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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
. 1980 Jul;77(7):4364–4368. doi: 10.1073/pnas.77.7.4364

Synthesis of enkephalins by adrenal medullary chromaffin cells: reserpine increases incorporation of radiolabeled amino acids.

S P Wilson, K J Chang, O H Viveros
PMCID: PMC349835  PMID: 6933489

Abstract

Adrenal medullary chromaffin cells maintained in a chemically defined, serum-free medium retain high levels of both catecholamines and opiate-like peptides. At least 25% of the opioid activity as measured by radioreceptor assay is present as [Met]- and [Leu]enkephalin. Incubation of chromaffin cells with [3H]tyrosine and [35S]methionine for 3 days results in incorporation of 3H and 35S into [Met]enkephalin and 3H incorporation into [Leu]enkephalin, demonstrating de novo enkephalin synthesis. Addition of 100 nM reserpine to the culture medium causes an 80% depletion of catecholamines and a 100% increase in opioid peptide content 3-4 days later. [Met]Enkephalin content is preferentially induced (290% above control) compared with [Leu]enkephalin (70%) Reserpine increases the fractional radiolabeling of [Met]- and [Leu]enkephalin in excess of control cultures, suggesting that the increase in opiate-like peptides results from an increased rate of synthesis rather than decreased degradation, decreased secretion, or increased processing of preformed precursor(s).

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Selected References

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

  1. ANTON A. H., SAYRE D. F. A study of the factors affecting the aluminum oxide-trihydroxyindole procedure for the analysis of catecholamines. J Pharmacol Exp Ther. 1962 Dec;138:360–375. [PubMed] [Google Scholar]
  2. Austen B. M., Smyth D. G., Snell C. R. gamma endorphin, alpha endorphin and Met-enkephalin are formed extracellularly from lipotropin C fragment. Nature. 1977 Oct 13;269(5629):619–621. doi: 10.1038/269619a0. [DOI] [PubMed] [Google Scholar]
  3. Chang K. J., Cuatrecasas P. Multiple opiate receptors. Enkephalins and morphine bind to receptors of different specificity. J Biol Chem. 1979 Apr 25;254(8):2610–2618. [PubMed] [Google Scholar]
  4. Fenwick E. M., Fajdiga P. B., Howe N. B., Livett B. G. Functional and morphological characterization of isolated bovine adrenal medullary cells. J Cell Biol. 1978 Jan;76(1):12–30. doi: 10.1083/jcb.76.1.12. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gillin J. C., Hong J. S., Yang H. Y., Costa E. [Met5]Enkephalin content in brain regions of rats treated with lithium. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2991–2993. doi: 10.1073/pnas.75.6.2991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hong J. S., Yang H. Y., Fratta W., Costa E. Rat striatal methionine-enkephalin content after chronic treatment with cataleptogenic and noncataleptogenic antischizophrenic drugs. J Pharmacol Exp Ther. 1978 Apr;205(1):141–147. [PubMed] [Google Scholar]
  7. Hughes J., Kosterlitz H. W., McKnight A. T. The incorporation of [3H]-tyrosine into the enkephalins of striatal slices of guinea-pig brain [proceedings]. Br J Pharmacol. 1978 Jun;63(2):396P–396P. [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Lewis R. V., Stein S., Gerber L. D., Rubinstein M., Udenfriend S. High molecular weight opioid-containing proteins in striatum. Proc Natl Acad Sci U S A. 1978 Aug;75(8):4021–4023. doi: 10.1073/pnas.75.8.4021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lewis R. V., Stern A. S., Rossier J., Stein S., Udenfriend S. Putative enkephalin precursors in bovine adrenal medulla. Biochem Biophys Res Commun. 1979 Aug 13;89(3):822–829. doi: 10.1016/0006-291x(79)91852-7. [DOI] [PubMed] [Google Scholar]
  11. Miller R. J., Chang K. J., Cooper B., Cuatrecasas P. Radioimmunoassay and characterization of enkephalins in rat tissues. J Biol Chem. 1978 Jan 25;253(2):531–538. [PubMed] [Google Scholar]
  12. Rossier J., French E., Gros C., Minick S., Guillemin R., Bloom F. E. Adrenalectomy, dexamethasone or stress alters opioid peptides levels in rat anterior pituitary but not intermediate lobe or brain. Life Sci. 1979 Dec 10;25(24-25):2105–2112. doi: 10.1016/0024-3205(79)90204-2. [DOI] [PubMed] [Google Scholar]
  13. Schultzberg M., Lundberg J. M., Hökfelt T., Terenius L., Brandt J., Elde R. P., Goldstein M. Enkephalin-like immunoreactivity in gland cells and nerve terminals of the adrenal medulla. Neuroscience. 1978;3(12):1169–1186. doi: 10.1016/0306-4522(78)90137-9. [DOI] [PubMed] [Google Scholar]
  14. Sosa R. P., McKnight A. T., Hughes J., Kosterlitz H. W. Incorporation of labelled amino acids into the enkephalins. FEBS Lett. 1977 Dec 1;84(1):195–198. doi: 10.1016/0014-5793(77)81088-0. [DOI] [PubMed] [Google Scholar]
  15. Stern A. S., Lewis R. V., Kimura S., Rossier J., Gerber L. D., Brink L., Stein S., Udenfriend S. Isolation of the opioid heptapeptide Met-enkephalin [Arg6,Phe7] from bovine adrenal medullary granules and striatum. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6680–6683. doi: 10.1073/pnas.76.12.6680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Thoenen H. Trans-synaptic enzyme induction. Life Sci. 1974 Jan 16;14(2):223–235. doi: 10.1016/0024-3205(74)90052-6. [DOI] [PubMed] [Google Scholar]
  17. Viveros O. H., Arqueros L., Connett R. J., Kirshner N. Mechanism of secretion from the adrenal medulla. IV. The fate of the storage vesicles following insulin and reserpine administration. Mol Pharmacol. 1969 Jan;5(1):69–82. [PubMed] [Google Scholar]
  18. Viveros O. H., Arqueros L., Kirshner N. Mechanism of secretion from the adrenal medulla. VI. Effect of reserpine on the dopamine -hydroxylase and catecholamine content and on the buoyant density of adrenal storage vesicles. Mol Pharmacol. 1971 Jul;7(4):434–443. [PubMed] [Google Scholar]
  19. Viveros O. H., Diliberto E. J., Jr, Hazum E., Chang K. J. Opiate-like materials in the adrenal medulla: evidence for storage and secretion with catecholamines. Mol Pharmacol. 1979 Nov;16(3):1101–1108. [PubMed] [Google Scholar]
  20. Yang H. Y., Hong J. S., Fratta W., Costa E. Rat brain enkephalins: distribution and biosynthesis. Adv Biochem Psychopharmacol. 1978;18:149–159. [PubMed] [Google Scholar]

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