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. 1986 May;77(5):1436–1440. doi: 10.1172/JCI112455

Enkephalins increase cyclic adenosine monophosphate content, calcium uptake, and contractile state in cultured chick embryo heart cells.

S Laurent, J D Marsh, T W Smith
PMCID: PMC424543  PMID: 2422207

Abstract

Peripheral vascular effects of opioid peptides are well known, but direct myocardial effects have not been established. We studied the inotropic response of spontaneously beating cultured chick embryo ventricular cells to the enkephalin analogue [D-Ala2]-enkephalin. Amplitude of cell motion increased in a concentration-dependent manner with 0.53 microM [D-Ala2]-enkephalin producing half-maximal response. The mechanism of this positive inotropic effect was investigated by examining alterations in 45Ca influx, cyclic AMP accumulation and adenylate cyclase activity in response to [D-Ala2]-enkephalin. At maximally inotropic concentrations, the 45Ca influx rate increased 39%, adenylate cyclase was stimulated by 30%, and cyclic AMP content rose more than twofold. Thus, in contrast to neural tissue, receptors for enkephalin in cultured heart cells are coupled to adenylate cyclase in a stimulatory manner. Occupancy of these receptors produces an increase in cyclic AMP levels and exerts a positive inotropic effect via a verapamil-sensitive enhancement of Ca influx.

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Selected References

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

  1. Barry W. H., Hasin Y., Smith T. W. Sodium pump inhibition, enhanced calcium influx via sodium-calcium exchange, and positive inotropic response in cultured heart cells. Circ Res. 1985 Feb;56(2):231–241. doi: 10.1161/01.res.56.2.231. [DOI] [PubMed] [Google Scholar]
  2. Biedert S., Barry W. H., Smith T. W. Inotropic effects and changes in sodium and calcium contents associated with inhibition of monovalent cation active transport by ouabain in cultured myocardial cells. J Gen Physiol. 1979 Oct;74(4):479–494. doi: 10.1085/jgp.74.4.479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bitar K. N., Makhlouf G. M. Specific opiate receptors on isolated mammalian gastric smooth muscle cells. Nature. 1982 May 6;297(5861):72–74. doi: 10.1038/297072a0. [DOI] [PubMed] [Google Scholar]
  4. Caffrey J. L., Gaugl J. F., Jones C. E. Local endogenous opiate activity in dog myocardium: receptor blockade with naloxone. Am J Physiol. 1985 Mar;248(3 Pt 2):H382–H388. doi: 10.1152/ajpheart.1985.248.3.H382. [DOI] [PubMed] [Google Scholar]
  5. Cardenas H. L., Ross D. H. Calcium depletion of synaptosomes after morphine treatment. Br J Pharmacol. 1976 Aug;57(4):521–526. doi: 10.1111/j.1476-5381.1976.tb10379.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chan T. M., Exton J. H. alpha-Adrenergic-mediated accumulation of adenosine 3':5' monophosphate in calcium-depleted hepatocytes. J Biol Chem. 1977 Dec 10;252(23):8645–8651. [PubMed] [Google Scholar]
  7. Clusin W. T., Hamilton W. E., Nelson D. V. The mechanical activity of chick embryonic myocardial cell aggregates. J Physiol. 1981 Nov;320:149–174. doi: 10.1113/jphysiol.1981.sp013941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Collier H. O., Roy A. C. Morphine-like drugs inhibit the stimulation of E prostaglandins of cyclic AMP formation by rat brain homogenate. Nature. 1974 Mar 1;248(5443):24–27. doi: 10.1038/248024a0. [DOI] [PubMed] [Google Scholar]
  9. Down J. A., Szerb J. C. Kinetics of morphine-sensitive [3H]-acetylcholine release from the guinea-pig myenteric plexus. Br J Pharmacol. 1980 Jan;68(1):47–55. doi: 10.1111/j.1476-5381.1980.tb10697.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Eiden L. E., Ruth J. A. Enkephalins modulate the responsiveness of rat atria in vitro to norepinephrine. Peptides. 1982 May-Jun;3(3):475–478. doi: 10.1016/0196-9781(82)90110-3. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Henderson G., North R. A. Depression by morphine of excitatory junction potentials in the vas deferens of the mouse. Br J Pharmacol. 1976 Jul;57(3):341–346. [PMC free article] [PubMed] [Google Scholar]
  13. Holaday J. W. Cardiovascular effects of endogenous opiate systems. Annu Rev Pharmacol Toxicol. 1983;23:541–594. doi: 10.1146/annurev.pa.23.040183.002545. [DOI] [PubMed] [Google Scholar]
  14. Howells R. D., Kilpatrick D. L., Bhatt R., Monahan J. J., Poonian M., Udenfriend S. Molecular cloning and sequence determination of rat preproenkephalin cDNA: sensitive probe for studying transcriptional changes in rat tissues. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7651–7655. doi: 10.1073/pnas.81.23.7651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Iwatsubo K., Clouet D. H. Dopamine-sensitive adenylate cyclase of the caudate nucleus of rats treated with morphine or haloperidol. Biochem Pharmacol. 1975 Aug 15;24(16):1499–1503. doi: 10.1016/0006-2952(75)90026-x. [DOI] [PubMed] [Google Scholar]
  16. KOSTERLITZ H. W., TAYLOR D. W. The effect of morphine on vagal inhibition of the heart. Br J Pharmacol Chemother. 1959 Jun;14(2):209–214. doi: 10.1111/j.1476-5381.1959.tb01385.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kosterlitz H. W., Lord J. A., Paterson S. J., Waterfield A. A. Effects of changes in the structure of enkephalins and of narcotic analgesic drugs on their interactions with mu- and delta-receptors. Br J Pharmacol. 1980 Feb;68(2):333–342. doi: 10.1111/j.1476-5381.1980.tb10422.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Krishna G., Weiss B., Brodie B. B. A simple, sensitive method for the assay of adenyl cyclase. J Pharmacol Exp Ther. 1968 Oct;163(2):379–385. [PubMed] [Google Scholar]
  19. 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]
  20. Lang R. E., Hermann K., Dietz R., Gaida W., Ganten D., Kraft K., Unger T. Evidence for the presence of enkephalins in the heart. Life Sci. 1983 Jan 24;32(4):399–406. doi: 10.1016/0024-3205(83)90086-3. [DOI] [PubMed] [Google Scholar]
  21. Laurent S., Marsh J. D., Smith T. W. Enkephalins have a direct positive inotropic effect on cultured cardiac myocytes. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5930–5934. doi: 10.1073/pnas.82.17.5930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lechner R. B., Gurll N. J., Reynolds D. G. Naloxone potentiates the cardiovascular effects of catecholamines in canine hemorrhagic shock. Circ Shock. 1985;16(4):347–361. [PubMed] [Google Scholar]
  23. Marsh J. D., Barry W. H., Neer E. J., Alexander R. W., Smith T. W. Evidence for uncoupling of the beta receptor-adenylate cyclase complex. Circ Res. 1980 Oct;47(4):493–501. doi: 10.1161/01.res.47.4.493. [DOI] [PubMed] [Google Scholar]
  24. Marsh J. D., Smith T. W. Receptors for beta-adrenergic agonists in cultured chick ventricular cells. Relationship between agonist binding and physiologic effect. Mol Pharmacol. 1985 Jan;27(1):10–18. [PubMed] [Google Scholar]
  25. Marsh J. D. The cultured heart cell: a useful model for physiological and biochemical investigation. Int J Cardiol. 1983 Jul;3(4):465–468. doi: 10.1016/0167-5273(83)90120-1. [DOI] [PubMed] [Google Scholar]
  26. Martin W. R. Pharmacology of opioids. Pharmacol Rev. 1983 Dec;35(4):283–323. [PubMed] [Google Scholar]
  27. Marvin W. J., Jr, Atkins D. L., Chittick V. L., Lund D. D., Hermsmeyer K. In vitro adrenergic and cholinergic innervation of the developing rat myocyte. Circ Res. 1984 Jul;55(1):49–58. doi: 10.1161/01.res.55.1.49. [DOI] [PubMed] [Google Scholar]
  28. Neer E. J. The vasopressin-sensitive adenylate cyclase of the rat renal medulla. J Biol Chem. 1973 Jul 10;248(13):4775–4781. [PubMed] [Google Scholar]
  29. Puri S. K., Cochin J., Volicer L. Effect of morphine sulfate on adenylate cyclase and phosphodiesterase activities in rat corpus striatum. Life Sci. 1975 Mar 1;16(5):759–767. doi: 10.1016/0024-3205(75)90352-5. [DOI] [PubMed] [Google Scholar]
  30. Reuter H. Ion channels in cardiac cell membranes. Annu Rev Physiol. 1984;46:473–484. doi: 10.1146/annurev.ph.46.030184.002353. [DOI] [PubMed] [Google Scholar]
  31. Rodbell M. The role of hormone receptors and GTP-regulatory proteins in membrane transduction. Nature. 1980 Mar 6;284(5751):17–22. doi: 10.1038/284017a0. [DOI] [PubMed] [Google Scholar]
  32. Ruth J. A., Cuizon J. V., Eiden L. E. Leucine-enkephalin increases norepinephrine-stimulated chronotropy and 45Ca++ uptake in guinea-pig atria. Neuropeptides. 1984 May;4(3):185–191. doi: 10.1016/0143-4179(84)90099-4. [DOI] [PubMed] [Google Scholar]
  33. Sabol S. L., Nirenberg M. Regulation of adenylate cyclase of neuroblastoma x glioma hybrid cells by alpha-adrenergic receptors. I. Inhibition of adenylate cyclase mediated by alpha receptors. J Biol Chem. 1979 Mar 25;254(6):1913–1920. [PubMed] [Google Scholar]
  34. Saunders W. S., Thornhill J. A. No inotropic action of enkephalins or enkephalin derivatives on electrically-stimulated atria isolated from lean and obese rats. Br J Pharmacol. 1985 Jun;85(2):513–522. doi: 10.1111/j.1476-5381.1985.tb08888.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sperelakis N., Pappano A. J. Physiology and pharmacology of developing heart cells. Pharmacol Ther. 1983;22(1):1–39. doi: 10.1016/0163-7258(83)90050-5. [DOI] [PubMed] [Google Scholar]
  36. Steinberg S. F., Drugge E. D., Bilezikian J. P., Robinson R. B. Acquisition by innervated cardiac myocytes of a pertussis toxin-specific regulatory protein linked to the alpha 1-receptor. Science. 1985 Oct 11;230(4722):186–188. doi: 10.1126/science.2994230. [DOI] [PubMed] [Google Scholar]
  37. Waterfield A. A., Smokcum R. W., Hughes J., Kosterlitz H. W., Henderson G. In vitro pharmacology of the opioid peptides, enkephalins and endorphins. Eur J Pharmacol. 1977 May 15;43(2):107–116. doi: 10.1016/0014-2999(77)90123-6. [DOI] [PubMed] [Google Scholar]
  38. Weihe E., McKnight A. T., Corbett A. D., Hartschuh W., Reinecke M., Kosterlitz H. W. Characterization of opioid peptides in guinea-pig heart and skin. Life Sci. 1983;33 (Suppl 1):711–714. doi: 10.1016/0024-3205(83)90601-x. [DOI] [PubMed] [Google Scholar]

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