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PubMed1996

Histochemical analysis of insulin-like growth factor-1 binding sites in mouse normal and experimentally induced arthritic articular cartilage.

Insulin-like growth factor-1 (IGF-1) plays a key role in regulation of chondrocyte metabolism. We examined the localization of IGF-1 binding sites on chondrocytes in cartilage from normal and experimentally induced arthritic mouse knee joints. Cryostat sections from patellar cartilage were incubated either with IGF-1 receptor antibody or biotinylated IGF-1. Subsequently confocal laser scanning microscopy was applied to compare the two staining procedures qualitatively and quantitatively. This approach allowed detailed analysis of membrane-associated and intracellular staining. Using IGF-1 receptor antibody, IGF-1 receptors were found on the cell membrane of chondrocytes in the middle and deeper cartilage zones, whereas intracellular staining was highest in chondrocytes of superficial zones. After incubation with biotinylated IGF-1, distinct membrane staining was not present and fluorescence was localized homogeneously in the middle and deeper zones but not in superficial zones. In cartilage from inflamed knee joints staining with the use of IGF-1 receptor antibody did not change significantly, whereas a pronounced increase in staining was noted with biotinylated IGF-1 in chondrocytes of the middle and deeper zones of cartilage. It is concluded that the staining patterns obtained with the use of IGF-1 receptor antibody and biotinylated IGF-1 are remarkably different, suggesting that the latter also detects IGF-binding proteins. The results suggest that joint inflammation has no consistent effect on IGF-1 receptor expression but may induce a significant upregulation of IGF-binding proteins in chondrocytes of the middle and deeper zones of cartilage.

Animals↗
PubMed1996

Alpha and epsilon isozymes of protein kinase C in the chondrocytes in normal and early osteoarthritic articular cartilage.

Protein kinase C (PKC) isozymes (alpha, beta I, beta II, gamma and, epsilon-PKC) were examined immunocytochemically in control and mechanically induced osteoarthritic knee joints in rats. beta I, beta II, and gamma PKC-positive cells were not observed in sham-operated or osteoarthritic knee joints. alpha-PKC, which was observed only in the cells in the subchondral bony layer in the controls, appeared in the chondrocytes in the superficial and columnar layers of the osteoarthritic knees. epsilon-PKC was observed in the control chondrocytes in the superficial portion of the columnar layers. In early osteoarthritic joints, however, epsilon-PKC-positive chondrocytes disappeared from the superficial portion of columnar layer and increased in number in the middle columnar layers. The appearance and changes in distribution of these PKC-positive chondrocytes, either under normal or osteoarthritic conditions, have not been reported previously. The role of the redistribution of PKC isozymes is still unclear as to whether they are involved in initiating destructive processes, reflect attempted cell repair mechanisms, or are simply a consequence of the cellular changes.

Animals↗