Ochratoxin A impairs “postproximal” nephron function in vivo and blocks plasma membrane anion conductance in Madin-Darby canine kidney cells in vitro

Pflügers Archiv volume 425, pages401–408(1993)Cite this article

Abstract

Ochratoxin A (OTA) is a widespread nephrotoxin which causes porcine nephropathy and is supposed to have caused the human Balkan endemic nephropathy. We performed experiments in vivo and in vitro to elucidate the mechanism of OTA action in renal epithelium. Application of OTA to male Wistar rats [1.25 μmol/ (kg · day)] for 6 days led to a reduction of glomerular filtration rate (to 63% of control), an increased fractional water (194% of control), Na+ (199% of control), K+ (147% of control) and Cl− (270% of control) excretion and an increased dependence of the osmole clearance on urine flow. Acute application of OTA to rats (3 μmol/ kg) increased urinary pH from 6.0±0.2 to 6.6±0.1 and urinary NaCl excretion, but decreased titratable acid excretion to 47% of control. As these in vivo findings may be the result of an action of OTA beyond the proximal tubule (“postproximal”) we investigated the effect of OTA on cultured Madin-Darby canine kidney (MDCK) cells, regarded as a model of collecting duct epithelium. In confluent monolayers formed by MDCK cells OTA reduced the number of domes in a dose-dependent manner and impaired the formation of a transepithelial Cl− gradient. Electrophysiological measurements in giant MDCK cells revealed that OTA blocks fractional anion conductance of the plasma membrane with an IC50 value of 30±5 nmol/l, unmasking OTA as a naturally occurring anion conductance blocker about 20-times more effective than the most potent synthetic blocker 5-nitro-2-(3-phenylpropyl-amino) benzoic acid (NPPB) (IC50=600±50 nmol/l). The effects of OTA and NPPB are not additive. We conclude that OTA acts acutely on “postproximal” parts of the nephron. This effect is probably located in the collecting duct and is due to its inhibitory action on plasma membrane anion conductance.

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References

  1. 1.

    Bauer J, Gareis M, Gedek B (1984) Zum Nachweis und Vorkommen von Ochratoxin A bei Schlachtschweinen. Berl Münch Tierärztl Wochenschr 97: 279–283

    Google Scholar 

  2. 2.

    Berndt WO, Hayes AW (1979) In vivo and in vitro changes in renal function caused by ochratoxin A in the rat. Toxicology 12: 5–17

    Google Scholar 

  3. 3.

    Creppy EE, Kern D, Steyn PS, Vleggaar R, Röschenthaler R, Dirheimer C (1983) Comparative study of the effect of ochratoxin A analogues on yeast aminoacyl-tRNA synthetases and on the growth and protein synthesis of hepatoma cells. Toxicol Lett 19: 217–224

    Google Scholar 

  4. 4.

    Creppy EE, Röschenthaler R, Dirheimer G (1984) Inhibition of protein synthesis in the mice by ochratoxin-A and its prevention by phenylalanine. Food Chem Toxicol 22: 883–886

    Google Scholar 

  5. 5.

    Creppy EE, Kane A, Giessen-Crouse E, Roth A, Röschenthaler R, Dirheimer G (1986) Effect of ochratoxin A on enzyme activities and macromolecules synthesis in MDCK cells. Arch Toxicol Suppl 9: 310–314

    Google Scholar 

  6. 6.

    Galtier P, Charpenteau JL, Alvinerie M, Labouche C (1979) The pharmacokinetic profile of ochratoxin A in the rat after oral and intravenous administration. Drug Metab Dispos 7: 429–434

    Google Scholar 

  7. 7.

    Gekle M, Silbernagl S (1993) The effect of ochratoxin A on renal hemodynamics. Pflügers Arch 422: R126

    Google Scholar 

  8. 8.

    Gekle M, Oberleithner H, Silbernagl S (1992) The nephrotoxin ochratoxin A is a powerful Cl− channel blocker. J Am Soc Nephrol 3: 724

    Google Scholar 

  9. 9.

    Gekle M, Oberleithner H, Silbernagl S (1992) Ochratoxin A blockiert Cl−-Kanäle: Ist diese Wirkung verantwortlich für seine Nephrotoxizität? Nieren-Hochdruckkr 9: 375–376

    Google Scholar 

  10. 10.

    Gekle M, Silbernagl S, Löwe H (1992) Where in the kidney is ochratoxin A nephrotoxic? Renal Physiol Biochem 15: 197

    Google Scholar 

  11. 11.

    Gekle M, Silbernagl S, Mildenberger S, Freudinger R (1993) Effect on dome formation and uptake of ochratoxin A in proximal tubule-derived Opossum kidney cell monolayers. Cell Physiol Biochem 3: 68–77

    Google Scholar 

  12. 12.

    Hagelberg S, Hult K, Fuchs R (1989) Toxokinetics of ochratoxin A in several species and its plasma-binding properties. J Appl Toxicol 9: 91–96

    Google Scholar 

  13. 13.

    Kersting U, Joha H, Steigner W, Gaßner B, Gstraunthaler G, Pfaller W, Oberleithner H (1988) Fusion of cultured dog kidney (MDCK) cells. I. Technique, fate of plasma membrane and cell nuclei. J Membr Biol 111: 37–48

    Google Scholar 

  14. 14.

    Krogh P, Axelsen NH, Elling F, Gryd-Hansen N, Hald B, Hyldgaard-Jensen J, Larsen AE, Madsen A, Mortensen HP, Moller T, Petersen OK, Ravnskov U, Rostgaard M, Aalund O (1974) Experimental porcine nephropathy. Acta Pathol Microbiol Scand Sect A Pathol [Suppl] 246: 1–21

    Google Scholar 

  15. 15.

    Krogh P, Hald B, Plestina R, Ceovic S (1977) Balkan (endemic) nephropathy and foodborn ochratoxin A: preliminary results of a survey of foodstuffs. Acta Pathol Microbiol Scand Sect B Microbiol 85: 238–240

    Google Scholar 

  16. 16.

    Kuiper-Goodman T, Scott PM (1989) Risk assessment of the mycotoxin ochratoxin A. Biomed Environ Sci 2: 179–248

    Google Scholar 

  17. 17.

    Mandal AK, Sindjic M, Sommers SC (1987) Kidney pathology in endemic nephrophathy. Clin Nephrol 27: 304–308

    Google Scholar 

  18. 18.

    Meisner H (1976) Energy-dependent uptake of ochratoxin A by mitochondria. Arch Biochem Biophys 173: 132–140

    Google Scholar 

  19. 19.

    Merwe KJ, Steyn PS, Fourie L, Scott DB, Theron JJ (1965) Ochratoxin A, a toxic metabolite produced by Aspergillus ochraceus Wilh. Nature 205: 1112–1113

    Google Scholar 

  20. 20.

    van der Merwe KJ, Steyn PS, Fourie L (1965) Mycotoxins. II. The constitution of ochratoxin A, B and C metabolites of Aspergillus ochraceus Wilh. J Chem Soc Perkin Trans I: 7083–7088

    Google Scholar 

  21. 21.

    NTP (1989) NTP technical report on the toxicology and carcinogenesis studies of ochratoxin A (CAS No. 303-47-9) in F344/N rats (Gavage studies). Research Triangle Park, NC. NHI Publication No. 89-2813. United States Department of Health and Human Services

  22. 22.

    Oberleithner H, Lang F, Messner G, Wang W (1984) Mechanism of hydrogen ion transport in the diluting segment of frog kidney. Pflügers Arch 402: 272–280

    Google Scholar 

  23. 23.

    Oberleithner H, Kersting U, Silbernagl S, Steigner W, Vogel U (1989) Fusion of cultured dog kidney (MDCK) cells: II. Relationship between cell pH and K+ conductance in response to aldosterone. J Membr Biol 111: 49–46

    Google Scholar 

  24. 24.

    Oberleithner H, Vogel U, Kersting U (1990) Madin-Darby canine kidney cells. I. Aldosterone-induced domes and their evaluation as a model system. Pflügers Arch 416: 526–532

    Google Scholar 

  25. 25.

    Petkova-Bocharova T, Castegnaro M (1985) Ochratoxin A contamination of cereals in an area of high incidence of Balkan endemic nephropathy in Bulgaria. Food Addit Contam 2: 267–270

    Google Scholar 

  26. 26.

    Sachs L (1984) Angewandte Statistik, 6th edn. Springer, Berlin Heidelberg New York

    Google Scholar 

  27. 27.

    Sothell M, Karlmark B, Ulfendahl H (1983) FITC-inulin as a kidney tubule marker in rat. Acta Physiol Scand 119: 313–316

    Google Scholar 

  28. 28.

    Stetson DL, Beauwens R, Palmisano J, Mitchell PP, Steinmetz PR (1985) A double-membrane model for urinary bicarbonate secretion. Am J Physiol 249: F546-F552

    Google Scholar 

  29. 29.

    Tanner C, Frambach DA, Misfeldt DS (1983) Transepithelial transport in cell culture. A theoretical and experimental analysis of the biophysical properties of domes. Biophys J 43: 183–190

    Google Scholar 

  30. 30.

    Tapia MO, Seawright AA (1984) Experimental ochratoxicosis A in pigs. Aust Vet J 61: 219–222

    Google Scholar 

  31. 31.

    Valentich JD (1981) Morphological similarities between the dog kidney cell line MDCK and the mammalian cortical collecting tubule. Ann NY Acad Sci 372: 384–405

    Google Scholar 

  32. 32.

    Wangemann P, Wittner M, Di Stefano A, Englert HC, Lang HJ, Schlatter E, Greger R (1986) Cl− channel blockers in the thick ascending limb of the loop of Henle. Structure activity relationship. Pflügers Arch 407: S128-S141

    Google Scholar 

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Affiliations

  1. Department of Physiology, University of Würzburg, Röntgenring 9, D-97070, Würzburg, Germany

    Michael Gekle, Hans Oberleithner & Stefan Silbernagl

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  1. Michael Gekle
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  2. Hans Oberleithner
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  3. Stefan Silbernagl
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Correspondence to Michael Gekle.

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With technical assistance of Ruth Freudinger and Sigrid Mildenberger.

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Gekle, M., Oberleithner, H. & Silbernagl, S. Ochratoxin A impairs “postproximal” nephron function in vivo and blocks plasma membrane anion conductance in Madin-Darby canine kidney cells in vitro. Pflugers Arch. 425, 401–408 (1993). https://doi.org/10.1007/BF00374865

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Key words

  • Ochratoxin A
  • Anion conductance
  • Chloride channel
  • Distal nephron function
  • Distal nephron toxicity