Abstract
Changes in glomerular anionic sites in puromycin aminonucleoside nephrosis (PAN) in the rat are controversial. The authors examined glomerular anionic sites in PAN by in vivo staining with polyethyleneimine (PEI). They also quantitated and characterized glomerular heparan sulfate (HS), which is known to be a major glomerular polyanion in PAN, using in vivo incorporation of 35S-sulfate. PAN rats had a mean protein excretion of 96 +/- 23 mg per 24 hours. Staining of anionic sites with PEI showed 15.3 +/- 2.8 sites per 1000-nm length of glomerular basement membrane in controls, 13.7 +/- 1.9 sites in PAN rats (P greater than 0.05), and 50% of rats with early PAN had absent staining. Total 35S-sulfate incorporation was similar in both the controls and established PAN rats (2900 +/- 150 dpm/mg dry wt of glomeruli versus 3005 +/- 260, P greater than 0.05) but decreased in early PAN rats (2025 +/- 148). The percentage of 35S-sulfate incorporated into chondroitin sulfate was similar in all three groups of animals. HS uronic acid was also similar (1.8 +/- 0.2 g/mg dry wt of glomeruli versus 1.7 +/- 0.3, P greater than 0.05) but decreased in early PAN (1.1 +/- 0.2). The distribution of 35S-sulfate activity within the HS subfractions was examined by ion-exchange chromatography and showed a shift in percent present from 1.0 M to 1.25 M fraction in established and early PAN animals (control 1.0 M 37% +/- 3.2% versus PAN 19% +/- 3.4%, P less than 0.01, and 1.25 M 36% +/- 2.9% versus 53% +/- 2.9%, P less than 0.01). These results demonstrate that glomerular heparan sulfate is unchanged in established PAN but decreased in early PAN. SO4 incorporation is unchanged in established PAN and diminished in early PAN. Thus, early in PAN HS synthesis is impaired, but in established PAN the HS is normal, and changes in glomerular HS cannot explain the increased permeability.
Full text
PDF






Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Blumenkrantz N., Asboe-Hansen G. New method for quantitative determination of uronic acids. Anal Biochem. 1973 Aug;54(2):484–489. doi: 10.1016/0003-2697(73)90377-1. [DOI] [PubMed] [Google Scholar]
- 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]
- Caulfield J. P., Farquhar M. G. Loss of anionic sites from the glomerular basement membrane in aminonucleoside nephrosis. Lab Invest. 1978 Nov;39(5):505–512. [PubMed] [Google Scholar]
- FRENK S., ANTONOWICZ I., CRAIG J. M., METCOFF J. Experimental nephrotic syndrome induced in rats by aminonucleoside; renal lesions and body electrolyte composition. Proc Soc Exp Biol Med. 1955 Jul;89(3):424–427. doi: 10.3181/00379727-89-21833. [DOI] [PubMed] [Google Scholar]
- Fishman J. A., Karnovsky M. J. Effects of the aminonucleoside of puromycin on glomerular epithelial cells in vitro. Am J Pathol. 1985 Mar;118(3):398–407. [PMC free article] [PubMed] [Google Scholar]
- Foidart J. B., Pirard Y. S., Winand R. J., Mahieu P. R. Tissue culture of normal rat glomeruli: glycosaminoglycan biosynthesis by homogeneous epithelial and mesangial cell populations. Ren Physiol. 1980;3(1-6):169–173. [PubMed] [Google Scholar]
- 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]
- Kanwar Y. S., Hascall V. C., Farquhar M. G. Partial characterization of newly synthesized proteoglycans isolated from the glomerular basement membrane. J Cell Biol. 1981 Aug;90(2):527–532. doi: 10.1083/jcb.90.2.527. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kanwar Y. S., Jakubowski M. L. Unaltered anionic sites of glomerular basement membrane in aminonucleoside nephrosis. Kidney Int. 1984 Apr;25(4):613–618. doi: 10.1038/ki.1984.65. [DOI] [PubMed] [Google Scholar]
- Kanwar Y. S., Linker A., Farquhar M. G. Increased permeability of the glomerular basement membrane to ferritin after removal of glycosaminoglycans (heparan sulfate) by enzyme digestion. J Cell Biol. 1980 Aug;86(2):688–693. doi: 10.1083/jcb.86.2.688. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kanwar Y. S., Veis A., Kimura J. H., Jakubowski M. L. Characterization of heparan sulfate-proteoglycan of glomerular basement membranes. Proc Natl Acad Sci U S A. 1984 Feb;81(3):762–766. doi: 10.1073/pnas.81.3.762. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Keller K. L., Keller J. M., Moy J. N. Heparan sulfates from Swiss mouse 3T3 and SV3T3 cells: O-sulfate difference. Biochemistry. 1980 May 27;19(11):2529–2536. doi: 10.1021/bi00552a035. [DOI] [PubMed] [Google Scholar]
- Kerjaschki D., Sharkey D. J., Farquhar M. G. Identification and characterization of podocalyxin--the major sialoprotein of the renal glomerular epithelial cell. J Cell Biol. 1984 Apr;98(4):1591–1596. doi: 10.1083/jcb.98.4.1591. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klein D. J., Dehnel P. J., Oegema T. R., Brown D. M. Alterations in proteoglycan metabolism in the nephrotic syndrome induced by the aminonucleoside of puromycin. Lab Invest. 1984 May;50(5):543–551. [PubMed] [Google Scholar]
- Linker A., Hovingh P. Structural studies of heparitin sulfates. Biochim Biophys Acta. 1975 Apr 7;385(2):324–333. doi: 10.1016/0304-4165(75)90360-8. [DOI] [PubMed] [Google Scholar]
- Linker A., Hovingh P. The heparitin sulfates (heparan sulfates). Carbohydr Res. 1973 Jul;29(1):41–62. doi: 10.1016/s0008-6215(00)82069-8. [DOI] [PubMed] [Google Scholar]
- Mahan J. D., Sisson-Ross S., Vernier R. L. Glomerular basement membrane anionic charge site changes early in aminonucleoside nephrosis. Am J Pathol. 1986 Nov;125(2):393–401. [PMC free article] [PubMed] [Google Scholar]
- 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]
- Mynderse L. A., Hassell J. R., Kleinman H. K., Martin G. R., Martinez-Hernandez A. Loss of heparan sulfate proteoglycan from glomerular basement membrane of nephrotic rats. Lab Invest. 1983 Mar;48(3):292–302. [PubMed] [Google Scholar]
- Olson J. L., Rennke H. G., Venkatachalam M. A. Alterations in the charge and size selectivity barrier of the glomerular filter in aminonucleoside nephrosis in rats. Lab Invest. 1981 Mar;44(3):271–279. [PubMed] [Google Scholar]
- Rennke H. G., Olson J. L., Venkatachalam M. A. Glomerular filtration of macromolecules: normal mechanisms and the pathogenesis of proteinuria. Contrib Nephrol. 1981;24:30–41. doi: 10.1159/000395227. [DOI] [PubMed] [Google Scholar]
- Rosenzweig L. J., Kanwar Y. S. Removal of sulfated (heparan sulfate) or nonsulfated (hyaluronic acid) glycosaminoglycans results in increased permeability of the glomerular basement membrane to 125I-bovine serum albumin. Lab Invest. 1982 Aug;47(2):177–184. [PubMed] [Google Scholar]
- Schurer J. W., Hoedemaeker J., Molenaar I. Polyethyleneimine as tracer particle for (immuno) electron microscopy. J Histochem Cytochem. 1977 May;25(5):384–387. doi: 10.1177/25.5.325123. [DOI] [PubMed] [Google Scholar]
- Shively J. E., Conrad H. E. Formation of anhydrosugars in the chemical depolymerization of heparin. Biochemistry. 1976 Sep 7;15(18):3932–3942. doi: 10.1021/bi00663a005. [DOI] [PubMed] [Google Scholar]
- Striker G. E., Killen P. D., Farin F. M. Human glomerular cells in vitro: isolation and characterization. Transplant Proc. 1980 Sep;12(3 Suppl 1):88–99. [PubMed] [Google Scholar]
- Trelstad R. L. Glycosaminoglycans: mortar, matrix, mentor. Lab Invest. 1985 Jul;53(1):1–4. [PubMed] [Google Scholar]
- Vernier R. L., Klein D. J., Sisson S. P., Mahan J. D., Oegema T. R., Brown D. M. Heparan sulfate--rich anionic sites in the human glomerular basement membrane. Decreased concentration in congenital nephrotic syndrome. N Engl J Med. 1983 Oct 27;309(17):1001–1009. doi: 10.1056/NEJM198310273091701. [DOI] [PubMed] [Google Scholar]

