Abstract
Epithelial cells lose their usual polarization during carcinogenesis. Although most malignant tumours are of epithelial origin little is known about ion channels in carcinoma cells. Previously, we observed that migration of transformed Madin-Darby canine kidney (MDCK-F) cells depended on oscillating K+ channel activity. In the present study we examined whether periodic K+ channel activity may cause changes of cell volume, and whether K+ channel activity is distributed in a uniform way in MDCK-F cells. After determining the average volume of MDCK-F cells (2013±270 μm3; n=8) by means of atomic force microscopy we deduced volume changes by calculating the K+ efflux during bursts of K+ channel activity. Therefore, we measured the membrane conductance of MDCK-F cells which periodically rose by 22.3±2.5 nS from a resting level of 6.5±1.4 nS (n=12), and we measured the membrane potential which hyperpolarized in parallel from −35.4±1.2 mV to −71.6±1.8 mV (n=11). The distribution of K+ channel activity was assessed by locally superfusing the front or rear end of migrating MDCK-F cells with the K+ channel blocker charybdotoxin (CTX). Only exposure of the rear end to CTX inhibited migration providing evidence for “horizontal” polarization of K+ channel activity in transformed MDCK-F cells. This is in contrast to the “vertical” polarization in parent MDCK cells. We propose that the asymmetrical distribution of K+ channel activity is a prerequisite for migration of MDCK-F cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Binnig G, Quate CF, Gerber CH (1986) Atomic force microscopy. Phys Rev Lett 56:930–933
Chen C, Corbley MJ, Roberts TM, Hess P (1988) Voltage-sensitive calcium channels in normal and transformed 3T3 fibroblasts. Science 239:1024–1026
Enomoto K, Furuya K, Maeno T, Edwards C, Oka T (1991) Oscillating activity of a calcium-activated K+ channel in normal and cancerous mammary cells in culture. J Membr Biol 119:133–139
Forbush B III, Haas M, Lyttle C (1992) Na-K-Cl cotransport in the shark rectal gland. I. Regulation in the intact perfused gland. Am J Physiol 262:C1000-C1008
Gekle M, Wünsch S, Oberleithner H, Silbernagl S (1994) Characterization of two MDCK-cell subtypes as a model system to study principal cell and intercalated cell properties. Pflügers Arch 428:157–162
Greger R, Allert N, Fröbe U, Normann C (1993) Increase in cytosolic Ca2+ regulates exocytosis and Cl− conductance in HT29 cells. Pflügers Arch 424:329–334
Grimstad IA (1987) Direct evidence that cancer cell locomotion contributes importantly to invasion. Exp Cell Res 173:515–523
Hahn K, DeBiasio R, Taylor DL (1992) Patterns of elevated free calcium and calmodulin activation in living cells. Nature 359:736–738
Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved techniques for high-resolution current recording from cell-free membrane patches. Pflügers Arch 391:85–100
Hammerton RW, Krzeminski KA, Mays RW, Ryan TA, Wollner DA, Nelson WJ (1991) Mechanism for regulating cell surface distribution of Na+, K+-ATPase in polarized epithelial cells. Science 254:847–850
Horowitz SB, Fenichel IR (1970) Analysis of sodium transport in the amphibian oocyte by extractive and radioautographic techniques. J Cell Biol 47:120–131
Huang Y, Rane SG (1993) Single channel study of a Ca2+-activated K+ current associated with ras-induced cell transformation. J Physiol (Lond) 461:601–618
Ito T, Zaner KS, Stossel TP (1987) Nonideality of volume flows and phase transitions of F-actin solutions in response to osmotic stress. Biophys J 51:745–753
Janson LW, Taylor DL (1993) In vitro model of tail contraction and cytoplasmic streaming in amoeboid cells. J Cell Biol 123:345–356
Kersting U, Wojnowski I, Steigner W, Oberleithner H (1991) Hypotonic stress-induced release of KHCO3 in fused renal epitheloid (MDCK) cells. Kidney Int 39:891–900
Lang F, Ritter M, Wöll E, Weiss H, Häussinger D, Hoflacher J, Maly K, Grunicke H (1992) Altered cell volume regulation in ras oncogene expressing NIH fibroblasts. Pflügers Arch 420:424–427
Lindau M, Neher E (1988) Patch-clamp techniques for time-resolved capacitance measurements in single cells. Pflügers Arch 411:137–146
Lyttle C, Forbush B III (1992) Na-K-Cl cotransport in the shark rectal gland. II. Regulation in isolated tubules. Am J Physiol 262:C1009-C1017
Matlin KS, Caplan MJ (1992) Epithelial cell structure and polarity. In: Seldin DW, Giebisch G (eds) The kidney: physiology and pathophysiology. Raven, New York, pp 447–473
Oberleithner H (1992) Endogenous pH oscillations in transformed epithelial cells. Cell Physiol Biochem 2:180–188
Oberleithner H, Westphale H-J, Gaßner B (1991) Alkaline stress transforms Madin-Darby canine kidney cells. Pflügers Arch 419:418–420
Oberleithner H, Brinckmann E, Schwab A, Krohne G (1994) Imaging nuclear pores of aldosterone sensitive kidney cells by atomic force microscopy. Proc Natl Acad Sci USA 91:9784–9788
Palfrey AC, O'Donnell ME (1992) Characteristics and regulation of the Na/K/2Cl cotransporter. Cell Physiol Biochem 2:293–307
Partin AW, Isaacs JT, Treiger B, Coffey DS (1988) Early cell motility changes associated with an increase in metastatic ability in rat prostatic cancer cells transfected with the v-Harvey-ras oncogene. Cancer Res 48:6050–6053
Repp H, Draheim H, Ruland J, Seidel G, Beise J, Presek P, Dreyer F (1993) Profound differences in potassium current properties of normal and Rous sarcoma virus-transformed-chicken embryo fibroblasts. Proc Natl Acad Sci USA 90:3403–3407
Rodriguez-Boulan E, Nelson WJ (1989) Morphogenesis of the polarized epithelial cell phenotype. Science 245:718–725
Ruggeri B, Caamano J, Slaga TJ, Conti CJ, Nelseon WJ, Klein-Szanto AJP (1992) Alterations in the expression of uvomorulin and Na+, K+-adenosine triphosphatase during mouse skin tumor progression. Am J Pathol 140:1179–1185
Schoenenberger C-A, Matlin KS (1991) Cell polarity and epithelial oncogenesis. Trends Cell Biol 1:87–92
Schwab A, Westphale H-J, Wojnowski L, Wünsch S, Oberleithner H (1993) Spontaneously oscillating K+ channels in alkali-transformed MDCK cells. J Clin Invest 92:218–223
Schwab A, Wojnowski L, Gabriel K, Oberleithner H (1994) Oscillating activity of a Ca2+-sensitive K+ cannel. A prerequisite for migration of alkali-transformed Madin-Darby canine kidney (MDCK-F) cells. J Clin Invest 93:1631–1636
Stossel TP (1992) The mechanical response of white blood cells. In. Gallin JI, Goldstein IM, Snyderman R (eds) Inflammation: basic principles and clinical correlates. Raven, New York, pp 459–475
Taylor DL, Fechheimer M (1982) Cytoplasmic structure and contractility: the solation-contraction coupling hypothesis. Philos Trans R Soc Lond Biol 299:185–197
Taylor DL, Blinks JR, Reynolds G (1980) Contractile basis of ameboid movement. VIII. Aequorin luminescence during ameboid movement, endocytosis, and capping. J Cell Biol 86:599–607
Westphale H-J, Wojnowski L, Schwab A, Oberleithner H (1992) Spontaneous membrane potential oscillations in Madin-Darby canine kidney cells transformed by alkaline stress. Pflügers Arch 421:218–223
Wojnowski L, Hoyland J, Mason WT, Schwab A, Westphale H-J, Oberleithner H (1994) Cell transformation induces a cytoplasmic Ca2+ oscillator in Madin-Darby canine kidney cells. Pflügers Arch 426:89–94
Zigmond SH, Levitsky HI, Kreel BJ (1981) Cell polarity: an examination of its behavioral expression and its consequences for polymorphonuclear leukocyte chemotaxis. J Cell Biol 89:585–592
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Schwab, A., Gabriel, K., Finsterwalder, F. et al. Polarized ion transport during migration of transformed Madin-Darby canine kidney cells. Pflugers Arch. 430, 802–807 (1995). https://doi.org/10.1007/BF00386179
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00386179