Effect of ouabain on electrolyte concentrations in principal and intercalated cells of the isolated perfused cortical collecting duct

  • Michael Sauer
  • Adolf Dörge
  • Klaus Thurau
  • Franz-X. Beck
Transport Processes, Metabolism and Endocrinology; Kidney, Gastroinstestinal Tract, and Exocrine Glands


Sodium, phosphorus, chloride and potassium concentrations were measured by a new method in individual principal and intercalated cells in the cortical collecting duct in vitro. Electron microprobe analysis was applied to freezedried cryosections of the isolated perfused rabbit cortical collecting duct. Cell analyses were performed under control conditions and after addition of ouabain to the bath. Under control conditions similar sodium, potassium, chloride, and phosphorus concentration (means±SEM) were observed in principal (10.0±0.6, 126.5±2.7, 24.6±1.0, and 121.5±3.5 mmol/kg wet weight, respectively) and intercalated cells (9.0±0.9, 127.1±4.2, 27.4±1.8, and 118.7±4.9 mmol/kg wet weight, respectively). In principal cells ouabain (10 min) caused an increase in sodium and chloride concentrations by 104 and 13 mmol/kg wet weight, and a decrease in potassium and phosphorus concentrations by 106 and 32 mmol/kg wet weight. These changes in cell element concentrations can be ascribed to an exchange of intracellular potassium against extracellular sodium and to cell swelling due to influx of extracellular fluid. The effects of ouabain on intercalated cells were far less pronounced than on principal cells. This different susceptibility to ouabain of principal and intercalated cells can be ascribed to differences in active and passive transmembrane ion transport pathways.

Key words

Cortical collecting duct Isolated perfused tubules Principal cells Intercalated cells Cell electrolyte concentrations Ouabain Electron microprobe analysis 


  1. 1.
    Atkins JL, Burg MB (1985) Bicarbonate transport by isolated perfused rat collecting ducts. Am J Physiol 249:F485–F489PubMedGoogle Scholar
  2. 2.
    Bauer R, Rick R (1978) Computer analysis of X-ray spectra from this biological specimens. X-ray Spectrom 7:63–69CrossRefGoogle Scholar
  3. 3.
    Beck F, Bauer R, Bauer U, Mason J, Dörge A, Rick R, Thurau K (1980) Electron microprobe analysis of intracellular elements in the rat kidney. Kidney Int 17:756–763CrossRefPubMedGoogle Scholar
  4. 4.
    Beck FX, Dörge A, Rick R, Schramm M, Thurau K (1987) Effect of potassium adaptation on the distribution of potassium, sodium and chloride across the apical membrane of renal tubular cells. Pflügers Arch 409:477–485CrossRefPubMedGoogle Scholar
  5. 5.
    Beck FX, Dörge A, Blümner E, Giebisch G, Thurau K (1988) Cell rubidium uptake: A method for studying functional heterogeneity in the nephron. Kidney Int 33:642–651CrossRefPubMedGoogle Scholar
  6. 6.
    Burg M, Grantham J, Abramow M, Orloff J (1966) Preparation and study of fragments of single rabbit nephrons. Am J Physiol 210:1293–1298PubMedGoogle Scholar
  7. 7.
    Chaillet JR, Lopes AG, Boron WF (1985) Basolateral Na−H exchange in the rabbit cortical collecting tubule. J Gen Physiol 86:795–812CrossRefPubMedGoogle Scholar
  8. 8.
    Dörge A, Rick R, Gehring K, Thurau K (1978) Preparation of freeze-dried cryosections for quantitative X-ray microanalysis of electrolytes in biological soft tissues. Pflügers Arch 373:85–97CrossRefPubMedGoogle Scholar
  9. 9.
    Dörge A, Wienecke P, Beck F, Wörndl B, Rick R, Thurau K (1988) Na transport compartment in rabbit urinary bladder. Pflügers Arch 411:681–687CrossRefPubMedGoogle Scholar
  10. 10.
    Doucet A, Barlet C (1986) Evidence for differences in the sensitivity to ouabain of NaK-ATPase along the nephrons of rabbit kidney. J Biol Chem 261:993–995PubMedGoogle Scholar
  11. 11.
    Grantham JJ, Burg MB, Orloff J (1970) The nature of transtubular Na and K transport in isolated rabbit renal collecting tubules. J Clin Invest 49:1815–1826CrossRefPubMedGoogle Scholar
  12. 12.
    Kashgarian M, Biemesderfer D, Caplan M, Forbush B III (1985) Monoclonal antibody to Na,K-ATPase: Immunocytochemical localization along nephron segments. Kidney Int 28:899–913CrossRefPubMedGoogle Scholar
  13. 13.
    Koeppen BM, Biagi BA, Giebisch GH (1983) Intracellular microelectrode characterization of the rabbit cortical collecting duct. Am J Physiol 244:F35–F47PubMedGoogle Scholar
  14. 14.
    Lombard WE, Kokko JP, Jacobson HR (1983) Bicarbonate transport in cortical and outer medullary collecting tubules. Am J Physiol 244:F289–F296PubMedGoogle Scholar
  15. 15.
    McKinney TD, Burg MB (1978) Bicarbonate secretion by rabbit cortical collecting tubules in vitro. J Clin Invest 61:1421–1427CrossRefPubMedGoogle Scholar
  16. 16.
    Muto S, Giebisch G, Sansom S (1987) Effect of adrenalectomy on electrical properties of the rabbit cortical collecting duct: evidence for differential response of two cell types. Am J Physiol 253:F742–F752PubMedGoogle Scholar
  17. 17.
    Natke E, Stoner LC (1982) Na+ transport properties of the peritubular membrane of cortical collecting tubule. Am J Physiol 242:F664–F671PubMedGoogle Scholar
  18. 18.
    O'Neil RG, Hayhurst RA (1985) Functional differentiation of cell types of cortical collecting duct. Am J Physiol 248:F449–F453PubMedGoogle Scholar
  19. 19.
    O'Neil RG, Sansom SC (1984) Characterization of apical cell membrane Na+ and K+ conductance of cortical collecting duct using microelectrode techniques. Am J Physiol 247:F14–F24PubMedGoogle Scholar
  20. 20.
    Rick R, Dörge A, Thurau K (1982) Quantitative analysis of electrolytes in frozen dried sections. J Microsc 125:239–247PubMedGoogle Scholar
  21. 21.
    Rick R, Roloff C, Dörge A, Beck FX, Thurau K (1984) Intracellular electrolyte concentrations in the frog skin epithelium: Effect of vasopressin and dependence on the Na concentration in the bathing media. J Membr Biol 78:129–145CrossRefPubMedGoogle Scholar
  22. 22.
    Sachs L (1984) Angewandte Statistik: Anwendung statistischer Methoden (6th edn) Springer, Berlin Heidelberg New YorkGoogle Scholar
  23. 23.
    Sansom SC, Weinman EJ, O'Neil RG (1984) Microelectrode assessment of chloride-conductive properties of cortical collecting duct. Am J Physiol 247:F291–F302PubMedGoogle Scholar
  24. 24.
    Sansom S, Agulian S, Giebisch G (1989) Mineralocorticoid regulation of intracellular K activity (Ki) of principal cells (PC) of isolated cortical collecting duct (CCD). Am J Physiol 256:F136–F142PubMedGoogle Scholar
  25. 25.
    Schlatter E, Schafer JA (1987) Electrophysiological studies in principal cells of rat cortical collecting tubules: ADH increases the apical Na+-conductance. Pflügers Arch 409:81–92CrossRefPubMedGoogle Scholar
  26. 26.
    Schlatter E, Schafer JA (1988) Intracellular chloride activity in principal cells of rat cortical collecting ducts (CCT). Pflügers Arch 411:R86 (abstr)Google Scholar
  27. 27.
    Schuster VL, Stokes JB (1987) Chloride transport by the cortical and outer medullary collecting duct. Am J Physiol 253:F203–F212PubMedGoogle Scholar
  28. 28.
    Stokes JB (1981) Potassium secretion by cortical collecting tubule: relation to sodium absorption, luminal sodium concentration, and transepithelial voltage. Am J Physiol 241:F395–F402PubMedGoogle Scholar
  29. 29.
    Stoner LC, Burg MB, Orloff J (1974) Ion transport in cortical collecting tubule; effect of amiloride. Am J Physiol 227:453–459PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • Michael Sauer
    • 1
  • Adolf Dörge
    • 1
  • Klaus Thurau
    • 1
  • Franz-X. Beck
    • 1
  1. 1.Physiologisches Institut der Universität MünchenMünchen 2Federal Republic of Germany

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