Immunochemical and immunocytochemical studies of the C-propeptide of type II procollagen in chondrocytes of the growth plate.
The C-propeptide of type II procollagen has previously been implicated in cartilage calcification. To further characterize this propeptide, we have investigated its molecular status and intracellular distribution in bovine fetal growth plate chondrocytes, particularly within the calcifying zone, using cell isolation, Western blotting, and localization with immunofluorescence and immunogold techniques. We found that in all cells freshly isolated by collagenase digestion the C-propeptide was a component of type II pro-alpha chains. No free C-propeptide was detected intracellularly. In situ localization of the C-propeptide by immunostaining employing immunofluorescence revealed the presence of procollagen in most growth plate cells, staining being most intense in hypertrophic cells. In the latter, large dilations of the rough endoplasmic reticulum were observed. These were not found in proliferating cells and had an approximate diameter of 5 microns. With immunogold localization these, together with Golgi-derived secretory granules, stained for the C-propeptide. These combined results suggest that in all cells of the growth plate the C-propeptide is a constituent part of type II collagen pro-alpha chains, and that it is usually segregated in the rough endoplasmic reticulum at a time when, according to other studies, collagen synthesis ceases in the lower hypertrophic zone and calcification of the extracellular matrix ensues. This suggests that the intracellular translocation of type II collagen pro-alpha chains may change in hypertrophic cells at this time.
Lee
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ER
Shriners Hospital for Crippled Children, Department of Experimental Surgery, McGill University, Montreal, Quebec, Canada.
Matsui
Y
Y
Poole
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AR
D000818:Animals; D000906:Antibodies; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002469:Cell Separation; D055598:Chemical Phenomena; D002621:Chemistry; D003094:Collagen / Q000378:metabolism*; D024043:Collagen Type II; D006132:Growth Plate / Q000166:cytology / Q000378:metabolism*; D007120:Immunochemistry; D007150:Immunohistochemistry; D008970:Molecular Weight; D013194:Staining and Labeling
D000906:Antibodies; D002135:Calcium-Binding Proteins; D024043:Collagen Type II; C040072:chondrocalcin; D003094:Collagen