Skip to main content
Log in

Staining of proteoglycans in mouse lung alveoli. II. Characterization of the Cuprolinic Blue-positive, anionic sites

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

Summary

The nature of Cuprolinic Blue-positive anionic filaments in mouse lung alveoli has been characterized. The contrast of filaments in the alveolar basement membrane of type I epithelial cells was lost on treatment with nitrous acid and pronase (without prefixation). In contrast, neither neuraminidase, chondroitinase ABC or AC, norStreptomyces hyaluronidase had any effect. Treatment with pronase (after prefixation) and 2.0m MgCl2 (after prefixation) also had no effect, indicating that the filaments are heparan sulphate proteoglycans. The filaments in the alveolar basement membrane of type II epithelial cells and in the capillary basement membrane of the endothelial cells were also nitrous acid sensitive, but chondroitinase ABC-insensitive. A model in which the whole alveolus contains a single layer of heparan sulphate-containing proteoglycan monomers is proposed. Furthermore, the collagen fibril associated filaments remained unaffected after treatment with nitrous acid, neuraminidase orStreptomyces hyaluronidase, or after digestion with pronase (after prefixation) and treatment with 2.0m MgCl2 (after prefixation). These filaments, however, could no longer be detected when digestion with chondroitinase ABC or pronase (without prefixation) was applied; chondroitinase AC treatment clearly affected the filaments, although they still were visible. These results indicate that the filaments are dermatan sulphate-containing proteoglycans. Some functional aspects of the proteoglycans are discussed.

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

  • Ausprunk, D. H., Boudreau, C. L. &Nelson, D. A. (1981) Proteoglycans in the microvasculature. I: Histochemical localization in microvessels of the rabbit eye.Am. J. Path. 103, 353–66.

    Google Scholar 

  • Cantor, J., Parshley, M. S., Darnule, A. T., Mandl, I. &Turino, G. M. (1977) Glycosaminoglycan synthesis by a clone of rat lung endothelial cells.J. Cell Biol. 75, 75a.

    Google Scholar 

  • Castor, C. W., Heiss, P. R., Gray, R. M. &Seidman, J. C. (1979) Connective tissue formation by lung fibroblasts in vitro.Am. Rev. Respir. Dis. 120, 107–19.

    Google Scholar 

  • Cheung, D. T. &Nimni, M. E. (1982), Mechanism of crosslinking of proteins by glutaraldhyde. I: Reaction with model compounds; II: Reaction with monomeric and polymeric collagen.Conn. Tiss. Res. 10, 187–216.

    Google Scholar 

  • Clowes, A. W., Clowes, M. M., Gown, A. M. &Wight, T. N. (1982) Distribution of a proteoheparan sulfate in rat aorta.Fedn. Proc. 41, 441.

    Google Scholar 

  • Danielsen, C. C. (1982) Mechanical properties of reconstituted collagen fibrils. Influence of a glycosaminoglycan: dermatan sulfate.Conn. Tiss. Res. 9, 219–25.

    Google Scholar 

  • Ehrlich, K. C. (1981) Proteoglycans synthesis by rat lung cells culturedin vitro.J. biol. Chem. 256, 73–80.

    Google Scholar 

  • Eyre, D. R. &Muir, H. (1975) The distribution of different molecular species of collagen in fibrous, elastic and hyaline cartilages of the pig.Biochem. J. 151, 595–602.

    Google Scholar 

  • Farin, F., Killen, P. &Striker, G. (1980) Biosynthesis of heparan sulfate (glycosaminoglycans) by glomerular cellsin vitro.Fedn. Proc. 39, 873.

    Google Scholar 

  • Farquhar, M. G. &Kanwar, Y. S. (1982) Functional organization of the glomerulus. InImmune Mechanism in Renal Disease (edited byCummings, N. B., Michael, A. F. andWilson, C. B.) New York: Plenum Press.

    Google Scholar 

  • Geyer, G. (1973)Ultrahistochemie, 2nd edn., pp. 13–22. Stuttgart: Gustav Fischer Verlag.

    Google Scholar 

  • Greenwald, R. A., Schwartz, C. E. &Cantor, J. O. (1975) Interaction of cartilage proteoglycans with collagen-substituted agarose gels.Biochem. J. 145, 601–05.

    Google Scholar 

  • Hassell, J. R., Robey, P. G., Barrach, H.-J., Wilczek, J., Rennard, S. I. &Martin, G. R. (1980) Isolation of a heparan sulfate-containing proteoglycan from basement membrane.Proc. Natn. Acad. Sci. U.S.A. 77, 4494–8.

    Google Scholar 

  • Horwitz, A. L. &Crystal, R. G. (1975) Content and synthesis of glycosaminoglycans in the developing lung.J. clin. Invest. 56, 1312–8.

    Google Scholar 

  • Horwitz, A. L., Elson, N. A. &Crystal, R. G. (1976) Proteoglycans and elastic fibres. InThe Biochemical Basis of Pulmonary Function; Lung Biology in Health and Disease (edited byCrystal, R. G.), Vol. 2, pp. 273–311. New York, Basel, Marcel Dekker, Inc.

    Google Scholar 

  • Junqueira, L. C. U., Toledo, O. M. S. &Montes, G. S. (1981) Correlation of specific sulfated glycosaminoglycans with collagen types I, II and III.Cell Tiss. Res. 217, 171–5.

    Google Scholar 

  • Kanwar, Y. S. &Farquhar, M. G. (1979) Presence of heparan sulfate in the glomerular basement membrane.Proc. Natn. Acad. Sci. U.S.A. 76, 1303–7.

    Google Scholar 

  • Kanwar, Y. S. &Farquhar, M. G. (1980) Role of glycosaminoglycans (GAG) in the permeability of the glomerular basement membrane. (GBM).Fedn. Proc. 39, 334.

    Google Scholar 

  • Kanwar, Y. S., Linker, A. &Farquhar, M. G. (1980) Increased permeability of the glomerular basement membrane to ferritin after removal of glycosaminoglycans (heparan sulfate) by enzyme digestion.J. Cell Biol. 86, 688–93.

    Google Scholar 

  • Kanwar, Y. S., Hascall, V. C. &Farquhar, M. G. (1981) Partial characterization of newly synthesized proteoglycans isolated from the glomerular basement membrane.J. Cell Biol. 90, 527–32.

    Google Scholar 

  • Kawamoto, T. &Nagai, Y. (1976) Developmental changes in glycosaminoglycans, collagen and collagenase activity in embryonic chick skin.Biochim. biophys. Acta 437, 190–9.

    Google Scholar 

  • Kosher, R. A. &Searls, R. L. (1973) Sulfated mucopolysaccharide synthesis during the development ofRana pipiens.Devl Biol. 32, 50–68.

    Google Scholar 

  • Lindahl, U. &Höök, M. (1978) Glycosaminoglycans and their binding to biological macromolecules.Ann. Rev. Biochem. 47, 385–417.

    Google Scholar 

  • Linker, A. &Hovingh, P. (1972) Heparinase and heparitinase from flavobacteria.Meth. Enzym. 28, 902–11.

    Google Scholar 

  • Maeda, H., Ishikawa, H. &Ohta, S. (1981) Circumscribed myxoedema of lichen myxoedematosus as a sign of faulty formation of the proteoglycan macromolecule.Br. J. Derm. 105, 239–45.

    Google Scholar 

  • Mathews, M. B. (1965) The interaction of collagen and acid mucopolysaccharides. A model for connective tissue.Biochem. J. 96, 710–6.

    Google Scholar 

  • Matthay, M. A., Landolt, C. C. &Staub, N. C. (1982) Differential liquid and protein clearance from the alveoli of anesthetized sheep.J. appl. Physiol. 53, 96–104.

    Google Scholar 

  • Nakamura, N., Kojima, J., Okamoto, S. &Kitamura, Y. (1981) Absence of heparin in glycosaminoglycan fractions isolated from the skin of genetically mast cell-depletedW/W V mice.Biochem. Internat. 3, 449–56.

    Google Scholar 

  • Öbrink, B. (1973a) A study of the interactions between monomeric tropocollagen and glycosaminoglycans.Eur. J. Biochem. 33, 387–400.

    Google Scholar 

  • Öbrink, B. (1973b) The influence of glycosaminoglycans on the formation of fibres from monomeric tropocollagenin vitro.Eur. J. Biochem. 34, 129–37.

    Google Scholar 

  • Olver, R. E., Ramsden, C. A. &Strong, L. B. (1981) Adrenaline-induced changes in net lung liquid volume flow across the pulmonary epithelium of the fetal lamb: evidence for active sodium transport.J. Physiol. 319, 38P-39P.

    Google Scholar 

  • Oohira, A., Wight, T. N., McPherson, J. &Bornstein, P. (1982) Biochemical and ultrastructural studies of proteoheparan sulfates synthesized by PYS-2, a basement membrane-producing cell line.J. Cell Biol. 92, 357–67.

    Google Scholar 

  • Parthasarathy, N. &Spiro, R. G. (1982) Basement membrane glycosaminoglycans: examination of several membranes and evaluation of the effect of sonic treatment.Archs Biochem. Biophys. 213, 504–11.

    Google Scholar 

  • Radhakrishnamurthy, B., Smart, F., Dalferes, E. R. &Berenson, G. S. (1980) Isolation and characterization of proteoglycans from bovine lung.J. biol. Chem. 255, 7575–82.

    Google Scholar 

  • Sampson, P. M., Jimenez, S. A. &Bashey, R. I. (1979) Isolation and partial characterization of proteoglycans from sheep lung parenchyma.Biochim. biophys. Acta 588, 129–41.

    Google Scholar 

  • Schmid, K., Grundboeck-Jusco, J., Kimura, A., Tschopp, F. A., Zollinger, R., Binette, J. P., Lewis, W. &Hayashi, S. (1982) The distribution of the glycosaminoglycans in the anatomic components of the lung and the changes in concentration of these macromolecules during development and aging.Biochim. biophys. Acta 716, 178–87.

    Google Scholar 

  • Scott, J. E. (1972) Histochemistry of Alcian Blue. III: The molecular biological basis of staining by Alcian Blue 8GX and analogous phthalocyanins.Histochemie 32, 191–212.

    Google Scholar 

  • Scott, J. E., Orford, C. R. &Huges, E. W. (1981) Proteoglycan-collagen arrangements in developing rat tail tendon. An electron-microscopical and biochemical investigation.Biochem. J. 195, 573–81.

    Google Scholar 

  • Snowden, J. McK. &Swann, D. A. (1980) Effects of glycosaminoglycans and proteoglycans on thein vitro assembly and thermal stability of collagen fibrils.Biopolymers 19, 767–80.

    Google Scholar 

  • Stefanovich, V. &Gore, I. (1967) A micromethod for the determination of acid mucopolysaccharides in vascular tissue.J. Chromat. 31, 473–8.

    Google Scholar 

  • Straus, A. H., Nader, H. B. &Dietrich, C. P. (1982) Absence of heparin-like compounds in mast-cell-free tissue and animals.Biochim. biophys. Acta 717, 478–85.

    Google Scholar 

  • Toole, B. P. &Lowther, D. A. (1968) Dermatan sulfate-protein: Isolation from and interaction with collagen.Archs Biochem. Biophys. 128, 567–78.

    Google Scholar 

  • Vaccaro, C. A. &Brody, J. S. (1979) Ultrastructural localization of proteoglycans in the pulmonary alveolus.Am. Rev. Respir. Dis. 120, 901–10.

    Google Scholar 

  • Vaccaro, C. A. &Brody, J. S. (1981) Structural features of alveolar wall basement membrane in the adult rat lung.J. Cell Biol. 91, 427–37.

    Google Scholar 

  • Vogel, K. G. &Peterson, D. W. (1981) Extracellular, surface, and intracellular proteoglycans produced by human embryo lung fibroblast in culture. (IMR-90).J. biol. Chem. 256, 13235–42.

    Google Scholar 

  • Wusteman, F. S. (1972) Glycosaminoglycans of bovine lung parenchyma and pleura.Experientia 28, 887–8.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Van Kuppevelt, T.H.M.S.M., Cremers, F.P.M., Domen, J.G.W. et al. Staining of proteoglycans in mouse lung alveoli. II. Characterization of the Cuprolinic Blue-positive, anionic sites. Histochem J 16, 671–686 (1984). https://doi.org/10.1007/BF01003394

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01003394

Keywords

Navigation