The Histochemical Journal

, Volume 25, Issue 5, pp 401–405 | Cite as

Cationic gold staining of glomerular anionic sites in archived tissue, reprocessed from paraffin wax into LR gold resin

  • N. P. Goode
  • M. Shires
  • S. R. Aparicio
  • A. M. Davison


Glomerular capillary wall anionic sites have been demonstrated by cationic gold staining of archived renal biopsy tissue (up to 10 years old), obtained from six patients, originally embedded in paraffin wax, and subsequently reprocessed into LR gold resin. The staining patterns at pH 2.5 and pH 7.0, demonstrating different glomerular basement membrane (GBM) anionic constituents, were compared in three patients from whom tissue directly processed into LR gold and reprocessed tissue was available. Ultrastructural preservation was poorer and shrinkage artefact greater in paraformaldehyde-lysine periodate (PLP) as opposed to formol saline-fixed reprocessed tissue. However, GBM anionic site expression was well preserved, or even enhanced (lamina rara externa, pH 7.0) in reprocessed tissue, using either fixative. Although it may not be possible to compare subtle changes in anionic site distribution in variously fixed and processed tissues, due to these artefacts, the technique enables retrospective study of charge status in archived material from disease groups in which there are distinct anionic site aberrations.


Renal Biopsy Periodate Glomerular Basement Membrane Capillary Wall Anionic Site 
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  1. Barnes, J. L., Radnik, R. A., Gilchrist, E. P. & Venkatachalam, M. A. (1984) Size and charge selectivity defects induced in glomerular basement membrane by a polycation. Kidney Int. 25, 11–19.Google Scholar
  2. Bendayan, M., Benhamou, N. & Desjardins, M. (1990) Ultrastructural distribution of lectin-binding sites in the glomerular wall of streptozotocin-induced diabetic rats. J. Submicrosc. Cytol. Pathol. 22, 173–84.Google Scholar
  3. Bertolatus, J. A. & Hunsicker, L. G. (1987) Polycation binding to glomerular basement membrane: effect of biochemical modification. Lab. Invest. 56, 170–9.Google Scholar
  4. Brenner, B. M., Hostetter, T. H. & Humes, H. D. (1978) Molecular basis of proteinuria of glomerular origin. New Eng. J. Med. 15, 826–33.Google Scholar
  5. Chen, K. & Wight, T. N. (1984) Proteoglycans in arterial smooth muscle cell cultures: an ultrastructural histochemical analysis. J. Histochem. Cytochem. 32, 347–57.Google Scholar
  6. Fox, C. H., Johnson, F. B., Whiting, J. & Roller, P. P. (1985) Formaldehyde fixation. J. Histochem. Cytochem. 33, 845–53.Google Scholar
  7. Goode, N. P., Shires, M., Crellin, D. M. & Davison, A. M. (1991a) Detection of glomerular anionic sites in post-embedded ultrathin sections using cationic colloidal gold. J. Histochem. Cytochem. 39, 965–72.Google Scholar
  8. Goode, N. P., Shires, M., Aparicio, S. R. & Davison, A. M. (1991b) Cationic colloidal gold - a novel marker for the demonstration of glomerular polyanion status in routine renal biopsies. Nephrol. Dial. Transplant. 6, 923–30.Google Scholar
  9. Goode, N. P., Shires, M., Dyson, E. H. & Davison, A. M. (1991c) Glomerular charge detection in rat and human using cationic colloidal gold. J. Pathol. 163, 166A.Google Scholar
  10. Goode, N. P., Shires, M. & Davison, A. M. (1992) Preparation and use of the poly-l-lysine-gold probe: a differential marker of glomerular anionic sites. Histochemistry 98, 67–72.Google Scholar
  11. Hunsicker, L. G., Shearer, T. P. & Shaffer, S. J. (1981) Acute reversible proteinuria induced by infusion of the polycation hexadimethrine. Kidney Int. 20, 7–17.Google Scholar
  12. Kanwar, Y. S., Linker, A. & Farquhar, M. G. (1980) Increased permeability of the glomerular basement membrane to ferritin after removal of glycosaminoglycans (heparan sulphate) by enzyme digestion. J. Cell Biol. 86, 688–93.Google Scholar
  13. Lelongt, B., Marino, H. & Kanwar, Y. S. (1987) Status of glomerula proteoglycans in aminonucleoside nephrosis. Kidney Int. 31, 1299–310.Google Scholar
  14. Londono, I. & Bendayan, M. (1988) High-resolution cytochemistry of neuraminic and hexuronic acid-containing macromolecules applying the enzyme-gold approach. J. Histochem. Cytochem. 36, 1005–14.Google Scholar
  15. Londono, L., Coulombe, P. A. & Bendayan, M. (1989) Brief review of progresses in enzyme gold cytochemistry. Scan. Microsc. (suppl) 3, 7–14.Google Scholar
  16. Mcclean, I. W. & Nakane, P. K. (1974) Periodate-lysineparaformaldehyde fixative: a new fixative for immunoelectron microscopy. J. Histochem. Cytochem. 22, 1077–83.Google Scholar
  17. Moss, J., Woodrow, D. F., Shore, I., Gower, P., Philips, M. & Spiro, R. G. (1990) Ultrastructural immunogold studies of heparan sulphate proteoglycan in normal human glomeruli and glomerulonephritis. J. Pathol. 161, 137–43.Google Scholar
  18. Pilia, P. A., Swain, R. P., Williams, A. V., Loadholt, C. B. & Ainsworth, S. K. (1985) Glomerular anionic site distribution in nonproteinuric rats: a computer assisted morphometric analysis. Am. J. Pathol. 121, 474–85.Google Scholar
  19. Rosenzweig, L. J. & Kanwar, Y. S. (1982) Removal of sulfated (heparan sulphate) or nonsulfated (hyaluronic acid) glycosaminoglycans results in increased permeability of the glomerular basement membrane to 125I-bovine albumin. Lab. Invest. 47, 177–84.Google Scholar
  20. Shires, M., Goode, N. P., Crellin, D. M. & Davison, A. M. (1990) Immunogold-silver staining of mesangial antigen in Lowicryl K4M- and LR gold-embedded renal tissue using epipolarization microscopy. J. Histochem. Cytochem. 38, 287–9.Google Scholar
  21. Spicer, S. S. & Schulte, B. A. (1992) Diversity of cell glycoconjugates, shown histochemically: a perspective (review). J. Histochem. Cytochem. 40, 1–38.Google Scholar
  22. Vehaskari, M., Root, E. R., Germuth, F. G. & Robson, A. M. (1982) Glomerular charge and urinary protein excretion: effects of systemic and intrarenal polycation infusion in the rat. Kidney Int. 22, 127–35.Google Scholar
  23. Vorbrodt, A. W. (1989) Ultracytochemical characterization of anionic sites in the wall of brain capillaries. J. Neurocytol. 18, 359–68.Google Scholar

Copyright information

© Chapman & Hall 1993

Authors and Affiliations

  • N. P. Goode
    • 1
  • M. Shires
    • 1
  • S. R. Aparicio
    • 2
  • A. M. Davison
    • 3
  1. 1.Renal Research UnitSt James's University NHS TrustLeedsUK
  2. 2.Department of PathologySt James's University NHS TrustLeedsUK
  3. 3.Department of Renal MedicineSt James's University NHS TrustLeedsUK

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