Skip to main content
SpringerLink
Log in

Effects of ouabain and temperature on cell membrane potentials in isolated perfused straight proximal tubules of the mouse kidney

  • Transport Processes, Metabolism and Endocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands
  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

In isolated perfused segments of the mouse proximal tubule, the potential difference across the basolateral cell membrane (PDbl) was determined with conventional microelectrodes. Under control conditions with symmetrical solutions it amounted to −62±1 mV (n=118). The potential difference across the epithelium (PDte) was −1.7±0.1 mV (n=45). Transepithelial resistance amounted to 1.82±0.09 kΩ cm (n=28), corresponding to 11.4±0.6 Ω cm2. Increasing bath potassium concentration from 5 to 20 mmol/l depolarized PDbl by +24±1 mV (n=103), and PDte by +1.6±0.1 mV (n=19). Thus, the basolateral cell membrane is preferably conductive to potassium. Rapid cooling of the bath perfusate from 38°C to 10°C led to a transient hyperpolarization of PDbl from −60±1 to −65±1 mV (n=21) within 40 s followed by gradual depolarization by +18±1% (n=14) within 5 min. The transepithelial resistance increased significantly from 1.78±0.11 kΩ cm to 2.20±0.21 kΩ cm (n=15). Rapid rewarming of the bath to 38°C caused a depolarization from −61±2 mV (n=17) to −43±2 mV (n=16) within 15 s followed by a repolarization to −59±2 mV (n=10) within 40 s. Ouabain invariably depolarized PDbl. During both, sustained cooling or application of ouabain, the sensitivity of PDbl to bath potassium concentration decreased in parallel to PDbl pointing to a gradual decrease of potassium conductance. Phlorizin hyperpolarized the cell membrane from −59±2 to −66±1 mV (n=13), virtually abolished the transient hyperpolarization under cooling, and significantly reduced the depolarization after rewarming from +17±2 mV (n=16) to +9±3 mV (n=9).

The present data indicate that the contribution of peritubular potassium conductance to the cell membrane conductance decreases following inhibition of basolateral (Na++K+)-ATPase. Apparently, cooling from 37° to 10°C does not only reduce (Na−+K+)-ATPase activity but in addition luminal sodium uptake mechanisms such as the sodium glucose cotransporter. As a result, cooling leads to an initial hyperpolarization of the cell followed by depolarization only after some delay.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Anner BM (1981) A K-selective cation channel formed by Na−K-ATPase in liposomes. Biochem Int 2:365–371

    Google Scholar 

  2. Bello-Reuss E (1982) Electrical properties of the basolateral membrane of the straight portion of the rabbit proximal renal tubule. J Physiol 326:49–63

    Google Scholar 

  3. Biagi B, Kubota T, Sohtell M, Giebisch G (1981) Intracellular potentials in rabbit proximal tubules perfused in vitro. Am J Physiol 240:F200-F210

    Google Scholar 

  4. Blum RM, Hoffman JF (1971) The membrane locus of Castimulated K transport in energy depleted human red blood cells. J Membr Biol 6:315–328

    Google Scholar 

  5. Boulpaep EL (1979) Electrophysiology of the kidney. In: Giebisch G, Tosteson DC, Ussing HH (eds) Membrane transport in biology. Springer, Berlin Heidelberg New York, pp 97–145

    Google Scholar 

  6. Burckhardt B-Ch, Sato K, Froemter E (1984) Electrophysiological analysis of bicarbonate permeation across the peritubular cell membrane of rat kidney proximal tubule. I. Basic observations. Pflügers Arch 401:34–42

    Google Scholar 

  7. Burckhardt B-Ch, Cassola AC, Froemter E (1984) Electrophysiological analysis of bicarbonate permeation across the peritubular cell membrane of rat kidney proximal tubule. II. Exclusion of HCO3-effects on other ion permeabilities and of coupled electroneutral HCO3-transport. Pflügers Arch 401:43–51

    Google Scholar 

  8. Burg M, Grantham J, Abramov M, Orloff J (1966) Preparation and study of fragments of single rabbit nephrons. Am J Physiol 210:1293–1298

    Google Scholar 

  9. Cardinal J, Lapointe JY, Laprade R (1984) Luminal and peritubular ionic substitutions and intracellular potential of the rabbit proximal convoluted tubule. Am J Physiol 247:F352-F364

    Google Scholar 

  10. Cohen B, Giebisch G (1984) Relationship between potassium conductance and transport in renal tubular epithelium. In: Passow H, Heinz E (eds) Biological membranes — Information and energy transduction, Alan R. Liss Inc. New York

    Google Scholar 

  11. Da Cruz MEM, Kinne R, Lin JT (1983) Temperature dependence ofd-glucose transport in reconstituted liposomes. Biochim Biophys Acta 732:691–698

    Google Scholar 

  12. Davis CW, Finn AL (1982) Sodium transport inhibition by amiloride reduces basolateral membrane potassium conductance in tight epithelia. Science 216:525–527

    Google Scholar 

  13. De Smedt H, Kinne R (1981) Temperature dependence of solute transport and enzyme activities in hog renal brush border membrane vesicles. Biochim Biophys Acta 648:247–253

    Google Scholar 

  14. Froemter E (1982) Electrophysiological analysis of rat renal sugar and amino acid transport. I. Basic phenomena. Pflügers Arch 393:179–189

    Google Scholar 

  15. Froemter E, Gebler B (1977) Electrical properties of amphibian urinary bladder epithelia: III. The cell membrane resistances and the effect of amiloride. Pflügers Arch 371:99–108

    Google Scholar 

  16. Grasset E, Gunter-Smith P, Schultz SG (1983) Effects of Na-coupled alanine transport on intracellular K activities and the K conductance of the basolateral membranes of necturus small intestine. J Membr Biol 71:89–94

    Google Scholar 

  17. Greger R (1981) Cation selectivity of the isolated perfused cortical thick ascending limb of Henle's loop of rabbit kidney. Pflügers Arch 390:30–37

    Google Scholar 

  18. Greger R (1985) Ion transport mechanisms in thick ascending limb of Henle's loop of mammalian nephron. Physiol Rev 65:760–797

    Google Scholar 

  19. Greger R, Hampel W (1981) A modified system for in vitro perfusion of isolated renal tubules. Pflügers Arch 389:175–176

    Google Scholar 

  20. Greger R, Schlatter E (1983) Properties of the lumen membrane of the cortical thick ascending limb of Henle's loop of rabbit kidney. Pflügers Arch 396:325–334

    Google Scholar 

  21. Greger R, Schlatter E (1984) Mechanism of NaCl secretion in rectal gland tubules of spiny dogfish (Squalus acanthias). II. Effect of inhibitors. Pflügers Arch 402:364–375

    Google Scholar 

  22. Helman SI (1972) Determination of electrical resistance of the isolated cortical collecting tubule and its possible anatomical location. Yale J Biol Med 45:339–345

    Google Scholar 

  23. Helman SI, Thompson SM (1982) Interpretation and use of electrical equivalent circuits in studies of epithelial tissues. Am J Physiol 243:F519–531

    Google Scholar 

  24. Joergensen PL (1980) Sodium and potassium ion pump in kidney tubules. Physiol Rev 60:864–917

    Google Scholar 

  25. Lang F, Messner G, Rehwald W (1986) Electrophysiology of sodium coupled transport in proximal renal tubules. Am J Physiol (in press)

  26. Lang F, Messner G, Wang W, Paulmichl M, Oberleithner H, Deetjen P (1984) The influence of intracellular sodium activity on the transport of glucose in proximal tubule of frog kidney. Pflügers Arch 401:14–21

    Google Scholar 

  27. Lang F, Messner G, Wang W, Oberleithner H (1983) Interaction of intracellular electrolytes and tubular transport. Klin Wschr 61:1029–1037

    Google Scholar 

  28. Matsumura Y, Cohen B, Guggino WB, Giebisch G (1984) Regulation of the basolateral potassium conductance of the necturus proximal tubule. J Membr Biol 79:153–161

    Google Scholar 

  29. Messner G, Wang W, Paulmichl M, Oberleithner H, Lang F (1985) Ouabain decreases apparent potassium-conductance in proximal tubules of the amphibian kidney. Pflügers Arch 404:131–137

    Google Scholar 

  30. Messner G, Paulmichl M, Lang F (1985) Correlation of pump activity and potassium conductance in the basolateral cell membrane of frog proximal tubules. Pflügers Arch 403:R15

    Google Scholar 

  31. Paulmichl M, Gstraunthaler G, Lang F (1985) Electrical properties of Madin-Darby-canine-kidney cells. Effects of extracellular potassium and bicarbonate. Pflügers Arch 405:102–107

    Google Scholar 

  32. Samarzija I, Hinton BT, Froemter E (1982) Electrophysiological analysis of rat renal sugar and amino acid transport. II. Dependence on various transport parameters and inhibitors. Pflügers Arch 393:190–197

    Google Scholar 

  33. Schultz SG (1981) Homocellular regulatory mechanism on sodium-transporting epithelia: Avoidance of extinction by “flush-through”. Am J Physiol 241:F579-F590

    Google Scholar 

  34. Schwarz W, Passow H (1983) Ca2+ activated K+ channels in erythrocytes and excitable cells. Ann Rev Physiol 45:359–374

    Google Scholar 

  35. Wang W, Messner G, Oberleithner H, Lang F, Deetjen P (1984) The effect of ouabain on intracellular activities of K+, Na−, Cl−, H+ and Ca2+ in proximal tubules of frog kidneys. Pflügers Arch 401:6–13

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Physiologie der Universität Innsbruck, Fritz-Pregl-Strasse 3, A-6010, Innsbruck, Austria

    H. Völkl & J. Geibel

  2. Max-Planck-Institut für Biophysik, D-6000, Frankfurt, Federal Republic of Germany

    R. Greger & F. Lang

Authors
  1. H. Völkl
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Geibel
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R. Greger
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. F. Lang
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Völkl, H., Geibel, J., Greger, R. et al. Effects of ouabain and temperature on cell membrane potentials in isolated perfused straight proximal tubules of the mouse kidney. Pflügers Arch. 407, 252–257 (1986). https://doi.org/10.1007/BF00585299

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00585299

Key words

Search

Navigation