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Suppression of GABAB receptor function in vivo by disulfide reducing agent, dl-dithiothreitol (DTT)

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Abstract

Rationale

A recent in-vitro study demonstrated that the potent disulfide reducing agent, dl-dithiothreitol (DTT), may alter the structural stability of the GABAB receptor, probably inactivating the disulfide bonds between four cysteine residues located in the GABAB1(a) receptor structure.

Objectives

The present study was designed to evaluate whether DTT treatment was capable of antagonizing some behavioral effects of pharmacological stimulation of the GABAB receptor.

Methods

Experiments on sedation/hypnosis induced by the GABAB receptor agonists baclofen, SKF 97541, CGP 44532 and γ-hydroxybutyric acid (GHB) in DBA mice and selectively bred GHB-sensitive (GHB-S) rats, and a GHB drug discrimination study in Long Evans rats were conducted. Specificity of the DTT action on the GABAB receptor was investigated by assessing its effect on the sedative/hypnotic effect induced by diazepam, ketamine and ethanol.

Results

DTT prevented the sedative/hypnotic effect of all GABAB receptor agonists tested and also reversed baclofen-induced sedation/hypnosis. In contrast, DTT had no effect on, or even potentiated, sedation/hypnosis produced by diazepam, ketamine or ethanol. DTT completely blocked the discriminative stimulus effects of GHB.

Conclusions

These results are discussed in terms of DTT altering the stability of the binding domain of the GABAB receptor, hindering the drug-receptor interaction.

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References

  • Aizenman E, Lipton SA, Loring RH (1989) Selective modulation of NMDA responses by reduction and oxidation. Neuron 2:1257–1263

    CAS  PubMed  Google Scholar 

  • Bockaert J, Pin JP (1999) Molecular tinkering of G protein-coupled receptors: an evolutionary success. EMBO J 18:1723–1729

    CAS  PubMed  Google Scholar 

  • Bolser DC, Blythin DJ, Chapman RW, Egan RW, Hey JA, Rizzo C, Kuo S-C, Kreutner W (1995) The pharmacology of SCH 50911: a novel, orally-active GABA-B receptor antagonist. J Pharmacol Exp Ther 274:1393–1398

    CAS  PubMed  Google Scholar 

  • Bowery NG, Enna SJ (2000) γ-Aminobutyric acidB receptors: first of the functional metabotropic heterodimers. J Pharmacol Exp Ther 292:2–7

    CAS  PubMed  Google Scholar 

  • Bowery NG, Bettler B, Froestl W, Gallagher JP, Marshall F, Raiteri M, Bonner TI, Enna SJ (2002) International Union of Pharmacology XXXIII. Mammalian γ-aminobutyric acidB receptors: structure and function. Pharmacol Rev 54:247–264

    Article  CAS  PubMed  Google Scholar 

  • Carai MAM, Colombo G, Brunetti G, Melis S, Serra S, Vacca G, Mastinu S, Pistuddi AM, Solinas C, Cignarella G, Minardi G, Gessa GL (2001) Role of GABAB receptors in the sedative/hypnotic effect of γ-hydroxybutyric acid. Eur J Pharmacol 428:315–321

    Article  CAS  PubMed  Google Scholar 

  • Carai MAM, Colombo G, Reali R, Serra S, Mocci I, Castelli MP, Cignarella G, Gessa GL (2002) Central effects of 1,4-butanediol are mediated by GABAB receptors via its conversion into γ-hydroxybutyric acid. Eur J Pharmacol 441:157–163

    Article  CAS  PubMed  Google Scholar 

  • Carter LP, Flores LR, Wu H, Chen W, Unzeitig AW, Coop A, France CP (2003) The role of GABAB receptors in the discriminative stimulus effects of γ-hydroxybutyrate in rats: time course and antagonism studies. J Pharmacol Exp Ther 305:668–674

    Article  CAS  PubMed  Google Scholar 

  • Cleland WW (1964) Dithiothreitol, a new protective reagent for SH groups. Biochemistry 3:480–482

    CAS  Google Scholar 

  • Colombo G, Agabio R, Lobina C, Reali R, Gessa GL (1998) Involvement of GABAA and GABAB receptors in the mediation of discriminative stimulus effects of γ-hydroxybutyric acid. Physiol Behav 64:293–302

    Article  CAS  PubMed  Google Scholar 

  • Colombo G, Lobina C, Agabio R, Brunetti G, Diaz G, Littera M, Melis S, Pani M, Reali R, Serra S, Vacca G, Carai MAM, Gessa GL (2001) Selective breeding of two rat lines differing in sensitivity to GHB and baclofen. Brain Res 902:127–130

    Article  CAS  PubMed  Google Scholar 

  • Couve A, Moss SJ, Pangalos MN (2000) GABAB receptors: a new paradigm in G protein signaling. Mol Cell Neurosci 16:296–312

    Article  CAS  PubMed  Google Scholar 

  • Dudek BC, Fanelli RJ (1980) Effects of γ-butyrolactone, amphetamine, and haloperidol in mice differing in sensitivity to alcohol. Psychopharmacology 68:87–97

    Google Scholar 

  • Galvez T, Parmentier ML, Joly C, Malitschek B, Kaupmann K, Kuhn R, Bittiger H, Froestl W, Bettler B, Pin JP (1999) Mutagenesis and modeling of the GABAB receptor extracellular domain support a Venus flytrap mechanism for ligand binding. J Biol Chem 274:13362–13369

    Article  CAS  PubMed  Google Scholar 

  • Goudie AJ, Leathley MJ (1993) Drug-discrimination assays. In: Sahgal A (ed) Behavioural neuroscience: a practical approach, vol II. Oxford University Press, Oxford, pp 145–167

  • Grant KA, Colombo G (1993) Discriminative stimulus effects of ethanol: effect of training dose on the substitution of N-methyl-d-aspartate antagonists. J Pharmacol Exp Ther 264:1241–1247

    PubMed  Google Scholar 

  • Kaupmann K, Huggel K, Heid J, Flor PJ, Bischoff S, Mickel SJ, McMaster G, Angst C, Bittiger H, Froestl W, Bettler B (1997) Expression cloning of GABAB receptors uncovers similarity to metabotropic glutamate receptors. Nature 386:239–246

    CAS  PubMed  Google Scholar 

  • Kuriyama K, Taguchi J (1987) Glycoprotein as a constituent of purified γ-aminobutyric acid/benzodiazepine receptor complex: structures and physiological roles of its carbohydrate chain. J Neurochem 48:1897–1903

    CAS  PubMed  Google Scholar 

  • Lingenhoehl K, Brom R, Heid J, Beck P, Froestl W, Kaupmann K, Bettler B, Mosbacher J (1999) γ-Hydroxybutyrate is a weak agonist at recombinant GABAB receptors. Neuropharmacology 38:1667–1673

    Article  PubMed  Google Scholar 

  • Lobina C, Colombo G, Brunetti G, Diaz G, Melis S, Pani M, Serra S, Vacca G, Gessa GL, Carai MAM (2001) Procedure of bidirectional selective outbreeding for production of two rat lines differing in sensitivity to the sedative/hypnotic effect of γ-hydroxybutyric acid. Brain Res Protoc 8:74–81

    Article  CAS  Google Scholar 

  • Marzullo G, Hine B (1980) Opiate receptor function may be modulated through an oxidation-reduction mechanism. Science 208:1171–1173

    CAS  PubMed  Google Scholar 

  • Simon EJ, Groth J (1975) Kinetics of opiate receptor inactivation by sulfhydryl reagents: evidence for conformational change in presence of sodium ions. Proc Natl Acad Sci USA 72:2404–2407

    CAS  PubMed  Google Scholar 

  • Skowronski R, Beaumont K, Fanestil DD (1987) Modification of the peripheral-type benzodiazepine receptor by arachidonate, diethylpyrocarbonate and thiol reagents. Eur J Pharmacol 143:305–314

    Article  CAS  PubMed  Google Scholar 

  • Tabakoff B, Hoffman PL (1987) Biochemical pharmacology of alcohol. In: Meltzer HY (ed) Psychopharmacology: the third generation of progress. Raven Press, New York, pp 1521–1526

  • Vauquelin G, Bottari S, Kanarek L, Strosberg AD (1979) Evidence for essential disulfide bonds in beta1-adrenergic receptors of turkey erythrocyte membranes. Inactivation by dithiothreitol. J Biol Chem 254:4462–4469

    CAS  PubMed  Google Scholar 

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Acknowledgements

The authors are grateful to Mrs. Anne Farmer for language editing of the manuscript.

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Authors and Affiliations

  1. “Bernard B. Brodie” Department of Neuroscience, University of Cagliari, Viale Diaz 182, 09126, Cagliari, Italy

    Mauro A. M. Carai, Giovanni Vacca, Salvatore Serra & Gian Luigi Gessa

  2. C.N.R. Institute of Neuroscience, Viale Diaz 182, 09126, Cagliari, Italy

    Giancarlo Colombo & Gian Luigi Gessa

  3. Research Department, Novartis Pharma AG, 4002, Basel, Switzerland

    Wolfgang Froestl

Authors
  1. Mauro A. M. Carai
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  2. Giovanni Vacca
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  3. Salvatore Serra
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  4. Giancarlo Colombo
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  5. Wolfgang Froestl
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  6. Gian Luigi Gessa
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Correspondence to Mauro A. M. Carai.

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Carai, M.A.M., Vacca, G., Serra, S. et al. Suppression of GABAB receptor function in vivo by disulfide reducing agent, dl-dithiothreitol (DTT). Psychopharmacology 174, 283–290 (2004). https://doi.org/10.1007/s00213-003-1737-y

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  • DOI: https://doi.org/10.1007/s00213-003-1737-y

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