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Anionic sites in glomerular basement membrane of rats with serum sickness nephritis: quick-freezing and deep-etching study

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Summary

Serum sickness nephritis was induced in male Fisher 344/JCL rats by injecting egg albumin into the foot pads and peritoneal cavity. The alteration of anionic sites in the glomerular basement membrane (GBM) of the rats with significant proteinuria was studied with a quick-freezing and deep-etching method using polyethyleneimine as a cationic probe. In control rats, anionic sites were located around the fibrils of the lamina rara externa, which radiated perpendicularly from the lamina densa to podocyte cell membranes. In the glomeruli of proteinuric rats, many electron-dense deposits were observed in the subepithelial side of the GBM, where the fibrils of the lamina rara externa were usually obscured and anionic sites around them could not be recognized. However, in some areas, a clear boundary could be observed between deposits and the lamina densa. Electron micrographs of freeze-fractured deposits showed that the fibrils radiated perpendicularly from the lamina densa and that anionic sites around them had been preserved. These results suggest that some of the deposits simply passed through the GBM and masked transiently the fibril structures of the GBM, but others probably destroyed these fibril structures, including anionic sites.

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References

  • Bolton, W. K. & Sturgill, B. C. (1978) Bovine serum albumin chronic serum sickness nephropathy. Br. J. Exp. Pathol. 59, 167–77.

    Google Scholar 

  • Camussi, G., Tetta, C., Meroni, M., Torri-Tarelli, L., Roffinello, C., Alberton, A., DeregibusC. & SessaA. (1986) Localization of cationic proteins derived from platelets and polymorphonuclear neutrophils and local loss of anionic sites in glomeruli of rabbits with experimentally-induced acute serum sickness. Lab. Invest. 55, 56–62.

    Google Scholar 

  • Caulfield, J. P. (1979) Alterations in the distribution of alcian blue-staining fibrillar anionic sites in glomerular basement membrane in aminonucleo side nephrosis. Lab. Invest. 40, 503–11.

    Google Scholar 

  • Caulfield, J. P. & Farquhar, M. G. (1976) Distribution of anionic sites in glomerular basement membranes: their possible role in filtration and attachment. Proc. Natl. Acad. Sci. USA 73, 1646–50.

    Google Scholar 

  • Duan, H-J. (1990) Sequential ultrastructural podocytic lesions and development of proteinuria in serum sickness nephritis in the rat. Virchows Arch [A] 417, 279–90.

    Google Scholar 

  • Duan, H-J., Nakazawa, K., Ishigame, H., Itoh, N. & Shigematsu, H. (1991a) Masking of anionic sites by deposits in lamina rara externa in immune complex nephritis in rats. Virchows Arch [B] 60, 155–71.

    Google Scholar 

  • Duan, H-J., Nakazawa, K., Ishigame, H., Itoh, N., Yamaguchi, N. & Shigematsu, H. (1991b) Influence of subepithelial deposits on size and charge selective permeability of glomerular filter in serum sickness nephritis in rats. J. Electron Microsc. 40, 392–8.

    Google Scholar 

  • Furness, P. N., Turner, D. R. & Cotton R. E. (1986) Basement membrane charge in human glomerular disease. J. Pathol. 150, 267–78.

    Google Scholar 

  • Heuser, J. E. & Kirschner, M. W. (1980) Filament organization revealed in platinum replicas of freeze-dried cytoskeletons. J. Cell Biol. 86, 212–34.

    Google Scholar 

  • Hirokawa, N. & Heuser, J. E. (1981) Quick-freeze, deep-etch visualization of the cytoskeleton beneath surface differentiations of intestinal epithelial cells. J. Cell Biol. 91, 399–409.

    Google Scholar 

  • Hora, K., Ohno, S., Oguchi, H., Furukawa, T. & Furuta, S. (1990) Three-dimensional study of glomerular slit diaphragm by the quick-freezing and deep-etching replica method. Eur. J. Cell Biol. 53, 402–6.

    Google Scholar 

  • KanwarY. S. & Farquhar, M. G. (1979) Anionic sites in the glomerular basement membrane: in vivo localization to laminae rarae by cationic probes. J. Cell Biol. 81, 137–53.

    Google Scholar 

  • Kingsbury, F. B., Clark, C. P., Williams, G. & Post, A. L. (1926) The rapid determination of albumin in urine. J. Lab. Clin. Med. 11, 981–9.

    Google Scholar 

  • KubosawaH. & KondoY. (1985) Ultrastructural organization of the glomerular basement membrane as revealed by a deep-etch replica method. Cell Tissue Res. 242, 33–9.

    Google Scholar 

  • Lelongt, B., Makino, H. & Kanwar, Y. S. (1987) Status of glomerular proteoglycan in aminonucleoside nephrosis. Kidney Int. 31, 1299–310.

    Google Scholar 

  • Mahan, J. D., Sisson-Ross, S. & Vernier, R. L. (1986) Glomerular basement membrane anionic charge site changes early in aminonucleoside nephrosis. Am. J. Pathol. 125, 393–401.

    Google Scholar 

  • Mynderse, L. A., Hassell, J. R., Kleinman, H. K., Martin, G. R. & Martine-Hernandez, A. (1983) Loss of heparan sulfate proteoglycan from glomerular basement membrane of nephrotic rats. Lab. Invest. 48, 292–302.

    Google Scholar 

  • Naramoto, A., Ohno, S., Takmai, H., Nakazawa, K., Kasahara, H., Duan, H-J., Itoh, N. & Shigematsu, H. (1991a) Ultrastructure of matrical changes in chronic phase of Masugi nephritis by quick-freezing and deepetching method. Vichows Arch [A] 418, 51–9.

    Google Scholar 

  • Naramoto, A., Ohno, S., NakazawaK., Takami, H., Itoh, N. & Shigematsu, H. (1991b) Three-dimensional ultrastructure of glomerular injury in serum sickness nephritis using the quick-freezing and deep-etching method. Virchow Arch [A] 418, 185–92.

    Google Scholar 

  • Naramoto, A., Ohno, S., Itoh, N., Shibata, N. & Shigematsu, H. (1992) Localization of laminin in nephritis glomeruli as revealed by a quick-freezing and deep-etching method with immunohistochemistry. Histochem J. (in press).

  • Ohno, S., Hora, K., Furukawa, T. & Oguchi, H. (1992) Ultrastructural study of the glomerular slit diaphragm in fresh unfixed kidney by quick-freezing method. Virchows Archiv [B] 61, 351–8.

    Google Scholar 

  • Rada, J. A. & Carlson, E. C. (1991a) Electron microscopic histochemical and immunochemical analyses of heparan sulfate proteoglycan distribution in renal glomerular basement membranes. Histo Histopath 61, 149–60.

    Google Scholar 

  • Rada, J. A. & Carlson, E. C. (199lb) Anionic sites and immunogold quantitation of heparan sulfate proteoglycans in glomerular basement membranes of puromycin aminonculeoside nephrotic rats. Anal. Rec. 231, 35–47.

    Google Scholar 

  • Rennke, H. G., Cotran, R. S. & Venkatachalam, M. A. (1975) Role of molecular charge in glomerular permeability: tracer studies with cationized ferritin. J. Cell Biol. 67, 638–46.

    Google Scholar 

  • Sauren, Y. M. H. F., Mieremet, R. H. P., Groot, C. G., Koerten, H. K. & Scherft, J. P. (1991) Polyethyleneimine as a contrast agent for ultrastructural localization and characterization of proteoglycans in the matrix of cartilage and bone. J. Histochem. Cytochem. 39, 331–40.

    Google Scholar 

  • Schiller, A. & Taugner, R. (1980) Freeze-fracture and deepetching with the volatile cryoprotectant ethanol reveals true membrane surfaces of kidney structures. Cell Tissue Res. 210, 57–69.

    Google Scholar 

  • Schneeberger, E. E., Stavrakis, G. & Mccarthy, K. (1983) Alterations in glomerular anionic sites in autologous immune complex nephritis. Lab. Invest. 49, 445–52.

    Google Scholar 

  • Schurer, J. W., Hoedemaeker, P. H. J. & Molenaar, I. (1977) Polyethyleneimine as tracer particle for (immuno) electron microscopy. J. Histochem. Cytochem. 25, 384–7.

    Google Scholar 

  • Schurer, J. W., Kalicharan, D., Hoedemaeker, P. H. J. & Molenaar, I. (1978) The use of polyethyleneimine for demonstration of anionic sites in basement membrane and collagen fibrils. J. Histochem. Cytochem. 26, 688–9.

    Google Scholar 

  • Shigematsu, H. & Yano, A. (1986) Participation of antigen presenting cells in glomerulonephritis. Acta Pathol. Jpn. 36, 489–97.

    Google Scholar 

  • Suzuki, Y., Maesawa, A., Matsui, K., Oite, T., Koda, Y. & Arakawa, M. (1983) Alteration of glomerular anionic sites by the development of subepithelial deposits in experimental glomerulonephritis in the rat. Virchows Arch [B] 44, 209–22.

    Google Scholar 

  • Takami, H., Naramoto, A., Nakazawa, K., Shigematsu, H. & Ohno, S. (1990) Ultrastructure of glomerular mesangial matrix by quick-freeze and deep-etch methods. Kidney Int 38, 1211–5.

    Google Scholar 

  • Takami, H., Naramoto, A., Shigematsu, H. & Ohno, S. (1991) Ultrastructure of glomerular basement membrane by quickfreeze and deep-etch methods. Kidney Int. 39, 659–64.

    Google Scholar 

  • Vernier, R. L., Klein, D. J., Sisson, S. P., Mahan, J. D., Oegema, T. R. & Brown, D. M. (1983) Heparan sulfate-rich anionic sites in the human glomerular basement membrane: decreased concentration in congenital nephrotic syndrome. N. Engl. J. Med. 309, 1001–9.

    Google Scholar 

  • Washizawa, K. (1990) Loss of anionic sites from the glomerular basement membrane in human glomerulonephritis. Acta Pathol Jpn. 40, 172–80.

    Google Scholar 

  • Washizawa, K., Ishii, K., Itoh, N., Mori, T., Akahane, T. & Shigematsu, H. (1989) Morphometric changes in glomerular anionic sites during aminonucleoside nephrosis. Acta Pathol. Jpn. 39, 558–65.

    Google Scholar 

  • Whiteside, C., Prutis, K., Cameroh, R. & Thompson, J. (1989) Glomerular epithelial detachment, not reduced charge density, correlates with proteinuria in adriamycin and pyromycin nephrosis. Lab. Invest. 61, 650–60.

    Google Scholar 

  • Yamamoto, T., Kihara, I., Morita, K. & Oite, T. (1978) Bovine serum albumin (BSA) nephritis in rats. I. Experimental model. Acta Pathol. Jpn. 28, 859–66.

    Google Scholar 

  • Yoshimura, A., Ohno, S., Nakano, K., Oniki, H., Inui, K., Ideura, T. & Koshikawa, S. (1991) Three-dimensional ultrastructure of anionic sites of the glomerular basement membrane by a quick-freezing an deepetching method using a cationic tracer. Histochemistry 96, 107–13.

    Google Scholar 

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Duan, HJ., Ohno, S. & Shigematsu, H. Anionic sites in glomerular basement membrane of rats with serum sickness nephritis: quick-freezing and deep-etching study. Histochem J 25, 367–375 (1993). https://doi.org/10.1007/BF00159501

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