Inducible Expression of G Protein-Coupled Receptors in Transfected Cells

  • Beryl Koener
  • Emmanuel HermansEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 746)


Biochemical or pharmacological studies of G protein-coupled receptors (GPCRs) are widely conducted in transfected mammalian cells. A variety of commercially available systems allow the generation of stable cell-lines in which expression of the recombinant receptor can be induced on addition of a defined chemical to the culture medium, which operates as a control switch for the transcription of the cloned sequence. Such systems offer the possibility to induce graded levels of receptor expression in the experimental model, or to induce an abrupt downregulation of receptor expression during the maintenance of the cell-line. This chapter provides an overview of the different systems available and provides methods for the generation and validation of stably transfected cell-lines expressing the GPCR of choice.

Key words

Transfection Inducible expression Tetracycline Receptor density 


  1. 1.
    Gossen, M., Bonin, A. L. and Bujard, H. (1993) Control of gene activity in higher eukaryotic cells by prokaryotic regulatory ­elements. Trends Biochem. Sci. 18, 471–475.PubMedCrossRefGoogle Scholar
  2. 2.
    Van Craenenbroeck, K., Vanhoenacker, P., Leysen, J. E. and Haegeman, G. (2001) Evaluation of the tetracycline- and ecdysone-inducible systems for expression of neurotransmitter receptors in mammalian cells. Eur. J. Neurosci. 14, 968–976.PubMedCrossRefGoogle Scholar
  3. 3.
    Downey, P. M., Lozza, G., Petro, R., Diodato, E., Foglia, C., Bottazzoli, F., Brusa, R., Asquini, T., Reggiani, A. and Grilli, M. (2005) Ecdysone-based system for controlled inducible expression of metabotropic glutamate receptor subtypes 2, 5, and 8. J. Biomol. Screen. 10, 841–848.PubMedCrossRefGoogle Scholar
  4. 4.
    Hermans, E., Challiss, R. A.J. and Nahorski, S. R. (1999) Effects of varying the expression level of recombinant human mGlu1a receptors on the pharmacological properties of agonists and antagonists. Br. J. Pharmacol. 126, 873–882.PubMedCrossRefGoogle Scholar
  5. 5.
    Theroux, T. L., Esbenshade, T. A., Peavy, R. D. and Minneman, K. P. (1996) Coupling efficiencies of human α1-adrenergic receptor subtypes: titration of receptor density and responsiveness with inducible and repressible expression vectors. Mol. Pharmacol. 50, 1376–1387.PubMedGoogle Scholar
  6. 6.
    Violin, J. D., Dewire, S. M., Barnes, W. G. and Lefkowitz, R. J. (2006) G protein-coupled receptor kinase and β-arrestin-mediated desensitization of the angiotensin II type 1A receptor elucidated by diacylglycerol dynamics. J. Biol. Chem. 281, 36411–36419.PubMedCrossRefGoogle Scholar
  7. 7.
    Urlinger, S., Baron, U., Thellmann, M., Hasan, M. T., Bujard, H. and Hillen, W. (2000) Exploring the sequence space for tetracycline-dependent transcriptional activators: novel mutations yield expanded range and sensitivity. Proc. Natl. Acad. Sci. USA 97, 7963–7968.PubMedCrossRefGoogle Scholar
  8. 8.
    Canals, M., Jenkins, L., Kellett, E. and Milligan, G. (2006) Up-regulation of the angiotensin II type 1 receptor by the MAS proto-oncogene is due to constitutive activation of Gq/G11 by MAS. J. Biol. Chem. 281, 16757–16767.PubMedCrossRefGoogle Scholar
  9. 9.
    Canals, M. and Milligan, G. (2008) Constitutive activity of the cannabinoid CB1 receptor regulates the function of co-expressed μ-opioid receptors. J. Biol. Chem. 283, 11424–11434.PubMedCrossRefGoogle Scholar
  10. 10.
    Williams, N. G., Zhong, H. and Minneman, K. P. (1998) Differential coupling of α1-, α2-, and β-adrenergic receptors to mitogen-­activated protein kinase pathways and differentiation in transfected PC12 cells. J. Biol. Chem. 273, 24624–24632.PubMedCrossRefGoogle Scholar
  11. 11.
    Zhong, H., Guerrero, S. W., Esbenshade, T. A. and Minneman, K. P. (1996) Inducible expression of β1- and β2-adrenergic receptors in rat C6 glioma cells: functional interactions between closely related subtypes. Mol. Pharmacol. 50, 175–184.PubMedGoogle Scholar
  12. 12.
    Corti, C., Crepaldi, L., Mion, S., Roth, A. L., Xuereb, J. H. and Ferraguti, F. (2007) Altered dimerization of metabotropic glutamate receptor 3 in schizophrenia. Biol. Psychiatry 62, 747–755.PubMedCrossRefGoogle Scholar
  13. 13.
    Law, P. Y., Kouhen, O. M., Solberg, J., Wang, W., Erickson, L. J. and Loh, H. H. (2000) Deltorphin II-induced rapid desensitization of δ-opioid receptor requires both phosphorylation and internalization of the receptor. J. Biol. Chem. 275, 32057–32065.PubMedCrossRefGoogle Scholar
  14. 14.
    Choi, D. S., Wang, D., Tolbert, L. and Sadee, W. (2000) Basal signaling activity of human ­dopamine D2L receptor demonstrated with an ecdysone-inducible mammalian expression system. J. Neurosci. Methods 94, 217–225.PubMedCrossRefGoogle Scholar
  15. 15.
    Kozak, M. (1987) At least six nucleotides ­preceding the AUG initiator codon enhance translation in mammalian cells. J. Mol. Biol. 196, 947–950.PubMedCrossRefGoogle Scholar
  16. 16.
    McDonald, R. L., Balmforth, A. J., Palmer, A. C., Ball, S. G., Peers, C. and Vaughan, P. F. (1995) The effect of the angiotensin II (AT1A) receptor stably transfected into human neuroblastoma SH-SY5Y cells on noradrenaline release and changes in intracellular calcium. Neurosci. Lett. 199, 115–118.PubMedCrossRefGoogle Scholar
  17. 17.
    Nash, M. S., Selkirk, J. V., Gaymer, C. E., Challiss, R. A. J. and Nahorski, S. R. (2001) Enhanced inducible mGlu1a receptor expression in Chinese hamster ovary cells. J. Neurochem. 77, 1664–1667.PubMedCrossRefGoogle Scholar
  18. 18.
    Pindon, A., van-Hecke, G., van-Gompel, P., Lesage, A. S., Leysen, J. E. and Jurzak, M. (2002) Differences in signal transduction of two 5-HT4 receptor splice-variants: ­compound specificity and dual coupling with Gαs- and Gαi/o-proteins. Mol. Pharmacol. 61, 85–96.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Institute of NeurosciencesUniversité Catholique de LouvainBrusselsBelgium

Personalised recommendations