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Potential mechanism of fibronectin deposits in acute renal failure induced by mercuric chloride

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Abstract

Many glomerular diseases are associated with changes in the expression and distribution in the components of extracellular matrix. A remarkable feature in acute renal failure induced by mercuric chloride in rats was large fibronectin (Fn) deposits in kidneys 1 h post-HgCl2 injection (5 mg/kg body wt., s.c.). Our study examined some mechanisms as potential explanation of the early Fn deposits in mercuric chloride induced acute renal failure. Total tissue mRNA of livers and kidneys of control and treated rats were used in Northern blot to determine whether accumulation of Fn in kidney is associated with increases in the expression of this protein in the kidney and/or in the liver. Analysis of Fn levels by Western blot were also performed. Northern blot did not show significant difference between control and treated rats, while the abundance of polymerized-Fn in kidney tissue was increased 1 h and 5 h post HgCl2 injection.HgCl2 influence on Fn folding was studied in vitro to detect possible conformational changes that could altered its normal pattern of matrix assembly and/or binding to different ligands. In this context HgCl2 binding to Fn was measured following native thryptophan fluorescence of Fn in the presence of HgCl2 (0.5–250 mM). Binding parameters for the HgCl2–Fn complex formation were Kd = (1.6 ± 0.2) 10–4 M; n = 1 ± 0.3, indicating a low apparent affinity and one type binding site. Thermal denaturation of Fn showed, between 30–60°C, a soft reversible conformational change, while between 75–80°C a highly and irreversible transition is produced suggesting a modification of the tertiary structure. HgCl2 abolished this transition. The kinetic of thermal unfolding of Fn was also measured and the effects observed due to HgCl2 presence reinforced the previous data. Finally, the effect of HgCl2 on Fn binding to denatured collagen (gelatin) was also measured as an index of the effect of this cation on biological properties of Fn. Fn binds gelatin strongest in the presence of HgCl2.Our results suggest that higher Fn deposits in kidney treated rats seems not to be associated to augmented mRNA-Fn neither in kidney nor in liver. On the other hand, increased levels of polymerized Fn abundance was observed in kidney tissue from mercury-treated rats. We also describe that HgCl2 promotes, in vitro, conformational changes on Fn structure, inducing its denaturation and increasing its binding to gelatin, all events that could be related to the Fn deposits in renal tissues of HgCl2-treated rats, and could be expected in other situations that promoted interstitial fibrosis, not associated to overexpression of matrix-proteins.

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References

  1. Oomura A, Nakamura T, Arakawa M, Ooshima A, Isemura M: Alterations in the extracellular matrix components in human glomerular diseases. Virchow Arch 415: 151–159, 1989

    Google Scholar 

  2. Buyükbabani N, Droz D: Distribution of the extracellular matrix components in human glomerular lesions. J Pathol 172: 199–207, 1994

    Google Scholar 

  3. Jones CL, Buch B, Post M, McCullogh L, Eddy AA: Renal extracellular matrix accumulation in acute purinomycin aminonucleoside nephrosis in rats. Am J Pathol 141: 1381–1396, 1992

    Google Scholar 

  4. Floege J, Johnson RJ, Gordon K, Yoshimura A, Campbell C, Iruela-Arispe L, Alpers CE, Couser WG: Altered glomerular extracellular matrix synthesis in experimental membranous nephropathy. Kidney Int 42: 573–585, 1992

    Google Scholar 

  5. Ziyadeh FN: The extracellular matrix in diabetic nephropathy. Am J Kidney Dis 23: 736–744, 1993

    Google Scholar 

  6. Kashtan CE, Michael AF: Alport syndrome. Kidney Int 501445–501463, 1996

  7. Slomowitz LA, Klair S, Schreiner GF, Ichikawa Y: Progression of renal disease. N Engl J Med 319: 1547–1548, 1988

    Google Scholar 

  8. Hynes RO: Molecular biology of fibronectin. Annu Rev Cell Biol 1: 67–90, 1985

    Google Scholar 

  9. Oyama F, Hirohashi S, Shimosato Y, Titani K, Sekiguchi K: Deregulation of alternative splicing of fibronectin pre-mRNA in malignant human liver tumors. J Biol Chem 264: 10331–10334, 1989

    Google Scholar 

  10. Brown L, Dubin D, Lavigne B, Logan B, Dvorak HF, Vandewater L: Macrophages and fibroblasts express embryonic fibronectin during cutaneous wound healing. Am J Pathol 142: 793–801, 1993

    Google Scholar 

  11. Barnes JL, Hastings RR, De La Garza MA: Sequential expression of cellular fibronectin by platelets, macrophages and mesangial cells in proliferative glomerulonephritis. Am J Pathol 145: 585–597, 1994

    Google Scholar 

  12. Glukhova MA, Frid MG, Shekhonin BV, Vasilevskaya TD, Grunwald J, Saginati M, Koteliansky VE: Expression of extra domain A fibronectin sequence in vascular smooth muscle cells is phenotype dependant. J Biol Chem 109: 357–366, 1989

    Google Scholar 

  13. Pawluczyk IZA, Harris KPG: Cytokine interactions promote synergistic fibronectin accumulation by mesangial cells. Kidney Int 54: 62–70, 1998

    Google Scholar 

  14. Benecky MJ, Wine RW, Kolvenbach CG, Mosesson MW: Ionic-strength and pH-dependent conformational states of human plasma fibronectin. Biochemistry 30: 4298–4306, 1991

    Google Scholar 

  15. Zuk A, Bonventre J, Brown D, Matlin K: Polarity, integrin and extracellular matrix dynamics in postischemic rat kidney. Am J Physiol 275: C711–C731, 1998

    Google Scholar 

  16. Girardi G, Saball DE, Salvarrey MS, Elías MM: Glomerular compromise in mercuric chloride-induced nephrotoxicity. J Biochem Toxicol 11: 189–196, 1996

    Google Scholar 

  17. Zalups RK: Molecular interactions with mercury in the kidney. Pharmacol Rev 52: 113–142, 2000

    Google Scholar 

  18. Girardi G, Elías MM: Effectiveness of N-acetylcysteine in protecting against mercuric chloride-induced nephrotoxicity. Toxicology 67: 155–164, 1991

    Google Scholar 

  19. Lai C, Wolff CE, Novello D, Griffone L, Cuniberti C, Molina F, Rocco M: Solution structure of human plasma fibronectin under different solvent conditions. J Mol Biol 230: 625–640, 1993

    Google Scholar 

  20. Engvall E, Ruoslahti E: Binding of soluble form fibroblast surface protein fibronectin to collagen. Int J Cancer 20: 1–4, 1977

    Google Scholar 

  21. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970

    Google Scholar 

  22. Girardi G, Torres AM, Elías MM: The implication of renal glutathione levels in mercuric chloride nephrotoxicity. Toxicology 58: 187–195, 1989

    Google Scholar 

  23. Jackson P, Blythe D: Immunolabelling techniques for light microscopy. In: J.E. Beesley (ed). Immunocytochemistry. Oxford University Press, New York, 1993, pp 33–37

    Google Scholar 

  24. Davis LG, Kuehl WM, Battey JF: Basic Methods in Molecular Biology, 2nd ed. Appleton and Lange, Norwalk, CT, 1994

    Google Scholar 

  25. Sambrook J, Fritsch E, Maniatis L: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 1989

    Google Scholar 

  26. Dignam JD: Preparation of extracts from higher eukaryotes. Meth Enzymol 182: 194–203, 1990

    Google Scholar 

  27. Farruggia B, García G, D'Angelo A, Picó G: Destabilization of human serum albumin by polyethylene glycol studied by thermodynamical equilibrium and kinetic approaches. Int J Biol Macromol 20: 43–51, 1997

    Google Scholar 

  28. Mosesson MW, Umflet RA: The cold-insoluble globulin of human plasma. Purification, primary characterization, and relationship to fibrinogen and adult cold insoluble fraction components. J Biol Chem 245: 5728–5736, 1970

    Google Scholar 

  29. Raghow R, Postlethwaite AE, Kecki-Oja J, Moses HL, Kang AH: Transforming growth factor-β increases steady state levels of type I procollagen and fibronectin messenger mRNAs postranscriptionally in culture human dermal fibroblast. J Clin Invest 79: 1285–1288, 1987

    Google Scholar 

  30. Nakamura T, Fukui M, Ebihara I, Osada S, Tomino Y, Koide H: Abnormal gene expression of matrix metalloproteinases and their inhibitor in glomeruli from diabetic rats. Renal Physiol Biochem 17: 316–325, 1994

    Google Scholar 

  31. Studer RK, Negrete H, Craven PA, DeRubertis FR: Protein kinase C signals thromboxane induced increases in fibronectin synthesis and TGF-β bioactivity in mesangial cells. Kidney Int 48: 422–430, 1995

    Google Scholar 

  32. Moud AP, Akiyama SK, Humphries MJ: Regulation of integrin α5β1-fibronectin interactions by divalent cations. Evidence for distinct classes of binding sites for Mn2+, Mg2+ and Ca2+. J Biol Chem 270: 26270–26277, 1995

    Google Scholar 

  33. Piotrowski JK, Trojanowska B, Wisniewska-Knypl JM, Bolanowska W: Mercury binding in the kidney and liver of rats repeatedly exposed to mercury chloride. Toxicol Appl Pharmacol 27: 11–19, 1974

    Google Scholar 

  34. Zalups RK, Cherian MG: Renal metallothionein metabolism after reduction of renal mass. I. Effect of unilateral nephrectomy and compensatory renal growth on basal and metal induced renal metallothionein metabolism. Toxicology 71: 83–102, 1992

    Google Scholar 

  35. Cherian MG, Clarkson TW: Biochemical changes in rat kidney on exposure to elemental mercury vapor. Effect on biosynthesis of metallothionein. Chem Biol Interact 12: 109–120, 1976

    Google Scholar 

  36. Setcher VJ, Kaplan JE, Connoly K, Mielens Z, Saelens JK: Fibronectin in acute and chronic inflammation. Arthritis Rheum 29: 394–399, 1986

    Google Scholar 

  37. Tapio V: Disulfide-bonded polymerization of plasma fibronectin in the presence of metal ions. J Biol Chem 261: 9433–9437, 1986

    Google Scholar 

  38. Mosher DF: Assembly of fibronectin into extracellular matrix. Curr Opin Struct Biol 3: 214–222, 1993

    Google Scholar 

  39. Litvinovich SV, Ingham KC: Interactions between type III domains in the 110 kDa cell-binding fragment of fibronectin. J Mol Biol 248: 611–626, 1995

    Google Scholar 

  40. Khan MY, Medow MS, Newman SA: Unfolding transitions of fibronectin and its domains. Stabilization and structural alteration of the Nterminal domain by heparin. Biochem J 270: 33–38, 1990

    Google Scholar 

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Authors and Affiliations

  1. Facultad de Ciencias Bioquímicas y Farmacéuticas –, Universidad Nacional de Rosario, Suipacha 531 –, 2000, Rosario, Argentina

    Ester Saball, Marcela Salvarrey, Esteban Serra, Guillermo Picó & María Mónica Elías

  2. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina

    Esteban Serra & Guillermo Picó

  3. Instituto de Biología Molecular y Celular de Rosario, Rosario, Argentina

    Esteban Serra

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  1. Ester Saball
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  2. Marcela Salvarrey
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  3. Esteban Serra
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  4. Guillermo Picó
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  5. María Mónica Elías
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Correspondence to María Mónica Elías.

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Saball, E., Salvarrey, M., Serra, E. et al. Potential mechanism of fibronectin deposits in acute renal failure induced by mercuric chloride. Mol Cell Biochem 226, 67–75 (2001). https://doi.org/10.1023/A:1012733719017

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