Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1983 Jul 15;214(1):29–35. doi: 10.1042/bj2140029

Mineralocorticoid-specificity of aldosterone-induced protein synthesis in giant-toad (Bufo marinus) urinary bladders.

M Geheb, R Alvis, E Hercker, M Cox
PMCID: PMC1152206  PMID: 6412695

Abstract

We have identified a group of proteins (Mr approximately 70000-80000; pI approximately 5.8-6.4) in giant-toad (Bufo marinus) urinary-bladder epithelial cells whose synthesis appears to be related to aldosterone-stimulated Na+ transport. To define this relationship further, we examined whether submaximal natriferic concentrations of aldosterone induced these proteins and whether spironolactone (a specific mineralocorticoid antagonist in renal epithelia) inhibited their synthesis. Short-circuit current was used to measure Na+ transport and epithelial-cell protein synthesis was detected with high-resolution two-dimensional polyacrylamide-gel electrophoresis and autoradiography. Submaximal natriferic concentrations of aldosterone (1.4 X 10(-8) M) induced the same proteins as maximal concentrations of the hormone (1.4 X 10(-7) M). In contrast, in previous experiments, similar proteins were not induced by subnatriferic concentrations (5.0 X 10(-8) M) of cortisol, a glucocorticoid. A spironolactone/aldosterone molar ratio of 2000:1 was required to inhibit aldosterone-stimulated Na+ transport completely; ratios of 200:1 and 500:1 produced partial inhibition. Concentrations of spironolactone that abolished aldosterone-stimulated Na+ transport also inhibited aldosterone-induced protein synthesis. We conclude that the synthesis of the proteins we have identified is specifically related to activation of the mineralocorticoid pathway.

Full text

PDF
29

Images in this article

33Fig. 3.
on p.33
34Fig. 4.
on p.34

Selected References

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

  1. Benjamin W. B., Singer I. Aldosterone-induced protein in toad urinary bladder. Science. 1974 Oct 18;186(4160):269–272. doi: 10.1126/science.186.4160.269. [DOI] [PubMed] [Google Scholar]
  2. Corvol P., Claire M., Oblin M. E., Geering K., Rossier B. Mechanism of the antimineralocorticoid effects of spirolactones. Kidney Int. 1981 Jul;20(1):1–6. doi: 10.1038/ki.1981.97. [DOI] [PubMed] [Google Scholar]
  3. Fanestil D. D. Mode of spirolactone action: competitive inhibition of aldosterone binding to kidney mineralocorticoid receptors. Biochem Pharmacol. 1968 Oct;17(10):2240–2242. doi: 10.1016/0006-2952(68)90203-7. [DOI] [PubMed] [Google Scholar]
  4. Farman N., Kusch M., Edelman I. S. Aldosterone receptor occupancy and sodium transport in the urinary bladder of Bufo marinus. Am J Physiol. 1978 Sep;235(3):C90–C96. doi: 10.1152/ajpcell.1978.235.3.C90. [DOI] [PubMed] [Google Scholar]
  5. Geheb M., Hercker E., Singer I., Cox M. Subcellular localization of aldosterone-induced proteins in toad urinary bladders. Biochim Biophys Acta. 1981 Mar 6;641(2):422–426. doi: 10.1016/0005-2736(81)90499-5. [DOI] [PubMed] [Google Scholar]
  6. Guzzo J., Cox M., Kelley A. B., Singer I. Tetracycline-induced inhibition of Na+ transport in the toad urinary bladder. Am J Physiol. 1978 Oct;235(4):F359–F366. doi: 10.1152/ajprenal.1978.235.4.F359. [DOI] [PubMed] [Google Scholar]
  7. Herman T. S., Fimognari G. M., Edelman I. S. Studies on renal aldosterone-binding proteins. J Biol Chem. 1968 Jul 25;243(14):3849–3856. [PubMed] [Google Scholar]
  8. Kusch M., Farman N., Edelman I. S. Binding of aldosterone to cytoplasmic and nuclear receptors of the urinary bladder epithelium of Bufo marinus. Am J Physiol. 1978 Sep;235(3):C82–C89. doi: 10.1152/ajpcell.1978.235.3.C82. [DOI] [PubMed] [Google Scholar]
  9. Law P. Y., Edelman I. S. Induction of citrate synthase by aldosterone in the rat kidney. J Membr Biol. 1978 Jun 22;41(1):41–64. doi: 10.1007/BF01873339. [DOI] [PubMed] [Google Scholar]
  10. Marver D., Stewart J., Funder J. W., Feldman D., Edelman I. S. Renal aldosterone receptors: studies with (3H)aldosterone and the anti-mineralocorticoid (3H)spirolactone (SC-26304). Proc Natl Acad Sci U S A. 1974 Apr;71(4):1431–1435. doi: 10.1073/pnas.71.4.1431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  12. Porter G. A. In vitro inhibition of aldosterone-stimulated sodium transport by steroidal spirolactones. Mol Pharmacol. 1968 May;4(3):224–237. [PubMed] [Google Scholar]
  13. Reich I. M., Skipski I. A., Scott W. N. Mechanisms of hormonal modulation of ion transport in the toad's urinary bladder. Subcellular localization of aldosterone-induced proteins. Biochim Biophys Acta. 1981 Sep 4;676(3):379–385. doi: 10.1016/0304-4165(81)90174-4. [DOI] [PubMed] [Google Scholar]
  14. Rossier B. C., Claire M., Rafestin-Oblin M. E., Geering K., Gäggeler H. P., Corvol P. Binding and antimineralocorticoid activities of spirolactones in toad bladder. Am J Physiol. 1983 Jan;244(1):C24–C31. doi: 10.1152/ajpcell.1983.244.1.C24. [DOI] [PubMed] [Google Scholar]
  15. Rossier B. C., Gäggeler H. P., Rossier M. Effects of 3'deoxyadenosine and actinomycin D on RNA synthesis in toad bladder: analysis of response to aldosterone. J Membr Biol. 1978 Jun 28;41(2):149–166. doi: 10.1007/BF01972630. [DOI] [PubMed] [Google Scholar]
  16. Rossier B. C. Role of RNA in the action of aldosterone on Na+ transport. J Membr Biol. 1978;40(Spec No):187–197. doi: 10.1007/BF02026005. [DOI] [PubMed] [Google Scholar]
  17. Rossier B. C., Wilce P. A., Edelman I. S. Kinetics of RNA labeling in toad bladder epithelium: effects of aldosterone and related steroids. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3101–3105. doi: 10.1073/pnas.71.8.3101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rossier B. C., Wilce P. A., Edelman I. S. Spironolactone antagonism of aldosterone action on Na+ transport and RNA metabolism in toad bladder epithelium. J Membr Biol. 1977 Apr 7;32(1-2):177–194. doi: 10.1007/BF01905216. [DOI] [PubMed] [Google Scholar]
  19. Sakauye C., Feldman D. Agonist and antimineralocorticoid activities of spirolactones. Am J Physiol. 1976 Jul;231(1):93–97. doi: 10.1152/ajplegacy.1976.231.1.93. [DOI] [PubMed] [Google Scholar]
  20. Scott W. N., Reich I. M., Brown J. A., Jr, Yang C. P. Comparison of toad bladder aldosterone-induced proteins and proteins synthesized in vitro using aldosterone-induced messenger RNA as template. J Membr Biol. 1978;40(Spec No):213–220. doi: 10.1007/BF02026007. [DOI] [PubMed] [Google Scholar]
  21. Scott W. N., Reich I. M., Goodman D. B. Inhibition of fatty acid synthesis prevents the incorporation of aldosterone-induced proteins into membranes. J Biol Chem. 1979 Jun 25;254(12):4957–4959. [PubMed] [Google Scholar]
  22. Sharp G. W., Leaf A. Mechanism of action of aldosterone. Physiol Rev. 1966 Oct;46(4):593–633. doi: 10.1152/physrev.1966.46.4.593. [DOI] [PubMed] [Google Scholar]
  23. USSING H. H., ZERAHN K. Active transport of sodium as the source of electric current in the short-circuited isolated frog skin. Acta Physiol Scand. 1951 Aug 25;23(2-3):110–127. doi: 10.1111/j.1748-1716.1951.tb00800.x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES