The Journal of Membrane Biology

, Volume 193, Issue 3, pp 153–170 | Cite as

Swelling-activated Chloride and Potassium Conductance in Primary Cultures of Mouse Proximal Tubules. Implication of KCNE1 Protein

  • H. Barrière
  • I. Rubera
  • R. Belfodil
  • M. Tauc
  • N. Tonnerieux
  • C. Poujeol
  • J. Barhanin
  • P. Poujeol
Article

Abstract

Volume-sensitive chloride and potassium currents were studied, using the whole-cell clamp technique, in cultured wild-type mouse proximal convoluted tubule (PCT) epithelial cells and compared with those measured in PCT cells from null mutant kcne1 −/− mice. In wild-type PCT cells in primary culture, a Cl conductance activated by cell swelling was identified. The initial current exhibited an outwardly rectifying current-voltage (I-V) relationship, whereas steady-state current showed decay at depolarized membrane potentials. The ion selectivity was I > Br > Cl >> gluconate. This conductance was sensitive to 1 mM 4,4′-Diisothiocyanostilbene-2,2′-disulfonic acid (DIDS), 0.1 mM 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) and 1 mM diphenylamine-2-carboxylate (DPC). Osmotic stress also activated K+ currents. These currents are time-independent, activated at depolarized potentials, and inhibited by 0.5 mM quinidine, 5 mM barium, and 10 µM clofilium but are insensitive to 1 mM tetraethylammonium (TEA), 10 nM charybdotoxin (CTX), and 10 µM 293B. In contrast, the null mutation of kcne1 completely impaired volume-sensitive chloride and potassium currents in PCT. The transitory transfection of kcne1 restores both Cl and K+ swelling-activated currents, confirming the implication of KCNE1 protein in the cell-volume regulation in PCT cells in primary cultures.

Keywords

Cell volume kcne1 Kidney Chloride conductance Potassium conductance 

References

  1. 1.
    Arrazola, A., Rota, R., Hannaert, P., Soler, A., Garay, R.P. 1993Cell volume regulation in rat thymocytes.J. Physiol.465403414PubMedGoogle Scholar
  2. 2.
    Attali, B., Guillemare, E., Lesage, F., Honore, E., Romey, G., Lazdunski, M., Barhanin, J. 1993The protein IsK is a dual activator of K+ and Cl channels.Nature365850852PubMedGoogle Scholar
  3. 3.
    Barhanin, J., Lesage, F., Guillemare, E., Fink, M., Lazdunski, M., Romey, G. 1996K(V)LQT1 and IsK (minK) proteins associate to form the I(Ks) cardiac potassium current.Nature3847880PubMedGoogle Scholar
  4. 4.
    Ben-Efraim, I., Shai, Y., Attali, B. 1996Cytoplasmic and extracellular IsK peptides activate endogenous K+ and Cl channels in Xenopus oocytes. Evidence for regulatory function.J. Biol. Chem.27187688771CrossRefPubMedGoogle Scholar
  5. 5.
    Bidet, M., Tauc, M., Rubera, I., de Renzis, G., Poujeol, C., Bohn, M.T., Poujeol, P. 1996Calcium-activated chloride currents in primary cultures of rabbit distal convoluted tubule.Am. J. Physiol.271F940F950PubMedGoogle Scholar
  6. 6.
    Bleich, M., Warth, R. 2000The very small-conductance K+ channel KvLQT1 and epithelial function.Pfluegers Arch.440202206Google Scholar
  7. 7.
    Boucherot, A., Schreiber, R., Kunzehmann, K. 2001Regulation and properties of KCNQ1 (K(V)LQT1) and impact of the cystic fibrosis transmembrane conductance regulator.J. Membrane Biol.1823947Google Scholar
  8. 8.
    Busch, A.E., Varnum, M., Adelman, J.P., North, R.A. 1992Hypotonic solution increases the slowly activating potassium current IsK expressed in Xenopus oocytes.Biochem. Biophys. Res. Commun.184804810PubMedGoogle Scholar
  9. 9.
    Chouabe, C., Neyroud, N., Guicheney, P., Lazdunski, M., Romey, G., Barhanin, J. 1997Properties of KvLQT1 K+ channel mutations in Romano-Ward and Jervell and Lange-Nielsen inherited cardiac arrhythmias.Embo J.1654725479CrossRefPubMedGoogle Scholar
  10. 10.
    Demolombe, S., Franco, D., de Boer, P., Kuperschmidt, S., Roden, D., Pereon, Y., Jarry, A., Moorman, A.F., Escande, D. 2001Differential expression of KvLQT1 and its regulator IsK in mouse epithelia.Am. J. Physiol.280C359C372Google Scholar
  11. 11.
    Eveloff, J.L., Warnock, D.G. 1987Activation of ion transport systems during cell volume regulation.Am. J. Physiol.252F1F10PubMedGoogle Scholar
  12. 12.
    Freeman, L.C., Kass, R.S. 1993Expression of a minimal K+ channel protein in mammalian cells and immunolocalization in guinea pig heart.Circ. Res.73968973PubMedGoogle Scholar
  13. 13.
    Frizzell, R., Morris, A. 1994Chloride conductance of salt-secreting epithelial cells.Curr. Top. Membr.: Chloride Channels173214Google Scholar
  14. 14.
    Greenwood, I.A., Miller, L.J., Ohya, S., Horowitz, B. 2002The large conductance potassium channel beta-subunit can interact with and modulate the functional properties of a calcium-activated chloride channel, CLCA1.J. Biol. Chem.2772211922222CrossRefPubMedGoogle Scholar
  15. 15.
    Greger, R., Schlatter, E. 1984Mechanism of NaCl secretion in the rectal gland of spiny dogfish (Squalus acanthias). I. Experiments in isolated in vitro perfused rectal gland tubules.Pfluegers Arch.4026375Google Scholar
  16. 16.
    Hoffmann, E.K. 2000Intracellular signalling involved in volume regulatory decrease.Cell Physiol. Biochem.10273288CrossRefPubMedGoogle Scholar
  17. 17.
    Hoffmann, E.K., Hougaard, C. 2001Intracellular signalling involved in activation of the volume-sensitive K+ current in Ehrlich ascites tumour cells.Comp. Biochem. Physiol. A Mol. Integr. Physiol.130355366CrossRefPubMedGoogle Scholar
  18. 18.
    Jurman, M.E., Boland, L.M., Liu, Y, Yellen, G. 1994Visual identification of individual transfected cells for electrophysiology using antibody-coated beads.Biotechniques17876881PubMedGoogle Scholar
  19. 19.
    Knoblauch, C., Montrose, M.H., Murer, H. 1989Regulatory volume decrease by cultured renal cells.Am. J. Physiol.256C252C259PubMedGoogle Scholar
  20. 20.
    le Maout, S., Tauc, M., Koechlin, N., Poujeol, P. 1990Polarized 86Rb+ effluxes in primary cultures of rabbit kidney proximal cells: role of calcium ana hypotonicity.Biochim. Biophys. Acta10262939CrossRefPubMedGoogle Scholar
  21. 21.
    Lock, H., Valverde, M.A. 2000Contribution of the IsK (MinK) potassium channel subunit to regulatory volume decrease in murine tracheal epithelial cells.J. Biol. Chem.2753484934852CrossRefPubMedGoogle Scholar
  22. 22.
    MacLeod, R.J., Hamilton, J.R. 1996Activation of Na+/H+ exchange is required for regulatory volume decrease after modest “physiological” volume increases in jejunal villus epithelial cells.J. Biol. Chem.2712313823145CrossRefPubMedGoogle Scholar
  23. 23.
    Mall, M., Wissner, A., Schreiber, R., Kuehr, J., Seydewitz, H.H., Brandis, M., Greger, R., Kunzelmann, K. 2000Role of K(V)LQT1 in cyclic adenosine monophosphate-mediated Cl secretion in human airway epithelia.Am. J. Respir. Cell Mol. Biol.23283289PubMedGoogle Scholar
  24. 24.
    Merot, J., Bidet, M., Gachot, B., Le Maout, S., Koechlin, N., Tauc, M., Poujeol, P. 1989Electrical properties of rabbit early distal convoluted tubule in primary culture.Am. J. Physiol.257F288F299PubMedGoogle Scholar
  25. 25.
    Merot, J., Poncet, V., Bidet, M., Tauc, M., Poujeol, P. 1991Apical membrane ionic channels in the rabbit cortical thick ascending limb in primary culture.Biochim. Biophys. Acta1070387400CrossRefPubMedGoogle Scholar
  26. 26.
    Moorman, J.R., Palmer, C.J., John 3rd, J.E., Durieux, M.E., Jones, L.R. 1992Phospholemman expression induces a hyperpolarization-activated chloride current in Xenopus oocytes.J. Biol. Chem.2671455114554PubMedGoogle Scholar
  27. 27.
    Niemeyer, M.I., Cid, L.P., Barros, L.F., Sepulveda, F.V. 2001Modulation of the two-pore domain acid-sensitive K+ channel TASK-2 (KCNK5) by changes in cell volume.J. Biol. Chem.2764316643174CrossRefPubMedGoogle Scholar
  28. 28.
    Poncet, V., Tauc, M., Bidet, M., Poujeol, P. 1994Chloride channels in apical membrane of primary cultures of rabbit distal bright convoluted tubule.Am. J. Physiol.266F543F553PubMedGoogle Scholar
  29. 29.
    Rubera, I., Tauc, M., Bidet, M., Poujeol, C., Cuiller, B., Watrin, A., Touret, N., Poujeol, P. 1998Chloride currents in primary cultures of rabbit proximal and distal convoluted tubules.Am. J. Physiol.275F651F663PubMedGoogle Scholar
  30. 30.
    Rubera, I., Tauc, M., Michel, F., Poujeol, C., Poujeol, P. 1997aSimultaneous functional expression of swelling and forskolin-activated chloride currents in primary cultures of rabbit distal convoluted tubule.C. R. Acad. Sci. III320223232Google Scholar
  31. 31.
    Rubera, I., Tauc., M., Poujeol, C., Bohn, M.T., Bidet, M., De Renzis, G., Poujeol, P. 1997bCl and K+ conductances activated by cell swelling in primary cultures of rabbit distal bright convoluted tubules.Am. J. Physiol.273F680F697Google Scholar
  32. 32.
    Shiga, N., Wangemann, P. 1995Ion selectivity of volume regulatory mechanisms present during a hypoosmotic challenge in vestibular dark cells.Biochim. Biophys. Acta12404854CrossRefPubMedGoogle Scholar
  33. 33.
    Strange, K. 1988RVD in principal and intercalated cells of rabbit cortical collecting tubule.Am. J. Physiol.255C612C621PubMedGoogle Scholar
  34. 34.
    Sugimoto, T., Tanabe, Y., Shigemoto, R., Iwai, M., Takumi, T., Ohkubo, H., Nakanishi, S. 1990Immunohistochemical study of a rat membrane protein which induces a selective potassium permeation: its localization in the apical membrane portion of epithelial cells.J. Membrane Biol.1133947Google Scholar
  35. 35.
    Takumi, T., Ohkubo, H., Nakanishi, S. 1988Cloning of a membrane protein that induces a slow voltage-gated potassium current.Science24210421045PubMedGoogle Scholar
  36. 36.
    Tauc, M., Bidet, M., Poujeol, P. 1996Chloride currents activated by calcitonin and cAMP in primary cultures of rabbit distal convoluted tubule.J. Membrane Biol.150255273CrossRefGoogle Scholar
  37. 37.
    Tauc, M., Le Maout, S., Poujeol, P. 1990Fluorescent video-microscopy study of regulatory volume decrease in primary culture of rabbit proximal convoluted tubule.Biochim. Biophys. Acta1052278284CrossRefPubMedGoogle Scholar
  38. 38.
    Vallon, V., Grahammer, F., Richter, K., Bleich, M., Lang, F., Barhanin, J., Volkl, H., Warth, R. 2001Role of KCNE1-dependent K+ fluxes in mouse proximal tubule.J. Am. Soc. Nephrol.1220032011PubMedGoogle Scholar
  39. 39.
    Vanoye, C.G., Reuss, L. 1999Stretch-activated single K+ channels account for whole-cell currents elicited by swelling.Proc. Natl. Acad. Sci. USA9665116516CrossRefPubMedGoogle Scholar
  40. 40.
    Vetter, D.E., Mann, J.R., Wangemann, P., Liu, J., McLaughlin, K.J., Lesage, F., Marcus, D.C., Lazdunski, M., Heinemann, S.F., Barhanin, J. 1996Inner ear defects induced by null mutation of the isk gene.Neuron1712511264PubMedGoogle Scholar
  41. 41.
    vom Dahl, S., Schliess, F., Graf, D., Haussinger, D. 2001Role of p38(MAPK) in cell volume regulation of perfused rat liver.Cell Physiol. Biochem.11285294CrossRefPubMedGoogle Scholar
  42. 42.
    Wangemann, P., Liu, J., Shen, Z., Shipley, A., Marcus, D.C. 1995Hypo-osmotic challenge stimulates transepithelial K+ secretion and activates apical IsK channel in vestibular dark cells.J. Membrane Biol.147263273Google Scholar
  43. 43.
    Warth, R., Riedemann, N., Bleich, M., Van Driessche, W., Busch, A.E., Greger, R. 1996The cAMP-regulated and 293B-inhibited K+ conductance of rat colonic crypt base cells.Pfluegers Arch.4328188CrossRefGoogle Scholar

Copyright information

© Springer-Verlag New York Inc. 2003

Authors and Affiliations

  • H. Barrière
    • 1
  • I. Rubera
    • 1
  • R. Belfodil
    • 1
  • M. Tauc
    • 1
  • N. Tonnerieux
    • 1
  • C. Poujeol
    • 1
  • J. Barhanin
    • 2
  • P. Poujeol
    • 1
  1. 1.UMR CNRS 6548Université de Nice-Sophia Antipolis, 06108 Nice Cedex 2France
  2. 2.UMR CNRS 6097IPMC, Sophia Antipolis, 06560 ValbonneFrance

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