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. 1970 Nov 30;132(6):1153–1167. doi: 10.1084/jem.132.6.1153

AN ULTRASTRUCTURAL STUDY OF GLOMERULAR PERMEABILITY USING CATALASE AND PEROXIDASE AS TRACER PROTEINS

M A Venkatachalam 1, M J Karnovsky 1, H D Fahimi 1, R S Cotran 1
PMCID: PMC2180510  PMID: 5511568

Abstract

Mice were injected intravenously with beef liver catalase (mol wt 240,000) and very small doses of horseradish peroxidase (mol wt 40,000) and the site of localization of these enzymes in the kidney was studied by ultrastructural cytochemistry. 1 min after injection, catalase was present in glomerular capillary lumina and there was minimal permeation of the basement membrane. After 5–180 min, staining of the basement membrane increased progressively but was usually less than that in capillary lumina. At all time intervals the inner (sub-endothelial) layer of the basement membrane contained more reaction product than the lamina densa and the outer (subepithelial) layer. Catalase permeated the entire thickness of the basement membrane and extended up to the slit pore but not beyond the level of the slit diaphragm and was not seen in the urinary space or tubular lumina. Horseradish peroxidase permeated the whole thickness of the basement membrane within 2 min after injection; however, gradients of staining from the inner to outer layers of the basement membrane were frequently seen. The findings with both enzymes indicate that (a) the basement membrane restricts the passage of proteins over a wide range of molecular size with increasing impediment for larger molecules and (b) the slit pore functions as an additional barrier for molecules that cross the basement membrane.

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Selected References

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

  1. ARTURSON G., WALLENIUS G. THE RENAL CLEARANCE OF DEXTRAN OF DIFFERENT MOLECULAR SIZES IN NORMAL HUMANS. Scand J Clin Lab Invest. 1964;16:81–86. doi: 10.3109/00365516409060486. [DOI] [PubMed] [Google Scholar]
  2. Beard M. E., Novikoff A. B. Distribution of peroxisomes (microbodies) in the nephron of the rat: a cytochemical study. J Cell Biol. 1969 Aug;42(2):501–518. doi: 10.1083/jcb.42.2.501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bunn H. F., Esham W. T., Bull R. W. The renal handling of hemoglobin. I. Glomerular filtration. J Exp Med. 1969 May 1;129(5):909–923. doi: 10.1084/jem.129.5.909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DEODHAR S. D., CUPPAGE F. E., GABLEMAN E. STUDIES ON THE MECHANISM OF EXPERIMENTAL PROTEINURIA INDUCED BY RENIN. J Exp Med. 1964 Oct 1;120:677–690. doi: 10.1084/jem.120.4.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DIRKS J. H., CLAPP J. R., BERLINER R. W. THE PROTEIN CONCENTRATION IN THE PROXIMAL TUBULE OF THE DOG. J Clin Invest. 1964 May;43:916–921. doi: 10.1172/JCI104977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. FISHER E. R., HELLSTROM H. R. Mechanism of proteinuria: Functional and ultrastructural correlation of effects of infusion of homologous and heterologous protein (bovine serum albumin) in the rat. Lab Invest. 1962 Aug;11:617–637. [PubMed] [Google Scholar]
  8. Graham R. C., Jr, Karnovsky M. J. Glomerular permeability. Ultrastructural cytochemical studies using peroxidases as protein tracers. J Exp Med. 1966 Dec 1;124(6):1123–1134. doi: 10.1084/jem.124.6.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Graham R. C., Jr, Karnovsky M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. doi: 10.1177/14.4.291. [DOI] [PubMed] [Google Scholar]
  10. Groniowski J., Biczyskowa W., Walski M. Electron microscope studies on the surface coat of the nephron. J Cell Biol. 1969 Mar;40(3):585–601. doi: 10.1083/jcb.40.3.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. HALL V. The protoplasmic basis of glomerular ultrafiltration. Am Heart J. 1957 Jul;54(1):1–9. doi: 10.1016/0002-8703(57)90073-x. [DOI] [PubMed] [Google Scholar]
  12. Jones D. B. Mucosubstances of the glomerulus. Lab Invest. 1969 Aug;21(2):119–125. [PubMed] [Google Scholar]
  13. Jorgensen F. Electron microscopic studies of normal glomerular basement membrane. Lab Invest. 1967 Oct;17(4):416–424. [PubMed] [Google Scholar]
  14. LAMBERT P. P., GREGOIRE F. Hémodynamique glomérulaire et excrétion de l'hémoglobine. Arch Int Physiol Biochim. 1955 Feb;63(1):7–34. doi: 10.3109/13813455509150857. [DOI] [PubMed] [Google Scholar]
  15. MEYER F., PUTNAM F. W. The fate of injected Bence Jones protein. J Exp Med. 1963 Apr 1;117:573–581. doi: 10.1084/jem.117.4.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. PAPPENHEIMER J. R. Uber die Permeabililität der Glomerulummembranen in der Niere. Klin Wochenschr. 1955 Apr 15;33(15-16):362–365. doi: 10.1007/BF01467967. [DOI] [PubMed] [Google Scholar]
  18. Rambourg A., Leblond C. P. Electron microscope observations on the carbohydrate-rich cell coat present at the surface of cells in the rat. J Cell Biol. 1967 Jan;32(1):27–53. doi: 10.1083/jcb.32.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. SAMEJIMA T., YANG J. T. RECONSTITUTION OF ACID-DENATURED CATALASE. J Biol Chem. 1963 Oct;238:3256–3261. [PubMed] [Google Scholar]
  20. SPECTOR W. G. The reabsorption of labelled proteins by the normal and nephrotic rat kidney. J Pathol Bacteriol. 1954 Jul;68(1):187–196. doi: 10.1002/path.1700680123. [DOI] [PubMed] [Google Scholar]
  21. Seligman A. M., Karnovsky M. J., Wasserkrug H. L., Hanker J. S. Nondroplet ultrastructural demonstration of cytochrome oxidase activity with a polymerizing osmiophilic reagent, diaminobenzidine (DAB). J Cell Biol. 1968 Jul;38(1):1–14. doi: 10.1083/jcb.38.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Venkatachalam M. A., Fahimi H. D. The use of beef liver catalase as a protein tracer for electron microscopy. J Cell Biol. 1969 Aug;42(2):480–489. doi: 10.1083/jcb.42.2.480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. YAMADA E. The fine structure of the renal glomerulus of the mouse. J Biophys Biochem Cytol. 1955 Nov 25;1(6):551–566. doi: 10.1083/jcb.1.6.551. [DOI] [PMC free article] [PubMed] [Google Scholar]

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