Skip to main content
Log in

Characterization of chloride and cation channels in cultured human keratinocytes

  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

Patch-clamp experiments on human cultured keratinocytes revealed the presence of three types of ion channel. The first type was a Cl-selective channel, the current/voltage relationship of which showed outward rectification, the mean conductance at positive and negative membrane potentials being 66 pS and 16 pS respectively. The second type of channel showed almost equal permeability to alkali ions but was impermeable to Cl and to the large organic cation N-methyl-d-glucamine. Its current/voltage relationship was linear with a mean unitary conductance of 18 pS in symmetrical 140 mmol/l NaCl. Finally, the third type was a large-conductance cation channel, which had in physiological ionic conditions a peculiar rectifying current/voltage relationship, the shape of which was strongly dependent on the concentration of divalent cations on both sides of the membrane. Lowering of Ca2+ and/or Mg2+ on either side of the patch led to a marked increase of the single-channel current. With identical solutions without Ca2+ and Mg2+ on both sides of the patch the current/voltage relationship became ohmic and reached a conductance of 150–200 pS. In addition, channel activity was reversibly affected by changes of the external Ca2+ concentration. In particular, open-channel probability strongly increased at negative membrane potentials when the external Ca2+ was lowered from millimolar to micromolar values. Whole-cell experiments confirm the role of the extracellular Ca2+ as a modulator of the cation conductance.

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

  • Barrandon Y, Green H (1985) Cell size as a determinant of the clone-forming ability of human keratinocytes. Proc Natl Acad Sci USA 82:5390–5394

    Google Scholar 

  • Bijman J, Kansen N, Hoogeveen AH, Scholte B, van der Kamp A, de Jonge H (1988) Electrolyte transport in normal and CF epithelia. In: Wong PYD, Young JA (eds) Exocrine secretion. University Press, Hong Kong, pp 17–20

    Google Scholar 

  • Boyce ST, Ham RG (1983) Calcium-regulated differentiation of normal human epidermal keratinocytes in chemically defined clonal culture and serum-free serial culture. J Invest Dermatol 81:33s-40s

    Google Scholar 

  • Cone CD, Cone CM (1976) Induction of mitosis in mature neurons in central nervous system by sustained depolarization. Science 192:155–158

    Google Scholar 

  • Das S, Palmer LG (1989) Extracellular Ca2+ controls outward rectification by apical cation channels in toad urinary bladder: patch-clamp and whole-bladder studies. J Membr Biol 107:157–168

    Google Scholar 

  • De Luca M, Cancedda R (1990) Culture of human epithelium. Burns (in press)

  • De Luca M, D'Anna F, Bondanza S, Franzi AT, Cancedda R (1988) Human epithelial cells induce human melanocyte growth in vitro but only skin keratinocytes regulate its proper differentiation in the absence of dermis. J Cell Biol 107:1919–1926

    Google Scholar 

  • De Luca M, Albanese E, Bondanza S, Megna M, Ugozzoli L, Molina F, Cancedda R, Santi PL, Bormioli M, Stella M, Magliancani G (1989) Multicentre experience in the treatment of burns with autologous and allogeneic cultured epithelium fresh or preserved in a frozen state. Burns 15:303–309

    Google Scholar 

  • Dempster J (1987) A range of computer programs for electrophysiological signal analysis using the IBM PC AT. J Physiol (Lond) 392:20P

  • Gallico GG, O'Connor NE, Compton CC, Kehinde O, Green H (1984) Permanent coverage of large burn wounds with autologous cultured human epithelium. N Engl J Med 311:448–451

    Google Scholar 

  • Gögelein H (1988) Chloride channels in epithelia. Biochim Biophys Acta 947:521–547

    Google Scholar 

  • Green H (1978) Cyclic AMP in relation to proliferation of the epidermal cell: a new view. Cell 15:801–811

    Google Scholar 

  • Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recordings from cells and cell-free membrane patches. Pflügers Arch 391:85–100

    Google Scholar 

  • Hennings H, Michael D, Cheng C, Steinert P, Holbrook K, Yuspa SH (1980) Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell 19:245–254

    Google Scholar 

  • 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 

  • Li M, McCann JD, Liedtke CM, Nairn AC, Greengard P, Welsh MJ (1988) Cyclic AMP-dependent protein kinase opens chloride channels in normal but not cystic fibrosis airway epithelium. Nature 331:358–360

    Google Scholar 

  • Menon GK, Grayson S, Elias PM (1985) Ionic calcium reservoirs in mammalian epidermis: ultrastructural localization by ioncapture cytochemistry. J Invest Dermatol 84:508–512

    Google Scholar 

  • Partridge LD, Swandulla D (1988) Calcium-activated non-specific cation channels. Trends Neurosci 11:69–72

    Google Scholar 

  • Schoumacher RA, Shoemaker RL, Halm DR, Tallant EA, Wallace RW, Frizzell RA (1987) Phosphorylation fails to activate chloride channels from cystic fibrosis airway cells. Nature 330:752–754

    Google Scholar 

  • Smith JB, Rozengurt E (1978) Serum stimulates the Na+, K+ pump in quiescent fibroblasts by increasing Na+ entry. Proc Natl Acad Sci USA 75:5560–5564

    Google Scholar 

  • Steele JA (1989) Voltage- and time-dependent chloride currents in chick skeletal muscle cells grown in tissue culture. Pflügers Arch 414:265–272

    Google Scholar 

  • Sturgess NC, Hales CN, Ashford MLJ (1987) Calcium and ATP regulate the activity of a non-selective cation channel in a rat insulinoma cell line. Pflügers Arch 409:607–615

    Google Scholar 

  • Toback FG (1980) Induction of growth in kidney epithelial cells in culture by Na+. Proc Natl Acad Sci USA 77:6654–6656

    Google Scholar 

  • Van Driessche W, Zeiske W (1985) Ca2+-sensitive, spontaneously fluctuating, cation channels in the apical membrane of the adult frog skin epithelium. Pflügers Arch 405:250–259

    Google Scholar 

  • Watt FM (1989) Terminal differentiation of epidermal keratinocytes. Curr Opinion Cell Biol 1:1107–1115

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Galietta, L.J.V., Barone, V., De Luca, M. et al. Characterization of chloride and cation channels in cultured human keratinocytes. Pflügers Archiv 418, 18–25 (1991). https://doi.org/10.1007/BF00370447

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00370447

Key words

Navigation