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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
. 1978 Nov;75(11):5520–5524. doi: 10.1073/pnas.75.11.5520

Temperature sensitivity of cyclic AMP production and catecholamine-induced refractoriness in a rat astrocytoma cell line

G Allen Nickols 1, Gary Brooker 1
PMCID: PMC392997  PMID: 82968

Abstract

Intracellular cyclic AMP was increased more than 100-fold when rat C6-2B astrocytoma cells were treated with isoproterenol in the cold (4°C). When the cells were treated with the phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine, and isoproterenol, cyclic AMP levels rose more than 150-fold. The levels achieved compared favorably with the 250-fold increase in cyclin AMP produced by (-)isoproterenol at 37°C.(-)Isoproterenol at 5 nM stimulated half-maximal cyclic AMP production at 4°C and at 37°C and was blocked by (-)propranolol at both temperatures. The concentrations of cyclic AMP attained by these cells after (-)isoproterenol stimulation in the cold may be accounted for, in part, by alterations in the efflux of the nucleotide from the cells since extracellular cyclic AMP, an indicator of cyclic AMP efflux, was found to be dramatically reduced in the cold.

The cells, when exposed to (-)isoproterenol for up to 6 hr at low temperature, maintained normal responsiveness to this agent when rechallenged at 4° or 37°C. Thus, they did not display agonist-induced refractoriness during that period of exposure at 4°C, although refractoriness is always seen within 90 min at 37°C. Refractoriness, once established by (-)isoproterenol treatment at 37°C, was not reversed by exposure of the cells to cold. These data suggest that the development of catecholamine refractoriness requires a temperature-sensitive step that lies distal to the hormone-receptor interaction and cyclic AMP generaton.

Keywords: isoproterenol, C6-2B cells, cyclic AMP efflux

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Selected References

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

  1. Barber R., Kelly L. A., McGuire R. F., Butcher R. W. Distortion of cyclic AMP responses to catecholamine due to destruction of the hormone. J Cyclic Nucleotide Res. 1977 Aug;3(4):249–261. [PubMed] [Google Scholar]
  2. Brooker G., Terasaki W. L., Price M. G. Gammaflow: a completely automated radioimmunoassay system. Science. 1976 Oct 15;194(4262):270–276. doi: 10.1126/science.184530. [DOI] [PubMed] [Google Scholar]
  3. Ciosek C. P., Jr, Fahey J. V., Ishikawa Y., Newcombe D. S. PGE1-mediated cyclic AMP refractoriness: effects of cycloheximide and indomethacin. J Cyclic Nucleotide Res. 1975;1(4):229–235. [PubMed] [Google Scholar]
  4. DeVellis J., Brooker G. Reversal of catecholamine refractoriness by inhibitors of RNA and protein synthesis. Science. 1974 Dec 27;186(4170):1221–1223. doi: 10.1126/science.186.4170.1221. [DOI] [PubMed] [Google Scholar]
  5. Doore B. J., Bashor M. M., Spitzer N., Mawe R. C., Saier M. H., Jr Regulation of adenosine 3' :5'-monophosphate efflux from rat glioma cells in culture*. J Biol Chem. 1975 Jun 10;250(11):4371–4372. [PubMed] [Google Scholar]
  6. Fichman M. P., Brooker G. Deficient renal cyclic adenosine 3'-5' monophosphate production in nephrogenic diabetes insipidus. J Clin Endocrinol Metab. 1972 Jul;35(1):35–47. doi: 10.1210/jcem-35-1-35. [DOI] [PubMed] [Google Scholar]
  7. Forte L. R., Nickols G. A., Anast C. S. Renal adenylate cyclase and the interrelationship between parathyroid hormone and vitamin D in the regulation of urinary phosphate and adenosine cyclic 3',5'-monophosphate excretion. J Clin Invest. 1976 Mar;57(3):559–568. doi: 10.1172/JCI108311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Harper J. F., Brooker G. Femtomole sensitive radioimmunoassay for cyclic AMP and cyclic GMP after 2'0 acetylation by acetic anhydride in aqueous solution. J Cyclic Nucleotide Res. 1975;1(4):207–218. [PubMed] [Google Scholar]
  9. Houslay M. D., Hesketh T. R., Smith G. A., Warren G. B., Metcalfe J. C. The lipid environment of the glucagon receptor regulates adenylate cyclase activity. Biochim Biophys Acta. 1976 Jun 17;436(2):495–504. doi: 10.1016/0005-2736(76)90211-x. [DOI] [PubMed] [Google Scholar]
  10. Kakiuchi S., Rall T. W. The influence of chemical agents on the accumulation of adenosine 3',5'-Phosphate in slices of rabbit cerebellum. Mol Pharmacol. 1968 Jul;4(4):367–378. [PubMed] [Google Scholar]
  11. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  12. Lesniak M. A., Roth J. Regulation of receptor concentration by homologous hormone. Effect of human growth hormone on its receptor in IM-9 lymphocytes. J Biol Chem. 1976 Jun 25;251(12):3720–3729. [PubMed] [Google Scholar]
  13. Mickey J., Tate R., Lefkowitz R. J. Subsensitivity of adenylate cyclase and decreased beta-adrenergic receptor binding after chronic exposure to (minus)-isoproterenol in vitro. J Biol Chem. 1975 Jul 25;250(14):5727–5729. [PubMed] [Google Scholar]
  14. Newcombe D. S., Ciosek C. P., Jr, Ishikawa Y., Fahey J. V. Human synoviocytes: activation and desensitization by prostaglandins and 1-epinephrine. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3124–3128. doi: 10.1073/pnas.72.8.3124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Orly J., Schramm M. Fatty acids as modulators of membrane functions: catecholamine-activated adenylate cyclase of the turkey erythrocyte. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3433–3437. doi: 10.1073/pnas.72.9.3433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rapoport B., Adams R. J. Induction of refractoriness to thyrotropin stimulation in cultured thyroid cells. Dependence on new protein synthesis. J Biol Chem. 1976 Nov 10;251(21):6653–6661. [PubMed] [Google Scholar]
  17. Su Y. F., Cubeddu L., Perkins J. P. Regulation of adenosine 3':5'-monophosphate content of human astrocytoma cells: desensitization to catecholamines and prostaglandins. J Cyclic Nucleotide Res. 1976 Jul-Aug;2(4):257–270. [PubMed] [Google Scholar]
  18. Su Y. F., Johnson G. L., Cubeddu L., Leichtling B. H., Ortmann R., Perkins J. P. Regulation of adenosine 3':5'-monophosphate content of human astrocytoma cells: mechanism of agonist-specific desensitization. J Cyclic Nucleotide Res. 1976 Jul-Aug;2(4):271–285. [PubMed] [Google Scholar]
  19. Terasaki W. L., Brooker G. [125I]Iodohydroxybenzylpindolol binding sites on intact rat glioma cells. Evidence for beta-adrenergic receptors of high coupling efficiency. J Biol Chem. 1978 Aug 10;253(15):5418–5425. [PubMed] [Google Scholar]
  20. Terasaki W. L., Brooker G., de Vellis J., Inglish D., Hsu C. Y., Moylan R. D. Involvement of cyclic amp and protein synthesis in catecholamine refractoriness. Adv Cyclic Nucleotide Res. 1978;9:33–52. [PubMed] [Google Scholar]

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