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Protein kinase C activation causes inhibition of Na/K-ATPase activity in Madin-Darby canine kidney epithelial (MDCK) cells

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Abstract

To evaluate the influence of protein kinase C (PKC) activation on Na/K-ATPase activity in MDCK cells, we studied the effect of phorbol myristate acetate (PMA) and two diacylglycerol analogues, oleoylacetylglycerol and dioctanoylglycerol, on the enzyme activity. Na/K-ATPase activity was determined by cytochemistry. PMA induced a time- and dose-dependent inhibition of Na/K-ATPase activity and at 100 ng/ml decreased the enzyme activity by 55% of the initial value. These effects were mimicked by oleoylacetylglycerol and dioctanoylglycerol, and were abolished by two inhibitors of PKC, 1-(5-isoquinolinylsulphonyl)-2-methylpiperazine (H7) and sphingosine. A phorbol ester that does not activate PKC, 4α-phorbol 12, 13-didecanoate, did not inhibit Na/ K-ATPase activity. PMA inhibition persisted in the presence of cycloheximide and actinomycin D but not in the presence of amiloride. Dopamine (10 μM) inhibition of Na/K-ATPase activity was abolished in a dose-dependent manner by sphingosine. Results suggest that in MDCK cells Na/K-ATPase is an effector protein for PKC and that dopamine inhibition of its activity may be mediated by PKC.

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References

  1. Ahn J, Chang EB, Field M (1985) Phorbol ester inhibition of Na-H exchange in rabbit proximal colon. Am J Physiol 249:C527-C530

    Google Scholar 

  2. Aperia A, Bertorello A, Seri I (1987) Dopamine causes inhibition of Na+-K+-ATPase activity in rat proximal convoluted tubule segments. Am J Physiol 252:F39-F45

    Google Scholar 

  3. Barlet-Bas C, Khadouri C, Marsy S, Doucet A (1990) Enhanced intracellular sodium concentration in kidney cells recruits a latent pool of Na-K-ATPase whose size is modulated by corticosteroids. J Biol Chem 265:7800

    Google Scholar 

  4. Baum M, Hays SR (1988) Phorbol myristate acetate and dioctanoylglycerol inhibit transport in rabbit proximal convoluted tubule. Am J Physiol 254:F9-F14

    Google Scholar 

  5. Bertorello A, Aperia A (1989) Na+-K+-ATPase is an effector protein for protein kinase C in renal proximal tubule cells. Am J Physiol 256:F370-F373

    Google Scholar 

  6. Carey RM, Hughes JM (1987) Selective renal dopamine-1 receptor stimulation in man. Clin Exp Hypertens [A] 9:1009–1020

    Google Scholar 

  7. Castagna M, Takai Y, Kaibuchi K, Sano K, Kikkawa U, Nishizuka Y (1982) Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem 257:7847–7851

    Google Scholar 

  8. Chayen, Frost GTB, Dodds RA, Bitensky L, Pitchfork J, Baylis PH, Barrnett RJ (1981) The use of a hidden metal-capture reagent for the measurement of Na+-K+-ATPase activity: a new concept in cytochemistry. Histochemistry 71:533–541

    Google Scholar 

  9. Fey EG, Penman S (1984) Tumour promoters induce a specific morphological signature in the nuclear matrix-intermediate filament scaffold of Madin-Darby canine kidney (MDCK) cell colonies. Proc Natl Sci USA 18:4409–4413

    Google Scholar 

  10. Friedlander G, Amiel C (1987) Protein kinase C activators and bradykinin selectively inhibit vasopressin-stimulated AMP synthesis in MDCK cells. Biochim Biophys Acta 929:311–317

    Google Scholar 

  11. Friedlander G, Amiel C (1989) Protein kinase C activation has dissimilar effects on sodium-coupled uptakes in renal proximal tubular cells in primary culture. J Biol Chem 264:3935–3941

    Google Scholar 

  12. Hannun YA, Loomis CR, Merrill AH, Jr, Bell RM (1986) Sphingosine inhibition of protein kinase C activity and phorbol dibutyrate binding in vitro and in human platelets. J Biol Chem 261:12604–12609

    Google Scholar 

  13. Hays SR, Baum M, Kokko JP 11987) Effects of protein kinase C activation on sodium, potassium, chloride, and total CO2 transport in rabbit cortical collecting tubule. J Clin Invest 80:1561–1570

  14. Hidaka H, Hagiwara M (1987) Pharmacology of the isoquinolinesulfonamide protein kinase C inhibitors. Trends Pharmacol Sci 8:162–163

    Google Scholar 

  15. Hidaka H, Inagaki M, Kawamoto S, Sasaki Y (1984) Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinase and protein kinase C. Biochemistry 23:5036–5041

    Google Scholar 

  16. Hughes JM, Ragsdale NV, Felder RA, Chevalier RL, King B, Carey RM (1988) Diuresis and natruresis during continuous dopamine-1 receptor stimulation. Hypertension 11 [Suppl I]: 169–174

    Google Scholar 

  17. Jorgenson PL (1986) Structure, function and regulation of Na, K-ATPase in the kidney. Kidney Int 29:10–20

    Google Scholar 

  18. Katz AI (1982) Renal Na-K-ATPase: its role in tubular sodium and potassium transport. Am J Physiol 242:F207-F219

    Google Scholar 

  19. Kawamoto S, Hidaka H (1984) 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7) is a selective inhibitor of protein kinase C in rabbit platelets. Biochem Biophys Res Commun 125:258–261

    Google Scholar 

  20. Milton H, Saier JR (1981) Growth differentiated properties of a kidney epithelial cell line (MDCK). Am J Physiol 240:C106-C109

    Google Scholar 

  21. Mohrmann, I, Mohrmann M, Biber J, Murer H (1986) Stimulation of Na+/phosphate cotransport in LLC-PK1 cells by 12-o-tetradecanoylphorbol 13-acetate (TPA). Biochim Biophys Acta 860:35–43

    Google Scholar 

  22. Nishizuka Y (1984) The role of protein kinase C in cell surface signal transduction and tumor promotion. Nature 308:693–698

    Google Scholar 

  23. Ohuchi K, Levine L (1978) Stimulation of prostaglandin synthesis by tumor-promoting phorbol-12, 13-diesters in canine kidney (MDCK) cells. J Biol Chem 253:4783–4790

    Google Scholar 

  24. Ojakian GK (1981) Tumour promoter-induced changes in the permeability of epithelial cell tight junctions. Cell 23:95–103

    Google Scholar 

  25. Rindler MJ, McRoberts JA, Milton H, Saier JR (1982) (Na+, K+)-cotransport in the Madin-Darby canine kidney cell line. J Biol Chem 257:2254–2259

    Google Scholar 

  26. Satoh T, Endou H (1990) Inhibitory effect of phorbol ester on sodium transport in frog urinary bladder. Am J Physiol 259:F425-F431

    Google Scholar 

  27. Seri I, Kone BC, Gullans SR, Aperia A, Brenner BM, Ballermann BJ (1988) Locally dopamine inhibits Na+-K+-ATPase activity in rat renal cortical tubule cells. Am J Physiol 255:F666-F673

    Google Scholar 

  28. Seri I, Kone BC, Gullans SR, Brenner BM, Ballermann BJ (1989) Inhibition of Na+-K+-ATPase cortical tubule cells (RCTC) by locally formed DA (DA) involves DA1 but not DA2, receptors (abstract). Kidney Int 35:320

    Google Scholar 

  29. Shahedi M, Laborde K, Lelong B, Oudar O, Sachs C (1991) A cytochemical procedure for determination of Na+-K+-ATPase activity in MDCK cells. Kidney Int (in press)

  30. Taub M, Chuman L, Saier MH, Sato G (1979) Growth of Madin-Darby canine kidney epithelial cell (MDCK) line in hormone-supplemented, serum-free medium. Proc Natl Acad Sci USA 76:3338–3342

    Google Scholar 

  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. Yanase M, Handler JS (1986) Activators of protein kinase C inhibit sodium transport in A6 epithelia. Am J Physiol 250:C517-C522

    Google Scholar 

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Shahedi, M., Laborde, K., Bussières, L. et al. Protein kinase C activation causes inhibition of Na/K-ATPase activity in Madin-Darby canine kidney epithelial (MDCK) cells. Pflügers Arch 420, 269–274 (1992). https://doi.org/10.1007/BF00374458

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  • DOI: https://doi.org/10.1007/BF00374458

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