LOCALIZATION OF PROCOLLAGENS IN DENTAL TISSUES

牙齿组织中原胶原的定位

• 批准号: 3219577
• 负责人: CHARLES P LEBLOND
• 金额: $ 5.99万
• 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 1988-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3219577
• 关键词: affinity chromatography; autoradiography; basement membrane; electron microscopy; fibroblasts; immunoglobulin G; radiotracer; tooth enamel

The collagens are elaborated within cells as procollagens along a complex biosynthetic pathway, which has been partly identified in the case of type I and IV collagens but is little known in the case of type III and V collagens. The proposal is to identify the pathway in the latter two and to measure the rates of passage of the precursors through various cell compartments in all four. In addition, the incidental findings of new structures in basement membrane had led to a study of their tridimensional architecture and immunocharacterization. For the intracellular biogenesis of the collagens, two main methods will be utilized: a) immunostaining of Lowicryl embedded tissues by protein A-gold; and counting gold particles per unit area over the structures under study; b) high resolution radioautography at various times after a 3H-proline injection. The results obtained by the two approaches will be combined to prepare specific activity time curves for each organelle in the producer cells and thus work out steps and rates of migration along the biogenetic pathway of the various collagens. In practice, the antibodies capable of reacting with the intracellular precursor of collagen will be used for two purposes: firstly, to identify the cells producing the collagen and secondly, to examine the intracellular steps in the biogenesis of this collagen. The cells selected to study biogenesis are cells which are clearly immunostained for only one type of collagen: odontoblasts for type I collagen, periodontal fibroblasts for type III collagen, and endodermal cells associated with a multilayered basement membrane of rat embryos (Reichert's membrane) for type IV collagen. As for type V collagen, exploratory work on rat placenta and other tissues will be needed to detect producer cells and use them for the study of biogenesis. Finally, incidental results on the structure of Reichert's membrane offer hope of clarifying the organization of type IV collagen within basement membranes. In Reichert's membrane, type IV collagen is present as an axial filament in cords arranged into a network. These obervations have to be extended to the basement membranes of tooth and other tissues. Moreover, the finding of rigid hollow rods called basotubules and dots called double-pegs has to be confirmed in common basement membranes. It is thus hoped to extend our knowledge of collagen biogenesis in general and basement membranes in particular.

胶原蛋白在细胞内以前胶原的形式沿着复杂的结构精加工而成 生物合成途径，在类型的情况下已部分确定 I 型和 IV 型胶原蛋白，但 III 型和 V 型胶原蛋白鲜为人知 胶原蛋白。 该提案是确定后两者的途径 测量前体细胞通过各种细胞的速率 所有四个隔间。 此外，偶然发现的新 基底膜的结构导致了对其三维空间的研究 结构和免疫表征。 对于胶原蛋白的细胞内生物合成，有两种主要方法： 使用：a)通过蛋白A-金对Lowicryl包埋的组织进行免疫染色； 计算所研究结构上每单位面积的金颗粒； b) 3H-脯氨酸后不同时间的高分辨率放射自显影 注射。 两种方法获得的结果将合并为 为生产者中的每个细胞器准备特定的活动时间曲线 细胞，从而计算出沿着生物遗传的迁移步骤和速率 各种胶原蛋白的途径。 在实践中，能够与细胞内的抗体发生反应的抗体 胶原蛋白前体将用于两个目的：首先，识别 产生胶原蛋白的细胞，其次检查细胞内 这种胶原蛋白生物合成的步骤。 选择进行研究的细胞 生物发生是仅对一种类型进行明确免疫染色的细胞 胶原蛋白：I型胶原蛋白为成牙本质细胞，I型胶原蛋白为牙周成纤维细胞 III 型胶原蛋白和与多层相关的内胚层细胞 IV 型大鼠胚胎基底膜（Reichert 膜） 胶原。 对于V型胶原蛋白，在大鼠胎盘和 需要其他组织来检测生产细胞并将其用于 生物发生的研究。 最后，关于 Reichert 膜产品结构的附带结果 希望澄清地下室内 IV 型胶原蛋白的组织 膜。 在 Reichert 膜中，IV 型胶原蛋白以轴向形式存在 排列成网络的绳索中的细丝。 这些观察必须是 延伸至牙齿和其他组织的基底膜。 而且， 发现称为基底小管的刚性空心棒和称为 双钉必须在常见的基底膜中得到证实。 因此，希望能够扩展我们对胶原蛋白生物发生的一般知识 特别是基底膜。