Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1988 Nov;85(21):7867–7871. doi: 10.1073/pnas.85.21.7867

A Ca2+-linked increase in coupled cAMP synthesis and hydrolysis is an early event in cholinergic and beta-adrenergic stimulation of parotid secretion.

M A Deeg 1, R M Graeff 1, T F Walseth 1, N D Goldberg 1
PMCID: PMC282298  PMID: 2460856

Abstract

The dynamics and compartmental characteristics of cAMP metabolism were examined by 18O labeling of cellular adenine nucleotide alpha phosphoryls in rat parotid gland stimulated to secrete with beta-adrenergic and cholinergic agents. The secretory response occurred in association with a rapidly increased rate of cAMP hydrolysis apparently coordinated with an equivalent increase in the rate of cAMP synthesis, since the cellular concentration of cAMP remained unchanged. The magnitude of this metabolic response was equivalent to the metabolism of 10-75 times the cellular content of cAMP within the first minute of stimulation. This increased metabolic rate occurred only during the early (1-3 min) period of stimulation, in what appeared to be an exclusive cellular compartment distinguished by a unique distribution of 18O among adenine nucleotide alpha phosphoryls. This 18O distribution contrasted with that produced by forskolin, which increased cellular cAMP concentration and elicited only a delayed response missing the early secretory component. The early acceleration of cAMP metabolism appeared linked to a stimulus-induced increase in intracellular Ca2+ concentration, since the Ca2+ ionophore ionomycin produced the same metabolic response in association with secretion. These observations suggest that cAMP metabolism is involved in stimulus-secretion coupling by a Ca2+-linked mechanism different from that in which cAMP plays the role of a second messenger.

Full text

PDF
7867

Images in this article

7868Image
on p.7868
7868Image
on p.7868

Selected References

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

  1. Ames A., 3rd, Walseth T. F., Heyman R. A., Barad M., Graeff R. M., Goldberg N. D. Light-induced increases in cGMP metabolic flux correspond with electrical responses of photoreceptors. J Biol Chem. 1986 Oct 5;261(28):13034–13042. [PubMed] [Google Scholar]
  2. Aub D. L., Putney J. W., Jr Properties of receptor-controlled inositol trisphosphate formation in parotid acinar cells. Biochem J. 1985 Jan 1;225(1):263–266. doi: 10.1042/bj2250263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bilezikian J. P., Dornfeld A. M., Gammon D. E. Structure-binding-activity analysis of beta-adrenergic amines--I. Binding to the beta receptor and activation of adenylate cyclase. Biochem Pharmacol. 1978 May 15;27(10):1445–1454. doi: 10.1016/0006-2952(78)90100-4. [DOI] [PubMed] [Google Scholar]
  4. Butcher F. R., Goldman J. A., Nemerovski Effect of adrenergic agents on alpha-amylase release and adenosine 3',5'-monophosphate accumulation in rat parotid tissue slices. Biochim Biophys Acta. 1975 May 5;392(1):82–94. doi: 10.1016/0304-4165(75)90168-3. [DOI] [PubMed] [Google Scholar]
  5. Butcher F. R., McBride P. A., Rudich L. Cholinergic regulation of cyclic nucleotide levels, amylase release, and K+ efflux from rat parotid glands. Mol Cell Endocrinol. 1976 Aug-Sep;5(3-4):243–254. doi: 10.1016/0303-7207(76)90087-3. [DOI] [PubMed] [Google Scholar]
  6. Butcher F. R., Putney J. W., Jr Regulation of parotid gland function by cyclic nucleotides and calcium. Adv Cyclic Nucleotide Res. 1980;13:215–249. [PubMed] [Google Scholar]
  7. Butcher F. R. Regulation of calcium efflux from isolated rat parotid cells. Biochim Biophys Acta. 1980 Jun 19;630(2):254–260. doi: 10.1016/0304-4165(80)90429-8. [DOI] [PubMed] [Google Scholar]
  8. Dawis S. M., Graeff R. M., Heyman R. A., Walseth T. F., Goldberg N. D. Regulation of cyclic GMP metabolism in toad photoreceptors. Definition of the metabolic events subserving photoexcited and attenuated states. J Biol Chem. 1988 Jun 25;263(18):8771–8785. [PubMed] [Google Scholar]
  9. Dreux C., Imhoff V., Huleux C., Busson S., Rossignol B. Forskolin, a tool for rat parotid secretion studies: 45Ca efflux is not related to cAMP. Am J Physiol. 1986 Nov;251(5 Pt 1):C754–C762. doi: 10.1152/ajpcell.1986.251.5.C754. [DOI] [PubMed] [Google Scholar]
  10. Gilman A. G. G proteins: transducers of receptor-generated signals. Annu Rev Biochem. 1987;56:615–649. doi: 10.1146/annurev.bi.56.070187.003151. [DOI] [PubMed] [Google Scholar]
  11. Goldberg N. D., Ames A. A., 3rd, Gander J. E., Walseth T. F. Magnitude of increase in retinal cGMP metabolic flux determined by 18O incorporation into nucleotide alpha-phosphoryls corresponds with intensity of photic stimulation. J Biol Chem. 1983 Aug 10;258(15):9213–9219. [PubMed] [Google Scholar]
  12. Goldberg N. D., Haddox M. K. Cyclic GMP metabolism and involvement in biological regulation. Annu Rev Biochem. 1977;46:823–896. doi: 10.1146/annurev.bi.46.070177.004135. [DOI] [PubMed] [Google Scholar]
  13. Graeff R. M., Walseth T. F., Goldberg N. D. The dynamics of cGMP metabolism in neuroblastoma N1E-115 cells determined by 18O labeling of guanine nucleotide alpha-phosphoryls. Neurochem Res. 1987 Jun;12(6):551–560. doi: 10.1007/BF01000240. [DOI] [PubMed] [Google Scholar]
  14. Harper J. F., Brooker G. Alcohol potentiation of isoproterenol-stimulated cyclic AMP accumulation in rat parotid. J Cyclic Nucleotide Res. 1980;6(1):51–62. [PubMed] [Google Scholar]
  15. Harper J. F., Brooker G. Amylase secretion from the rat parotid: refractoriness to muscarinic and adrenergic agonists. Mol Pharmacol. 1978 Nov;14(6):1031–1045. [PubMed] [Google Scholar]
  16. Kanagasuntheram P., Lim S. C. Calcium-dependent inhibition of protein synthesis in rat parotid gland. Biochem J. 1978 Oct 15;176(1):23–29. doi: 10.1042/bj1760023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kikuchi A., Kozawa O., Kaibuchi K., Katada T., Ui M., Takai Y. Direct evidence for involvement of a guanine nucleotide-binding protein in chemotactic peptide-stimulated formation of inositol bisphosphate and trisphosphate in differentiated human leukemic (HL-60) cells. Reconstitution with Gi or Go of the plasma membranes ADP-ribosylated by pertussis toxin. J Biol Chem. 1986 Sep 5;261(25):11558–11562. [PubMed] [Google Scholar]
  18. Ku K. Y., Butcher F. R. Detection of a calmodulin-sensitive cyclic nucleotide phosphodiesterase in rat parotid gland. Biochim Biophys Acta. 1980 Aug 1;631(1):70–78. doi: 10.1016/0304-4165(80)90054-9. [DOI] [PubMed] [Google Scholar]
  19. Merritt J. E., Rink T. J. Rapid increases in cytosolic free calcium in response to muscarinic stimulation of rat parotid acinar cells. J Biol Chem. 1987 Apr 15;262(11):4958–4960. [PubMed] [Google Scholar]
  20. Oron Y., Kellogg J., Larner J. Alpha adrenergic and cholinergic-muscarinic regulation of adenosine cyclic 3',5'-monophosphate levels in the rat parotid. Mol Pharmacol. 1978 Nov;14(6):1018–1030. [PubMed] [Google Scholar]
  21. Piascik M. T., Babich M., Jacobson K. L., Watson E. L. Calmodulin activation and calcium regulation of parotid gland adenylate cyclase. Am J Physiol. 1986 Apr;250(4 Pt 1):C642–C645. doi: 10.1152/ajpcell.1986.250.4.C642. [DOI] [PubMed] [Google Scholar]
  22. Putney J. W., Jr Biphasic modulation of potassium release in rat parotid gland by carbachol and phenylephrine. J Pharmacol Exp Ther. 1976 Aug;198(2):375–384. [PubMed] [Google Scholar]
  23. Seamon K. B., Wetzel B. Interaction of forskolin with dually regulated adenylate cyclase. Adv Cyclic Nucleotide Protein Phosphorylation Res. 1984;17:91–99. [PubMed] [Google Scholar]
  24. Spearman T. N., Durham J. P., Butcher F. R. The role of cyclic AMP in the regulation of exocytosis in the rat parotid gland: evidence obtained with the isoproterenol analog PI-39. J Cyclic Nucleotide Res. 1982;8(4):225–234. [PubMed] [Google Scholar]
  25. Takemura H. Changes in cytosolic free calcium concentration in isolated rat parotid cells by cholinergic and beta-adrenergic agonists. Biochem Biophys Res Commun. 1985 Sep 30;131(3):1048–1055. doi: 10.1016/0006-291x(85)90196-2. [DOI] [PubMed] [Google Scholar]
  26. Walseth T. F., Gander J. E., Eide S. J., Krick T. P., Goldberg N. D. 18O labeling of adenine nucleotide alpha-phosphoryls in platelets. Contribution by phosphodiesterase-catalyzed hydrolysis of cAMP. J Biol Chem. 1983 Feb 10;258(3):1544–1558. [PubMed] [Google Scholar]
  27. Walseth T. F., Graeff R. M., Goldberg N. D. Monitoring cyclic nucleotide metabolism in intact cells by 18O labeling. Methods Enzymol. 1988;159:60–74. doi: 10.1016/0076-6879(88)59008-0. [DOI] [PubMed] [Google Scholar]
  28. Watanabe A. M., McConnaughey M. M., Strawbridge R. A., Fleming J. W., Jones L. R., Besch H. R., Jr Muscarinic cholinergic receptor modulation of beta-adrenergic receptor affinity for catecholamines. J Biol Chem. 1978 Jul 25;253(14):4833–4836. [PubMed] [Google Scholar]
  29. Zeilig C. E., Goldberg N. D. Cell-cycle-related changes of 3':5'-cyclic GMP levels in Novikoff hepatoma cells. Proc Natl Acad Sci U S A. 1977 Mar;74(3):1052–1056. doi: 10.1073/pnas.74.3.1052. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES