Skip to main content
Log in

Antisense oligonucleotides suppress cell-volume-induced activation of chloride channels

  • Original Article
  • Transport Processes, Metabolism and Endocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands
  • Published:
Pflügers Archiv Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. 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

    Google Scholar 

  2. Cahalan MD (1994) Volume-regulated Cl channels in lymphocytes. Biophys J 66:A3

    Google Scholar 

  3. 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

    Article  PubMed  Google Scholar 

  4. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Ana Biochem 162:156–159

    Article  Google Scholar 

  5. 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

    Article  PubMed  Google Scholar 

  6. 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

    PubMed  Google Scholar 

  7. 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

    Article  PubMed  Google Scholar 

  8. 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

    Article  PubMed  Google Scholar 

  9. 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

    PubMed  Google Scholar 

  10. 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

    Article  PubMed  Google Scholar 

  11. 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

    Article  PubMed  Google Scholar 

  12. 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

    Article  PubMed  Google Scholar 

  13. 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

    Google Scholar 

  14. 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)

  15. Lallier T, Bronner-Fraser M (1993) Inhibition of neural crest cell attachment by integrin antisense oligonucleotides. Science 259:692–695

    PubMed  Google Scholar 

  16. Lang F, Völkl H, Häussinger D (1990) General principles in cell volume regulation. Comp Physiol 4:1–25

    Google Scholar 

  17. Maniatis T, Fritsch E, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  18. 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

    Google Scholar 

  19. 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

    Article  PubMed  Google Scholar 

  20. 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

    PubMed  Google Scholar 

  21. 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

    Google Scholar 

  22. 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

    Article  PubMed  Google Scholar 

  23. 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

    Google Scholar 

  24. 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

    PubMed  Google Scholar 

  25. 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

    PubMed  Google Scholar 

  26. Tabares L, Mazzanti M, Clapham DE (1991) Chloride channels in the nuclear membrane. J Membr Biol 123:49–54

    PubMed  Google Scholar 

  27. Valverde MA, Diaz M, Sepulveda FV (1992) Volume-regualted chloride channels associated with the human multidrug-resistance P-glycoprotein. Nature 355:830–833

    Article  PubMed  Google Scholar 

  28. 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

    Article  PubMed  Google Scholar 

  29. 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

    PubMed  Google Scholar 

  30. 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

    Google Scholar 

  31. Weiss H, Lang F (1992) Ion channels activated by swelling of Madin Darby canine kidney (MDCK) cells. J Membr Biol 126:109–114

    PubMed  Google Scholar 

  32. 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

    PubMed  Google Scholar 

  33. 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

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints 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

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00373882

Key words

Navigation