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. 1981 Feb;78(2):1057–1061. doi: 10.1073/pnas.78.2.1057

Reduction of K+ efflux in cultured mouse fibroblasts, by mutation or by diuretics, permits growth in K+-deficient medium.

D W Jayme, E A Adelberg, C W Slayman
PMCID: PMC319945  PMID: 6940122

Abstract

The mouse fibroblastic cell line LM(TK-) is unable to grow at external K+ concentrations below a threshold value of 0.4 mM. At subthreshold K+ concentrations, LM(TK-) cells rapidly lose intracellular K+ and eventually lyse. We have analyzed the pathway primarily responsible for K+ efflux under these experimental conditions and reports its specific inhibition by two diuretics, furosemide and bumetanide. Bumetanide, an analog of furosemide, was a more potent inhibitor (by several orders of magnitude) than was furosemide itself. The effects of ouabain and bumetanide were additive, suggesting independence of diuretic-sensitive K+ efflux from Na+/K+ pump-mediated fluxes. Characterization of K+ efflux in LTK-5, a mutant derived from LM(TK-) and selected for its ability to grow at 0.2 mM K+ indicated that the mutant had lost the diuretic-sensitive K+ efflux pathway. Net cation fluxes, steady-state intracellular cation concentrations, and growth at reduced K+ concentrations were comparable for LM(TK-) cells maximally inhibited by diuretics and for the LTK-5 mutant grown either in the presence or absence of diuretics. Thus, reduction in K+ efflux, either by diuretic addition diuretics. Thus, reduction in K+ efflux, either by diuretic addition or by genetic alteration, can permit the cell to maintain normal cation gradients and to grow at otherwise subthreshold external K+ concentrations.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Frederiksen O. Functional distinction between two transport mechanisms in rabbit gall-bladder epithelium by use of ouabain, ethacrynic acid and metabolic inhibitors. J Physiol. 1978 Jul;280:373–387. doi: 10.1113/jphysiol.1978.sp012389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Gargus J. J., Miller I. L., Slayman C. W., Adelberg E. A. Genetic alterations in potassium transport in L cells. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5589–5593. doi: 10.1073/pnas.75.11.5589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gargus J. J., Slayman C. W. Mechanism and role of furosemide-sensitive K+ transport in L cells: a genetic approach. J Membr Biol. 1980;52(3):245–256. doi: 10.1007/BF01869193. [DOI] [PubMed] [Google Scholar]
  4. Geck P., Heinz E. Coupling in secondary transport. Effect of electrical potentials on the kinetics of ion linked co-transport. Biochim Biophys Acta. 1976 Aug 4;443(1):49–63. doi: 10.1016/0005-2736(76)90490-9. [DOI] [PubMed] [Google Scholar]
  5. Geck P., Pietrzyk C., Burckhardt B. C., Pfeiffer B., Heinz E. Electrically silent cotransport on Na+, K+ and Cl- in Ehrlich cells. Biochim Biophys Acta. 1980 Aug 4;600(2):432–447. doi: 10.1016/0005-2736(80)90446-0. [DOI] [PubMed] [Google Scholar]
  6. Gunn R. B. Co- and counter-transport mechanisms in cell membranes. Annu Rev Physiol. 1980;42:249–259. doi: 10.1146/annurev.ph.42.030180.001341. [DOI] [PubMed] [Google Scholar]
  7. HODGKIN A. L., KATZ B. The effect of sodium ions on the electrical activity of giant axon of the squid. J Physiol. 1949 Mar 1;108(1):37–77. doi: 10.1113/jphysiol.1949.sp004310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hoffman J. F., Kregenow F. M. The characterization of new energy dependent cation transport processes in red blood cells. Ann N Y Acad Sci. 1966 Jul 14;137(2):566–576. doi: 10.1111/j.1749-6632.1966.tb50182.x. [DOI] [PubMed] [Google Scholar]
  9. KIT S., DUBBS D. R., PIEKARSKI L. J., HSU T. C. DELETION OF THYMIDINE KINASE ACTIVITY FROM L CELLS RESISTANT TO BROMODEOXYURIDINE. Exp Cell Res. 1963 Aug;31:297–312. doi: 10.1016/0014-4827(63)90007-7. [DOI] [PubMed] [Google Scholar]
  10. Kregenow F. M. The response of duck erythrocytes to nonhemolytic hypotonic media. Evidence for a volume-controlling mechanism. J Gen Physiol. 1971 Oct;58(4):372–395. doi: 10.1085/jgp.58.4.372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lamb J. F., Lindsay R. Transient effects of ethacrynic acid on Na and K movements in cultured cells. Q J Exp Physiol Cogn Med Sci. 1973 Oct;58(4):345–355. doi: 10.1113/expphysiol.1973.sp002228. [DOI] [PubMed] [Google Scholar]
  12. Mills B., Tupper J. T. Cation permeability and ouabain-insensitive cation flux in the Ehrlich ascites tumor cell. J Membr Biol. 1975;20(1-2):75–97. doi: 10.1007/BF01870629. [DOI] [PubMed] [Google Scholar]
  13. Mills B., Tupper J. T. Cell cycle dependent changes in potassium transport. J Cell Physiol. 1976 Sep;89(1):123–132. doi: 10.1002/jcp.1040890112. [DOI] [PubMed] [Google Scholar]
  14. Nelson P. G., Peacock J. H. Transmission on an active electrical response between fibroblasts (L cells) in cell culture. J Gen Physiol. 1973 Jul;62(1):25–36. doi: 10.1085/jgp.62.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Okada Y., Doida Y., Roy G., Tsuchiya W., Inouye K., Inouye A. Oscillations of membrane potential in L cells. I. Basic characteristics. J Membr Biol. 1977 Aug 4;35(4):319–335. doi: 10.1007/BF01869957. [DOI] [PubMed] [Google Scholar]
  16. Okada Y., Roy G., Tsuchiya W., Doida Y., Inouye A. Oscillations of membrane potential in L cells. II. Effect of monovalent ion concentrations and conductance changes associated with oscillations. J Membr Biol. 1977 Aug 4;35(4):337–350. doi: 10.1007/BF01869958. [DOI] [PubMed] [Google Scholar]
  17. Palfrey H. C., Feit P. W., Greengard P. cAMP-stimulated cation cotransport in avian erythrocytes: inhibition by "loop" diuretics. Am J Physiol. 1980 Mar;238(3):C139–C148. doi: 10.1152/ajpcell.1980.238.3.C139. [DOI] [PubMed] [Google Scholar]
  18. Rindler M. J., Taub M., Saier M. H., Jr Uptake of 22Na+ by cultured dog kidney cells (MDCK). J Biol Chem. 1979 Nov 25;254(22):11431–11439. [PubMed] [Google Scholar]
  19. Schmidt W. F., 3rd, McManus T. J. Ouabain-insensitive salt and water movements in duck red cells. I. Kinetics of cation transport under hypertonic conditions. J Gen Physiol. 1977 Jul;70(1):59–79. doi: 10.1085/jgp.70.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schultz S. G., Curran P. F. Coupled transport of sodium and organic solutes. Physiol Rev. 1970 Oct;50(4):637–718. doi: 10.1152/physrev.1970.50.4.637. [DOI] [PubMed] [Google Scholar]
  21. Spaggiare S., Wallach M. J., Tupper J. T. Potassium transport in normal and transformed mouse 3T3 cells. J Cell Physiol. 1976 Nov;89(3):403–416. doi: 10.1002/jcp.1040890306. [DOI] [PubMed] [Google Scholar]
  22. TOSTESON D. C., HOFFMAN J. F. Regulation of cell volume by active cation transport in high and low potassium sheep red cells. J Gen Physiol. 1960 Sep;44:169–194. doi: 10.1085/jgp.44.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tupper J. T. Cation flux in the ehrlich ascites tumor cell. Evidence for Na+-for-Na+ and K+-for-K+ exchange diffusion. Biochim Biophys Acta. 1975 Jul 18;394(4):586–596. doi: 10.1016/0005-2736(75)90144-3. [DOI] [PubMed] [Google Scholar]
  24. Tupper J. T., Zorgniotti F., Mills B. Potassium transport and content during G1 and S phase following serum stimulation of 3T3 cells. J Cell Physiol. 1977 Jun;91(3):429–440. doi: 10.1002/jcp.1040910313. [DOI] [PubMed] [Google Scholar]
  25. Wiley J. S., Cooper R. A. A furosemide-sensitive cotransport of sodium plus potassium in the human red cell. J Clin Invest. 1974 Mar;53(3):745–755. doi: 10.1172/JCI107613. [DOI] [PMC free article] [PubMed] [Google Scholar]

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