Fleischmajer R, Olsen B R, Timpl R, Perlish J S, Lovelace O
Proc Natl Acad Sci U S A. 1983 Jun;80(11):3354-8. doi: 10.1073/pnas.80.11.3354.
Chicken embryo skin of different ages and adult skin were labeled with antibodies against the amino propeptide and carboxyl propeptide of type I collagen and processed for indirect immunoelectron microscopy by the ferritin technique. The results indicate that the formation of thin collagen fibrils involves polymerization of pN-collagen. Fibrils that are thicker than 35-40 nm do not appear to contain the amino propeptide. How fibrils increase in size is not clear, but growth may involve mechanisms such as lateral aggregation of subfibril structures or fusion of thin fibrils. Carboxyl propeptides were localized near or in contact with thin collagen fibrils, but they did not appear to be arranged in a periodic manner along the fibrils. In experiments using antibodies against the amino propeptides of type III collagen, fibrils 20-40 nm in diameter were also labeled in a periodic fashion. pN-Collagen chains were extracted from embryonic skin and identified by NaDodSO4/polyacrylamide gel electrophoresis and by immunoblotting. The presence of significant amounts of pN-collagen in skin from 10- and 12-day chicken embryos agreed well with the labeling of amino propeptides by immunoelectron microscopy. This study provides evidence for the role of the amino propeptide in collagen fibrillogenesis in embryonic skin.
用抗I型胶原氨基前肽和羧基前肽的抗体标记不同年龄的鸡胚皮肤和成年皮肤,并采用铁蛋白技术进行间接免疫电子显微镜处理。结果表明,细胶原纤维的形成涉及pN-胶原的聚合。直径大于35 - 40nm的纤维似乎不含有氨基前肽。纤维大小如何增加尚不清楚,但生长可能涉及亚纤维结构的横向聚集或细纤维融合等机制。羧基前肽定位于细胶原纤维附近或与之接触,但它们似乎不是沿纤维呈周期性排列。在使用抗III型胶原氨基前肽抗体的实验中,直径20 - 40nm的纤维也呈周期性标记。从胚胎皮肤中提取pN-胶原链,并通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳和免疫印迹进行鉴定。10日龄和12日龄鸡胚皮肤中存在大量pN-胶原,这与免疫电子显微镜对氨基前肽的标记结果非常吻合。本研究为氨基前肽在胚胎皮肤胶原纤维形成中的作用提供了证据。