Skip to main content
The Journal of General Physiology logoLink to The Journal of General Physiology
. 1990 May 1;95(5):791–818. doi: 10.1085/jgp.95.5.791

Maxi K+ channels and their relationship to the apical membrane conductance in Necturus gallbladder epithelium

PMCID: PMC2216345  PMID: 2362182

Abstract

Using the patch-clamp technique, we have identified large-conductance (maxi) K+ channels in the apical membrane of Necturus gallbladder epithelium, and in dissociated gallbladder epithelial cells. These channels are more than tenfold selective for K+ over Na+, and exhibit unitary conductance of approximately 200 pS in symmetric 100 mM KCl. They are activated by elevation of internal Ca2+ levels and membrane depolarization. The properties of these channels could account for the previously observed voltage and Ca2+ sensitivities of the macroscopic apical membrane conductance (Ga). Ga was determined as a function of apical membrane voltage, using intracellular microelectrode techniques. Its value was 180 microS/cm2 at the control membrane voltage of -68 mV, and increased steeply with membrane depolarization, reaching 650 microS/cm2 at -25 mV. We have related maxi K+ channel properties and Ga quantitatively, relying on the premise that at any apical membrane voltage Ga comprises a leakage conductance and a conductance due to maxi K+ channels. Comparison between Ga and maxi K+ channels reveals that the latter are present at a surface density of 0.09/microns 2, are open approximately 15% of the time under control conditions, and account for 17% of control Ga. Depolarizing the apical membrane voltage leads to a steep increase in channel steady-state open probability. When correlated with patch-clamp studies examining the Ca2+ and voltage dependencies of single maxi K+ channels, results from intracellular microelectrode experiments indicate that maxi K+ channel activity in situ is higher than predicted from the measured apical membrane voltage and estimated bulk cytosolic Ca2+ activity. Mechanisms that could account for this finding are proposed.

Full Text

The Full Text of this article is available as a PDF (1.5 MB).

Selected References

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

  1. Bello-Reuss E., Grady T. P., Reuss L. Mechanism of the effect of cyanide on cell membrane potentials in Necturus gall-bladder epithelium. J Physiol. 1981 May;314:343–357. doi: 10.1113/jphysiol.1981.sp013712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bjorkman D. J., Allan C. H., Hagen S. J., Trier J. S. Structural features of absorptive cell and microvillus membrane preparations from rat small intestine. Gastroenterology. 1986 Dec;91(6):1401–1414. doi: 10.1016/0016-5085(86)90194-0. [DOI] [PubMed] [Google Scholar]
  3. Bolívar J. J., Cereijido M. Voltage and Ca2+-activated K+ channel in cultured epithelial cells (MDCK). J Membr Biol. 1987;97(1):43–51. doi: 10.1007/BF01869613. [DOI] [PubMed] [Google Scholar]
  4. Christensen O. Mediation of cell volume regulation by Ca2+ influx through stretch-activated channels. Nature. 1987 Nov 5;330(6143):66–68. doi: 10.1038/330066a0. [DOI] [PubMed] [Google Scholar]
  5. Christensen O., Zeuthen T. Maxi K+ channels in leaky epithelia are regulated by intracellular Ca2+, pH and membrane potential. Pflugers Arch. 1987 Mar;408(3):249–259. doi: 10.1007/BF02181467. [DOI] [PubMed] [Google Scholar]
  6. Cook D. L., Ikeuchi M., Fujimoto W. Y. Lowering of pHi inhibits Ca2+-activated K+ channels in pancreatic B-cells. Nature. 1984 Sep 20;311(5983):269–271. doi: 10.1038/311269a0. [DOI] [PubMed] [Google Scholar]
  7. Eisenman G., Latorre R., Miller C. Multi-ion conduction and selectivity in the high-conductance Ca++-activated K+ channel from skeletal muscle. Biophys J. 1986 Dec;50(6):1025–1034. doi: 10.1016/S0006-3495(86)83546-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Frindt G., Palmer L. G. Ca-activated K channels in apical membrane of mammalian CCT, and their role in K secretion. Am J Physiol. 1987 Mar;252(3 Pt 2):F458–F467. doi: 10.1152/ajprenal.1987.252.3.F458. [DOI] [PubMed] [Google Scholar]
  9. Frindt G., Palmer L. G. Low-conductance K channels in apical membrane of rat cortical collecting tubule. Am J Physiol. 1989 Jan;256(1 Pt 2):F143–F151. doi: 10.1152/ajprenal.1989.256.1.F143. [DOI] [PubMed] [Google Scholar]
  10. García-Díaz J. F., Nagel W., Essig A. Voltage-dependent K conductance at the apical membrane of Necturus gallbladder. Biophys J. 1983 Sep;43(3):269–278. doi: 10.1016/S0006-3495(83)84350-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Golowasch J., Kirkwood A., Miller C. Allosteric effects of Mg2+ on the gating of Ca2+-activated K+ channels from mammalian skeletal muscle. J Exp Biol. 1986 Sep;124:5–13. doi: 10.1242/jeb.124.1.5. [DOI] [PubMed] [Google Scholar]
  12. Grasset E., Gunter-Smith P., Schultz S. G. Effects of Na-coupled alanine transport on intracellular K activities and the K conductance of the basolateral membranes of Necturus small intestine. J Membr Biol. 1983;71(1-2):89–94. doi: 10.1007/BF01870677. [DOI] [PubMed] [Google Scholar]
  13. Greger R., Gögelein H. Role of K+ conductive pathways in the nephron. Kidney Int. 1987 May;31(5):1055–1064. doi: 10.1038/ki.1987.108. [DOI] [PubMed] [Google Scholar]
  14. Guggino S. E., Guggino W. B., Green N., Sacktor B. Ca2+-activated K+ channels in cultured medullary thick ascending limb cells. Am J Physiol. 1987 Feb;252(2 Pt 1):C121–C127. doi: 10.1152/ajpcell.1987.252.2.C121. [DOI] [PubMed] [Google Scholar]
  15. Gunter-Smith P. J. Apical membrane potassium conductance in guinea pig gallbladder epithelial cells. Am J Physiol. 1988 Dec;255(6 Pt 1):C808–C815. doi: 10.1152/ajpcell.1988.255.6.C808. [DOI] [PubMed] [Google Scholar]
  16. Gögelein H., Van Driessche W. Noise analysis of the K+ current through the apical membrane of Necturus gallbladder. J Membr Biol. 1981;60(3):187–198. doi: 10.1007/BF01992557. [DOI] [PubMed] [Google Scholar]
  17. HODGKIN A. L., KEYNES R. D. The potassium permeability of a giant nerve fibre. J Physiol. 1955 Apr 28;128(1):61–88. doi: 10.1113/jphysiol.1955.sp005291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hille B., Schwarz W. Potassium channels as multi-ion single-file pores. J Gen Physiol. 1978 Oct;72(4):409–442. doi: 10.1085/jgp.72.4.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hunter M., Lopes A. G., Boulpaep E. L., Giebisch G. H. Single channel recordings of calcium-activated potassium channels in the apical membrane of rabbit cortical collecting tubules. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4237–4239. doi: 10.1073/pnas.81.13.4237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hunter M., Lopes A. G., Boulpaep E., Giebisch G. Regulation of single potassium ion channels from apical membrane of rabbit collecting tubule. Am J Physiol. 1986 Oct;251(4 Pt 2):F725–F733. doi: 10.1152/ajprenal.1986.251.4.F725. [DOI] [PubMed] [Google Scholar]
  21. Kawahara K., Hunter M., Giebisch G. Potassium channels in Necturus proximal tubule. Am J Physiol. 1987 Sep;253(3 Pt 2):F488–F494. doi: 10.1152/ajprenal.1987.253.3.F488. [DOI] [PubMed] [Google Scholar]
  22. Kolb H. A., Brown C. D., Murer H. Characterization of a Ca-dependent maxi K channel in the apical membrane of a cultured renal epithelium (JTC-12.P3). J Membr Biol. 1986;92(3):207–215. doi: 10.1007/BF01869389. [DOI] [PubMed] [Google Scholar]
  23. Lang D. G., Ritchie A. K. Large and small conductance calcium-activated potassium channels in the GH3 anterior pituitary cell line. Pflugers Arch. 1987 Dec;410(6):614–622. doi: 10.1007/BF00581321. [DOI] [PubMed] [Google Scholar]
  24. Latorre R., Miller C. Conduction and selectivity in potassium channels. J Membr Biol. 1983;71(1-2):11–30. doi: 10.1007/BF01870671. [DOI] [PubMed] [Google Scholar]
  25. Latorre R., Vergara C., Hidalgo C. Reconstitution in planar lipid bilayers of a Ca2+-dependent K+ channel from transverse tubule membranes isolated from rabbit skeletal muscle. Proc Natl Acad Sci U S A. 1982 Feb;79(3):805–809. doi: 10.1073/pnas.79.3.805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Marty A. Blocking of large unitary calcium-dependent potassium currents by internal sodium ions. Pflugers Arch. 1983 Feb;396(2):179–181. doi: 10.1007/BF00615524. [DOI] [PubMed] [Google Scholar]
  27. Maruyama Y., Matsunaga H., Hoshi T. Ca2+- and voltage activated K+ channel in apical cell membrane of gallbladder epithelium from Triturus. Pflugers Arch. 1986 Jun;406(6):563–567. doi: 10.1007/BF00584021. [DOI] [PubMed] [Google Scholar]
  28. McManus O. B., Magleby K. L. Kinetic states and modes of single large-conductance calcium-activated potassium channels in cultured rat skeletal muscle. J Physiol. 1988 Aug;402:79–120. doi: 10.1113/jphysiol.1988.sp017195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Merot J., Bidet M., Le Maout S., Tauc M., Poujeol P. Two types of K+ channels in the apical membrane of rabbit proximal tubule in primary culture. Biochim Biophys Acta. 1989 Jan 16;978(1):134–144. doi: 10.1016/0005-2736(89)90508-7. [DOI] [PubMed] [Google Scholar]
  30. Methfessel C., Boheim G. The gating of single calcium-dependent potassium channels is described by an activation/blockade mechanism. Biophys Struct Mech. 1982;9(1):35–60. doi: 10.1007/BF00536014. [DOI] [PubMed] [Google Scholar]
  31. Moczydlowski E., Alvarez O., Vergara C., Latorre R. Effect of phospholipid surface charge on the conductance and gating of a Ca2+-activated K+ channel in planar lipid bilayers. J Membr Biol. 1985;83(3):273–282. doi: 10.1007/BF01868701. [DOI] [PubMed] [Google Scholar]
  32. Moczydlowski E., Latorre R. Gating kinetics of Ca2+-activated K+ channels from rat muscle incorporated into planar lipid bilayers. Evidence for two voltage-dependent Ca2+ binding reactions. J Gen Physiol. 1983 Oct;82(4):511–542. doi: 10.1085/jgp.82.4.511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Montrose M. H., Lester R., Zimniak P., Anwer M. S., Murer H. Bile acids increase cellular free calcium in cultured kidney cells (LLC-PK). Pflugers Arch. 1988 Jul;412(1-2):164–171. doi: 10.1007/BF00583746. [DOI] [PubMed] [Google Scholar]
  34. Morris A. P., Gallacher D. V., Lee J. A. A large conductance, voltage- and calcium-activated K+ channel in the basolateral membrane of rat enterocytes. FEBS Lett. 1986 Sep 29;206(1):87–92. doi: 10.1016/0014-5793(86)81346-1. [DOI] [PubMed] [Google Scholar]
  35. Neyton J., Miller C. Discrete Ba2+ block as a probe of ion occupancy and pore structure in the high-conductance Ca2+ -activated K+ channel. J Gen Physiol. 1988 Nov;92(5):569–586. doi: 10.1085/jgp.92.5.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Palant C. E., Kurtz I. Measurement of intracellular Ca2+ activity in Necturus gallbladder. Am J Physiol. 1987 Aug;253(2 Pt 1):C309–C315. doi: 10.1152/ajpcell.1987.253.2.C309. [DOI] [PubMed] [Google Scholar]
  37. Pallotta B. S. Calcium-activated potassium channels in rat muscle inactivate from a short-duration open state. J Physiol. 1985 Jun;363:501–516. doi: 10.1113/jphysiol.1985.sp015724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Pallotta B. S., Hepler J. R., Oglesby S. A., Harden T. K. A comparison of calcium-activated potassium channel currents in cell-attached and excised patches. J Gen Physiol. 1987 Jun;89(6):985–997. doi: 10.1085/jgp.89.6.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Palmer L. G. Patch-clamp technique in renal physiology. Am J Physiol. 1986 Mar;250(3 Pt 2):F379–F385. doi: 10.1152/ajprenal.1986.250.3.F379. [DOI] [PubMed] [Google Scholar]
  40. Parent L., Cardinal J., Sauvé R. Single-channel analysis of a K channel at basolateral membrane of rabbit proximal convoluted tubule. Am J Physiol. 1988 Jan;254(1 Pt 2):F105–F113. doi: 10.1152/ajprenal.1988.254.1.F105. [DOI] [PubMed] [Google Scholar]
  41. Petersen K. U., Reuss L. Cyclic AMP-induced chloride permeability in the apical membrane of Necturus gallbladder epithelium. J Gen Physiol. 1983 May;81(5):705–729. doi: 10.1085/jgp.81.5.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Reinhart P. H., Chung S., Levitan I. B. A family of calcium-dependent potassium channels from rat brain. Neuron. 1989 Jan;2(1):1031–1041. doi: 10.1016/0896-6273(89)90227-4. [DOI] [PubMed] [Google Scholar]
  43. Reuss L., Cheung L. Y., Grady T. P. Mechanisms of cation permeation across apical cell membrane of Necturus gallbladder: effects of luminal pH and divalent cations on K+ and Na+ permeability. J Membr Biol. 1981 Apr 30;59(3):211–224. doi: 10.1007/BF01875426. [DOI] [PubMed] [Google Scholar]
  44. Reuss L., Finn A. L. Electrical properties of the cellular transepithelial pathway in Necturus gallbladder. I. Circuit analysis and steady-state effects of mucosal solution ionic substitutions. J Membr Biol. 1975 Dec 4;25(1-2):115–139. doi: 10.1007/BF01868571. [DOI] [PubMed] [Google Scholar]
  45. Reuss L., Finn A. L. Electrical properties of the cellular transepithelial pathway in Necturus gallbladder. II. Ionic permeability of the apical cell membrane. J Membr Biol. 1975 Dec 4;25(1-2):141–161. doi: 10.1007/BF01868572. [DOI] [PubMed] [Google Scholar]
  46. Reuss L. Ion transport across gallbladder epithelium. Physiol Rev. 1989 Apr;69(2):503–545. doi: 10.1152/physrev.1989.69.2.503. [DOI] [PubMed] [Google Scholar]
  47. Reuss L. Potassium transport mechanisms by amphibian gallbladder. Soc Gen Physiol Ser. 1981;36:109–128. [PubMed] [Google Scholar]
  48. Sackin H., Palmer L. G. Basolateral potassium channels in renal proximal tubule. Am J Physiol. 1987 Sep;253(3 Pt 2):F476–F487. doi: 10.1152/ajprenal.1987.253.3.F476. [DOI] [PubMed] [Google Scholar]
  49. Sheppard D. N., Giraldez F., Sepúlveda F. V. Kinetics of voltage- and Ca2+ activation and Ba2+ blockade of a large-conductance K+ channel from Necturus enterocytes. J Membr Biol. 1988 Oct;105(1):65–75. doi: 10.1007/BF01871107. [DOI] [PubMed] [Google Scholar]
  50. Singer J. J., Walsh J. V., Jr Characterization of calcium-activated potassium channels in single smooth muscle cells using the patch-clamp technique. Pflugers Arch. 1987 Feb;408(2):98–111. doi: 10.1007/BF00581337. [DOI] [PubMed] [Google Scholar]
  51. Stoddard J. S., Reuss L. Dependence of cell membrane conductances on bathing solution HCO3-/CO2 in Necturus gallbladder. J Membr Biol. 1988 May;102(2):163–174. doi: 10.1007/BF01870454. [DOI] [PubMed] [Google Scholar]
  52. Stoddard J. S., Reuss L. Voltage- and time dependence of apical membrane conductance during current clamp in Necturus gallbladder epithelium. J Membr Biol. 1988 Jul;103(2):191–204. doi: 10.1007/BF01870949. [DOI] [PubMed] [Google Scholar]
  53. Tsien R. Y., Rink T. J. Neutral carrier ion-selective microelectrodes for measurement of intracellular free calcium. Biochim Biophys Acta. 1980 Jul;599(2):623–638. doi: 10.1016/0005-2736(80)90205-9. [DOI] [PubMed] [Google Scholar]
  54. Vergara C., Latorre R. Kinetics of Ca2+-activated K+ channels from rabbit muscle incorporated into planar bilayers. Evidence for a Ca2+ and Ba2+ blockade. J Gen Physiol. 1983 Oct;82(4):543–568. doi: 10.1085/jgp.82.4.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Weinman S. A., Reuss L. Na+-H+ exchange at the apical membrane of Necturus gallbladder. Extracellular and intracellular pH studies. J Gen Physiol. 1982 Aug;80(2):299–321. doi: 10.1085/jgp.80.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Wills N. K., Zweifach A. Recent advances in the characterization of epithelial ionic channels. Biochim Biophys Acta. 1987 Apr 27;906(1):1–31. doi: 10.1016/0304-4157(87)90003-7. [DOI] [PubMed] [Google Scholar]
  57. Ziomek C. A., Schulman S., Edidin M. Redistribution of membrane proteins in isolated mouse intestinal epithelial cells. J Cell Biol. 1980 Sep;86(3):849–857. doi: 10.1083/jcb.86.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of General Physiology are provided here courtesy of The Rockefeller University Press

RESOURCES