Skip to main content
SpringerLink
Log in

Western Blot Analysis of the Delta (δ)-Opioid Receptor in Activated Murine T Cells

  • Chapter
Drugs of Abuse, Immunomodulation, and Aids

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 437))

Abstract

Although it has been known for many years that opioids are immunomodulatory in vivo and can modulate immune related functions in isolated cells in vitro (reviews include 1–3) it has proven difficult to confirm the absence or presence of neural type opioid receptors in the immune system. Using sequence information made available by the recent cloning of all of the known major opioid receptor types (4) it has been possible to document mRNA expression for δ, K, and ε opioid receptors, as well as for an orphan opioid receptor, in different immune cell populations (5–11). In this laboratory (12), δ opioid receptor mRNA that was apparently identical to the δ opioid receptor cDNA originally cloned from NG108-15 cells (13,14), was found to be induced in CD41 and CD8+ T cells purified from concanavalin A (Con A) stimulated splenocyte cultures, δ opioid receptor mRNA was not detected in unstimulated T cells (12).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
eBook
USD 16.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 54.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Sibinga, N.E.S. and Goldstein, A. (1988) Opioid peptides and opioid receptors in cells of the immune system. Annu. Rev. Immunol. 6, 219–249

    Article  PubMed  CAS  Google Scholar 

  2. Stefano, G.B. (1989) Role of opioid neuropeptides in immune regulation. Prog. Neurobiol. 33, 149–159

    Article  PubMed  CAS  Google Scholar 

  3. Bhargava, H.N. (1990) Opioid peptides, receptors, and immune function. NIDA Res. Monogr. 96, 220–233

    PubMed  CAS  Google Scholar 

  4. Knapp, R.J., Malatynska, E., Collins, N., Fang, L., Wang, J.Y., Hruby, V.J., Roeske, W.R., and Yamamura, H.I. (1995) Molecular biology and pharmacology of cloned opioid receptors. FASEB J. 9, 516–525

    PubMed  CAS  Google Scholar 

  5. Chuang, L.F., Chuang, T.K., Killam, K.F., Jr., Chuang, A.J., Kung, H., Yu, L, and Chuang, Y.R. (1994) Delta opioid receptor gene expression in lymphocytes. Biochem. Biophys. Res. Comm. 202, 1291–1299

    Article  PubMed  CAS  Google Scholar 

  6. Chuang, L.F., Chuang, T.K., Killam, K.F. Jr., Qiu, Q., Wang, X.R., Lin, J.J., Kung, H.F., Sheng, W., Chao, C., and Yu, L. (1995) Expression of kappa opioid receptors in human and monkey lymphocytes. Biochem. Biophys. Res. Comm. 209, 1003–1010

    Article  PubMed  CAS  Google Scholar 

  7. Belkowski, S.M., Zhu, J., Liu-Chen, L.Y., Eisenstein, T.K., Adler, M.W., and Rogers, T.J. (1995) Detection of kappa-opioid receptor mRNA in immature T cells. Adv. Exptl. Med. & Biol. 373, 11–16

    Article  CAS  Google Scholar 

  8. Haiford, W.P., Gebhardt, B.M., and Carr, D.J.J. (1995) Functional role and sequence analysis of a lymphocyte orphan opioid receptor. J. Neuroimmunol. 59, 91–101

    Article  Google Scholar 

  9. Gaveriaux, C.J., Paluso, J., Simonin, F., Laforet, J., and Kieffer, B. (1995) Identification of kappa-and delta-opioid receptor transcripts in immune cells. FEBS Lett. 369, 272–276

    Article  PubMed  CAS  Google Scholar 

  10. Sedqi, M., Roy, S, Ramakrishnan, S., Elde, R., and Loh, H.H. (1995) Complementary DNA cloning of a mu-opioid receptor from rat peritoneal macrophages. Biochem. Biophys. Res. Comm. 209, 563–74

    Article  PubMed  CAS  Google Scholar 

  11. Sharp, B., McKean, D., Shahabi, N., McAllen, K., and Heagy, W. (1996) Expression and regulation of delta opioid receptor mRNAs in murine lymphoid tissues. J. Neuroimmunol. 69, 4–5

    Google Scholar 

  12. Miller, B.C. (1996) δ Opioid Receptor Expression Is Induced by Concanavalin A in CD4+ T Cells. J. Immunology 157, 5324–5328

    CAS  Google Scholar 

  13. Evans, C.J., Keith, D.E., Jr., Magendzo, K., Morrison, H. and Edwards, R.H. (1992) Cloning of a delta opioid receptor by functional expression. Science 258, 1952–1955

    Article  PubMed  CAS  Google Scholar 

  14. Kieffer, B.L., Befort, K., Gaveriaux-Ruff, C. and Hirth, C.G. (1992) The δ-opioid receptor: Isolation of a cDNA by expression cloning and pharmacological characterization. Proc. Natl. Acad. Sci. USA 89, 12048–12052

    Article  PubMed  CAS  Google Scholar 

  15. Dado, R.J., Law, P.Y., Loh, H.H., and Elde, R. (1993) Immunofluorescent identification of a delta (δ) opioid receptor on primary afferent nerve terminals Neuroreport 5, 341–344

    Article  PubMed  CAS  Google Scholar 

  16. Arvidsson, U., Dado, R.J., Riedl, M., Lee, J.-H., Law, P.Y., Loh, H.H., Elde, R., and Wessendorf, M.W. (1995) δ-opioid receptor immunoreactivity: Distribution in brainstem and spinal cord, and relationship to biogenic amines and enkephalin J. Neuroscience 15, 1215–1235

    CAS  Google Scholar 

  17. Elde, R., Arvidsson, U., Riedl, M., Vulchanova, L., Lee, J.H., Daoa, R., Nakano, A., Chakrabarti, S., Zhang, X., and Loh, H.H. (1995) Distribution of neuropeptide receptors. New views of peptidergic neurotransmission made possible by antibodies to opioid receptors Ann. New York Acad. Sci. 757, 390–404

    Article  CAS  Google Scholar 

  18. Lang, P., Guizani, L., Vitte-Mony, I., Stancou, R., Dorseuil, O., Gacon, G., and Bertoglio, J. (1992) ADP-ribosylation of the ras-related, GTP-binding protein RhoA inhibits lymphocyte mediated cytotoxicity. J. Biol. Chem. 267, 11677–11680

    PubMed  CAS  Google Scholar 

  19. Laemmli, U.K. (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4 Nature (London) 227, 680–685

    Article  CAS  Google Scholar 

  20. Dunn, S.D. (1986) Effects of the Modification of Transfer Buffer Composition and the Renaturation of Proteins in Gels on the Recognition of Proteins on Western Blots by Monoclonal Antibodies Analytical Biochemistry 157, 144–153

    Article  PubMed  CAS  Google Scholar 

  21. Towbin, H., Staehelin, T., and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications Proc. Natl. Acad. Sci. USA 76, 4350–4354

    Article  PubMed  CAS  Google Scholar 

  22. Gaudriault, G., Nouel, D., Dal Farra, C., Beaudet, A., and Vincent, J.-P. (1996) Receptor induced internali-zation of selective peptidic ε and δ opioid ligands J. Biol. Chem. 272, 2880–2888

    Google Scholar 

  23. Simonds, W.F., Burke, T.R., Jr., Rice, K.C., Jacobson, A.E., and Klee, W.A. (1985) Purification of the opiate receptor of NG108-15 neuroblastoma-glioma hybrid cells. Proc. Natl. Acad. Sci. USA 82, 4974–4979

    Article  PubMed  CAS  Google Scholar 

  24. Carr, D.J.J., DeCosta, B.R., Jacobson, A.E., Bost, K.L., Rice, K.C., and Blalock, J.E. (1987) Immunoaffmity-purified opiate receptor specifically binds the δ-class opiate receptor ligand, cis-(+)-3-methylfen-tanylisothiocyanate, SUPERFIT. FEBS Lett. 224, 272–276

    Article  PubMed  CAS  Google Scholar 

  25. Cucumel, K. Cupo, A. (1995) Anti-idiotypic antibodies: a useful alternative for studying the biochemical expression of mu/delta opioid binding sites in mammalian brain. J. Neuroimmunol. 62, 183–195

    Article  PubMed  CAS  Google Scholar 

  26. Standifer, K.M., Jenab, S., Su, W., Chien, C-C, Pan, Y-X, Inturrisi, C.E., and Pasternak, G.W. (1995) Antisense oligodeoxynucleotides to the cloned δ receptor DOR-1: Uptake, stability, and regulation of gene expression J. Neurochemistry 65, 1981–1987

    Article  CAS  Google Scholar 

  27. Cho, T.M., Yamato, C., Cho, J.S., and Loh, H.H. (1981) Solubilization of membrane bound opiate receptor from rat brain Life Science 28, 2651–2657

    Article  CAS  Google Scholar 

  28. Harada, J., Ueda, H., Iso, Y, and Satoh, M. (1992) Affinity cross-linked δ-opioid receptor in NG108-15 cells is low molecular weight (25 kda) and coupled to GTP-binding proteins Eur. J. Pharmacol. 227, 301–307

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Blvd., Dallas, Texas, 75235-9038, USA

    Bonnie C. Miller

Authors
  1. Bonnie C. Miller
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. University of South Florida College of Medicine, Tampa, Florida, USA

    Herman Friedman  & Thomas W. Klein  & 

  2. Emory University, Atlanta, Georgia, USA

    John J. Madden

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Springer Science+Business Media New York

About this chapter

Cite this chapter

Miller, B.C. (1998). Western Blot Analysis of the Delta (δ)-Opioid Receptor in Activated Murine T Cells. In: Friedman, H., Madden, J.J., Klein, T.W. (eds) Drugs of Abuse, Immunomodulation, and Aids. Advances in Experimental Medicine and Biology, vol 437. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-5347-2_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-4615-5347-2_17

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-7439-8

  • Online ISBN: 978-1-4615-5347-2

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation