Age-related changes in collagenase expression in cultured embryonic and fetal human skin fibroblasts.
Since skin collagenase is required for initiation of the degradation of types I and III collagens, the major collagens of the human dermis, we examined its expression during embryonic and fetal development. When using skin fibroblasts cultured from human embryos and fetuses, immunoreactive collagenase concentrations were strongly correlated with estimated gestational age (p less than 0.001), with levels at 7-8 weeks of gestation that were about one-twentieth of those in the 29-week cell cultures. In crude culture medium, the apparent catalytic efficiency (activity per unit immunoreactive protein) was variable, an observation attributable in part to variable expression of a collagenase-inhibitory protein. Following chromatographic purification, four of ten fetal collagenases were found to have greater than or equal to 4-fold decrease in specific activity, suggesting that these particular fetal collagenases may be structurally and/or catalytically altered. Since the decreased levels of immunoreactive protein suggested that decreased enzyme synthesis was the major mechanism, we examined collagenase synthesis in a cell-free translation system. Here, we quantitated collagenase expression in the culture medium of intact cells prior to harvesting mRNA. Compared with the intact adult cells, the fetal cells had 3-17 times less collagenase activity in the medium, while in cell-free translation there was a 2- to 3-fold decrease in collagenase synthesis. These data suggest that decreased in vitro expression is correlated with decreased levels of translatable collagenase mRNA but that other factors, such as the collagenase inhibitor and altered specific activity of the enzyme, may be important in modulating collagenase activity.
D000375:Aging; D002474:Cell-Free System; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D005786:Gene Expression Regulation; D005865:Gestational Age; D006801:Humans; D003012:Microbial Collagenase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000378:metabolism; D012867:Skin / Q000196:embryology / Q000201:enzymology*
D012333:RNA, Messenger; D003094:Collagen; D003012:Microbial Collagenase