Skip to main content
NCBI home page
The Journal of Physiology logoLink to The Journal of Physiology
. 1990 May;424:109–131. doi: 10.1113/jphysiol.1990.sp018058

Cation transport by sweat ducts in primary culture. Ionic mechanism of cholinergically evoked current oscillations.

E H Larsen 1, I Novak 1, P S Pedersen 1
PMCID: PMC1189804  PMID: 2167967

Abstract

1. The coiled reabsorptive segment of human sweat ducts was cultured in vitro. Cells were then harvested and plated onto a dialysis membrane which was glued over a hole in a small disc. Cultures were maintained in a low serum, hormone-supplemented medium that allowed the cells to grow to confluency. The disc was then placed as a partition between two compartments of a miniature Ussing chamber. The chamber was mounted on the stage of an inverted microscope and intracellular potentials were recorded under transepithelial open-circuit or voltage clamp conditions. All values are given as means +/- S.E.M. and n refers to the number of preparations or duct cells. 2. Under control conditions, the cultured epithelia developed mucosa-negative transepithelial potentials (Vt) ranging from -2.5 to -38 mV (-13.5 +/- 1.5 mV, n = 36). The basolateral membrane potential (Vb) was -39.4 +/- 0.7 mV (n = 50 cells), and the apical membrane potential (Va) was linearly correlated with Vt:Va = 1.0 Vt -39.3 mV (r = -0.78, n = 50). 3. The epithelium generated inwardly directed short-circuit currents (Isc) of 12-95 microA cm-2 (45 +/- 4 microA cm-2, n = 36) with a steady-state intracellular potential. Vc = -31.1 +/- 0.6 mV and a fractional resistance of the apical membrane, fR = 0.59 +/- 0.01 (n = 115 cells). 4. The Na+ channel blocker amiloride (mucosal bath, 10 microM) abolished Isc -0.8 +/- 0.6 microA cm-2), the cells hyperpolarized to -61.0 +/- 1.2 mV, and fR increased to 0.85 +/- 0.01 (n = 44). These effects were fully reversible. 5. During initial stimulation with the cholinergic agonist, methacholine (serosa, 5 or 10 microM), the short-circuit current increased to 80 +/- 10 microA cm-2, the cells hyperpolarized to -55.8 +/- 1.2 mV, and fR increased to 0.82 +/- 0.01 (n = 35). 6. In short-circuited preparations stimulated with methacholine an increase in mucosal potassium concentration ([K+]m) from 5 to 25 mM had no significant effect, while a similar increase in the serosal K+ concentration ([K+]s) produced a change in Vc of 44 +/- 3 mV per log10[K+]s (n = 9). In non-stimulated preparations this change was only 16 +/- 2 mV per log10[K+]s (n = 13). After blocking the apical Na+ channels with amiloride the slope was 24 +/- 5 mV per log10[K+]s in unstimulated preparations.(ABSTRACT TRUNCATED AT 400 WORDS)

Full text

PDF
109

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berridge M. J., Cobbold P. H., Cuthbertson K. S. Spatial and temporal aspects of cell signalling. Philos Trans R Soc Lond B Biol Sci. 1988 Jul 26;320(1199):325–343. doi: 10.1098/rstb.1988.0080. [DOI] [PubMed] [Google Scholar]
  2. Bijman J., Frömter E. Direct demonstration of high transepithelial chloride-conductance in normal human sweat duct which is absent in cystic fibrosis. Pflugers Arch. 1986;407 (Suppl 2):S123–S127. doi: 10.1007/BF00584941. [DOI] [PubMed] [Google Scholar]
  3. Boucher R. C., Larsen E. H. Comparison of ion transport by cultured secretory and absorptive canine airway epithelia. Am J Physiol. 1988 Apr;254(4 Pt 1):C535–C547. doi: 10.1152/ajpcell.1988.254.4.C535. [DOI] [PubMed] [Google Scholar]
  4. Brayden D. J., Cuthbert A. W., Lee C. M. Human eccrine sweat gland epithelial cultures express ductal characteristics. J Physiol. 1988 Nov;405:657–675. doi: 10.1113/jphysiol.1988.sp017354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Collie G., Buchwald M., Harper P., Riordan J. R. Culture of sweat gland epithelial cells from normal individuals and patients with cystic fibrosis. In Vitro Cell Dev Biol. 1985 Oct;21(10):597–602. doi: 10.1007/BF02620892. [DOI] [PubMed] [Google Scholar]
  6. Doughney C., Pedersen P. S., McPherson M. A., Dormer R. L. Formation of inositol polyphosphates in cultured human sweat duct cells in response to cholinergic stimulation. Biochim Biophys Acta. 1989 Mar 6;1010(3):352–356. doi: 10.1016/0167-4889(89)90061-x. [DOI] [PubMed] [Google Scholar]
  7. Gray P. T. Oscillations of free cytosolic calcium evoked by cholinergic and catecholaminergic agonists in rat parotid acinar cells. J Physiol. 1988 Dec;406:35–53. doi: 10.1113/jphysiol.1988.sp017367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Higgins J. T., Jr, Gebler B., Frömter E. Electrical properties of amphibian urinary bladder epithelia. II. The cell potential profile in necturus maculosus. Pflugers Arch. 1977 Oct 19;371(1-2):87–97. doi: 10.1007/BF00580776. [DOI] [PubMed] [Google Scholar]
  9. Jones C. J., Bell C. L., Quinton P. M. Different physiological signatures of sweat gland secretory and duct cells in culture. Am J Physiol. 1988 Jul;255(1 Pt 1):C102–C111. doi: 10.1152/ajpcell.1988.255.1.C102. [DOI] [PubMed] [Google Scholar]
  10. KOEFOED-JOHNSEN V., USSING H. H. The nature of the frog skin potential. Acta Physiol Scand. 1958 Jun 2;42(3-4):298–308. doi: 10.1111/j.1748-1716.1958.tb01563.x. [DOI] [PubMed] [Google Scholar]
  11. Lee C. M., Carpenter F., Coaker T., Kealey T. The primary culture of epithelia from the secretory coil and collecting duct of normal human and cystic fibrotic eccrine sweat glands. J Cell Sci. 1986 Jul;83:103–118. doi: 10.1242/jcs.83.1.103. [DOI] [PubMed] [Google Scholar]
  12. Maruyama Y., Gallacher D. V., Petersen O. H. Voltage and Ca2+-activated K+ channel in baso-lateral acinar cell membranes of mammalian salivary glands. Nature. 1983 Apr 28;302(5911):827–829. doi: 10.1038/302827a0. [DOI] [PubMed] [Google Scholar]
  13. Nagel W. The intracellular electrical potential profile of the frog skin epithelium. Pflugers Arch. 1976 Sep 30;365(2-3):135–143. doi: 10.1007/BF01067010. [DOI] [PubMed] [Google Scholar]
  14. Pedersen P. S. Cellular mechanism of cholinergic action in epithelial cell cultures derived from the human reabsorptive sweat duct. Prog Clin Biol Res. 1987;254:151–165. [PubMed] [Google Scholar]
  15. Pedersen P. S. Chloride permeability regulation via a cyclic AMP pathway in cultured human sweat duct cells. J Physiol. 1990 Feb;421:379–397. doi: 10.1113/jphysiol.1990.sp017950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pedersen P. S. Human sweat duct cells in primary culture. Basic bioelectric properties of cultures derived from normals and patients with cystic fibrosis. In Vitro Cell Dev Biol. 1989 Apr;25(4):342–352. doi: 10.1007/BF02624597. [DOI] [PubMed] [Google Scholar]
  17. Petersen O. H., Gallacher D. V. Electrophysiology of pancreatic and salivary acinar cells. Annu Rev Physiol. 1988;50:65–80. doi: 10.1146/annurev.ph.50.030188.000433. [DOI] [PubMed] [Google Scholar]
  18. Quinton P. M., Bijman J. Higher bioelectric potentials due to decreased chloride absorption in the sweat glands of patients with cystic fibrosis. N Engl J Med. 1983 May 19;308(20):1185–1189. doi: 10.1056/NEJM198305193082002. [DOI] [PubMed] [Google Scholar]
  19. Quinton P. M. Chloride impermeability in cystic fibrosis. Nature. 1983 Feb 3;301(5899):421–422. doi: 10.1038/301421a0. [DOI] [PubMed] [Google Scholar]
  20. Quinton P. M. Effects of some ion transport inhibitors on secretion and reabsorption in intact and perfused single human sweat glands. Pflugers Arch. 1981 Oct;391(4):309–313. doi: 10.1007/BF00581513. [DOI] [PubMed] [Google Scholar]
  21. SCHWARTZ I. L., THAYSEN J. H. Excretion of sodium and potassium in human sweat. J Clin Invest. 1956 Jan;35(1):114–120. doi: 10.1172/JCI103245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schulz I. J. Micropuncture studies of the sweat formation in cystic fibrosis patients. J Clin Invest. 1969 Aug;48(8):1470–1477. doi: 10.1172/JCI106113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Stoddard J. S., Jakobsson E., Helman S. I. Basolateral membrane chloride transport in isolated epithelia of frog skin. Am J Physiol. 1985 Sep;249(3 Pt 1):C318–C329. doi: 10.1152/ajpcell.1985.249.3.C318. [DOI] [PubMed] [Google Scholar]
  24. Ussing H. H. Volume regulation of frog skin epithelium. Acta Physiol Scand. 1982 Mar;114(3):363–369. doi: 10.1111/j.1748-1716.1982.tb06996.x. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Physiology are provided here courtesy of The Physiological Society

RESOURCES