Summary
In the mammalian distal colon, the surface epithelium is responsible for electrolyte absorption, while the crypts are the site of secretion. This study examines the properties of electrical potential-driven86Rb+ fluxes through K+ channels in basolateral membrane vesicles of surface and crypt cells of the rabbit distal colon epithelium. We show that Ba2+-sensitive, Ca2+-activated K+ channels are present in both surface and crypt cell derived vesicles with half-maximal activation at 5×10−7 m free Ca2+. This suggests an important role of cytoplasmic Ca2+ in the regulation of the bidirectional ion fluxes in the colon epithelium.
The properties of K+ channels in the surface cell membrane fraction differ from those of the channels in the crypt cell derived membranes. The peptide toxin apamin inhibits Ca2+-activated K+ channels exclusively in surface cell vesicles, while charybdotoxin inhibits predominantely in the crypt cell membrane fraction. Titrations with H+ and tetraethylammonium show that both high-and low-sensitive86Rb+ flux components are present in surface cell vesicles, while the high-sensitive component is absent in the crypt cell membrane fraction. The Ba2+-sensitive, Ca2+-activated K+ channels can be solubilized in CHAPS and reconstituted into phospholipid vesicles. This is an essential step for further characterization of channel properties and for identification of the channel proteins in purification procedures.
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Asher, C., Cragoe, E.J., Jr., Garty, H. 1987. Effects of amiloride analogues on Na+ transport in toad bladder membrane vesicles.J. Biol. Chem. 262:8566–8573
Bardsley, W.G., McGinlay, P.B. 1987. The use of non-linear regression analysis and the F test for model discrimination with dose-response curves and ligand binding data.J. Theor. Biol. 126:183–201
Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Anal. Biochem. 72:248–254
Burgess, G.M., Claret, M., Jenkinson, D.H. 1981. Effects of quinine and apamin on the calcium-dependent potassium permeability of mammalian hepatocytes and red cells.J. Physiol. (London) 317:67–90
Burnham, C., Braw, R., Karlish, S.J.D. 1986. A Ca-dependent K channel in “luminal” membranes from the renal outer medulla.J. Membrane Biol. 93:177–186
Chang, W.W.L., Leblond, C.P. 1971. Renewal of the epithelium in the descending colon of the mouse.Am. J. Anat. 131:73–100
Chase, H.S., Jr. 1984. Does calcium couple the apical and basolateral membrane permeabilities in epithelia?Am. J. Physiol. 247:F869-F876
Christensen, O., Zeuthen, T. 1987. Maxi K+ channels in leaky epithelia are regulated by intracellular Ca2+, pH and membrane potential.Pfluegers Arch. 408:249–259
Cook, N.S., Haylett, D.G. 1985. Effects of apamin, quinine and neuromuscular blockers on calcium-activated potassium channels in guinea pig hepatocytes.J. Physiol. (London) 358:373–394
DeCoursey, T.E., Chandy, K.G., Gupta, S., Cahalan, M.D. 1985. Voltage-dependent ion channels in T-lymphocytes.J. Neuroimmunol. 10:71–95
DeCoursey, T.E., Chandy, K.G., Gupta, S., Cahalan, M.D. 1987. Two types of potassium channels in murine T lymphocytes.J. Gen. Physiol. 89:379–404
Frizzell, R.A., Schultz, S.G. 1978. Effect of aldosterone on ion transport by rabbit colon in vitro.J. Membrane Biol. 39:1–26
Fukushima, Y., Hagiwara, S. 1985. Currents carried by monovalent cations through calcium channels in mouse neoplastic B lymphocytes.J. Physiol. (London) 358:255–284
Garty, H., Asher, C., Yeger, O. 1987. Direct inhibition of epithelial Na+ channels by a pH-dependent interaction with calcium, and by other divalent ions.J. Membrane Biol. 95:151–162
Garty, H., Benos, D.J. 1988. Characteristics and regulatory mechanisms of the amiloride-blockable Na+ channel.Physiol. Rev. 68:309–373
Gustin, M.C., Goodman, D.B.P. 1981. Isolation of brush-border membrane from the rabbit descending colon epithelium.J. Biol. Chem. 256:10651–10656
Halm, D.R., Frizzell, R.A. 1986. Active K+ transport across rabbit distal colon: Relation to Na+ absorption and Cl− secretion.Am. J. Physiol. 251:C252-C267
Heintze, K., Stewart, C.P., Frizzell, R.A. 1983. Sodium-dependent chloride secretion across rabbit descending colon.Am. J. Physiol. 244:G357-G365
Hermann, A., Gorman, A.L.F. 1981. Effects of 4-aminopyridine on potassium currents in a molluscan neuron.J. Gen. Physiol. 78:63–86
Hugues, M., Duval, D., Schmid, H., Kitabgi, P., Lazdunski, M., Vincent, J.P. 1982. Specific binding and pharmacological interactions of apamin, the neurotoxin from bee venom, with guinea pig colon.Life Sci. 31:437–443
Hugues, M., Romey, G., Duval, D., Vincent, J.P., Lazdunski, M. 1982. Apamin as a selective blocker of the calcium-dependent potassium channel in neuroblastoma cells: Volt-age-clamp and biochemical characterization of the toxin receptor.Proc. Natl. Acad. Sci. USA 79:1308–1312
Jørgensen, P.L. 1986. Structure, function and regulation of Na,K-ATPase in the kidney.Kidney Int. 29:10–20
Klaerke, D.A., Karlish, S.J.D., Jørgensen, P.L. 1987. Reconstitution in phospholipid vesicles of calcium-activated potassium channel from outer renal medulla.J. Membrane Biol. 95:105–112
Latorre, R., Miller, C. 1983. Conduction and selectivity in potassium channels.J. Membrane Biol. 71:11–30
Latorre, R., Oberhauser, A., Labarca, P., Alvarez, O. 1989. Varieties of calcium-activated potassium channels.Annu. Rev. Physiol. 51:385–399
Loo, D.D.F., Kaunitz, J.D. 1989. Ca2+ and cAMP activate K+ channels in the basolateral membrane of crypt cells isolated from rabbit distal colon.J. Membrane Biol. 110:19–28
MacKinnon, R., Miller, C. 1988. Mechanism of charybdotoxin block of the high-conductance, Ca2+-activated K+ channel.J. Gen. Physiol. 91:335–349
McCabe, R.D., Smith, P.L. 1985. Colonic potassium and chloride secretion: Role of cAMP and calcium.Am. J. Physiol. 248:G103-G109
Moczydlowski, E., Alvarez, O., Vergara, C., Latorre, R. 1985. Effect of phospholipid surface charge on the conductance and gating of a Ca2+-activated K+ channel in planar lipid bilayers.J. Membrane Biol. 83:273–282
Moczydlowski, E., Lucchesi, K., Ravindran, A. 1988. An emerging pharmacology of peptide toxins targeted against potassium channels.J. Membrane Biol. 105:95–111
Petersen, O.H., Maruyama, Y. 1984. Calcium-activated potassium channels and their role in secretion.Nature (London) 307:693–696
Plass, H., Gridl, A., Turnheim, K. 1986. Absorption and secretion of potassium by rabbit descending colon.Pfluegers Arch. 406:509–519
Potter, G.D., Tran, T., Sellin, J.H. 1989. Colonic epithelial cell calcium response to bile acid in vitro.Gastroenterology 96:A398
Schmid-Antomarchi, H., De Weille, J., Fosset, M., Lazdunski, M. 1987. The receptor for antidiabetic sulfonylureas controls the activity of the ATP-modulated K+ channel in insulin-secreting cells.J. Biol. Chem. 262:15840–15844
Schultz, S.G. 1984. A cellular model for active sodium absorption by mammalian colon.Annu. Rev. Physiol. 46:435–451
Seagar, M.J., Marqueze, B., Couraud, F. 1987. Solubilization of the apamin receptor associated with a calcium-activated potassium channel from rat brain.J. Neurosci. 7:565–570
Smith, P.L., McCabe, R.D. 1984. Mechanism and regulation of transcellular potassium transport by the colon.Am. J. Physiol. 247:G445-G456
Turnheim, K., Costantin, J., Chan, S., Schultz, S.G. 1989. Reconstitution of a calcium-activated potassium channel in basolateral membranes of rabbit colonocytes into planar lipid bilayers.J. Membrane Biol. 112:247–254
Turnheim, K., Plass, H., Grasl, M., Krivanek, P., Wiener, H. 1986. Sodium absorption and potassium secretion in rabbit colon during sodium deficiency.Am. J. Physiol. 250:F235-F245
Welsh, M.J., Smith, P.L., Fromm, M., Frizzell, R.A. 1982. Crypts are the site of intestinal fluid and electrolyte secretion.Science 218:1219–1221
Wiener, H., Turnheim, K., van Os, C.H. 1989. Rabbit distal colon epithelium: I. Isolation and characterization of basolateral plasma membrane vesicles from surface and crypt cells.J. Membrane Biol. 110:147–162
Wiener, H., van Os, C.H. 1989. Rabbit distal colon epithelium: II. Characterization of (Na+,K+,Cl−)-cotransport and [3H]bumetanide binding.J. Membrane Biol. 110:163–174
Wills, N.K. 1984. Mechanisms of ion transport by the mammalian colon revealed by frequency domain analysis techniques.Curr. Top. Membrane Transp. 20:61–85
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Wiener, H., Klaerke, D.A. & Jørgensen, P.L. Rabbit distal colon epithelium: III. Ca2+-activated K+ channels in basolateral plasma membrane vesicles of surface and crypt cells. J. Membrain Biol. 117, 275–283 (1990). https://doi.org/10.1007/BF01868457
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DOI: https://doi.org/10.1007/BF01868457