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Chloride channels on epithelial cells cultured from human fetal epididymis

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Summary

Using single-channel recording techniques, we have detected two types of outwardly rectifying chloride channel on epithelial cells cultured from human fetal epididymis. A small-conductance channel (2.8–5.0 pS) was spontaneously active in 29% of cell-attached patches but rapidly disappeared on patch excision. This channel often occurred in clusters and exhibited slow kinetics with open and closed times of the order of tens or hundreds of msec; an open-state probability that was essentially independent of voltage; and a very low permeability to bicarbonate relative to chloride. Exposing epididymal cells to either forskolin (3 μm) or adrenaline (1 μm) activated this channel (up to 350-fold), suggesting that it may be involved in cyclic AMP-mediated anion secretion by the male reproductive tract. The large-conductance channel (14 to 29 pS) was never detected in cell-attached patches but could be activated by depolarization (40 mV) in 3% of excised, inside-out patches. Once activated, opening of this ‘large’ channel was voltage independent, and it had a relatively high permeability to both gluconate (P gluconate/P chloride=0.24) and bicarbonate (P bicarbonate/P chloride=0.4). The proportion of excised patches that contained this channel was increased 2.5-fold by prior stimulation of the epididymal cells; however, because the channel was never observed in cell-attached patches its physiological role must remain uncertain.

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References

  1. Anderson, M.P., Rich, D. P., Gregory, R.J., Smith, A.E., Welsh, M.J. 1981. Generation of cAMP-activated chloride currents by expression of CFTR.Science 251:679–682

    Google Scholar 

  2. Ashford, M.L.J. 1986. Single chloride channels in cultured rat epididymal cells.J. Physiol. 371:142P

    Google Scholar 

  3. Bubien, J.K., Kirk, K.L., Rado, T.A., Frizzell, R.A. 1990. Cell cycle dependence of chloride permeability in normal and cystic fibrosis lymphocytes.Science 248:1416–1419

    PubMed  Google Scholar 

  4. Champigny, G., Verrier, B., Gérard, C., Mauchamp, J., Lazdunski, M. 1990. Small conductance chloride channels in the apical membrane of thyroid cells.FEBS Lett. 259:263–268

    PubMed  Google Scholar 

  5. Christensen, O., Simon, M., Randlev, T. 1989. Anion channels in a leaky epithelium. A patch clamp study of choroid plexus.Pfluegers Arch. 415:37–46

    Google Scholar 

  6. Cliff, W.H., Frizzell, R.A. 1990. Separate Cl conductances activated by cAMP and Ca2+ in Cl-secreting epithelial cells.Proc. Natl. Acad. Sci. USA 87:4956–4960

    PubMed  Google Scholar 

  7. Coleman, L., Harris, A. 1991. Immortalization of male genital duct epithelium: An assay system for the cystic fibrosis gene.J. Cell Sci. 98:85–89

    PubMed  Google Scholar 

  8. Cook, D.I., Huang, S.J., Wilson, S.M., Wong, P.Y.D., Young, J.A. 1990. Ion channels in the apical plasma membrane of the rat cauda epididymis.J. Physiol. 423:57P

    Google Scholar 

  9. Cuthbert, A.W., Wong, P.Y.D. 1986. Anion secretion in cultured rat epididymal epithelium.J. Physiol. 378:335–345

    PubMed  Google Scholar 

  10. Diener, M., Rummel, W., Mestres, P., Lindemann, B. 1989. Single chloride channels in colon mucosa and isolated colonic enterocytes of the rat.J. Membrane Biol. 108:21–30

    Google Scholar 

  11. Drumm, M.L., Pope, H.A., Cliff, W. H., Rommens, J.M., Marvin, S.A., Tsui, L.-S., Collins, F.S., Frizzell, R.A., Wilson, J.M. 1990. Correction of the cystic fibrosis defect in vitro by retrovirus-mediated gene transfer.Cell 62:1227–1233

    PubMed  Google Scholar 

  12. Giraldez, F., Murray, K.J., Sepúlveda, F.V., Sheppard, D.N. 1989. Characterization of a phosphorylation-activated Cl-selective channel in isolatedNecturus enterocytes.J. Physiol. 416:517–537

    PubMed  Google Scholar 

  13. Gögelein, H., Schlatter, E., Greger, R. 1987. The ‘small’ conductance chloride channel in the luminal membrane of the rectal gland of the dogfish.Pfluegers Arch. 409:122–125

    Google Scholar 

  14. Gray, M.A., Greenwell, J.R., Argent, B.E. 1988. Secretin-regulated chloride channel on the apical plasma membrane of pancreatic duct cells.J. Membrane Biol. 105:131–142

    Google Scholar 

  15. Gray, M.A., Harris, A., Coleman, L., Greenwell, J.R., Argent, B.E. 1989. Two types of chloride channel on duct cells cultured from human fetal pancreas.Am. J. Physiol. 257:C240-C251

    PubMed  Google Scholar 

  16. Gray, M.A., Pollard, C.E., Harris, A., Coleman, L., Greenwell, J.R., Argent, B.E. 1990. Anion selectivity and block of the small conductance chloride channel on pancreatic duct cells.Am. J. Physiol. 259:752–761

    Google Scholar 

  17. Halm, D.R., Rechkemmer, G.R., Schoumacher, R.A., Frizzell, R.A. 1988. Apical membrane chloride channels in a colonic cell line activated by secretory agonists.Am. J. Physiol. 254:C505-C511

    PubMed  Google Scholar 

  18. Hamill, O.P., Marty, A., Neher, E., Sakmann, B., Sigworth, F.J. 1981. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.Pfluegers Arch. 391:85–100

    Google Scholar 

  19. Harris, A., Coleman, L. 1989. Ductal epithelial cells cultured from human fetal epididymis and vas deferens: Relevance to sterility in cystic fibrosis.J. Cell Sci. 92:687–690

    PubMed  Google Scholar 

  20. Hayslett, J.P., Gögelein, H., Kunzelmann, K., Greger, R. 1987. Characteristics of apical chloride channels in human colonic cells (HT29).Pfluegers Arch. 410:487–494

    Google Scholar 

  21. Hinton, B.T., Turner, T.T. 1988. Is the epididymis a kidney analogue?.News Physiol. Sci. 3:28–31

    Google Scholar 

  22. Hwang, T.-C., Lu, L., Zeitlin, P.L., Gruenert, D.C., Huganir, R., Guggino, W.B. 1989. Cl channels in CF: Lack of activation by protein kinase C and cAMP-dependent protein kinase.244:1351–1353

  23. Kartner, N., Hanrahan, J.W., Jensen, T.J., Naismith, A.L., Sun, S., Ackerley, C.A., Reyes, E.F., Tsui, L.-C., Rommens, J.M., Bear, C.E., Riordan, J.R. 1991. Expression of the cystic fibrosis gene in non-epithelial invertebrate cells produces a regulated anion conductance.Cell 64:681–691

    PubMed  Google Scholar 

  24. Li, M., McCann, D., Liedtke, C.M., Nairn, A.C., Greengard, P., Welsh, M.J. 1988. Cyclic AMP-dependent protein kinase opens chloride channels in normal but not cystic fibrosis airway epithelium.Nature 331:358–360

    PubMed  Google Scholar 

  25. Li, M., McCann, J.D., Anderson, M.P., Clancy, J.P., Liedtke, C.M., Nairn, A.C., Greengard, P., Welsh, M.J. 1989. Regulation of chloride channels by protein kinase C in normal and cystic fibrosis airway epithelia.Science 244:1353–1356

    PubMed  Google Scholar 

  26. Marty, A., Tan, Y.P., Trautmann, A. 1984. Three types of calcium-dependent channel in rat lacrimal glands.J. Physiol. 357:293–325

    PubMed  Google Scholar 

  27. Okamura, N., Tajima, Y., Sugita, Y. 1988. Decrease in bicarbonate transport activities during epididymal maturation of porcine sperm.Biochem. Biophys. Res. Commun. 157:1280–1287

    PubMed  Google Scholar 

  28. Pollard, C.E., Harris, A., Coleman, L., Argent, B.E. 1990. Chloride channels on human epididymal cellsin vitro.J. Physiol. 426:72P

    Google Scholar 

  29. Schoumacher, R.A., Shoemaker, R.L., Halm, D.R., Tallant, E.A., Wallace, R.W., Frizzell, R.A. 1987. Phosphorylation fails to activate chloride channels from cystic fibrosis airway cells.Nature 330:752–754

    PubMed  Google Scholar 

  30. Tabcharani, J.A., Jensen, T.J., Riordan, J.R., Hanrahan, J.W. 1989. Bicarbonate permeability of the outwardly rectifying anion channel.J. Membrane Biol. 112:109–122

    Google Scholar 

  31. Tabcharani, J.A., Low, W., Elie, D., Hanrahan, J.W. 1990. Low-conductance chloride channel activated by cAMP in the epithelial cell line T84.FEBS Lett. 270:157–164

    PubMed  Google Scholar 

  32. Wong, P.Y.D. 1988. Mechanism of adrenergic stimulation of anion secretion in cultured rat epididymal epithelium.Am. J. Physiol. 254:F121-F133

    PubMed  Google Scholar 

  33. Wong, P.Y.D. 1988. Inhibition by chloride channel blockers of anion secretion in cultured epididymal epithelium and intact epididymis of rats.Br. J. Pharmacol. 94:155–163

    PubMed  Google Scholar 

  34. Wong, P.Y.D., Chan, T.P.T. 1988. Adrenergic control of electrogenic anion secretion in primary cultures of rat epididymal cells.In: Gastrointestinal and hepatic Secretions: Mechanism and Control. J.S. Davison and E.A. Shaffer, editors. pp. 216–219. University of Calgary Press, Calgary

    Google Scholar 

  35. Wong, P.Y.D., Huang, S.J. 1989. Intracellular pH measurement in primary monolayer cultures of rat epididymal cells.Pfluegers Arch. 413:414–421

    Google Scholar 

  36. Wong, P.Y.D., Huang, S.J. 1990. Secretory agonists stimulate a rise in intracellular cyclic AMP but nor Ca2+ and inositol phosphates in cultured rat epididymal epithelium.Exp. Physiol. 75:321–337

    PubMed  Google Scholar 

  37. Wong, P.Y.D., Yeung, C.H. 1978. Absorptive and secretory functions of the perfused rat cauda epididymidis.J. Physiol. 275:13–26

    PubMed  Google Scholar 

  38. Worrell, R.T., Butt, A.G., Cliff, W.H., Frizzell, R.A. 1989. A volume-sensitive chloride conductance in human colonic cell line T84.Am. J. Physiol. 256:C1111-C1119

    PubMed  Google Scholar 

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Pollard, C.E., Harris, A., Coleman, L. et al. Chloride channels on epithelial cells cultured from human fetal epididymis. J. Membrain Biol. 124, 275–284 (1991). https://doi.org/10.1007/BF01994360

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  • DOI: https://doi.org/10.1007/BF01994360

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