Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 299, Issue 3, pp 273–275 | Cite as

Tissue levels of cyclic nucleotides in the rat kidney after administration of furosemide

  • H. Osswald
  • A. Hawlina
  • U. Clasen
  • O. Heidenreich


In Sprague Dawlwy rats left kidneys were quickly frozen 6–35 min after intravenous injection of furosemide (0.5, 5 and 50 mg/kg) using a clamp precooled with liquid nitrogen. Cyclic adenosine-3′:5′-monophosphate (cAMP) extracted from the tissue was assayed by a binding test with cAMP dependent protein kinase from beef heart and cyclic guanosine-3′:5′-monophosphate (cGMP) with a cGMP dependent protein kinase from lobster tail (humarus vulgaris). Glomerular filtration rate (as inulin clearance) and sodium excretion were measured. Within the first 8 min 0.5 mg/kg furosemide increased urine volume and sodium excretion 3-fold. cGMP levels rose two and a half-fold whereas the cAMP content was not significantly changed. Higher doses of 5 and 50 mg/kg furosemide increased urine volume and sodium excretion further. These doses now increased cAMP levels from 3.12±0.31 to 11.3±0.91 and 12.1±0.13 pmol/mg dry weight, respectively (P<0.001). cGMP levels were elevated from 0.44 ±0.05 to 1.97±0.25 and 1.12±0.22 pmol/mg dry weight (P<0.001). 26 min after the administration of the diuretic, tissue levels of both nucleotides had returned to control values.

These data do not support the idea that the cAMP-adenylcyclase-system is involved in the mechanism of diuretic action of furosemide.

Key words

cAMP cGMP Furosemide Diuretic action 


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  1. Baumann, K., Chan, Y. L., Bode, F., Papavassilion, F.: Effect of parathyroid hormone and cyclic nucleotides on the isotonic sodium reabsorption in the proximal convoluted tubule of rat kidney. Pfügers Arch. 347, 142, abstract (1974)Google Scholar
  2. Bentley, P. J.: Amiloride: a potent inhibitor of sodium transport across the toad bladder. J. Physiol. (Lond.) 195, 317–330 (1968)Google Scholar
  3. Duggan, D. E., Noll, R. M.: Effect of ethacrynic acid and cardiac glycosides upon a membrane adenosine triphosphatase of renal cortex. Arch. Biochem. 109, 388–396 (1965)Google Scholar
  4. Ebel, H.: Effect of diuretics on renal Na-K-ATPase and adenylcyclase. Naunyn-Schmiedeberg's Arch. Pharmacol. 281, 301–314 (1974)Google Scholar
  5. Ebel, H., De Santo, N. G., Hierholzer, K.: Plasma cell membranes of the rat kidney. I. Purification and properties of cell membranes ATPase. Pflügers Arch. 324, 1–25 (1971)Google Scholar
  6. Ebel, H., Ehrlich, J., De Santo, N. G., Doerken, U.: Plasma membranes of the kidney. III. Influence of diuretics on ATPase activity. Pflügers Arch. 335, 224–234 (1972)Google Scholar
  7. Ferguson, D. R., Twite, B. R.: Furosemide-Pharmacology and cellular mode of action. Naunyn-Schmiedeberg's Arch. Pharmacol. 281, 295–300 (1974)Google Scholar
  8. Ferguson, D. R., Twite, B. R.: Effect of diuretics on Na-K-ATPase and cAMP levels in toad bladder epithelial cells. Naunyn-Schmiedeberg's Arch. Pharmacol. 287, 111–116 (1975)Google Scholar
  9. Fitzpatrick, D. F., Davenport, G. R., Forte, L., Landon, F. J.: Characterization of plasma membrane proteins in mammalian kidney. I. Preparation of a membrane fraction and separation of the protein. J. Biol. Chem. 244, 3561–3569 (1969)Google Scholar
  10. Führ, J., Kaczmarczyk, J., Krüttgen, C. D.: Eine einfache Methode zur Inulinbestimmung für Nieren-Clearance-Untersuchungen bei Stoffwechselgesunden und Diabetikern. Klin. Wochenschr. 33, 729–730 (1955)Google Scholar
  11. Fülgraff, G., Meiforth, A.: Effects of cyclic nucleotide monophosphates (cAMP, cGMP, CIMP, dibutyryl cAMP) on transtubular net fluxes in rat kidney. Naunyn-Schmiedeberg's Arch. Pharmacol. 283, 425–429 (1974)Google Scholar
  12. Gilman, A. G.: A protein binding assay for adenosine-3′:5′-monophosphate. Proc. Natl. Acad. Sci. (U.S.A.) 67, 305–312 (1970)Google Scholar
  13. Kinne, R., Schmitz, J. E., Kinne-Saffran, E.: The localization of the Na+−K+-ATPase in the cells of rat kidney cortex. A study on isolated plasma membranes. Pflügers Arch. 329, 191–206 (1971)Google Scholar
  14. Kuo, J. F., Lee, T. P., Reyes, P. L., Walton, K. G., Donnelly, T. E., Greengard, P.: Cyclic nucleotide-dependent proteinkinases. An assay method for measurement of guanosine-3′:5′-monophosphate in various biological materials and a study of agents regulating its levels in heart and brain. J. Biol. Chem. 247, 16–22 (1972)Google Scholar
  15. Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with folinphenol reagent. J. Biol. Chem. 193, 265–275 (1951)Google Scholar
  16. Murad, F., Manganiello, V., Vaughan, M.: A simple, sensitive protein binding assay for 3′:5′-guanosine monophosphate. Proc. Natl. Acad. Sci. (U.S.A.) 68, 736–739 (1971)Google Scholar
  17. Nechay, B. R., Nelson, I. A.: Renal ouabain-sensitive adenosine triphosphatase activity and Na+-reabsorption. J. Pharmacol. Exp. Ther. 175, 717–726 (1970)Google Scholar
  18. Osswald, H.: Cyclic guanosine-3′:5′-monophosphate induced diuresis in rats. Naunyn-Schmiedeberg's Arch. Pharmacol. 284, 207–214 (1974)Google Scholar
  19. Radtke H. W., Rumrich, G., Kinne-Saffran, E., Ullrich, J. J.: Dual action of acetazolamide and furosemide on proximal volume absorption in the rat kidney. Kidney Int. 1, 100–105 (1972)Google Scholar
  20. Senft, G., Schultz, G., Munske, K., Hoffmann, M.: Der Einfluß von Hydrochlorothiazid und anderen sulfonamidierten Diuretica auf die 3′:5′-AMP Phosphodiesterase-Aktivität in der Rattenniere. Naunyn-Schmiedeberg's Arch. Pharmacol. 259, 344–359 (1968)Google Scholar
  21. Steiner, A. L., Whitley, T. H., Ong, S.-H. H., Stowe, N. W.: Cyclic AMP and cyclic GMP: Studies utilizing immunobistochemical techniques for the localization of the nucleotides in tissue. Metabolism 24, 419–428 (1975)Google Scholar
  22. Taylor, C. B.: Effect of mercurial diuretics on adenosine triphosphatase of rabbit kidney in vitro. Biochem. Pharmacol. 12, 539–550 (1963)Google Scholar

Copyright information

© Springer-Verlag 1977

Authors and Affiliations

  • H. Osswald
    • 1
  • A. Hawlina
    • 1
  • U. Clasen
    • 1
  • O. Heidenreich
    • 1
  1. 1.Abteilung Pharmakologie der Medizinischen Fakultät der RWTH AachenAachenGermany

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