hormones; phosphorylation; proteoglycan; tissue /cell culture

The specific aims of this proposal are 1) to analyze in detail the chemistry and biosynthesis of bone proteoglycans, 2) to use antibodies in immunolocalization experiments, and 3) to evaluate the molecular mechanism by which 1,25-dihydroxyvitamin D3 appears to regulate osteopontin biosynthesis. The first two aims depend on classical biochemical and immunological methods while the last aim relies, to some extent, on the techniques of molecular biology. The applicant has some experience using these techniques, however, the funding of this Research Career Development Award would greatly enhance his research career development by allowing more extensive training in this area. The major hypothesis upon which this research is based is that non-collagenous components of mineralized connective tissues are intimately involved in the formation of the tissues. Furthermore, an understanding of the chemial composition, localization and regulatory mechanisms of biosynthesis are required before specific, functional roles can be elucidated. In this proposal we will address the aforementioned areas as they relate to proteoglycans and osteopontin. The approach taken is to isolate chemical amounts of these components, verify their purity and use them as antigens. The antibodies will be used in immunolocalization studies and as reagents in immunoassays. We will explore the kinetics of proteoglycan biosynthesis in rat bone, in ROS 17/2.8 cells and in cultures of rat calvarial osteoblasts. We will evaluate the role and mechanism that 1,25- dihydroxyvitamin D3 may have in the regulation of osteopontin biosynthesis.

该提案的具体目的是1）详细分析 骨蛋白聚糖的化学和生物合成，2） 免疫定位实验中的抗体，3）评估 1,25-二羟基维生素D3的分子机制 似乎调节骨桥蛋白生物合成。 前两个目标 取决于经典的生化和免疫学方法 最后的目标在某种程度上依赖于 分子生物学。 申请人有一些经验 但是，这项研究生涯的资金 发展奖将大大提高他的研究生涯 通过在该领域进行更广泛的培训来开发。 这项研究基于的主要假设是 矿化结缔组织的非胶原成分是 密切参与组织的形成。 此外， 了解化学组成，定位和 需要生物合成的调节机制 可以阐明特定的功能作用。 在这个建议中，我们 将解决与上述区域有关的问题 蛋白聚糖和骨桥。 采用的方法是隔离 这些成分的化学量，验证其纯度和 将它们用作抗原。 抗体将用于 免疫定位研究和作为免疫测定中的试剂。 我们 将探索大鼠骨中蛋白聚糖生物合成的动力学， 在ROS 17/2.8细胞中，在大鼠钙钙细胞的培养物中。 我们将评估1,25-的作用和机制 二羟基维生素D3在骨桥蛋白的调节中可能具有 生物合成。