Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1980 Aug 1;86(2):688–693. doi: 10.1083/jcb.86.2.688

Increased permeability of the glomerular basement membrane to ferritin after removal of glycosaminoglycans (heparan sulfate) by enzyme digestion

PMCID: PMC2111494  PMID: 6447156

Abstract

Glomerular basement membranes (GBM's) were subjected to digestion in situ with glycosaminoglycan-degrading enzymes to assess the effect of removing glycosaminoglycans (GAG) on the permeability of the GBM to native ferritin (NF). Kidneys were digested by perfusion with enzyme solutions followed by perfusion with NF. In controls treated with buffer alone, NF was seen in high concentration in the capillary lumina, but the tracer did not penetrate to any extent beyond the lamina rara interna (LRI) of the GBM, and litte or no NF reached the urinary spaces. Findings in kidneys perfused with Streptomyces hyaluronidase (removes hyaluronic acid) and chondroitinase-ABC (removes hyaluronic acid, chondroitin 4- and 6-sulfates, and dermatan sulfate, but not heparan sulfate) were the same as in controls. In kidneys digested with heparinase (which removes most GAG including heparan sulfate), NF penetrated the GBM in large amounts and reached the urinary spaces. Increased numbers of tracer molecules were found in the lamina densa (LD) and lamina rara externa (LRE) of the GBM. In control kidneys perfused with cationized ferritin (CF), CF bound to heparan- sulfate rich sites demonstrated previously in the laminae rarae; however, no CF binding was seen in heparinase-digested GBM's, confirming that the sites had been removed by the enzyme treatment. The results demonstrated that removal of heparan sulfate (but not other GAG) leads to a dramatic increase in the permeability of the GBM to NF.

Full Text

The Full Text of this article is available as a PDF (847.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brenner B. M., Hostetter T. H., Humes H. D. Molecular basis of proteinuria of glomerular origin. N Engl J Med. 1978 Apr 13;298(15):826–833. doi: 10.1056/NEJM197804132981507. [DOI] [PubMed] [Google Scholar]
  2. Caulfield J. P., Farquhar M. G. The permeability of glomerular capillaries to graded dextrans. Identification of the basement membrane as the primary filtration barrier. J Cell Biol. 1974 Dec;63(3):883–903. doi: 10.1083/jcb.63.3.883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. FARQUHAR M. G., WISSIG S. L., PALADE G. E. Glomerular permeability. I. Ferritin transfer across the normal glomerular capillary wall. J Exp Med. 1961 Jan 1;113:47–66. doi: 10.1084/jem.113.1.47. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Farquhar M. G. Editorial: The primary glomerular filtration barrier--basement membrane or epithelial slits? Kidney Int. 1975 Oct;8(4):197–211. doi: 10.1038/ki.1975.103. [DOI] [PubMed] [Google Scholar]
  5. Kanwar Y. S., Farquhar M. G. Anionic sites in the glomerular basement membrane. In vivo and in vitro localization to the laminae rarae by cationic probes. J Cell Biol. 1979 Apr;81(1):137–153. doi: 10.1083/jcb.81.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kanwar Y. S., Farquhar M. G. Isolation of glycosaminoglycans (heparan sulfate) from glomerular basement membranes. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4493–4497. doi: 10.1073/pnas.76.9.4493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kanwar Y. S., Farquhar M. G. Presence of heparan sulfate in the glomerular basement membrane. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1303–1307. doi: 10.1073/pnas.76.3.1303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kelley V. E., Cavallo T. Glomerular permeability: transfer of native ferritin in glomeruli with decreased anionic sites. Lab Invest. 1978 Dec;39(6):547–553. [PubMed] [Google Scholar]
  9. Laurent T. C. In vitro studies on the transport of macromolecules through the connective tissue. Fed Proc. 1966 May-Jun;25(3):1128–1134. [PubMed] [Google Scholar]
  10. Michael A. F., Blau E., Vernier R. L. Glomerular polyanion. Alteration in aminonucleoside nephrosis. Lab Invest. 1970 Dec;23(6):649–657. [PubMed] [Google Scholar]
  11. Mohos S. C., Skoza L. Glomerular sialoprotein. Science. 1969 Jun 27;164(3887):1519–1521. doi: 10.1126/science.164.3887.1519. [DOI] [PubMed] [Google Scholar]
  12. Reeves W. H., Kanwar Y. S., Farquhar M. G. Assembly of the glomerular filtration surface. Differentiation of anionic sites in glomerular capillaries of newborn rat kidney. J Cell Biol. 1980 Jun;85(3):735–753. doi: 10.1083/jcb.85.3.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Rennke H. G., Cotran R. S., Venkatachalam M. A. Role of molecular charge in glomerular permeability. Tracer studies with cationized ferritins. J Cell Biol. 1975 Dec;67(3):638–646. doi: 10.1083/jcb.67.3.638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ryan G. B., Hein S. J., Karnovsky M. J. Glomerular permeability to proteins. Effects of hemodynamic factors on the distribution of endogenous immunoglobulin G and exogenous catalase in the rat glomerulus. Lab Invest. 1976 Apr;34(4):415–427. [PubMed] [Google Scholar]
  15. Ryan G. B., Karnovsky M. J. Distribution of endogenous albumin in the rat glomerulus: role of hemodynamic factors in glomerular barrier function. Kidney Int. 1976 Jan;9(1):36–45. doi: 10.1038/ki.1976.5. [DOI] [PubMed] [Google Scholar]
  16. Seiler M. W., Venkatachalam M. A., Cotran R. S. Glomerular epithelium: structural alterations induced by polycations. Science. 1975 Aug 1;189(4200):390–393. doi: 10.1126/science.1145209. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES