Skip to main content
NCBI home page
Cell Proliferation logoLink to Cell Proliferation
. 2003 Jan 31;32(4):215–229. doi: 10.1046/j.1365-2184.1999.3240215.x

Differential effects of opioid and adrenergic agonists on proliferation in a cultured cell line

D Agarwal 1, JA Glasel 2
PMCID: PMC6726328  PMID: 10614711

Abstract

Abstract. A novel clonal cell line transfected with the δ‐opioid receptor (δ‐OR) encoding gene was used to study agonist‐activated regulation of cell proliferation. In this cell line, endogenous β2‐adrenergic receptors (β2‐ARs) are coexpressed with the exogenous δ‐ORs. Upon individual acute treatments with morphine and procaterol (a selective β2‐AR agonist), both the δ‐OR and β2‐AR are coupled to differential modulation of cyclic AMP (cAMP) levels in accord with the classical second messenger response patterns to these agonists in the normal cellular settings of the receptors. But chronic morphine activation of the δ‐OR inhibits cellular proliferation, while chronic procaterol activation of the δ2‐AR stimulates it. Chronic treatment with the individual agonists is accompanied by differential activation of the mitogen‐activated protein kinase (MAPK) isozymes, extracellular‐regulated kinase (ERK) and c‐Jun N‐terminal kinase (JNK). The findings suggest that chronic β2‐AR activation stimulates proliferation by interacting with the ERK signalling cascade independent of a cAMP‐mediated pathway. In contrast to treatment with individual agonists, chronic dual agonist treatment suppresses procaterol‐induced stimulation of ERK activity and stimulation of proliferation indicating that a cross‐regulatory interaction occurs between the δ‐OR and β2‐AR signalling systems in the cells under these conditions.

Full Text

The Full Text of this article is available as a PDF (1.8 MB).

REFERENCES

  1. Allen JM, Baetge EE, Abrass IB, Palmiter RD. (1988) Isoproterenol response following transfection of the mouse beta 2‐adrenergic receptor gene into Y1 cells. EMBO J. 7, 133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradford MM. (1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein‐dye binding. Anal. Biochem. 72, 248. [DOI] [PubMed] [Google Scholar]
  3. Evans CJ, Keith DEJ, Morrison H, Magendzo K, Edwards RH. (1992) Cloning of a delta opioid receptor by functional expression. Science 258, 1952. [DOI] [PubMed] [Google Scholar]
  4. Fukuda K, Kato S, Morikawa H, Shoda T, Mori K. (1996. Functional coupling of the δ‐, μ‐, and k‐opioid receptors to mitogen‐activated protein kinase and arachodonate release in chinese hamster ovary cells. J. Neurochem. 67, 1309. [DOI] [PubMed] [Google Scholar]
  5. Gardner AM, Lange‐carter CA, Vaillancourt RR, Johnson GL. (1994. Measuring activation of kinases in mitogen‐activated protein kinase regulatory network. Meth. Enzymol. 238, 258. [DOI] [PubMed] [Google Scholar]
  6. Glasel JA, Agarwal D. (1997a) A hormone‐defined cell system for studying G‐protein coupled receptor agonist‐activated growth modulation: δ‐opioid and serotonin‐5HT2C receptor activation show opposite mitogenic effects. J. Cell. Physiol. 171, 61. [DOI] [PubMed] [Google Scholar]
  7. Glasel JA, Agarwal D. (1997b) Theoretical analysis of a morphine withdrawal phenotype in a cultured cell line. Pharmacol. Lett. 61, PL305. [DOI] [PubMed] [Google Scholar]
  8. Kieffer B. (1999) Opioids: first lessons from knockout mice. Trends Pharmacol. Sci. 20, 19–26. [DOI] [PubMed] [Google Scholar]
  9. Lazar DF, Wiese RJ, Brady MJ et al. (1995) Mitogen‐activated protein kinase kinase inhibition does not block the stimulation of glucose utilization by insulin. J. Biol. Chem. 270, 20801. [DOI] [PubMed] [Google Scholar]
  10. Leslie FM, Chen Y, Winzer‐serhan UH. (1998. Opioid receptor and peptide mRNA expression in proliferative zones of fetal rat central nervous system. Can J. Physiol. Pharmacol. 76, 284. [PubMed] [Google Scholar]
  11. Li L, Chang K. (1996. The stimulatory effect of opioids on mitogen‐activated protein kinase in chinese hamster ovary cells transfected to express δ‐opioid receptors. Mol. Pharmacol. 50, 599. [PubMed] [Google Scholar]
  12. Libert F, Parmentier AL, Van Dinsart CSJ et al. (1989) Selective amplification and cloning of four new members of the G protein‐coupled receptor family. Science 244, 569. [DOI] [PubMed] [Google Scholar]
  13. Martin WR, Jasinski DR. (1969) Physiological parameters of morphine dependence in man — tolerance, early abstinence, protracted abstinence. J. Psychiat. Res. 7, 9. [DOI] [PubMed] [Google Scholar]
  14. Miller BS, Shankavaram UT, Horney MJ, Gore AC, Kurtz DT, Rosenzweig SA. (1996) Activation of cJun NH2‐terminal kinase/stress‐activated protein kinase by insulin. Biochemistry 35, 8769. [DOI] [PubMed] [Google Scholar]
  15. Minden A, Lin A, Mcmahon M et al. (1994) Differential activation of ERK and JNK mitogen‐activated protein kinases by Raf‐1 and MEKK. Science 266, 1719. [DOI] [PubMed] [Google Scholar]
  16. Minneman KP, Hedberg A, Molinoff PB. (1979) Comparison of beta‐adrenergic receptor subtypes in mammalian tissues. J. Pharmacol. Exptl. Ther. 211, 502. [PubMed] [Google Scholar]
  17. Mitchell FM, Russell M, Johnson GL. (1995) Differential calcium dependence in the activation of c‐jun kinase and mitogen‐activated protein kinase by muscarinic acetylcholine receptors in rat 1a cells. Biochem. J. 309, 381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mosmann T. (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Meth. 65, 55. [DOI] [PubMed] [Google Scholar]
  19. Prasad MV, Dermott JM, Heasley LE, Johnson GL, Dhanasekaran N. (1995) Activation of jun kinase/stress‐activated protein kinase by GTPase‐deficient mutants of G alpha 12 and G alpha 13. J. Biol. Chem. 270, 18655. [DOI] [PubMed] [Google Scholar]
  20. Selbie LA, Hill SJ. (1998) G Protein‐coupled‐receptor cross‐talk: the fine tuning of multiple receptor‐signalling pathways. Trends Pharmacol. Sci. 19, 87. [DOI] [PubMed] [Google Scholar]
  21. Sharma SK, Nirenberg M, Klee WA. (1975) Morphine receptors as regulators of adenylate cyclase activity. Proc. Natl. Acad. Sci. (USA) 72, 590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Staehelin M, Simons P, Jaeggi K, Wigger N. (1983) CGP‐12177. A hydrophilic β‐adrenergic receptor radioligand reveals high affinity binding of agonists to intact cells. J. Biol. Chem. 258, 3496. [PubMed] [Google Scholar]
  23. Widnell KL, Chen J‐S, Iredale PA et al. (1996) Transcriptional regulation of CREB (cyclic AMP response element‐binding protein) expression in CATH.a cells. J. Neurochem. 66, 1770. [DOI] [PubMed] [Google Scholar]
  24. Zadina JE, Hackler L, Ge LJ, Kastin AJ. (1997) A potent and selective endogenous agonist for the μ‐opiate receptor. Nature 386, 499. [DOI] [PubMed] [Google Scholar]
  25. Zhu Y, Pintar JE. (1998) Expression of opioid receptors and ligands in pregnant mouse uterus and placenta. Biol. Reprod. 59, 925. [DOI] [PubMed] [Google Scholar]

Articles from Cell Proliferation are provided here courtesy of Wiley

RESOURCES