Differential localization of mRNAs of collagen types I and II in chick fibroblasts, chondrocytes, and corneal cells by in situ hybridization using cDNA probes.
We have employed a highly specific in situ hybridization protocol that allows differential detection of mRNAs of collagen types I and II in paraffin sections from chick embryo tissues. All probes were cDNA restriction fragments encoding portions of the C-propeptide region of the pro alpha-chain, and some of the fragments also encoded the 3'-untranslated region of mRNAs of either type I or type II collagen. Smears of tendon fibroblasts and those of sternal chondrocytes from 17-d-old chick embryos as well as paraffin sections of 10-d-old whole embryos and of the cornea of 6.5-d-old embryos were hybridized with 3H-labeled probes for either type I or type II collagen mRNA. Autoradiographs revealed that the labeling was prominent in tendon fibroblasts with the type I collagen probe and in sternal chondrocytes with the type II collagen probe; that in the cartilage of sclera and limbs from 10-d-old embryos, the type I probe showed strong labeling of fibroblast sheets surrounding the cartilage and of a few chondrocytes in the cartilage, whereas the type II probe labeled chondrocytes intensely and only a few fibroblasts; and that in the cornea of 6.5-d-old embryos, the type I probe labeled the epithelial cells and fibroblasts in the stroma heavily, and the endothelial cells slightly, whereas the type II probe labeled almost exclusively the epithelial cells except for a slight labeling in the endothelial cells. These data indicate that embryonic tissues express these two collagen genes separately and/or simultaneously and offer new approaches to the study of the cellular regulation of extracellular matrix components.
D000818:Animals; D002356:Cartilage / Q000378:metabolism*; D002642:Chick Embryo; D003094:Collagen / Q000235:genetics / Q000378:metabolism*; D003315:Cornea / Q000378:metabolism*; D004247:DNA; D004262:DNA Restriction Enzymes; D004848:Epithelium / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D010006:Osteoblasts / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*
D012333:RNA, Messenger; D003094:Collagen; D004247:DNA; D004262:DNA Restriction Enzymes