The expression of various extracellular matrix genes is examined by skin fivroblast cultures and placental tissues using Northern and slot-blot hybridization. Total RNA from 3-6 passaged fibrobasts cultures was isolated.
In Northern blot analysis, t...
The expression of various extracellular matrix genes is examined by skin fivroblast cultures and placental tissues using Northern and slot-blot hybridization. Total RNA from 3-6 passaged fibrobasts cultures was isolated.
In Northern blot analysis, type Ⅰ and type Ⅲprocollagen-specific mRNA transcripts from cultured skin fibroblasts demostrated the specificity of hybridization with pro α1(Ⅰ) and pro α1(Ⅲ) collagen. specifically, pro α1(Ⅰ) or pro α1(Ⅲ) collagen cDNA probes hybridized to two distinct mRNA species with apparen sizes of 5.8 and 4.8 kilobases(kb), or 5.4 and 4.8 kb, respectively. The molecular size of fibronectin and β-actin mRNA revealed 8.0 kb and 2.0 kb, respectively. Therefore, it showed no changes of sizes of mRNAs between skin fibroblasts and placental tissues.
In slot-bolt analysis, the levels of type Ⅰ,Ⅲ collagen and fibronectin mRNA in skin fibroblasts is higher each 18.8, 3.2, and 1.7 folds than in placental tissues. And determination of type Ⅰ/Ⅲ procollagen ratios revealed 4.4in fibroblast cultures, and 0.75 in placental tissues.
This results indicate that type Ⅰ collgaen is predominant extracelluar matrix produced by skin fibroblasts when compaired to placental tissues.