Pflügers Archiv

, Volume 427, Issue 1–2, pp 24–32 | Cite as

Intracellular Cl concentration in striated intralobular ducts from rabbit mandibular salivary glands

  • K. R. Lau
  • R. L. Evans
  • R. M. Case
Transport Processes, Metabolism and Endocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands


Intralobular striated ducts have been isolated from rabbit mandibular salivary glands and maintained in primary culture for up to 2 days. Such ducts were loaded with the Cl-sensitive fluorescent dyeN-(ethoxycarbonylmethyl)-(6-methoxyquinolinium bromide) (MQAE) and intracellular Cl concentration ([Cl]i monitored using a fluorescence microscope. Intracellular Cl could be rapidly and reversibly emptied from striated duct cells by replacing Cl in the superfusing solution with NO 3 . [Cl]i could be lowered by removal of external Na+, exposure to 10 μM amiloride or to 10 μM 4,4′-diisothiocyanatostilbene-2,2′-disulphonic acid (DIDS). Both amiloride and DIDS were able to inhibit the recovery of [Cl]i after an initial exposure to Na+- or Cl-free solution. The amiloride derivatives, benzamil (2 μM) and N-isobutyl-N-methylamiloride (MIBA), (10 μM) also lowered [Cl]i by similar amounts as 10 μM amiloride. Varying external K+ concentration ([K+]o) also affected [Cl]i. Increasing [K+]o increased [Cl]i, but decreasing [K+]o did not decrease [Cl]i. Instead, [Cl]i was also increased when [K+]o was lowered below the control value. Bumetanide (0.1 mM) lowered [Cl]i by only a small amount, while ouabain (1 mM) had no significant effect on [Cl]i. These data are consistent with current models of electrolyte transport in salivary ducts which include Cl channels, Na+ channels, and Na+/H+ exchangers in the apical membrane. The effects of low [K+]o can be interpreted in terms of a K+-dependent exit mechanism for Cl.

Key words

Salivary gland Striated ducts Intralobular ducts Intracellular Cl Fluorescence MQAE DIDS Amiloride 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Augustus J, Bijman J, Os CH van (1978) Electrical resistance of rabbit submaxillary main duct: a tight epithelium with leaky cell membranes. J Membr Biol 43:203–226Google Scholar
  2. 2.
    Becq F, Fanjul M, Mahieu I, Berger Z, Gola M, Hollande E (1992) Anion channels in a human pancreatic cancer cell line (Capan-1) of ductal origin. Pflügers Arch 420:46–53Google Scholar
  3. 3.
    Bijman J (1982) Transport parameters of the main duct of the rabbit mandibular salivary gland. PhD Thesis, University of Nijmegen, The NetherlandsGoogle Scholar
  4. 4.
    Bijman J, Cook DI, Os CH van (1983) Effect of amiloride on electrolyte transport parameters of the main duct of the rabbit mandibular salivary gland. Pflügers Arch 398:96–102Google Scholar
  5. 5.
    Bridges RJ, Worrell RT, Frizzell RA, Benos DJ (1989) Stilbene disulfonate blockade of colonic secretory Cl channels in planar lipid bilayers. Am J Physiol 256:C 902-C 912Google Scholar
  6. 6.
    Cabantchik ZI, Greger R (1992) Chemical probes for anion transporters of mammalian cell membranes. Am J Physiol 262:C 803-C 827Google Scholar
  7. 7.
    Case RM, Conigrave AD, Novak I, Young JA (1980) Electrolyte and protein secretion by the perfused rabbit mandibular gland stimulated with acetylcholine or catecholamines. J Physiol (Lond) 300:467–487Google Scholar
  8. 8.
    Case RM, Conigrave AD, Favaloro EJ, Novak I, Thompson CH, Young JA (1982) The role of buffer anions and protons in secretion by the rabbit mandibular salivary gland. J Physiol (Lond) 322:273–286Google Scholar
  9. 9.
    Cliff WH, Schoumacher RA, Frizzell RA (1992) cAMP-activated Cl channels in CFTR-transfected cystic fibrosis pancreatic epithelial cells. Am J Physiol 262:C 1154-C 1160Google Scholar
  10. 10.
    Dinudom A, Young JA, Cook DI (1993) Amiloride-sensitive Na+ current in the granular duct cells of mouse mandibular glands. Pflügers Arch 423:164–166Google Scholar
  11. 11.
    Evans RL, Lau KR, Case RM (1992) The effect of isoprenaline on intracellular Cl concentration in isolated rabbit mandibular gland ducts. J Physiol (Lond) 446:351 PGoogle Scholar
  12. 12.
    Evans RL, Lau KR, Case RM (1993) Structural and functional characterization of striated ducts isolated from the rabbit mandibular salivary gland. Exp Physiol 78:49–64Google Scholar
  13. 13.
    Foskett JK (1990) [Ca2+]i modulation of Cl content controls cell volume in single salivary acinar cells during fluid secretion. Am J Physiol 259:C 998-C 1004Google Scholar
  14. 14.
    Frömter E, Gebler B, Schopow K, Pockrandt-Hemstedt J (1974) Cation and anion permeability of rabbit submaxillary main duct. In: Thorn NA, Petersen OH (eds) Secretory mechanisms of exocrine glands. Munksgaard, Copenhagen, pp 469–513Google Scholar
  15. 15.
    Garty H, Benos DJ (1988) Characteristics and regulatory mechanisms of the amiloride-blockable Na+ channel. Physiol Rev 68:309–373Google Scholar
  16. 16.
    Greger R, Schlatter R, Wang F, Forrest JN (1984) Mechanism of NaCl secretion in rectal gland tubules of spiny dogfish (Squalus acanthias). III. Effect of stimulation of secretion by cyclic AMP. Pflügers Arch 411:670–675Google Scholar
  17. 17.
    Karniski LP, Aronson PS (1987) Anion exchange pathways for Cl transport in rabbit renal microvillus membranes. Am J Physiol 253:F 513-F 521Google Scholar
  18. 18.
    Kleyman TR, Cragoe Jr EJ (1990) Cation transport probes: the amiloride series. Methods Enzymol 191:739–755Google Scholar
  19. 19.
    Knauf H (1972) The isolated salivary duct as a model for electrolyte transport studies. Pflügers Arch 333:82–94Google Scholar
  20. 20.
    Knauf H, Lübcke R, Kreutz W, Sachs G (1982) Interrelationships of ion transport in rat submaxillary duct epithelium. Am J Physiol 242:F 132-F 139Google Scholar
  21. 21.
    Knickelbein RG, Dobbins JW (1990) Sulfate and oxalate exchange for bicarbonate across the basolateral membrane of rabbit ileum. Am J Physiol 259:G 807-G 813Google Scholar
  22. 22.
    Krapf R, Berry CA, Verkman AS (1988) Estimation of intracellular chloride activity in isolated perfused rabbit proximal convoluted tubules using a fluorescent indicator. Biophys J 53:955–962Google Scholar
  23. 23.
    Kuo S-M, Aronson PS (1988) Oxalate transport via the sulfate/HCO3 exchanger in rabbit renal basolateral membrane vesicles. J Biol Chem 263:9710–9717Google Scholar
  24. 24.
    Lau KR, Howorth AJ, Case RM (1990) The effects of bumetanide, amiloride and Ba2+ on fluid and electrolyte secretion in rabbit salivary gland. J Physiol (Lond) 425:407–427Google Scholar
  25. 25.
    Lau KR, Evans RL, Case RM (1993) Amiloride and DIDS cause intracellular Cl to fall in intralobular striated ducts isolated from rabbit mandibular salivary glands. J Physiol (Lond) 467:218 PGoogle Scholar
  26. 26.
    Mangos JA, Braun G, Hamann KF (1966) Micropuncture study of sodium and potassium excretion in the rat parotid saliva. Pflügers Arch 291:99–106Google Scholar
  27. 27.
    Marino CR, Matovcik LM, Gorelick FS, Cohn JA (1991) Localization of the cystic fibrosis transmembrane conductance regulator in pancreas and salivary glands. In: The pancreatic duct cell: physiology and pathophysiology. NIH, Baltimore, pp 13–14Google Scholar
  28. 28.
    Martin CJ, Young JA (1971) Electrolyte concentrations in primary and final saliva of the rat sublingual gland studied by micropuncture and catheterization techniques. Pflügers Arch 324:344–360Google Scholar
  29. 29.
    Martin CJ, Fromter E, Gebler B, Knauf H, Young JA (1973) The effects of carbachol on water and electrolyte fluxes and transepithelial electrical potential differences of the rabbit submaxillary main duct perfused in vitro. Pflügers Arch 341: 131–142Google Scholar
  30. 30.
    Matalon S, Bauer ML, Benos DJ, Kleyman TR, Lin C, Cragoe EJ Jr, O'Brodovich H (1993) Fetal lung epithelial cells contain two populations of amiloride-sensitive Na+ channels. Am J Physiol 264:L 357-L 364Google Scholar
  31. 31.
    Novak I, Young JA (1986) Two independent anion transport systems in rabbit mandibular salivary glands. Pflügers Arch 407:649–656Google Scholar
  32. 32.
    Novak I, Pedersen PS, Larsen EH (1992) Chloride and potassium conductances of cultured human sweat ducts. Pflügers Arch 422:151–158Google Scholar
  33. 33.
    Ram SJ, Kirk KL (1989) Cl permeability of human sweat duct cells monitored with fluorescence-digital imaging microscopy: evidence for reduced plasma membrane Cl permeability in cystic fibrosis. Proc Natl Acad Sci USA 86: 10 166–10 170Google Scholar
  34. 34.
    Reddy MM, Quinton PM (1989) Altered electrical potential profile of human reabsorptive sweat duct cells in cystic fibrosis. Am J Physiol 257:C 722-C 726Google Scholar
  35. 35.
    Reddy MM, Quinton PM (1989) Localization of Cl conductance in normal and Cl impermeability in cystic fibrosis sweat duct epithelium. Am J Physiol 257:C 727-C 735Google Scholar
  36. 36.
    Reddy MM, Quinton PM (1991) Intracellular potassium activity and the role of potassium in transepithelial salt transport in the human reabsorptive sweat duct. J Membr Biol 119: 199–210Google Scholar
  37. 37.
    Silva P, Stoff J, Field M, Fine L, Forrest JN, Epstein FH (1977) Mechanism of active chloride secretion by shark rectal gland: role of Na-K-ATPase in chloride transport. Am J Physiol 233: F 298-F 306Google Scholar
  38. 38.
    Slegers JFG, Moons WM, Idzerda PP, Stadhouders AM (1975) The contribution of a chloride shunt to the transmucosal potential of the rabbit submaxillary duct. J Membr Biol 25:213–236Google Scholar
  39. 39.
    Thaysen JH, Thorn NA, Schwartz IL (1954) Excretion of sodium, potassium, chloride and carbon dioxide in human parotid saliva. Am J Physiol 178:155–159Google Scholar
  40. 40.
    Tilmann M, Kunzelmann K, Frobe U, Cabantchik ZI, Lang HJ, Englert HC, Greger R (1991) Different types of blockers of the intermediate conductance outwardly rectifying chloride channel (ICOR) of epithelia. Pflügers Arch 418:556–563Google Scholar
  41. 41.
    Trezise AEO, Buchwald M (1991) In vivo cell-specific expression of the cystic fibrosis transmembrane conductance regulator. Nature 353:434–437Google Scholar
  42. 42.
    Vasseur M, Frangne R, Alvarado F (1993) Buffer-dependent pH sensitivity of the fluorescent chloride-indicator dye SPQ. Am J Physiol 264:C 27-C 31Google Scholar
  43. 43.
    Verkman AS, Sellers MC, Chao AC, Leung T, Ketcham R (1989) Synthesis and characterization of improved chloridesensitive fluorescent indicators for biological applications. Anal Biochem 178:355–361Google Scholar
  44. 44.
    Young JA, Lennep EW van (1979) Transport in salivary and salt glands. In: Giebisch G, Tosteson DC, Ussing HH (eds) Membrane transport in biology IVB. Springer, Berlin Heidelberg New York, pp 563–574Google Scholar
  45. 45.
    Young JA, Schogel E (1966) Micropuncture investigation of sodium and potassium excretion in rat submaxillary saliva. Pflügers Arch 291:85–98Google Scholar
  46. 46.
    Young JA, Cook DI, Lennep EW van, Roberts M (1987) Secretion by the major salivary glands. In: Johnson LR (ed) Physiology of the gastrointestinal tract, 2nd edn. Raven Press, New York, pp 773–815Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • K. R. Lau
    • 1
  • R. L. Evans
    • 2
  • R. M. Case
    • 1
  1. 1.Department of Physiological SciencesUniversity of ManchesterManchesterUK
  2. 2.Clinical Investigation and Patient Care Branch, National Institute of Dental ResearchNational Institutes of HealthBethesdaUSA

Personalised recommendations