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. 1993 Sep;110(1):133–138. doi: 10.1111/j.1476-5381.1993.tb13782.x

Characterization of P2-purinoceptor mediated cyclic AMP formation in mouse C2C12 myotubes.

R H Henning 1, M Duin 1, A den Hertog 1, A Nelemans 1
PMCID: PMC2176000  PMID: 8220873

Abstract

1. The formation of adenosine 3':5'-cyclic monophosphate (cyclic AMP) and inositol(1,4,5)trisphosphate (Ins(1,4,5)P3), induced by ATP and other nucleotides was investigated in mouse C2C12 myotubes. 2. ATP (100 microM) and ATP gamma S (100 microM) caused a sustained increase in cyclic AMP content of the cells, reaching a maximum after 10 min. The cyclic AMP content reached a maximum in the presence of 100 microM ATP, followed by a decline at higher ATP concentrations. ATP-induced cyclic AMP formation was inhibited by the P2-purinoceptor antagonist, suramin. 3. Myotubes hydrolysed ATP to ADP at a rate of 9.7 +/- 1.0 nmol mg-1 protein min-1. However, further hydrolysis of ADP to AMP and adenosine was negligible. 4. The cyclic AMP formation induced by ADP (10 microM-1 mM) showed similar characteristics to that induced by ATP, but a less pronounced decline was observed than with ATP. ADP-induced cyclic AMP formation was blocked by suramin, while cyclic AMP formation elicited by adenosine (10 microM-1 mM) was insensitive to suramin. 5. The ATP analogue, alpha,beta-methylene-ATP also induced a suramin-sensitive cyclic AMP formation, while 2-methylthio-ATP and the pyrimidine, UTP, did not affect cyclic AMP levels. 6. Stimulation of the myotubes with ATP or UTP (10 microM-1 mM) caused a concentration-dependent increase in the Ins(1,4,5)P3 content of the cells. ADP (100 microM-1 mM) was less effective. Adenosine did not affect Ins(1,4,5)P3 levels.(ABSTRACT TRUNCATED AT 250 WORDS)

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Selected References

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

  1. Abou-Samra A. B., Harwood J. P., Manganiello V. C., Catt K. J., Aguilera G. Phorbol 12-myristate 13-acetate and vasopressin potentiate the effect of corticotropin-releasing factor on cyclic AMP production in rat anterior pituitary cells. Mechanisms of action. J Biol Chem. 1987 Jan 25;262(3):1129–1136. [PubMed] [Google Scholar]
  2. Albuquerque E. X., Deshpande S. S., Aracava Y., Alkondon M., Daly J. W. A possible involvement of cyclic AMP in the expression of desensitization of the nicotinic acetylcholine receptor. A study with forskolin and its analogs. FEBS Lett. 1986 Apr 7;199(1):113–120. doi: 10.1016/0014-5793(86)81235-2. [DOI] [PubMed] [Google Scholar]
  3. Bell J. D., Brunton L. L. Enhancement of adenylate cyclase activity in S49 lymphoma cells by phorbol esters. Withdrawal of GTP-dependent inhibition. J Biol Chem. 1986 Sep 15;261(26):12036–12041. [PubMed] [Google Scholar]
  4. Betz H., Changeux J. P. Regulation of muscle acetylcholine receptor synthesis in vitro by cyclic nucleotide derivatives. Nature. 1979 Apr 19;278(5706):749–752. doi: 10.1038/278749a0. [DOI] [PubMed] [Google Scholar]
  5. Birnbaumer L. Receptor-to-effector signaling through G proteins: roles for beta gamma dimers as well as alpha subunits. Cell. 1992 Dec 24;71(7):1069–1072. doi: 10.1016/s0092-8674(05)80056-x. [DOI] [PubMed] [Google Scholar]
  6. Boyajian C. L., Cooper D. M. Potent and cooperative feedback inhibition of adenylate cyclase activity by calcium in pituitary-derived GH3 cells. Cell Calcium. 1990 Apr;11(4):299–307. doi: 10.1016/0143-4160(90)90007-h. [DOI] [PubMed] [Google Scholar]
  7. Brown B. L., Albano J. D., Ekins R. P., Sgherzi A. M. A simple and sensitive saturation assay method for the measurement of adenosine 3':5'-cyclic monophosphate. Biochem J. 1971 Feb;121(3):561–562. doi: 10.1042/bj1210561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Changeux J. P. Compartmentalized transcription of acetylcholine receptor genes during motor endplate epigenesis. New Biol. 1991 May;3(5):413–429. [PubMed] [Google Scholar]
  9. Convents A., De Backer J. P., André C., Vauquelin G. Desensitization of alpha 2-adrenergic receptors in NG 108 15 cells by (-)-adrenaline and phorbol 12-myristate 13-acetate. Biochem J. 1989 Aug 15;262(1):245–251. doi: 10.1042/bj2620245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cusack N. J., Hourani S. M. Subtypes of P2-purinoceptors. Studies using analogues of ATP. Ann N Y Acad Sci. 1990;603:172–181. doi: 10.1111/j.1749-6632.1990.tb37671.x. [DOI] [PubMed] [Google Scholar]
  11. Den Hertog A., Hoiting B., Molleman A., Van den Akker J., Duin M., Nelemans A. Calcium release from separate receptor-specific intracellular stores induced by histamine and ATP in a hamster cell line. J Physiol. 1992 Aug;454:591–607. doi: 10.1113/jphysiol.1992.sp019281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dunn P. M., Blakeley A. G. Suramin: a reversible P2-purinoceptor antagonist in the mouse vas deferens. Br J Pharmacol. 1988 Feb;93(2):243–245. doi: 10.1111/j.1476-5381.1988.tb11427.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fontaine B., Klarsfeld A., Changeux J. P. Calcitonin gene-related peptide and muscle activity regulate acetylcholine receptor alpha-subunit mRNA levels by distinct intracellular pathways. J Cell Biol. 1987 Sep;105(3):1337–1342. doi: 10.1083/jcb.105.3.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Garte S. J., Belman S. Tumour promoter uncouples beta-adrenergic receptor from adenyl cyclase in mouse epidermis. Nature. 1980 Mar 13;284(5752):171–173. doi: 10.1038/284171a0. [DOI] [PubMed] [Google Scholar]
  15. Goureau O., Tanfin Z., Harbon S. Prostaglandins and muscarinic agonists induce cyclic AMP attenuation by two distinct mechanisms in the pregnant-rat myometrium. Interaction between cyclic AMP and Ca2+ signals. Biochem J. 1990 Nov 1;271(3):667–673. doi: 10.1042/bj2710667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Green W. N., Ross A. F., Claudio T. cAMP stimulation of acetylcholine receptor expression is mediated through posttranslational mechanisms. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):854–858. doi: 10.1073/pnas.88.3.854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Henning R. H., Nelemans A., van den Akker J., den Hertog A. The nucleotide receptors on mouse C2C12 myotubes. Br J Pharmacol. 1992 Aug;106(4):853–858. doi: 10.1111/j.1476-5381.1992.tb14424.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Houslay M. D. 'Crosstalk': a pivotal role for protein kinase C in modulating relationships between signal transduction pathways. Eur J Biochem. 1991 Jan 1;195(1):9–27. doi: 10.1111/j.1432-1033.1991.tb15671.x. [DOI] [PubMed] [Google Scholar]
  19. Huganir R. L., Greengard P. cAMP-dependent protein kinase phosphorylates the nicotinic acetylcholine receptor. Proc Natl Acad Sci U S A. 1983 Feb;80(4):1130–1134. doi: 10.1073/pnas.80.4.1130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Häggblad J., Heilbronn E. P2-purinoceptor-stimulated phosphoinositide turnover in chick myotubes. Calcium mobilization and the role of guanyl nucleotide-binding proteins. FEBS Lett. 1988 Aug 1;235(1-2):133–136. doi: 10.1016/0014-5793(88)81248-1. [DOI] [PubMed] [Google Scholar]
  21. Irvine F., Pyne N. J., Houslay M. D. The phorbol ester TPA inhibits cyclic AMP phosphodiesterase activity in intact hepatocytes. FEBS Lett. 1986 Nov 24;208(2):455–459. doi: 10.1016/0014-5793(86)81068-7. [DOI] [PubMed] [Google Scholar]
  22. Katada T., Gilman A. G., Watanabe Y., Bauer S., Jakobs K. H. Protein kinase C phosphorylates the inhibitory guanine-nucleotide-binding regulatory component and apparently suppresses its function in hormonal inhibition of adenylate cyclase. Eur J Biochem. 1985 Sep 2;151(2):431–437. doi: 10.1111/j.1432-1033.1985.tb09120.x. [DOI] [PubMed] [Google Scholar]
  23. Londos C., Cooper D. M., Wolff J. Subclasses of external adenosine receptors. Proc Natl Acad Sci U S A. 1980 May;77(5):2551–2554. doi: 10.1073/pnas.77.5.2551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Miles K., Anthony D. T., Rubin L. L., Greengard P., Huganir R. L. Regulation of nicotinic acetylcholine receptor phosphorylation in rat myotubes by forskolin and cAMP. Proc Natl Acad Sci U S A. 1987 Sep;84(18):6591–6595. doi: 10.1073/pnas.84.18.6591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Miles K., Greengard P., Huganir R. L. Calcitonin gene-related peptide regulates phosphorylation of the nicotinic acetylcholine receptor in rat myotubes. Neuron. 1989 May;2(5):1517–1524. doi: 10.1016/0896-6273(89)90198-0. [DOI] [PubMed] [Google Scholar]
  26. Moss S. J., Harkness P. C., Mason I. J., Barnard E. A., Mudge A. W. Evidence that CGRP and cAMP increase transcription of AChR alpha-subunit gene, but not of other subunit genes. J Mol Neurosci. 1991;3(2):101–108. doi: 10.1007/BF02885531. [DOI] [PubMed] [Google Scholar]
  27. Mulle C., Benoit P., Pinset C., Roa M., Changeux J. P. Calcitonin gene-related peptide enhances the rate of desensitization of the nicotinic acetylcholine receptor in cultured mouse muscle cells. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5728–5732. doi: 10.1073/pnas.85.15.5728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. O'Connor S. E., Dainty I. A., Leff P. Further subclassification of ATP receptors based on agonist studies. Trends Pharmacol Sci. 1991 Apr;12(4):137–141. doi: 10.1016/0165-6147(91)90530-6. [DOI] [PubMed] [Google Scholar]
  29. Okajima F., Tokumitsu Y., Kondo Y., Ui M. P2-purinergic receptors are coupled to two signal transduction systems leading to inhibition of cAMP generation and to production of inositol trisphosphate in rat hepatocytes. J Biol Chem. 1987 Oct 5;262(28):13483–13490. [PubMed] [Google Scholar]
  30. Silinsky E. M. On the association between transmitter secretion and the release of adenine nucleotides from mammalian motor nerve terminals. J Physiol. 1975 May;247(1):145–162. doi: 10.1113/jphysiol.1975.sp010925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tada S., Okajima F., Mitsui Y., Kondo Y., Ui M. P2 purinoceptor-mediated cyclic AMP accumulation in bovine vascular smooth muscle cells. Eur J Pharmacol. 1992 Sep 1;227(1):25–31. doi: 10.1016/0922-4106(92)90138-l. [DOI] [PubMed] [Google Scholar]
  32. Takami K., Hashimoto K., Uchida S., Tohyama M., Yoshida H. Effect of calcitonin gene-related peptide on the cyclic AMP level of isolated mouse diaphragm. Jpn J Pharmacol. 1986 Nov;42(3):345–350. doi: 10.1254/jjp.42.345. [DOI] [PubMed] [Google Scholar]
  33. Yaffe D., Saxel O. Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle. Nature. 1977 Dec 22;270(5639):725–727. doi: 10.1038/270725a0. [DOI] [PubMed] [Google Scholar]
  34. Yoshimasa T., Sibley D. R., Bouvier M., Lefkowitz R. J., Caron M. G. Cross-talk between cellular signalling pathways suggested by phorbol-ester-induced adenylate cyclase phosphorylation. Nature. 1987 May 7;327(6117):67–70. doi: 10.1038/327067a0. [DOI] [PubMed] [Google Scholar]
  35. den Hertog A., Nelemans A., Van den Akker J. The inhibitory action of suramin on the P2-purinoceptor response in smooth muscle cells of guinea-pig taenia caeci. Eur J Pharmacol. 1989 Aug 3;166(3):531–534. doi: 10.1016/0014-2999(89)90370-1. [DOI] [PubMed] [Google Scholar]

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