Pflügers Archiv - European Journal of Physiology

, Volume 413, Issue 3, pp 287–298 | Cite as

The nonselective cation channel in the basolateral membrane of rat exocrine pancreas

Inhibition by 3′,5-dichlorodiphenylamine-2-carboxylic acid (DCDPC) and activation by stilbene disulfonates
  • Heinz Gögelein
  • Bernd Pfannmüller
Transport Processes, Metabolism and Bendocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands

Abstract

Nonselective Ca2+-sensitive cation channels in the basolateral membrane of isolated cells of the rat exocrine pancreas were investigated with the patch clamp technique. With 1.3 mmol/l Ca2+ on the cytosolic side, the mean openstate probabilityPo of one channel was about 0.5. In insideout oriented cell-excised membrane patches the substances diphenylamine-2-carboxylic acid (DPC), 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB) and 3′,5-dichlorodiphenylamine-2-carboxylic acid (DCDPC) were applied to the cytosolic side. These compounds inhibited the nonselective cation channels by increasing the mean channel closed time (slow block). 100 μmol/l of NPPB or DPC decreasedPo from 0.5 (control conditions) to 0.2 and 0.04, respectively, whereas 100 μmol/l of DCDPC blocked the channel completely. All effects were reversible. 1 mmol/l quinine also reducedPo, but in contrast to the abov mentioned substances, it induced fast flickering. Ba2+ (70 mmol/l) and tetraethylammonium (TEA+; 20 mmol/l) had no effects. We investigated also the stilbene disulfonates 4-acetamido-4′-isothiocyanatostilbene-2,2′-disulfonic acid (SITS), 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS) and 4,4′-dinitro-2,2′-stilbenedisulfonate (DNDS). 10 μmol/l SITS applied to the cytosolic side increasedPo from 0.5 to 0.7 and with 100 μmol/l SITS the channels remained nearly permanently in its open state (Po≅1). A similar activation of the channels was also observed with DIDS and DNDS. These effects were poorly reversible. The stilbene disulfonates acted by increasing the channel mean open time. When the channel was inactivated by decreasing bath Ca2+ concentration to 0.1 μmol/l, addition of 100 μmol/l of SITS had no effect. Similarly, reducing bath Ca2+ concentration from 1.3 mmol/l in presence of 100 μmol/l SITS (channels are maximally activated) to 0.1 μmol/l, inactivated the channels completely. These results demonstrate, that SITS can only activate the channels in the presence of Ca2+. SITS had no effects, when applied to the extracellular side in outside out patches. In summary, the substances DPC, NPPB and DCDPC inhibit nonselective cation channels, where DCDPC has the most potent and NPPB the smallest effect; whereas SITS, DIDS and DNDS activate the channel when applied from the cytosolic side in the presence of Ca2+ ions.

Key words

Nonselective cation channels Rat exocrine pancreas Diphenylamine-2-carboxylic acid (DPC) 3′,5-Dichlorodiphenylamine-2-carboxylic acid (DCDPC) 4-Acetamino-4′-isothiocyanatostilbene-2,2′-disulfonic acid (SITS) 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Argent BE, Arkle S, Gray MA, Greenwell JR (1987) Two types of calcium-sensitive cation channels in isolated rat pancreatic duct cells. J Physiol 386:82PGoogle Scholar
  2. 2.
    Barzilay M, Ship S, Cabantchik ZI (1979) Anion transport in red blood cells. I. Chemical properties of anion recognition sites as revealed by structure-activity relationships of aromatic sulfonic acids. Membr Biochem 2:227–254CrossRefPubMedGoogle Scholar
  3. 3.
    Benzanilla F (1985) A high capacity data recording device based on a digital audio processor and video cassette recorder. Biophys J 47:437–441CrossRefGoogle Scholar
  4. 4.
    Bevan S, Gray PTA, Ritchie JM, (1984) A calcium-activated cation-selective channel in rat culured Schwann cells. Proc R Soc Lond [Biol] 412:224–232Google Scholar
  5. 5.
    Boron WF, Boupaep EL (1983) Intracellular pH regulation in the renal proximal tubule of the salamander. Basolateral HCO3 transport. J Gen Physiol 81:53–94CrossRefPubMedGoogle Scholar
  6. 6.
    Cabantchik ZI, Rothstein A (1972) The nature of the membrane sites controlling anion permeability of human red blood cells as determined by studies with disulfonic stilbene derivatives. J Membr Biol 10:311–330CrossRefPubMedGoogle Scholar
  7. 7.
    Cabantchik ZI, Rothstein A (1974) Membrane proteins related to anion permeability of human red blood cells. J Membr Biol 15:207–226CrossRefPubMedGoogle Scholar
  8. 8.
    Colquhoun D, Hawkes AG (1981) On the stochastic properties of single ion channels. Proc R Soc Lond [Biol] 211:205–235CrossRefGoogle Scholar
  9. 9.
    Colquhoun D, Sigworth F (1983) Fitting and statistical analysis of single-channel recording. In: Sakmann B, Neher E (eds) Single channel recording. Plenum Press. New York London, pp 191–264CrossRefGoogle Scholar
  10. 10.
    Colquhoun D, Neher E, Reuter H, Stevens CF (1981) Inward current channels activated by intracellular Ca in cultured cardiac cells. Nature 294:752–754CrossRefPubMedGoogle Scholar
  11. 11.
    Cooper KE, Tang JM, Rae JL, Eisenberg RS (1986) A cation channel in frog lens epithelia responsive to pressure and calcium. J Membr Biol 93:259–269CrossRefPubMedGoogle Scholar
  12. 12.
    Di Stefano A, Wittner M, Schlatter E, Lang HJ, Englert H, Greger R (1985) Diphenylamine-2-carboxylate, a blocker of the Cl-conductive pathway in Cl-transporting epithelia. Pflügers Arch 405:S95-S100CrossRefPubMedGoogle Scholar
  13. 13.
    Dreinhöfer J, Gögelein H, Greger R (1988) Blocking effect of 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB) on Cl channels in the colon carcinoma cells HT29. Pflügers Arch 411:R76CrossRefGoogle Scholar
  14. 14.
    Eaton DC, Brodwick S (1979) Amino group reagents affect inactivation in squid axon. Biophys J 25:305aGoogle Scholar
  15. 15.
    Eaton DC, Brodwick MS, Oxford GS, Rudy B (1978) Arginine-specific reagents remove sodium channel inactivation. Nature 271:473–476CrossRefPubMedGoogle Scholar
  16. 16.
    Findlay I, Dunne MJ, Ullrich S, Wollheim CB, Petersen OH (1985) Quinine inhibits Ca2+-independent K+ channels whereas tetraethylammonium inhibits Ca2+-activated K+ channels in insulin-secreting cells. FEBS Lett 185:4–8CrossRefPubMedGoogle Scholar
  17. 17.
    Fröhlich O (1982) The external anion binding site of the human erythrocyte anion transporter: DNDS binding and competition with chloride. J Membr Biol 65:111–123CrossRefPubMedGoogle Scholar
  18. 18.
    Frömter E, Ullrich KJ (1980) Effect of inhibitiors and the mechanisms of anion transport in the proximal renal tubule of rats. Ann NY Acad Sci 341:97–110CrossRefPubMedGoogle Scholar
  19. 19.
    Gögelein H, Greger R (1986) A voltage-dependent ionic channel in the basolateral membrane of late proximal tubules of the rabbit kidney. Pflügers Arch 407:S142-S148CrossRefPubMedGoogle Scholar
  20. 20.
    Gögelein H, Greger R (1987) Properties of single K+ channels in the basolateral membrane of rabbit proximal straight tubules. Pflügers Arch 410:288–295CrossRefPubMedGoogle Scholar
  21. 21.
    Gögelein H, Greger R (1988) Patch clamp analysis of ionic channels in renal proximal tubules. In: Davidson AM (ed) Nephrology, vol I, Proceedings of the Xth International Congress on Nephrology. Bailliere Tindall & Co, London, pp 159–178Google Scholar
  22. 22.
    Gögelein H, Pfannmüller B (1988) Diphenylamine-2-carboxylate inhibits and SITS activates nonselective cation channels in the rat exocrine pancreas. Pflügers Arch 411:R108Google Scholar
  23. 23.
    Gögelein H, Greger R, Schlatter E (1987) Potassium channels in the basolateral membrane of the rectal gland ofSqualus acanthias. Regulation and inhibitors. Pflügers Arch 409:107–113CrossRefPubMedGoogle Scholar
  24. 24.
    Greger R (1988) Chloride channel blockers. In: Fleischer B, Fleischer S (eds) Methods in enzymology, vol 5. Academic Press, New York London (in press)Google Scholar
  25. 25.
    Greger R, Gögelein H (1987) Role of K+ conductive pathways in the nephron. Kidney Int 31:1055–1064CrossRefPubMedGoogle Scholar
  26. 26.
    Greger R, Lang HJ, Englert HC, Wangemann P (1987) Blockers of the Na+2ClK+ carrier and of chloride channels in the thick ascending limb of the loop of Henle. In: Puschett JB, Greenberg A (eds) Diuretics II. Elsevier, Amsterdam New York, pp 131–137Google Scholar
  27. 27.
    Greger R, Schlatter E, Gögelein H (1987) Chloride channels in the luminal membrane of the rectal gland of the dogfish (Squalus acanthias). Properties of the “larger” conductance channel. Pflügers Arch 409:114–121CrossRefPubMedGoogle Scholar
  28. 28.
    Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100CrossRefPubMedGoogle Scholar
  29. 29.
    Hanrahan JW, Alles WP, Lewis SA (1985) Single anion-selective channels in basolateral membrane of a mammalian tight epithelium. Proc Natl Acad Sci USA 82:7791–7795CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Hayslett JP, Gögelein H, Kunzelmann K, Greger R (1987) Characteristics of apical chloride channels in human colon cells (HT29). Pflügers Arch 410:487–494CrossRefPubMedGoogle Scholar
  31. 31.
    Hess P, Lansman JB, Tsien RW (1984) Different modes of Ca channel gating behaviour favoured by dihydropyridine Ca agonists and antagonists. Nature 311:538–544CrossRefPubMedGoogle Scholar
  32. 32.
    Hille B (1984) Ionic channels of excitable membranes. Sinauer Associates Inc., Sunderland, MA, p 275Google Scholar
  33. 33.
    Horn R, Brodwick MS, Eaton DC (1980) Effect of protein cross-linking reagents on membrane currents of squid axon. Am J Physiol 238:C127-C132PubMedGoogle Scholar
  34. 34.
    Iwatsuki N, Petersen OH (1985) Inhibition of Ca2+-activated K+ channels in pig pancreatic acinar cells by Ba2+, Ca2+, quinine and quinidine. Biochim Biophys Acta 819:249–257CrossRefPubMedGoogle Scholar
  35. 35.
    Kokubun S, Reuter H (1984) Dihydropyridine derivatives prolong the open state of Ca channels in cultured cardiac cells. Proc Natl Acad Sci USA 81:4824–4827CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Lepke S, Fasold H, Pring M, Passow H (1976) A study of the relationship between inhibition of anion exchange and binding to the red blood cell membrane of 4,4′-diisothiocyano stilbene-2,2′-disulfonic acid (DIDS) and its dihydro derivative (H2DIDS). J Membr Biol 29:147–177CrossRefPubMedGoogle Scholar
  37. 37.
    Lindau M, Fernandez JM (1986) A patch-clamp study of histamine-secreting cells. J Gen Physiol 88:349–368CrossRefPubMedGoogle Scholar
  38. 38.
    Lipton SA (1986) Antibody activates cationic channels via second messenger Ca2+. Biochim Biophys Acta 856:59–67CrossRefPubMedGoogle Scholar
  39. 39.
    Löw I, Friedrich T, Burckhardt G (1984) Properties of an anion exchanger in rat renal basolateral membrane vesicles. Am J Physiol 246:F334-F342PubMedGoogle Scholar
  40. 40.
    Magleby KL, Pallotta BS (1983) Calcium dependence of open and shut interval distributions from calcium-activated potassium channels in cultured rat muscle. J Physiol 344:585–604CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Marty A, Tan YP, Trautmann A (1984) Three types of calciumdependent channel in rat lacrimal glands. J Physiol 357:293–325CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Maruyama Y, Petersen OH (1982) Single-channel currents in isolated patches of plasma membrane from basal surface of pancreatic acini. Nature 299:159–161CrossRefPubMedGoogle Scholar
  43. 43.
    Maruyama Y, Petersen OH (1982) Cholecystokinin activation of single-channel currents is mediated by internal messenger in pancreatic acinar cells. Nature 300:61–63CrossRefPubMedGoogle Scholar
  44. 44.
    Maruyama Y, Petersen OH (1984) Single calcium-dependent cation channels in mouse pancreatic acinar cells. J Membr Biol 81:83–87CrossRefPubMedGoogle Scholar
  45. 45.
    Maruyama Y, Gallacher DV, Petersen OH (1983) Voltage and Ca2+-activated K+ channel in basolateral acinar cell membranes of mammalian salivary glands. Nature 302:827–829CrossRefPubMedGoogle Scholar
  46. 46.
    Maruyama Y, Moore D, Petersen OH (1985) Calcium-activated cation channel in rat thyroid follicular cells. Biochim Biophys Acta 821:229–232CrossRefPubMedGoogle Scholar
  47. 47.
    Miller C, White MC (1984) Dimeric structure of single chloride channels fromTorpedo electroplax. Proc natl Acad Sci USA 81:2772–2775CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Moczydlowski E Latorre R (1983) Gating kinetics of Ca2+-activated K+ channels from rat muscle incorporated into lipid bilayers. Evidence for two voltage-dependent Ca2+ binding reactions. J Gen Physiol 82:511–542CrossRefPubMedGoogle Scholar
  49. 49.
    Nonner W, Spalding BC, Hille B (1980) Low intracellular pH and chemical agents slow inactivation gating in sodium channels of muscle. Nature 284:360–363CrossRefPubMedGoogle Scholar
  50. 50.
    Oxford GS, Wu CH, Narahashi T (1978) Removal of sodium channel inactivation in squid giant axons by n-bromoacetamide. J Gen Physiol 71:227–247CrossRefPubMedGoogle Scholar
  51. 51.
    Partridge LD, Swandulla D (1987) Single Ca-activated cation channels in bursting neurons of Helix. Pflügers Arch 410:627–631CrossRefPubMedGoogle Scholar
  52. 52.
    Partidge LD, Swandulla D (1988) Calcium-activated nonspecific cation channels. TINS 11:69–72Google Scholar
  53. 53.
    Passow H (1986) Molecular aspects of band 3 protein-mediated anion transport across the red blood cell membrane. Rev Physiol Biochem Pharmacol 103:61–203PubMedGoogle Scholar
  54. 54.
    Patlak J, Horn R (1982) Effect of n-bromoacetamide on single sodium channel currents in excised membrane patches. J Gen Physiol 79:333–351CrossRefPubMedGoogle Scholar
  55. 55.
    Reuss L, Constantin JL, Bazile JE (1987) Diphenylamine-2-carboxylate blocks Cl−HCO3 exchange in Necturus gallbladder epithelium. Am J Physiol 253:C79-C89PubMedGoogle Scholar
  56. 56.
    Schwarz W, Passow H (1983) Ca2+-activated K+ channels in erythrocytes and excitable cells. Annu Rev Physiol 45:359–374CrossRefPubMedGoogle Scholar
  57. 57.
    Siemen D, Reuhl T (1987) Non-selective cationic channel in primary cultured cells of brown adipose tissue. Pflügers Arch 408:534–536CrossRefPubMedGoogle Scholar
  58. 58.
    Simonneau, M., Distasi C, Tauc L, Barbin G (1987) Potassium channels in mouse neonatal root ganglion cells: a patch-clamp study. Brain Res 412:224–232CrossRefPubMedGoogle Scholar
  59. 59.
    Skydsgaard JM (1987) Inhibition of chloride self-exchange with stilbene disulphonates in depolarized skeletal muscle ofRana temporaria. J Physiol 397:433–447CrossRefGoogle Scholar
  60. 60.
    Streb H, Schulz I (1983) Regulation of cytosolic free Ca2+ concentration in acinar cells of rat pancreas. Am J Physiol 245:G347-G357PubMedGoogle Scholar
  61. 61.
    Sturgess NC, Hales N, Ashford MLJ (1986) Inhibition of a calcium-activated, non-selective cation channel, in a rat insulinoma cell line, by adenine derivates. FEBS Lett 208:397–400CrossRefPubMedGoogle Scholar
  62. 62.
    Sturgess NC, Hales CN, Ashford MLJ (1987) Calcium and ATP regulate the activity of a non-selective cation channel in a rat insolinoma cell line. Pflügers Arch 409:607–615CrossRefPubMedGoogle Scholar
  63. 63.
    Teulon J, Paulais M, Bouthier M (1987) A Ca2+-activated cation-selective channel in the basolateral membrane of the cortical thick ascending limb of Henle's loop of the mouse. Biochim Biophys Acta 905:125–132CrossRefPubMedGoogle Scholar
  64. 64.
    Vergara C, Latorre R (1983) Kinetics of Ca2+-activated K+ channels from rabbit muscle incorporated into planar bilayer. J Gen Physiol 82:543–568CrossRefPubMedGoogle Scholar
  65. 65.
    von Tscharner V, Prod'hom B, Baggiolini M, Reuter H (1986) Ion channels in human neutrophils activated by a rise in free cytosolic calcium concentration. Nature 324:369–372CrossRefGoogle Scholar
  66. 66.
    Wangemann P (1987) Chlorid-Kanal-Blocker an der dicken aufsteigenden Henle-Schleife der Säugerniere. Struktur-Wirkungsbeziehungen. Thesis, Albert-Ludwigs-Universität Freiburg, FRGGoogle Scholar
  67. 67.
    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-S141CrossRefPubMedGoogle Scholar
  68. 68.
    Weston AH, Abbot A (1987) New class of antihypertensive acts by opening K+ channels. TIPS 8:283–284Google Scholar
  69. 69.
    Yellen G (1982) Single Ca2+-activated nonselective cation channels in neuroblastoma. Nature 296:357–359CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • Heinz Gögelein
    • 1
  • Bernd Pfannmüller
    • 1
  1. 1.Max-Planck-Institut für BiophysikFrankfurt/Main 70Germany

Personalised recommendations