Abstract
The patch-clamp technique was used to characterize K+ channel activity in the basolateral membrane of isolated crypts from rat distal colon. In cell-attached patches with KCl in the pipette, channels with conductances ranging from 6 pS to 80 pS appeared. With NaCl in the pipette and KCl in the bath in excised inside-out membrane patches a small-conductance channel with a mean conductance of 12±6 pS (n=18) was observed. The channel has been identified as K+ channel by its selectivity for K+ over Na+ and by its sensitivity to conventional K+ channel blockers, Ba2+ and tetraethylammonium (TEA+). Changes of cytosolic pH did not attenuate channel activity. Activity of the 12-pS channel was increased by membrane depolarization and elevated cytosolic Ca2+ concentration. In addition, a maxi K+ channel with a mean conductance of 187±15 pS (n=4) in symmetrical KCl solutions was only occasionally recorded. The maxi K+ channel could be blocked by Ba2+ (5 mmol/l) on the cytosolic side. Using the slow-whole cell recording technique under control conditions, a cell membrane potential of −70±10mV (n=18) was measured. By application of various K+ channel blockers such as glibenclamide, charybdotoxin, apamin, risotilide, Ba2+ and TEA+ in the bath, only Ba2+ and TEA+ depolarized the cell membrane. The present data suggest that the small K+ channel (12 pS) is involved in the maintenance of the cell membrane resting potential.
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Beubler E, Lembeck F (1980) Inhibition by morphine of prostaglandin E1-stimulated secretion and cyclic adenosine 3′,5′-monophosphate formation in rat jenunum in vivo. Br J Pharmacol 68:513–518
Binder HJ, Foster ES, Budinger ME, Hayslett JP (1987) Mechanism of electroneutral sodium chloride adsorption in distal colon of the rat. Gastroenterology 93:449–455
Bleich M, Schlatter E, Greger R (1990) The luminal K+ channel of the thick ascending limb of Henle's loop. Pflügers Arch 415:449–460
Burckhardt BC, Gögelein H (1991) Basolateral K+ channels in intact crypts of the rat. Pflügers Arch 418 [Suppl 1]:R 70
Cheng H, Leblond CP (1974) Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine: V. Unitarian theory of the origin of the four epithelial cell types. Am J Anat 141:537–562
Colatsky TJ, Follmer CH, Starmer CF (1991) Channel specificity in antiarrhythmic drug action. Circulation 82:2235–2242
Cook NS (1988) The pharmacology of potassium channels and their therapeutic potential. Trends Pharmacol Sci 9:21–28
Dawson DC (1991) Ion channels and colonic salt transport. Annu Rev Physiol 53:321–329
Dawson DC, Richards NW (1990) Basolateral K conductance: role in regulation of NaCl absorption and secretion. Am J Physiol 259:C181-C195
Dharmasthaphorn K, Pandol SJ (1986) Mechanism of chloride secretion induced by carbachol in a colonic epithelial cell line. J Clin Incet 77:348–354
Dharmsathaphorn K, Mandel KG, Masui H, McRoberts JA (1985) Vasoactive intestinal polypeptide-induced chloride secretion by a colonic epithelial cell line. J Clin Invest 75:462–471
Gögelein H, Capek K (1990) Quinine inhibits chloride and nonselective cation channels in isolated rat distal colon cells. Biochim Biophys Acta 1027:191–198
Gögelein H, Greger R, Schlatter E (1987) Potassium channels in the basolateral membrane of the rectal gland of Squalus acanthias. Regulation and inhibitors. Pflügers Arch 409:107–113
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 391:85–100
Holtug K, Shipley A, Dantzer A, Sten-Knudsen O, Skadhauge E (1991) Localization of sodium absorption and chloride secretion in an intestinal epithelium. J Membr Biol 122:215–229
Horn R, Marty A (1988) Muscarinic activation of ionic currents measured by a new whole-cell recording method. J Gen Physiol 92:145–159
Kleinberg ME, Finkelstein A (1984) Single-length and double-length channels formed by nystatin in lipid bilayer membranes. J Membr Biol 80:257–269
Loo DDF, Kaunitz JD (1989) Ca2+ and cAMP activate K+ channels in the basolateral membrane of crypt cells isolated from rabbit distal colon. J Membr Biol 110:19–28
Mintenig GM, Monaghan AS, Sepulveda FV (1991) Cl− and K+ channels of enterocytes isolated from guinea-pig small intestinal villi. J Physiol (Lond) 438:333P
Morris AP, Gallacher AP, Lee JAC (1986) A large conductance, voltage- and calcium-activated K+ channel in the basolateral membrane of rat enterocytes. FEBS L 206:87–92
Ohno-Shosaku T, Kubota T, Yamaguchi J, Fujimoto M (1990) Regulation of inwardly rectifying K + channels by intracellular pH in opossum kidney cells. Pflügers Arch 416:138–143
Sepulveda FV, Mason WT (1985) Single channels recordings obtained from basolateral membranes of isolated rabbit enterocytes FEBS L 191:87–91
Sheppard DN, Giraldez F, Sepulveda FV (1988) Kinetics of voltage- and Ca2+ activation and BA2+ blockade of a large-conductance K+ channel from Necturus enteroctes. J Membr Biol 105:65–75
Sheppard DN, Giraldez F, Sepulveda FV (1988) K+ channels activaed by L-alanine transport in isolated Necturus enterocytes FEBS L 234:446–448
Sheppard DN, Valverde MA, Giraldez F, Sepulveda FV (1991) Potassium currents of isolated Necturus enterocytes: a whole-cell patch-clamp study. J Physiol (Lond) 433:663–676
Siemer C, Gögelein H (1991) Prostaglandin E2 activates nonselective cation channels in basal crypt cells from rat distal colon. Pflügers Arch 418 [Suppl 1]:R70
Specian RD, Neutra MR (1988) Mechanism of rapid mucus secretion in goblet cells stimulated by acetylcholine. J Cell Biol 85:626–640
Specian RD, Oliver MG (1991) Functional biology of intestinal goblet cells. Am J Physiol 260:C183-C193
Terakawa S, Suzuki Y (1991) Direct visualisation of exocytosis and water secretion in gastrointestinal exocrine cells. In: Murakami M, Seo Y, Kuwahara A, Watari H (eds) Ionic basis and energy metabolism of epithelial transport. Hot apsects from exocrine secretion. Kenbun Printing, Okazaki, Japan, pp 53–55
Wang W, Giebisch G (1991) Dual effect of adenosine triphosphate on the apical small conductance K+ channel of the rat cortical collecting duct. J Physiol (Lond) 98:35–61
Wang W, Schwab A, Giebisch G (1990) Regulation of small-conductance K+ channel in apical membrane of rat cortical collecting tubule. Am J Physiol 259:F494-F502
Wang W, White S, Geibel J, Giebisch G (1990) A potassium channel in the apical membrane of rabbit thick ascending limb of Henle's loop. Am J Physiol 258:F244–253
Welsh MJ, Smith PL, Fromm M, Frizzell RM (1982) Crypts are the site of intestinal fluid and electrolyte secretion. Nature 218:1219–1221
Weymer A, Huott P, Liu W, McRoberts JA, Dharmasathaporn K (1985) Chloride secretory mechanism induced by prostaglandin E1 in a colonic epithelial cell line. J Clin Invest 76:1828–1836
Wiener H, Klaerke DA, Jorgensen PL (1990) Rabbit distal colon epithelium: III. Ca2+-activated K+ channels in basolateral plasma membrane vesicles of surface and crypt cells. J Membr Biol 117:275–2283
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Burckhardt, BC., Gögelein, H. Small and maxi K+ channels in the basolateral membrane of isolated crypts from rat distal colon: single-channel and slow whole-cell recordings. Pflugers Arch. 420, 54–60 (1992). https://doi.org/10.1007/BF00378641
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DOI: https://doi.org/10.1007/BF00378641