Laboratory Investigation

Kidney International (1975) 7, 161–169; doi:10.1038/ki.1975.24

Effect of hypotonicity on cyclic adenosine monophosphate formation and action in vasopressin target cells

Patrick Eggena1, John Christakis1 and Ludwig Deppisch1

1Departments of Physiology and Pathology, the Mount Sinai Medical and Graduate Schools of the City University of New York, New York City, New York

Correspondence: Dr Patrick Eggena, Department of Physiology, Mount Sinai School of Medicine, Fifth Avenue and 100th Street, New York, New York 10029, U.S.A.

Received 2 May 1974; Revised 14 October 1974.

Top

Abstract

Effect of hypotonicity on cyclic adenosine monophosphate formation and action in vasopressin target cells. We have shown previously that overhydration of toads renders their urinary bladders less responsive to the antidiuretic action of vasopressin (AVP). The present study investigates the relationship between osmotic swelling of vasopressin target cells and their sensitivity to AVP and dibutyryl cyclic adenosine monophosphate (db-cAMP). Conditions which engender osmotic swelling of toad bladder epithelial cells, such as immersing bladders on both surfaces in hypotonic Ringer's fluid or subjecting them to a net mucosal-to-serosal volume flux, markedly inhibited the effectiveness of db-cAMP in raising bladder permeability to water. This inhibitory phenomenon was seen both with serosal and mucosal applications of the nucleotide. Examination of isolated epithelial cells by phase contrast microscopy showed them to behave as osmometers, doubling their volume as the effective osmolality of the incubation medium was halved. AVP was found to increase the total content of cAMP about 3.5-fold both in the swollen and the normal cells, so that the actual concentration of cAMP may have diminished as the cell volume increased. Consistent with this suggestion was the observation that increasing exogenous db-cAMP abolished, in part, the inhibitory effects of hypotonicity. These observations indicate that homeostasis of body fluids in the toad depends in part upon the osmotic regulation of antidiuretic hormone action, and that intracellular cAMP may participate in coupling changes in cell volume to the altered state of responsiveness of the vasopressin target cell.

Effet de l'hypotonicité sur la formation et l'action de l'AMP cyclique dans les cellules cibles de la vasopressine. Nous avons montré antérieurement que l'hyperhydratation chez le crapaud rend leurs vessies moins sensibles à l'action de la vasopressine (AVP). Ce travail étudie les relations entre le gonflement osmotique des cellules cibles de la vasopressine et leur sensibilité à l'AVP et au dibutyryl AMP cyclique (db-cAMP). Les conditions qui engendrent le gonflement osmotique des cellules épithéliales de la vessie de crapaud, telles l'immersion des deux faces des vessies dans du Ringer hypotonique ou l'induction d'un flux net de la muqueuse vers la séreuse, inhibent fortement l'effet de db-cAMP d'augmentation de la perméabilité à l'eau. Ce phénomène inhibiteur a été observé aussi bien après l'application séreuse que muqueuse du nucléotide. L'examen microscopique de cellules épithélilaes isolées, en contraste de phase, montre qu'elles se comportent comme des osmomètres, doublant leur volume quand l'osmolalité efficace du milieu d'incubation est diminuée de moitié. L'AVP augmente le contenu total en AMP cyclique d'un facteur 3,5 aussi bien dans les cellules gonflées que dans les cellules normales, de telle sorte que la concentration réelle d'AMP cyclique peut avoir diminué quand le volume cellulaire augmente. Dans le sens de cette suggestion va l'observation que l'augmentation de db-cAMP exogène abolit en partie les effets inhibiteurs de l'hypotonicité. Ces observations indiquent que l'homéostasie des liquides corporels chez le crapaud dépend en partie de la régulation osmotique de l'action de l'hormone antidiurétique et que l'AMP cyclique intracellulaire peut participer au couplage des modifications du volume cellularie avec les altérations du mode de réponse des cellules cibles de la vasopressine.

Top

References

  1. Bentley PJ: The effect of neurohypophyseal extracts on water transfer across the wall of the isolated urinary bladder of the toad Bufo marinus. J Endocrinol 17:201–209, 1958 | PubMed | ISI | ChemPort |
  2. Grantham J, Burg M: Effect of vasopressin and cyclic AMP on permeability of isolated collecting tubules. Am J Physiol 211:255–259, 1966 | PubMed | ISI | ChemPort |
  3. Orloff J, Handler JS, Preston AS: The similarity of effects of vasopressin, adenosine-3',5'-phosphate (cyclic AMP) and theophylline on the toad bladder. J Clin Invest 41:702–709, 1962 | PubMed | ISI | ChemPort |
  4. Peachey LD, Rasmussen H: Structure of the toad's urinary bladder as related to its physiology. J Biochem Biophys Cytol 10:529–553, 1961
  5. Leaf A, Hays RM: Permeability of the isolated toad bladder to solutes and its modification by vasopressin. J Gen Physiol 45:921–932, 1962 | Article | PubMed | ISI | ChemPort |
  6. Parisi M, Piccinni ZF: The penetration of water into the epithelium of the toad urinary bladder and its modification by oxytocin. J Membr Biol 12:227–246, 1973
  7. DeLorenzo RJ, Walton KG, Curran PF, Greengard P: Regulation of phosphorylation of a specific protein in toadbladder membrane by antidiuretic hormone and cyclic AMP, and its possible relationship to membrane permeability changes. Proc Natl Acad Sci USA 70:880–884, 1973
  8. Eggena P: Glutaraldehyde-fixation method for determining the permeability to water of the toad urinary bladder. Endocrinology 91:240–246, 1972
  9. DiBona DR, Civan MM, Leaf A: The cellular specificity of the effect of vasopressin on toad urinary bladder. J Membr Biol 1:79–91, 1969 | ChemPort |
  10. Jard S, Bourget J, Favard P, Carasso N: The role of intercellular channels in the transepithelial transfer of water and sodium in the frog urinary bladder. J Membr Biol 4:124–147, 1971
  11. Grantham JJ, Ganote CE, Burg MB, Orloff J: Paths of transtubular water flow in isolated renal collecting tubules. J Cell Biol 41:562–576, 1969 | Article | PubMed | ChemPort |
  12. Hays RM, Leaf A: The problem of clinical vasopressin resistance: In vitro studies. Ann Intern Med 54:700–709, 1961 | PubMed | ISI | ChemPort |
  13. Eggena P, Walter R, Schwartz IL: Relationship between hydroosmotic flow and the inhibited response of the toad bladder to vasopressin. Life Sci 7:59–63, 1968
  14. Eggena P: Osmotic regulation of toad bladder responsiveness to neurohypophyseal hormones. J Gen Physiol 60:665–678, 1972
  15. Meienhofer JA, Trzeciak A, Havran RT, Walter R: A solid-phase synthesis of (8-arginine)-vasopressin through crystalline protected nonapeptide intermediate and biological properties of the hormone. J Am Chem Soc 92:7199–7202, 1970
  16. Steiner AL, Kipnis DM, Utiger R, Parker C: Radioimmunoassay for the measurement of adenosine 3',5'-cyelic phosphate. Proc Natl Acad Sci 64:367–373, 1969
  17. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275, 1951 | PubMed | ISI | ChemPort |
  18. Kirchberger MA, Schwartz IL, Walter R: Cyclic 3',5'-AMP-dependent protein kinase activity in toad bladder epithelium. Proc Soc Expt Biol Med 140:657–660, 1972
  19. Dousa TP, Sands H, Hechter O: Cyclic AMP-dependent reversible phosphorylation of renal medullary plasma membrane protein. Endocrinology 91:757–763, 1972
  20. Dousa TP, Walter R, Schwartz IL, Sands H, Hechter O: Role of cyclic AMP in the action of neurohypophyseal hormones on kidney, in Advances in Cyclic Nucleotide Research, edited by Greengard P, Robinson GP, Paoletti R, New York, Raven Press, 1972, vol. 1, pp. 121–135
  21. Schwartz IL: Structure-activity relationships as a tool for the analysis of sequential events in the action of neurohypophyseal hormones on membrane permeability, in Structure Activity Relationships of Protein and Polypeptide Hormones, Part I: Proc of the 2nd Int Symp on Protein and Polypeptide Hormones, edited by Margoulies M, Greenwood FC. Amsterdam, Excerpta Medica Foundation, 1971, pp. 31–37
  22. Shlatz LJ, Kinne R, Kinne-Saffran E, Schwartz IL: Plasma membrane polarity of proximal tubule and collecting duct cells of rat and bovine kidney in relation to the mechanism of action of PTH and ADH. Physiologist 16:451, 1973
  23. De Lorenzo RJ, Greengard P: Activation by adenosine 3'5'-monophosphate of a membrane-bound phosphoprotein phosphatase from toad bladder. Proc Natl Acad Sci USA 70:1831–1835, 1973
  24. Eggena P, Polson AX: Osmotic stimulation of vasotocin secretion by the toad's hypothalamo-neurohypophyseal system in vitro. Endocrinology 94:35–44, 1974

Extra navigation

.
ADVERTISEMENT