Binding of wheat germ agglutinin in the matrix of rat tracheal cartilage
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The binding of wheat germ agglutinin (WGA) to the extracellular matrix of rat tracheal cartilage was studied at both the light and electron microscopic levels. The detection of binding sites was accomplished by a postembedment method using the direct fluorescence technique for light microscopy and the avidin—biotin bridging system for electron microscopy. Distinct fluorescence was observed in the pericellular region of chondrocytes, and this fluorescence was completely removed after treatment with 4 M guanidine hydrochloride. By electron microscopy, the reaction products as found in the pericellular region were not observed in the interterritorial collagenous matrix, confirming a similar distribution as found by fluorescence microscopy. These results show that WGA-binding sites are present in pericellular matrical substances, which are known to be rich in proteoglycan and glycosaminoglycan complexes and which exhibit similar staining with antibodies to proteoglycans or with cationic dyes. As WGA binds toN-acetyl-glucosamine andN-acetyl-neuraminic acid residues, the pericellular matrix of rat tracheal cartilage appears to consist of proteoglycan having a high concentration of these saccharides.
KeywordsHydrochloride Fluorescence Microscopy Biotin Guanidine Glycosaminoglycan
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- FARNUM, C. E. & WILSMAN, N. J. (1984) Lectin-binding histochemistry of non-decalcified growth plate cartilage.J. Histochem. Cytochem. 32, 593–607.Google Scholar
- HSU, S. M., RAINE, L. & FANGER, H. (1981) Use of avidin—biotin—peroxidase complex (ABC) in immunoperoxidase techniques.J. Histochem. Cytochem. 29, 577–80.Google Scholar
- HUNZIKER, E. B., HERRMANN, W. & SHENK, R. K. (1982) Improved cartilage fixation by ruthenium hexammine trichloride (RHT).J. Ultrastruct. Res. 81, 1–12.Google Scholar
- KIMATA, K., OIKE, Y., ITO, K., KARASAWA, K. & SUZUKI, S., (1978) The occurrence of low buoyant density proteoglycans in embryonic chick cartilage.Biochem. Biophys. Res. Commun. 85, 1431–9.Google Scholar
- POOLE, A. R., PIDOUX, A., REINER, A., TANG, L. H., CHOL, H. & ROSENBERG, L. (1980) Localization of proteoglycan monomer and link protein in the matrix of bovine articular cartilage.J. Histochem. Cytochem. 28, 621–35.Google Scholar
- SHEPARD, N. & MITCHELL, N. (1976) The localization of proteoglycan by light and electron microscopy using Safranin O.J. Ultrastruct. Res. 54, 451–60.Google Scholar
- YAMADA, K., FUJITA, Y. & SHIMIZU, S. (1982) The effect of digestion with keratanase (Pseudomonas sp.) on certain histochemical reactions for glycosaminoglycans in cartilaginous and corneal tissues.Histochem. J. 14, 897–910.Google Scholar