Abstract
Cell volume regulation is an essential feature of most cells. After swelling in hypotonic media, the simultaneous activation of potassium and chloride channels is believed to be the initial, time-determining step in cell volume regulation. The activation of both pathways is functionally linked and enables the cells to lose ions and water, subsequently leading to cell shrinkage and readjustment of the initial volume. NIH 3T3 fibroblasts efficiently regulate their volume after swelling and bear chloride channels that are activated by decreasing extracellular osmolarity. The chloride current elicited in these cells after swelling is reminiscent of the current found in oocytes expressing an outwardly rectifying chloride current termed ICln. Introduction of antisense oligodeoxynucleotides complementary to the first 30 nucleotides of the coding region of the ICln channel into NIH 3T3 fibroblasts suppresses the activation of the swelling-induced chloride current. The experiments directly demonstrate an unambiguous link between a volume-activated chloride current and a cloned protein involved in chloride transport.
Similar content being viewed by others
References
Alton EWFW, Manning SD, Schlatter PJ, Geddes DM, Williams AJ (1991) Characterization of a Ca2+-dependent anion channel from sheep tracheal epithelium incorporated into planar lipid bilayers. J Physiol (Lond) 443:137–159
Cahalan MD (1994) Volume-regulated Cl− channels in lymphocytes. Biophys J 66:A3
Cheng SH, Rich DP, Marshall J, Gregory RJ, Welsh MJ, Smith AE (1991) Phosphorylation of the R domain by cAMP-dependent protein kinase regulates the CFTR chloride channel. Cell 66:1027–1036
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Ana Biochem 162:156–159
Deng H, Wang C, Acsadi G, Wolff JA (1991) High-efficiency protein synthesis from T7 RNA polymerase transcripts in 3T3 fibroblasts. Gene 109:193–201
Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Northrop JP, Ringold GM, Danielsen M (1987) Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci USA 84:7413–7417
Gill DR, Hyde SC, Higgins CF, Valverde MA, Mintenig GM, Sepulveda FV (1992) Separation of drug transport and chloride channel functions of the human multidrug resistance P-glycoprotein. Cell 71:23–32
van der Goot FG, Gonzales-Manas JM, Lakey JH, Pattus F (1991) A ‘molten-globule’ membrane-insertion intermediate of the pore-forming domain of colicin. A. Nature 354:408–410
van der Goot FG, Ausio J, Wong KR, Pattus F, Buckley JT (1993) Dimerization stabilizes the pore-forming toxin aerolysin in solution. J Biol Chem 268:18272–18279
Gründer S, Thiemann A, Pusch M, Jentsch TJ (1992) Regions involved in the opening of C1C-2 chloride channels by voltage and cell volume. Nature 360:759–762
Kleuss C, Hescheler J, Ewel C, Rosenthal W, Schultz G, Wittig B (1991) Assignment of G-protein subtypes to specific receptors inducing inhibition of calcium currents. Nature 353:43–48
Krapivinsky GB, Ackerman MJ, Gordon EA, Krapivinsky LD, Clapham DE (1994) Molecular characterization of a swelling-induced chloride conductance regulatory protein, pICln. Cell 76:439–448
Kunzelmann K, Pavenstädt H, Greger R (1989) Properties and regulation of chloride channels in cystic fibrosis and normal airway cells. Pflügers Arch 415:172–182
Kunzelmann K, Slotki IN, Klein P, Koslowsky T, Ausiello DA, Greger R, Cabantchik ZI (1995) Effects of P-glycoprotein expression on cyclic AMP and volume activated ion fluxes and conductances in HT-29 colon adenocarcinoma cells. J Cell Physiol (in press)
Lallier T, Bronner-Fraser M (1993) Inhibition of neural crest cell attachment by integrin antisense oligonucleotides. Science 259:692–695
Lang F, Völkl H, Häussinger D (1990) General principles in cell volume regulation. Comp Physiol 4:1–25
Maniatis T, Fritsch E, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, New York
Marty A, Neher E (1983) Tight-seal whole-cell recording. In: Sakmann B, Neher E (eds) Single-channel recording. Plenum, New York, pp 107–122
McEwan GTA, Hunter J, Hirst BH, Simmons NL (1992) Volume-activated Cl− secretion and transepithelial vinblastine secretion mediated by P-glycoprotein are not correlated in cultured human T48 intestinal epithelial layers. FEBS Lett 304:233–236
Nilius B, Sehrer J, Droogmans G (1994) Permeation and modulation of volume-activated Cl−-currents in human endothelial cells. Br J Pharmacol 112:1049–1056
Paulmichl M, Friedrich F, Maly K, Lang F (1989) The effect of hypoosmolarity on the electrical properties of Madin Darby canine kidney cells. Pflügers Arch 413:456–462
Paulmichl M, Li Y, Wickman K, Ackerman M, Peralta E, Clapham D (1992) New mammalian chloride channel identified by expression cloning. Nature 356:238–241
Paulmichl M, Gschwentner M, Wöll E, Schmarda A, Ritter M, Kanin G, Ellemunter H, Waitz W, Deetjen P (1993) Insight into the structure-function of chloride channels. Cell Physiol Biochem 3:374–387
Rasola A, Galietta LJV, Gruenert DC, Romeo G (1994) Volume-sensitive chloride currents in four epithelial cell lines are not directly correlated to the expression of the MDR-1 gene. J Biol Chem 269:1432–1436
Solc CK, Wine JJ (1991) Swelling-induced and depolarization-induced Cl− channels in normal and cystic fibrosis epithelial cells. Am J Physiol 261:C658-C674
Tabares L, Mazzanti M, Clapham DE (1991) Chloride channels in the nuclear membrane. J Membr Biol 123:49–54
Valverde MA, Diaz M, Sepulveda FV (1992) Volume-regualted chloride channels associated with the human multidrug-resistance P-glycoprotein. Nature 355:830–833
Vanglarik CJ, Singh AK, Wang R, Bridges RJ (1993) Trinitrophenyl-ATP blocks colonic Cl− channels in planar phospholipid bilayers. Evidence for two nucelotide binding sites. J Gen Physiol 101:545–569
Wahlestedt C, Pich EM, Koob GF, Yee F, Heilig M (1993) Modulation of anxiety and neuropeptide Y-Y1 receptors by antisense oligodeoxynucleotides. Science 259:528–531
Wangemann P, Wittner M, DiStefano 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-S141
Weiss H, Lang F (1992) Ion channels activated by swelling of Madin Darby canine kidney (MDCK) cells. J Membr Biol 126:109–114
Worrell RT, Butt AG, Cliff WH, Frizzell RR (1989) A volume-sensitive chloride conductance in human colonic cell line T84. Am J Physiol 256:C1111-C1119
Zhang J, Smith T, Lobaugh L, Hall S, Lieberman M (1992) Cyclic AMP inhibits the swelling activated I Cl associated with cardiac cell volume regualtion (abstract). Physiologist 35:18
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gschwentner, M., Nagl, U.O., Wöll, E. et al. Antisense oligonucleotides suppress cell-volume-induced activation of chloride channels. Pflugers Arch. 430, 464–470 (1995). https://doi.org/10.1007/BF00373882
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00373882