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Immunohistochemical localization of articular cartilage proteoglycan and link protein in situ using monoclonal antibodies and lectin-binding methods

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Abstract

Lectins have specificity for certain carbohydrate structures in macromolecules. Lectins are, therefore, useful histochemical tools for demonstrating the composition and localization of components of connective tissue matrices, such as articular cartilage. In order to assess the significance of observed lectin-binding patterns, experiments were performed in which monoclonal antibodies against chondroitin sulphate- and keratan sulphate-containing proteolgycans and link proteins were applied to sections of bovine articular cartilage after enzymatic digestion with chondroitinase ABC and keratanase. The following conclusions were made: (1) Binding of peanut agglutinin (PNA) in the interterritorial matrix predominantly indicates the presence of keratan sulphate, but may also detectO-linked oligosaccharides of proteoglycans. (2) In normal cartilage wheat germ agglutinin (WGA) binds nearly exclusively to keratan sulphate. In cartilage degraded with chondroitinase ABC and keratanase this lectin may also detect carbohydrates in link protein due to enhanced accessibility. Binding of WGA toO-linked oligosaccharides may eventually occur. (3) In enzymatically digested cartilage matrix, staining with soybean agglutinin (SBA) may be due to link protein, but not to chondroitin sulphate, because specific breakdown of the glycosaminoglycan chain is required for binding of SBA. (4)Ulex europaeus agglutinin I (UEA I) binding sites are only detectable in digested cartilage matrix.

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Authors and Affiliations

  1. Institute of Pharmacology and Toxicology, University of Bonn, Reuterstrasse 2, W-5300, Bonn 1, Germany

    S. Hoedt-Schmidt & D. A. Kalbhen

  2. Department of Pathological Sciences, University of Manchester, The Medical School, Stopford Building, M13 9PT, Manchester, UK

    J. McClure

  3. Pharmaceutical Division, Ciba-Geigy Corporation, 07901, Summit, NJ, USA

    M. K. Jasani

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  1. S. Hoedt-Schmidt
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  2. J. McClure
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  3. M. K. Jasani
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  4. D. A. Kalbhen
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Hoedt-Schmidt, S., McClure, J., Jasani, M.K. et al. Immunohistochemical localization of articular cartilage proteoglycan and link protein in situ using monoclonal antibodies and lectin-binding methods. Histochemistry 99, 391–403 (1993). https://doi.org/10.1007/BF00717052

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