Skip to main content
Log in

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

  • Papers
  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Summary

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 themiddle 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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Adem, A., Jossan, S. S., D'argy, R., Gillberg, P. G., Nordberg, A., Winbald, B., &Sara, V. (1989) IGF-1 receptors in the human brain: quantitative autoradiographic localization.Brain Res. 503, 299–303.

    Article  CAS  PubMed  Google Scholar 

  • Altman, F. P. (1987) Gelatin embedding technique as an aid in the preparation of unfixed cryostat sections.Histochem. J. 10, 617.

    Google Scholar 

  • Aydelotte, M. B. & Keuttner, K. E. (1991) Heterogeneity of articular chondrocytes and cartilage matrix. InJoint Cartilage Degradation: Basic and Clinical Aspects (edited byWoessner, J. F. & Howell, D. S), pp. 37.

  • Baumick, B. &Bala, R. M. (1991) Differential effects of insulinlike growth factors 1 and 2 on growth, differentiation and glucoregulation in differentiating chondrocyte cells in culture.Acta Endocrinol. 125, 201–11.

    Google Scholar 

  • Baxter, R. C. (1993) Circulating binding proteins for the insulin-like growth factors.Trends Endocrinol. Metabol. 4, 91–6.

    CAS  Google Scholar 

  • Byers, J., Maroudas, A., Oztop, F., Stockwell, R. A. &Venn, M. F. (1977) Histological and biochemical studies on cartilage from osteoarthritic femoral heads with special reference to surface characteristics.Connect. Tissue Res. 5, 41–9.

    CAS  PubMed  Google Scholar 

  • Chin, E., Michels, K. &Bondy, C. A. (1994) Partition of insulin-like growth factor binding sites between the insulin-like growth factor-1 and insulin-like growth factor-2 receptors and IGF-binding proteins in the human kidney.J. Endocrinal. Metab. 78, 156–164.

    CAS  Google Scholar 

  • Dore, S., Peletier, J.-P., Dibattista, J. A., Tardif, G., Brazeau, P. &Martel-Pelletier, J. (1994) Human osteoarthritic chondrocytes possess an increased number of insulin-like growth factor 1 binding sites but are unresponsive to its stimulation.Arthritis Rheum. 37, 235–63.

    Google Scholar 

  • Hardingham, T. &Bayliss, M. (1990) Proteoglycans of articular cartilage: Changes in aging and jo joint disease.Sec. Arthritis Rheum. 20, 12–33.

    CAS  Google Scholar 

  • Hardingham, T. E., Fosang, A. J. &Dudhia, J. (1992a) Aggrecan, the chondroitin sulfate/keratan sulfate proteoglycan from cartilage. InArticular Cartilage and Osteoarthritis (edited byKuetiner, K. E.), pp. 5–20. New York: Raven Press.

    Google Scholar 

  • Hardingham, T. E., Bayliss, M. T., Rayan, V. &Noble, D. P.(1992b) Effects of growth factors and cytokines on proteoglycan turnover in articular cartilage.Br. J. Rheumatol. 31, (Suppl. 1), 1–6.

    PubMed  Google Scholar 

  • Hascall, V. C., Handley, C. J. McQuillan, D. J., Hacall, G. K. Robinson, H. C., &Lowther, D. A. (1983). The effect of serum on biosynthesis of proteoglycans by bovine articular cartilage in culture.Arch. Biochem. Biophys. 224, 206–23.

    Article  CAS  PubMed  Google Scholar 

  • Isaksson, O. G., Ohlsson, C., Nilsson, A., Isgaard, J. &Lindahl, A. (1991) Regulation of cartilage growth by growth hormone and insulin-like growth factor 1.Pathology 5, 451–3.

    CAS  Google Scholar 

  • Jansen, J., Van Buul-Offers, S. C., Hoogerbrugge, C. M., Poorter, T. L., Corvol, M. T. &van den Brande, J. L. (1989) Characterization of specific insulin-like growth factor [IGF]-1 and IGF-2 receptors on cultured rabbit articular chondrocyte membranes.Endocrinology 129, 245–9.

    Google Scholar 

  • Johnstone, J., Bitensky, L. &Chayen, J. (1972) Cryostat sections of unmineralized bone.J. Clin. Pathol. 25, 742.

    CAS  PubMed  Google Scholar 

  • Lafeber, F. P. J. G., Van der Kraan, P. M., Vanroy, H. L. A. M., Vitters, E. L., Huber-Bruning, O., Van den Berg, W. B. &Bijlsma, J. W. J. (1992) Local changes in proteoglycan synthesis during culture are different for normal and osteoarthritic cartilage.Am. J. Pathol. 140, 1421–9.

    CAS  PubMed  Google Scholar 

  • Lafeber, F. P. J. G., Vander Kraan, P. M., Roy, J. L. A. M., Huber-Bruning, O. &Bijlsma, J. W. J. (1993) Articular cartilage explant culture; an appropriatein vitro system to compare osteoarthritic and normal human cartilage.Connect. Tissue Res. 29, 287–99.

    CAS  PubMed  Google Scholar 

  • LeRoith, D. &Raizada, M. K. (1989)Molecular and Cellular Biology of Insulin-like Growth Factors and their Receptors. New York: Plenum Press.

    Google Scholar 

  • Lersniak, M. A., Hill, J. M., Kiess, W., Rojeski, M., Pert, C. B.,&Roth, J. (1988) Receptors for insulin-like growth factor-1 and 2; autoradiographic localization in rat brain and comparison to receptor for insulin.Endocrinology 123, 2089–99.

    Google Scholar 

  • Luyten, F. P., Hacall, V. C., Nissley, S. P., Morales, T. I. &Reddi, A. H. (1988) Insulin-like growth factors maintain steadystate metabolism of proteoglycans in bovine articular cartilage explants.Arch. Biochem. Biophys. 267, 416–25.

    Article  CAS  PubMed  Google Scholar 

  • McQuilland, D. J., Handley, C. J., Campbell, M. A., Bolis, S., Millway, V. E. &Herington, A. C. (1986) Stimulation of proteoglycan synthesis by serum and insulin-like growth factor-1 in culture bovine articular cartilage.Biochem. J. 240, 424–30.

    Google Scholar 

  • Meachim, G., Ghadially, F. N. & Collins, D. H. (1965) Regressive changes in the superficial layer of human articular cartilage.Ann. Rheum. Dis. 24, 23–30.

    CAS  PubMed  Google Scholar 

  • Middleton, J. F. S. &Tyler, J. A. (1992) Upnegulation of insulin-like growth factor 1 gene expression in the lesions of osteoarthritic human articular cartilage.Ann. Rheum. Dis. 51, 440–7.

    CAS  PubMed  Google Scholar 

  • Nefly, E. K., Beukers, M. W., Oh. Y., Cohen, P. &Rosenfeld, R. G. (1991) Insulin-like growth factor receptors.Acta Paediat. Scand. 372, 116–23.

    Google Scholar 

  • Oemar, B. S., Foellmer, H. G., Hodgedon-ananant, L. &Rosenzweig, S. A. (1991) Regulation of insulin-like growth factor receptor in diabetic mesangial cells.J. Biol. Chem. 266, 2369–73.

    CAS  PubMed  Google Scholar 

  • Osborn, K. D., Trippel, S. B. &Mankin, H. J. (1989) Growth factor stimulation of adult articular cartilage.J. Orthop. Res. 7, 35–42

    Article  CAS  PubMed  Google Scholar 

  • Pearse, A. D. &Gardner, D. L. (1972) Preparation of unfixed undermineralized bone sections: The Bright bone cryostat.J. Clin. Pathol. 25, 26–9.

    CAS  PubMed  Google Scholar 

  • Rechler, H. M. &Nissley, S. P. (1990) InPeptide Growth Factors and their Receptors (edited bySporn, M. B. &Robers, A. B.), pp.263–376. New York: Springer-Verlag.

    Google Scholar 

  • Rijntjes, N. V. M., Van de Putte, L. B. A., Van der Pol, M. &Guelen, P. J. M. (1979) Cryosectioning of undecalcified tissues for immunofluorescence.J. Immunol. Meth. 30, 263–8.

    Article  CAS  Google Scholar 

  • Rosenzweig, S. A., Zetterstrom, C. &Benjamin, A. (1990) Identification of retinal insulin receptors using site-specific antibodies to a carboxyl-terminal peptide of the human insulin receptor alpha-subunit. Upregulation of neuronal insulin receptors in diabetes.J. Biol. Chem. 265, 18030–4.

    CAS  PubMed  Google Scholar 

  • Sampson, H. W. &Cannon, M. S. (1986) Zonal analysis of metabolic profiles of articular-epiphyseal cartilage chondrocytes.Histochem. J. 18, 265–70.

    Article  Google Scholar 

  • Schalch, D. S., Sessions, C.M., Farley, A. C., Masakawa, A., Emler, C. A., &Dills, D. G. (1986) Interaction of insulin-like growth 1/Somatomedin with cultured rat chondrocytes: Receptor binding and internalization.Endocrinology 118, 1590–7.

    CAS  PubMed  Google Scholar 

  • Schalkwijk, J., Joosten, L. A. B., van den Berg, W. B., Wijk, J. J. &van de Putte, L. B. A. (1989a) Insulin-like growth factor stimulation of chondrocyte proteoglycan synthesis by human synovial fluid.Arthritis Rheum. 32, 66–71.

    CAS  PubMed  Google Scholar 

  • Schalkwijk, J., Joosten, L. A. B., Van den Berg, W. B. &Van de Putte, L. B. A. (1989b) Chondrocyte nonresponsiveness to insulin-like growth factor 1 in experimental arthritis.Arthritis Rheum. 32, 894–900.

    CAS  PubMed  Google Scholar 

  • Siczwkowski, M. &Watt, F. M. (1990) Subpopulations of chondrocytes from different zones of pig articular cartilage: Isolation, growth and proteoglycan synthesis in culture.J. Cell Sci. 97, 349–60.

    Google Scholar 

  • Stevenson, K. R., Gilmour, R. S. &Wathes, D. C. (1994). Localization and quantification of insulin-like growth factor-1 and-2 mRNA and type 1 receptors in the ovine uterus during the estrous cycle and early pregnancy.Endocrinol. 134, 1655–64.

    Article  CAS  Google Scholar 

  • Tesch, H. G., Handley, C. J., Cornell, H. J. &Herrington, A. C. (1992) Effects of free and bound insulin-like growth factors on proteoglycan metabolism in articular cartilage explants.J. Orthop. Res. Soc. 10, 14–22.

    CAS  Google Scholar 

  • Trippel, S. B., Van Wijk, J. J., Poster, M. B. &Svoboda, M. E. (1983) Characterization of specific somatomedim C receptors on isolated bovine growth plate chondrocytes.Endocrinol. 112, 2128–34.

    CAS  Google Scholar 

  • Trippel, S. B., Chernausek, S. D., van Wijk, J. J., Moses, A. C. &Mankin, H. J. (1988) Demonstration of type 1 and type 2 somatomedin receptors in bovine growth plate chondrocytes.J. Orthop. Res. 6, 817–26.

    Article  CAS  PubMed  Google Scholar 

  • Tyler, J. A. (1989) Insulin-like growth factor 1 can decrease degradation and promote synthesis of proteoglycan in cartilage exposed to cytokines.Biochem. J. 260, 543–8.

    CAS  PubMed  Google Scholar 

  • Ullrich, A., Gray, A., Tam, A. W., Yang-Feng, T., Tsubokawa, M., Collins, C., Henrel, W., Le Bon, T., Kathuria, S., Chen, E., Jacons, S., Francke, U., Ranchandran, J. &Fujita-Yamaguchi, Y. (1986) Insulin-like growth factor 1 receptor primary structure: Comparison with insulin receptor suggests structural determinants that define functional specificity.EMBO J. 5, 2503–12.

    CAS  PubMed  Google Scholar 

  • Van den Berg, W. B., Kruijsen, M. W. M., Van de Putte, L. B. A. &Zwarts, W. A. (1981) Antigen-induced arthritis and zymosan-induced arthritis in mice: Studies onin vivo cartilage proteoglycan synthesis and chondrocyte death.Br. J. Exp. Pathol. 62, 308–16.

    PubMed  Google Scholar 

  • Van den Berg, W. B., Kruijsen, M. W. M. &Van de Putte, L. B. A. (1982) The mouse patella assay: An easy method of quantitating articular cartilage chondrocyte functionin vivo andin vitro.Rheum. Int. 1, 165–9.

    Google Scholar 

  • Van den Berg, W. B., Joosten, L. A. B., Schalkwijk, J., van de Loo, F. A. J. &Van Beuningen, H. M. (1989) Mechanism of cartilage destruction in experimental arthritis: Lack of IGF-1 responsiveness. InTherapeutic Approaches to Inflammatory Diseases (edited byLewis, A. J., Doherty, N. S. andAckeman, N. R.), pp. 47–54. New York: Elsevier.

    Google Scholar 

  • Van Noorden, C. J. F. &Vogels, I. M. C. (1986) Enzyme histochemical reactions in unfixed and undecalcified cryostat sections of mouse knee joints with special reference to arthritic lesions.Histochemistry 86, 127–33.

    Article  PubMed  Google Scholar 

  • Verschure, P. J., Joosten, L. A. B., Van der Kraan, P. M. &Van den Berg, W. B. (1994a) Responsiveness of articular cartilage from normal and inflamed mouse knee joints to various growth factors.Ann. Rheum. Dis. 53, 455–60.

    CAS  PubMed  Google Scholar 

  • Verschure, P. J., Van der Kraan, P. M., Vitters, E. L. &Van den Berg, W. B. (1994b) Stimulation of proteoglycan synthesis by triamcinolone acetonide and insulin like growth factor in normal and arthritic murine articular cartilage.J. Rheumatol. 21, 920–5.

    CAS  PubMed  Google Scholar 

  • Verschure, P. J., Van Marle, J., Joosten, L. A. B. &Van den Berg, W. B. (1994c) Localization and quantification of the insulin-like growth factor-1 receptor in mouse articular cartilage by confocal laser scanning microscopy.J. Histochem. Cytochem. 42, 765–73.

    CAS  PubMed  Google Scholar 

  • Watanabe, N., Rosenfeld, R. G., Hintz, R. L., Hintz, R. L., Dollar, L. A., &Smith R. L. (1985) Characterization of a specific insulin-like growth factor-1/ Somatomedin-C receptor on high density primary monolayer cultures of bovine articular chondrocytes: regulation of receptor concentration by somatomedin, insulin and growth hormone.J. Endocrinol. 107, 275–83.

    CAS  PubMed  Google Scholar 

  • Wells, S. K., Sandison, D., Strickler, J. &Webb, W. W. (1989) Quantitative fluorescence imaging with laser scanning confocal microscopy. InThe Handbook of Confocal Microscopy (edited byPawley, S.), pp. 27–39. Madison: IMR Press.

    Google Scholar 

  • Wilson, T. (1989) The role of the pinhole in confocal imaging systems. InThe Handbook of Confocal Microscopy (edited byPawley, J.), pp. 113–26. Madison: IMR Press.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Verschure, P.J., Van Marle, J., Joosten, L.A.B. et al. Histochemical analysis of insulin-like growth factor-1 binding sites in mouse normal and expermmentally induced arthritic articular cartilage. Histochem J 28, 13–23 (1996). https://doi.org/10.1007/BF02331423

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02331423

Keywords

Navigation