LOCALIZATION OF PROCOLLAGENS IN DENTAL TISSUES

牙齿组织中原胶原的定位

• 批准号: 3219572
• 负责人: CHARLES P LEBLOND
• 金额: $ 7.53万
• 依托单位: MCGILL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-08-01 至 1991-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3219572
• 关键词: Golgi apparatus; amyloid proteins; autoradiography; basement membrane; chondrocytes; electron microscopy; fibroblasts; fibronectins; glycoproteins; histochemistry /cytochemistry; immunochemistry; laboratory mouse; laboratory rabbit; laboratory rat; odontoblasts; organelles; procollagen; proline; proteoglycan; radiotracer; tooth enamel; tritium

This application is designed to continue studies aimed at a clarification of the intracellular formation of various collagens. Enough progress has been made recently in the techniques of radioautography and immunohistochemistry at the electron microscopic level to make it possible to combine these methods on the same sections. Thus, at various times after in injection of 3H-proline, radioautographic silver grains (which are spherical and about 50 nm wide) and immunogold particles (15 nm) can be counted separately over the same organelle (rER cisterna, Golgi saccule, secretory granule). The ratio of the two counts will provide the specific activity of collagen precursors in each organelle and its variation with time. The turnover rate of the precursors will when be calculated at each step along the intracellular pathway of collagen. Specific activity measurements will be applied to the study of the biogenesis of type IV collagen in active chondrocytes, type I collagen in odontoblasts, type IV collagen in endodermal cells and type III collagen in periodontal fibroblasts. A second aim of the application is to continue the investigations of basement membrane and related structures. The major component of basement membrane is shown at high magnification of the electron microscope to be a tridimensional network of anastomosing strands, referred to as "cords", whose average width is 4 nm. It is planned to combine purified type IV collagen with other basement membrane components in vitro in the hope of reproducibly obtaining 50-100 nm sheets made up of a network of 4-nm cords, that is, sheets similar to the lamina densa of authentic basement membranes. A minor component of some basement membrane, the basotuble - a 9 nm wide rod of indefinite length - has been shown to have the same dimension and ultrastructure as the microfibrils of connective tissues. The main structural element of both is a core consisting of stacked molecules of amyloid P component (a glycoprotein known to be present in normal serum and to accumulate in amyloid degeneration). Three substances, i.e. fibronectin, fibrillin and a proteoglycan, have been shown to be associated with microfibrils. The aim of our work is to utilize immunolabeling for these substances using 5 nm gold particles to determine how they related to the amyloid P core of microfibrils and basotubules.

该应用程序旨在继续针对的研究 阐明各种胶原蛋白的细胞内形成。 最近在技术方面取得了足够的进步 电子的放射学和免疫组织化学 微观水平使这些方法在 相同的部分。 因此，在注射后的不同时间 3h proline，放射性银颗粒（球形和 可以计算约50 nm宽）和免疫颗粒（15 nm） 分别在同一细胞器上（rer cisterna，golgi accule， 分泌颗粒）。 这两个计数的比率将提供 每个细胞器及其胶原前体的特定活性 随时间变化。 前体的周转率将于 在沿着细胞内通路的每个步骤计算 胶原。 特定的活动测量将应用于 研究活性软骨细胞中IV型胶原蛋白的生物发生， I型I胶原蛋白胶原蛋白细胞，内胚层IV胶原蛋白 细胞和牙周成纤维细胞中的III型胶原蛋白。 该申请的第二个目的是继续调查 地下室膜和相关结构。 主要组成部分 在电子的高放大倍率时显示了地下膜的 显微镜是吻合链的三维网络， 称为“绳索”，其平均宽度为4 nm。 它是计划的 将纯化的IV型胶原蛋白与其他地下膜结合 体外的组件，希望可重复获得50-100 NM纸由4纳米电线的网络组成，即 类似于正宗基底膜的薄片。 某些地下膜的次要组成部分，basotuble -a 9 NM宽的无限长度 - 已证明具有相同的 尺寸和超微结构作为结缔组织的微纤维 组织。 两者的主要结构元素是组成的核心 淀粉样蛋白P成分的堆叠分子（已知糖蛋白 在正常血清中存在并积聚在淀粉样蛋白中 退化）。 三种物质，即纤连蛋白，原纤维蛋白和 蛋白聚糖已被证明与微纤维有关。 我们工作的目的是利用免疫标签 使用5 nm黄金颗粒来确定它们如何相关的物质 至微纤维和basotubules的淀粉样蛋白P核。