Abstract
We have followed the deposition and maturation of the pericellular matrix of amniotic epithelial cell cultures for up to eight weeks using metabolic labeling and immunoelectron microscopy. This matrix contains mainly collagen type III and fibronectin. Cleavage of the carboxypropeptide occurred after secretion of the procollagen molecules into the medium but was not accompanied by a significant release of the aminopropeptide. The early matrix, as isolated from the cultures by a deoxycholate procedure, contained collagenous proteins predominantly composed of pN alpha 1(III) chains, which still possessed the aminopropeptide, and only little material in the form of alpha 1(III) chains. The relative amount of alpha 1(III) chains increased during subsequent days of culture. Electron microscopy showed two types of structures in the matrix: thin fibrils, ranging from 10 to 30 nm in diameter, with no apparent cross-striation, and 50-500 nm thick bundles composed of filamentous and amorphous material. In the fibrils, immunoferritin electron microscopy showed a regular staining for the aminopropeptide of procollagen type III with a periodicity of 71 nm. These collagenous fibrils did not stain for fibronectin which was found in the bundles. Since most of the aminopropeptide in the matrix appeared covalently linked as pN-collagen, we conclude that the deposition of this intermediate form of procollagen is a general mechanism in collagen type III fibrillogenesis.