Vigilin和Ⅰ型、Ⅱ型前胶原mRNA在鸡胚中分布的原位杂交研究

Localization of Vigilin and Type Ⅰ,Ⅱ Procollagen mRNAs in Chick Embryos by In Situ Hybridization

用^(35)S标记的RNA-RNA原位杂交技术,在鸡胚组织中观察到vigilin mRNA在骨母细胞中含量较高,随骨母细胞成熟为骨细胞而显著减少。在软骨细胞不同分化阶段,增生区(后期)及肥大区的软骨细胞中vigilin mRNA含量最高。纤维母细胞含中等量vigilin mRNA。在骨母细胞和纤维母细胞中,vigilin mRNA与Ⅰ型前胶原mRNA分布相似,在软骨细胞中,与Ⅲ型前胶原mRNA分布一致。增生区(后期)及肥大区软骨细胞也有明显的proα1(Ⅰ)mRNA信号,并不被proα1(Ⅲ)反义RNA所阻断,但无proα2(Ⅰ)基因的表达。提示鸡胚软骨细胞确有proα1(Ⅰ)基因的表达,产物可能是α1(Ⅰ)三聚体。

Using a ^(3)5S-labeled RNA-RNA in situ hybridization technique,we found that vigilin mRNA was abundent in osteoblasts.It disappeared following the transition of osteoblasts to osteocytes.The amount of vigilin mRNA increased with proliferation and hypertrophy of chondrocytes.Fibroblasts contained noderate vigilin mRNA signals.Weak signals were observed in early chick embryos. Type Ⅰ procollagen mRNAs were abundent in osteoblasts and fibroblasts of the periosteum,perichondrium,tendon and ligament.Type Ⅱ procollagen mRNA was restricted to chondrocytes.Interestingly,a significant amount of proα1 (Ⅰ) mRNA was observed in late proliferative and hypertrophic chondrocytes, which could not be blocked by an excess amount of unlabeled pro α1 (Ⅱ) antisense RNA.No signal was found in chondrocytes of human fetal cartilage.Vigilin mRNA had a similar distribution to type Ⅱ procollagen mRNA in chondrocytes,and in other instances it showed a codistribution with type Ⅰ procollagen mRNAs.