PROTEOGLYCANS AND OSTEOPONTIN IN BONE FORMATION

本提案的具体目的是：（1）详细分析 骨蛋白聚糖的化学和生物合成，2）使用 免疫定位实验中的抗体，以及3）评估 1，25-二羟维生素D3 似乎调节骨桥蛋白的生物合成。 前两个目标 依赖于经典的生物化学和免疫学方法， 最后一个目标在某种程度上依赖于以下技术： 分子生物学 申请人有一定的使用经验， 技术，然而，这项研究事业的资金 发展奖将大大提高他的研究生涯 通过在这一领域进行更广泛的培训来促进发展。 这项研究所依据的主要假设是， 矿化结缔组织的非胶原成分是 与组织的形成密切相关。 此外，委员会认为， 了解化学成分、定位和 生物合成的调节机制是必需的， 可以阐明具体的职能作用。 在本提案中，我们 将处理上述领域，因为它们涉及 蛋白聚糖和骨桥蛋白。 采取的方法是隔离 这些成分的化学量，验证其纯度， 将它们用作抗原。 这些抗体将用于 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细胞和大鼠颅骨成骨细胞培养物中。 我们将评估1，25- 二羟维生素D3对骨桥蛋白的调节作用 生物合成

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.