Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Endothelin-1 blunts transepithelial transport and differentiation of Madin-Darby canine kidney cells

  • 25 Accesses

  • 6 Citations


We investigated the effects of endothelin-1 (ET-1) on Madin-Darby canine kidney (MDCK) cells, a cell line originating from the renal collecting duct. The activity of transepithelial transport was assessed as the rate of dome formation in monolayers grown on solid support. The pH value of the dome fluid (dome pH) was measured by means of pH-selective microelectrodes. Differentiation of monolayer cells was estimated as the peanut-lectin(PNA)-binding capacity of the apical membrane. Confluent monolayers were incubated for 12–72 h in serum-free medium at various concentrations of ET-1. Exposure to 1 nmol/l ET-1 reduced dome formation by a maximum of 41±8% (n=4; P<0.02) after 24 h. ET-1 (10 nmol/l; 24 h) decreased dome pH from 7.52±0.02 (n=53) to 7.36±0.03 (n=51; P<0.02). Apical application of amiloride (1 mmol/l) reduced dome pH in both ET-1-treated and non-treated domes to essentially the same level, 7.25±0.03 (n=19) and 7.23±0.03 (n=17) respectively. ET-1 (10 nmol/l; 24 h) reduced PNA-binding capacity by 19±3% (n=5; P < 0.02). Moreover, ET-1 prevented the increase in PNA binding (+53±7%; n=5) induced by 0.1 μmol/l aldosterone. We conclude that ET-1 inhibits transepithelial transport and PNA binding via inhibition of apical Na+/H+ exchange, thus antagonizing aldosterone action in MDCK cells.

This is a preview of subscription content, log in to check access.


  1. 1.

    Chabrier PE, Braquet P (1990) Endothelin. Horm Res 34:169–174

  2. 2.

    Claira J, Jimenez W, La Villa G, Asbert M, Castro A, Llibre J, Arroyo V, Rivera F (1991) Effects of endothelin on renal haemodynamics and segmental sodium handling in conscious rats. Acta Physiol Scand 141:305–308

  3. 3.

    Fried TA, Walker K, Ayon MA (1990) Immunohistochemical and autoradiographic localization of endothelin in the rat kidney. JASN 1:415

  4. 4.

    Gstraunthaler G, Pfaller W, Kotanko P (1985) Biochemical characterization of renal epithelial cell cultures (LLC-PK1 and MDCK). Am J Physiol 248:F536-F544

  5. 5.

    Guntupalli J, Phelps R, DuBose TD (1991) Effect(s) of endothelin-1 (ET) on Na-Pi cotransport and Na-H exchange in rat renal brush border membranes (BBM). JASN 2:402

  6. 6.

    Harris PJ, Zhuo J, Mendelsohn FAO, Skinner SL (1991) Haemodynamic and renal tubular effects of low doses of endothelin in anaesthetized rats. J Physiol (Lond) 433:25–39

  7. 7.

    Heilig CW, Brenner RM, Yu ASL, Kone BC, Gullans SL (1990) Modulation of osmolytes in MDCK cells by solutes, inhibitors, and vasopressin. Am J Physiol 259:F653-F659

  8. 8.

    Hoffman A, Grossman E, Goldstein DS, Gill JR, Keiser HR (1990) Low urinary levels of endothelin-1 in patients with essential hypertension. JASN 1

  9. 9.

    Iijima K, Lin L, Nasjletti A, Goligorsky MS (1991) Intracellular ramification of endothelin signal. Am J Physiol 260:C982-C992

  10. 10.

    Katoh T, Chang H, Uchida S, Okuda T, Kurokawa K (1990) Direct effects of endothelin in the rat kidney. Am J Physiol 258:F397-F402

  11. 11.

    Kennedy G, Lever JE (1984) Regulation of Na+, K+-ATPase activity in MDCK kidney epithelial cell cultures: role of growth state, cyclic AMP, and chemical inducers of dome formation and differentiation. J Physiol (Lond) 121:151–163

  12. 12.

    Kersting U, Schwab A, Treidtel M, Pfaller W, Gstraunthaler G, Steigner W, Oberleithner H (1991) Cell differentiation of renal epitheloid (MDCK) cells depends on cell culture conditions. Am J Physiol (in press)

  13. 13.

    Kersting U, Wojnowski L, Steigner W, Oberleithner H (1991) Hypotonic stress-induced release of KHCO3 in fused renal epitheloid (MDCK) cells. Kidney Int 39:891–900

  14. 14.

    King AJ, Brenner BM (1991) Endothelium-derived vasoactive factors and the renal vasculature. Am J Physiol 260:R653- R662

  15. 15.

    Kohan DE (1990) Endothelin (ET) synthesis by renal tubule cells. JASN 1:419–419

  16. 16.

    Kon V, Badr KF (1991) Biological actions and pathophysiological significance of endothelin in the kidney. Kidney Int 40:1–12

  17. 17.

    L'Allemain G, Franchi A, Gragoe E, Pouyssegur J (1984) Blockade of the Na+/H+ antiport abolishes growth factor-induced DNA synthesis in fibroblasts. J Biol Chem 259:4313–4319

  18. 18.

    LeHir M, Kaissling B, Koeppen BM, Wade JB (1982) Binding of peanut lectin to specific epithelial cell types in kidney. Am J Physiol 242:C117-C120

  19. 19.

    Madsen KM, Tisher CC (1986) Structural-functional relationship along the distal nephron. Am J Physiol 250:F1-F15

  20. 20.

    Minuth WW, Gilbert P, Gross P (1988) Appearance of specific proteins in the apical plasma membrane of cultured renal collecting duct principal cell epithelium after chronic administration of aldosterone and arginine vasopressin. Differentiation 38:194–202.

  21. 21.

    Minuth WW, Gilbert P, Rudolph U, Spielman WS (1989) Successive histochemical differentiation steps during postnatal development of the collecting duct in rabbit kidney. Histochemistry 93:19–25

  22. 22.

    Molenaar WH, Tsien RY, Van der Saag PT, De Laat WS (1983) Na+/H+ exchange and cytoplasmic pH in the action of growth factors in human fibroblasts. Nature 304:645–648

  23. 23.

    Mortensen LH, Pawloski CM, Kanagy NL, Fink GD (1990) Chronic hypertension produced by infusion of endothelin in rats. Hypertension 15:729–733

  24. 24.

    Neuser D, Zaiss S, Stasch J-P (1990) Endothelin receptors in cultured renal epithelial cells. Eur J Pharmacol 176:241–243

  25. 25.

    Oberleithner H (1991) Acute aldosterone action in renal target cells. Cell Physiol Biochem 1:2–12

  26. 26.

    Oberleithner H, Weigt M, Westphale HJ, Wang W (1987) Aldosterone activates Na+/H+ exchange and raises cytoplasmic pH in target cells of the amphibian kidney. Prog Natl Acad Sci USA 84:1464–1468

  27. 27.

    Oberleithner H, Steigner W, Silbernagl S, Vogel U, Gstraunthaler G, Pfaller W (1990) Madin-Darby canine kidney cells: III. Aldosterone stimulates an apical H+/K+ pump. Pflügers Arch 416:540–547

  28. 28.

    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

  29. 29.

    Oberleithner H, Vogel U, Kersting U, Steigner W (1990) Madin-Darby canine kidney cells: II. Aldosterone stimulates Na+/H+ and Cl/HCO 3 exchange. Pflügers Arch 416:533–539

  30. 30.

    Pfaller W, Gstraunthaler G, Kersting U, Oberleithner H (1989) Carbonic anhydrase activity in Madin-Darby canine kidney cells. Renal Physiol Biochem 12:328–337

  31. 31.

    Rindler MJ, Chuman LM, Schaffer L, Saier MH (1979) Retention of differentiated properties in an established dog kidney epithelial cell line MDCK. J Cell Biol 81:635–648

  32. 32.

    Rubanyi GM, Parker Bothello LH (1991) Endothelins. FASEB J 5:2713–2720

  33. 33.

    Schuster VL, Bonsib SM, Jennings ML (1986) Two types of collecting duct mitochondria-rich (intercalated) cells: lectin and band 3 cytochemistry. Am J Physiol 251:C347-C355

  34. 34.

    Schwartz GJ, Satlin LM, Bergmann JE (1988) Fluorescent characterization of collecting duct cells: a second H+-secreting type. Am J Physiol 255:F1003-F1014

  35. 35.

    Shichiri M, Hirata Y, Emori T, Ohta K, Nakajima T, Sato K, Sato A, Marumo F (1989) Secretion of endothelin and related peptides from renal epithelial cell lines. FEBS Lett 253:203–206

  36. 36.

    Uchida S, Horie M, Yanagisawa M, Matsushita Y, Ogata E, Kurokawa K (1990) Polarized secretion of endothelin-1 (ET-1) and big ET-1 in MDCK is disrupted by NH4Cl. JASN 1:428–428

  37. 37.

    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

  38. 38.

    Waeber Ch, Hoyer D, Palacios J-M (1990) Similar distribution of [125I]sarafotoxin-6b and [125I]endothelin-1, -2, -3 binding sites in the human kidney. Eur J Pharmacol 176:233–236

  39. 39.

    Webb DJ (1991) Endothelin receptors cloned, endothelin converting enzyme characterized and pathophysiological roles for endothelin proposed. Trends Physiol Sci 12:43–46

  40. 40.

    Yamada K, Yoshida S (1991) Role of endogenous endothelin on renal functions in rats. Am J Physiol 260:F34-F38

  41. 41.

    Yanagisawa M, Masaki T (1989) Molecular biology and biochemistry of the endothelins. Trends Physiol Sci 10:374–378

  42. 42.

    Yanagisawa M, Kurihara H, Kimura S, Tomobe Y, Kobayashi M, Mitusi Y, Yazaki Y, Goto K, Masaki T (1988) A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature 332:411–415

  43. 43.

    Zeidel ML, Brady HR, Kone BC, Gullans SR, Brenner BM (1989) Endothelin, a peptide inhibitor of Na+ -K+ -ATPase in intact renal tubular epithelial cells. Am J Physiol 257:C1101-C1107

Download references

Author information

Correspondence to L. Wojnowski.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Wojnowski, L., Gaßner, B., Steigner, W. et al. Endothelin-1 blunts transepithelial transport and differentiation of Madin-Darby canine kidney cells. Pflugers Arch. 420, 508–514 (1992). https://doi.org/10.1007/BF00374626

Download citation

Key words

  • Endothelin
  • MDCK cells
  • Cell differentiation
  • Peanut lectin
  • Na+/H+ antiporter
  • Renal medulla