Abstract
1.The D2-type dopamine receptors are thought to inhibit adenylyl cyclase (AC), via coupling to pertussis toxin (PTX)-sensitive G proteins of the Gi family. We examined whether and to what extent the various D2 receptors (D2S, D2L, D3S, D3L, and D4) couple to the PTX-insensitive G protein Gz, to produce inhibition of AC activity.
2.COS-7 cells were transiently transfected with the individual murine dopamine receptors alone, as well as together with the α subunit of Gz. PTX treatment was employed to inactivate endogenous αi, and coupling to Gi and Gz was estimated by measuring the inhibition of cAMP accumulation induced by quinpirole, in forskolin-stimulated cells.
3.D2S or D2L receptors can couple to the same extent to Gi and to Gz. The D4 dopamine receptor couples preferably to Gz, resulting in about 60% quinpirole-induced inhibition of cAMP accumulation. The D3S and D3L receptor isoforms couple slightly to Gz and result in 15 and 30% inhibition of cAMP accumulation, respectively.
4.We have demonstrated for the first time that the two D3 receptor isoforms, and not any of the other D2 receptor subtypes, also couple to Gs in both COS-7 and CHO transfected cells, in the presence of PTX.
5.Thus, the differential coupling of the D2 dopamine receptor subtypes to various G proteins may add another aspect to the diversity of dopamine receptor function.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
REFERENCES
Avidor-Reiss, T., Bayewitch, M., Levy, R., Matus-Leibovitch, N., Nevo, I., and Vogel, Z. (1995). Adenylyl cyclase supersensitization in μ-opioid receptor-transfected Chinese Hamster ovary cells following chronic opioid treatment. J. Biol. Chem. 270:29732–29738.
Burford, N. T., Tobin, A. B., and Nahorski, S. R. (1995). Differential coupling of m1, m2 and m3 muscarinic receptor subtypes to inositol 1,4,5-trisphosphate and adenosine 3′,5′-cyclic monophosphate accumulation in Chinese Hamster ovary cells. J. Pharmacol. Exp. Ther. 274:134–142.
Caine, S. B., and Koob, G. F. (1993). Effects of mesolimbic dopamine depletion on responding maintained by cocaine and food. Science 260:1814–1816.
Casey, P. J., Fong, H. K. W., Simon, M. I., and Gilman, A. G. (1990). Gz, a guanine nucleotide-binding protein with unique biochemical properties. J. Biol. Chem. 265:2383–2390.
Chan, J. S., Chiu, T. T., and Wong, Y. H. (1995). Activation of type II adenylyl cyclase by the cloned μ-opioid receptor: Coupling to multiple G proteins. J. Neurochem. 65:2682–2689.
Chio, C. L., Drong, R. F., Riley, D. T., Gill, G. S., Slightom, J. L., and Huff, R. (1994a). D4 dopamine receptor-mediated signaling events determined in transfected Chinese Hamster ovary cells. J. Biol. Chem. 269:11813–11819.
Chio, C. L., Lajiness, M. E., and Huff, R. M. (1994b). Activation of heterologously expressed D3 dopamine receptors: comparison with D2 dopamine receptors. Mol. Pharmacol. 45:51–60.
Cox, B. A., Rosser, M. P., Kozlowski, M. R., Duwe, K. M., Neve, R. L., and Neve, K. A. (1995). Regulation and functional characterization of a rat recombinant dopamine D3 receptor. Synapse 21:1–9.
David, C., and Fuchs, S. (1991). Antipeptide antibodies localize N-(4-azido-3-[125I]iodophenethyl)spiperone binding to the carboxyl-terminal portion of the D2 dopamine receptor. Mol. Pharmacol. 40:712–716.
Dearry, A., Falardeau, P., Shores, C., and Caron, M. G. (1991). D2 dopamine receptors in the human retina: Cloning of cDNA and localization of mRNA. Cell. Mol. Neurobiol. 11:437–452.
Eason, M. G., Kurose, H., Holt, B. D., Raymond, J. R., and Liggett, S. B. (1992). Simultaneus coupling of α 2-adrenergic receptors to two G-proteins with oposing effects. J. Biol. Chem. 267:15795–15801.
Fields, A. T., and Casey, P. J. (1997). Signalling functions and biochemical properties of pertussis toxin-resistant G-protein. Biocem. J. 321:561–571.
Fishburn, C. S., Belleli, D., David, C., Carmon, S., and Fuchs, S. (1993). A novel short isoform of the D3 dopamine receptor generated by alternative splicing in the third cytoplasmic loop. J. Biol. Chem. 268:5872–5878.
Fishburn, C. S., Elazar, Z., and Fuchs, S. (1995a). Differential glycosylation and intracellular trafficking for the long and short isoforms of the D2 dopamine receptor. J. Biol. Chem. 270:29819–29824.
Fishburn, C. S., Carmon, S., and Fuchs, S. (1995b). Molecular cloning and characterisation of the gene encoding the murine D4 dopamine receptor. FEBS Lett. 361:215–219.
Kenakin, T. (1996). The classification of seven transmembrane receptors in recombinant expression systems. Pharmacol. Rev. 48:413–463.
Keown, W. A., Campbell, C. R., and Kucherlapati, R. S. (1990). Methods for introducing DNA into mammalian cells. Methods Enzymol. 185:527–537.
Lai, H. W. L., Minami, M., Satoh, M., and Wong, Y. H. (1995). Gz coupling to the rat κ-opioid receptor. FEBS Lett. 360:97–99.
LeCam, A. M., Gottesman, M., and Pastan, I. (1980). Mechanism of cyclic AMP on nutrient transport in Chinese hamster ovary cells. J. Biol. Chem. 255:8103–8108.
Lim, K., and Chae, C. B. (1989). A simple assay for DNA transfection by incubation of the dishes with substrates for beta-galactosidase. Biotechniques 7:576–579.
MacKenzie, R. V., VanLeeuwen, D., Pugsley, T. A., Shih, Y. H., Demattos, S., Tang, L, Todd, R. D., and O'Malley, K. L. (1994). Characterization of the human dopamine D3 receptor expressed in transfected cell lines. Eur. J. Pharmacol. 266:79–85.
Mansour, A., Meador-Woodruff, H. J., Bunzow, J. R., Civelli, O., Akil, H., and Watson, S. J. (1990). Localization of dopamine D2 receptor mRNA and D1 and D2 receptor binding in the rat brain and pituitary: An in situ hybridization-receptor autoradiographic analysis. J. Neurosci. 10:2587–2600.
Matsuoka, M., Itoh, H., Kozas, T., and Kaziro, Y. (1988). Sequence analysis of cDNA and genomic DNA for a putative pertussis toxin-insensitive guanine nucleotide-binding regulatory protein α subunit. Proc. Natl. Acad. Sci. USA 85:5384–5388.
Mauger, C., Brockhaus, M., Sivan, B., Civelli, O., Fuchs, S., and Monsma, F. (1996). Immunocytochemical localization of the D4 dopamine receptor in the mouse brain. Soc. Neurosci. Abstr. 22:1580.
Monsama, F. J., McVittie, L. D., Gerfen, C. R., Mahan, L. C., and Sibley, D. R. (1989). Multiple D2 dopamine receptors produced by alternative RNA splicing. Nature 342:926–929.
Potenza, M. N., Graminski, G. F., Schmauss, C., and Lerner, M. R. (1994). Functional expression and characterization of human D2 and D3 dopamine receptors. J. Neurosci. 14:1463–1476.
Salomon, Y. (1991). Cellular responsivness to hormones and neurotransmitters: conversion of [3H] adenine to [3H]cAMP in cell monolayers, cell suspensions, and tissue slices. Methods Enzymol. 195:22–28.
Sander, T., Harms, H., Podschus, J., Finckh, U., Nickel, B., Rolfs, A., Rommelspacher, H., and Schmidt, L. G. (1995). Dopamine D1, D2 and D3 receptor genes in alcohol dependence. Psychiatr. Genet. 5:171–176.
Seeman, P., Guan, H. C., and Van Tol, H. H. (1993). Dopamine D4 receptors elevated in schizophrenia. Nature 365:441–445.
Sibley, D. R., Monsma, F. J. J., and Shen, Y. (1993). Molecular neurobiology of dopamine receptors. Int. Rev. Neurobiol. 35:391–415.
Simon, M. I., Strathmann, M. P., and Gautman, N. (1991). Diversity of G protein in signal transduction. Science 252:802–808.
Sokoloff, P., Giros, B., Martres, M. P., Bouthenet, M. L., and Schwarz, J. C. (1990). Molecular cloning and characterization of a novel dopamine receptor (D3) as a target for neuroleptics. Nature 347:146–151.
Tsu, R. C., Allen, R. A., and Wong, Y. H. (1995a). Stimulation of type II adenylyl cyclase by chemoattractant formyl peptide and C5a receptors. Mol. Pharmacol. 47:835–841.
Tsu, R. C., Lai, H. W. L., Allen, R. A., and Wong, Y. H. (1995b). Differential coupling of the formyl peptide receptor to adenylate cyclase and phospholipase C by the pertussis toxin-insensitive Gz protein. Biochem. J. 309:331–339.
Tsu, R. C., Chan, J. S. C., and Wong, Y. H. (1995c). Regulation of multiple effects by the cloned δ-opioid receptor: stimulation of phospholipase C and type II adenylyl cyclase. J. Neurochem. 64:2700–2707.
Vallar, V., and Meldolesi, J. (1989). Mechanism of signal transduction at the dopamine D2 receptor. Trends Pharmacol. Sci. 10:74–77.
Wang, J., Tu, Y., Woodson, J., Song, X., and Ross, E. M. (1997). A GTPase-activating protein for the G protein Gα z. J. Biol. Chem. 272:5732–5740.
Wong, Y. H., Conklin, B. R., and Bourne, H. R. (1992). Gz-mediated hormonal inhibition of cyclic AMP accumulation. Science 255:339–342.
Yamaguchi, I., Harmon, S. K., Todd, R. D., and O'Malley, K. L. (1997). The rat D4 dopamine receptor couples to cone transducin (Gα t2) to inhibit forskolin-stimulated cAMP accumulation. J. Biol. Chem. 272:16599–16602.
Yung, L. Y., Tsim, S.-T., and Wong, Y. H. (1995). Stimulation of cAMP accumulation by the cloned Xenopus melatonin receptor trough Gi and Gz proteins. FEBS Lett. 372:99–102.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Obadiah, J., Avidor-Reiss, T., Fishburn, C.S. et al. Adenylyl Cyclase Interaction with the D2 Dopamine Receptor Family; Differential Coupling to Gi, Gz, and Gs. Cell Mol Neurobiol 19, 653–664 (1999). https://doi.org/10.1023/A:1006988603199
Issue Date:
DOI: https://doi.org/10.1023/A:1006988603199