Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 325, Issue 2, pp 102–115 | Cite as

Selection of dopamine antagonists discriminating various behavioral responses and radioligand binding sites

  • M. P. Martres
  • P. Sokoloff
  • M. Delandre
  • J. -C. Schwartz
  • P. Protais
  • J. Costentin


In order to document the hypothesis that anti-psychotics may interact with more than one class of cerebral dopamine receptors, the relative potencies of a series of compounds were compared in three behavioral tests and in binding studies with two radioligands.

Apomorphine (0.6 mg/kg) simultaneously clicited in rat two kinds of facial stereotypies (sniffing and licking) and a stereotyped climbing behavior, allowing to compare in the same animals the relative potencies of various antipsychotics against the three behaviors. Only some substituted benzamides (Sulpiride, LUR 2366 and DAN 2163) antagonised at significantly lower dosages climbing than sniffing (or licking).

The possibility that this discriminant potency might be related to a distinct affinity for two classes of dopamine receptors was investigated by binding studies on striatal membranes with 3H-apomorphine and 3H-domperidone.

From lesion and subcellular fractionation studies, two classes of binding sites both labeled with 3H-domperidone but distinguished by apomorphine i.e. D-2 sites with nM affinity and D-4 sites with μM affinity for the dopamine agonist (according to the nomenclature of Sokoloff et al. 1980b) appear to be differently localised in striatum. Thus D-2 sites, whose number decreases after kainate lesion, are not significantly modified following cortical lesions and preferentially sediment with heavy primary subcellular fractions. In contrast D-4 sites, less affected by kainate lesions, are significantly decreased following cortical lesions (−30%) and preferentially sediment with the light subcellular fractions. In addition the apparently heterogenous recognition of total 3H-domperidone binding sites (i.e. the sum of D-2 and D-4 sites) by dopamine and apomorphine persists in the presence of guanosine-5′-triphosphate (pseudo-Hill coefficient of 0.60 instead of 0.55). This suggests that D-2 and D-4 sites cannot be considered as two discrete states of the same receptor strictly convertible one into the other by guanylnucleotides.

Whereas most dopamine antagonists inhibited D-2 and D-4 site binding with similar affinities, the three benzamide derivatives with the largest selectivity in behavioral tests displayed 2–3-fold higher affinity for D-4 than for D-2 sites and the ratios of ID50 values of the whole series of antagonists against sniffing (or licking) and climbing behaviors were correlated (P<0.01) with the ratios of Ki values regarding D-2 and D-4 site binding. Also, sulpiride and LUR 2366 unlike haloperidol and metoclopramide, inhibited the total specific 3H-domperidone binding in a biphasic manner. However, the distinction by sulpiride and LUR 2366 of low and high affinity sites did not superimpose that of D-2 and D-4 sites, as distinguished by agonists. In this test the relative proportion of low affinity sites was 2-fold higher than that of D-2 sites and Ki values for high affinity sites were lower than that for D-4 sites. Also the heterogeneity of 3H-domperidone sites regarding affinity of LUR 2366 persisted in the presence of low concentrations of apomorphine. Hence low affinity sites for discriminant benzamide derivatives may exist in two forms, distinguished by agonists and possibly interconvertible by GTP. Thus the hypothesis that two classes of central dopamine receptor can be distinguished by some substituted benzamides, but perhaps display no great difference in affinity of agonists in their physiological state, fits partic-ularly well with behavioral data.

Key words

Dopamine receptors Climbing, sniffing and licking behaviors Radioligand studies 


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  1. Amsler C (1923) Beiträge zur Pharmakologie des Gehirns. Naunyn-Schmiedeberg's Arch Pharmacol 97:1–14Google Scholar
  2. Andén NE, Stock G (1973) Effect of clozapine on the turnover of dopamine in the corpus striatum and in the limbic system. J Pharm Pharmacol 25:346–348Google Scholar
  3. Arbilla S, Langer SZ (1981) Stereoselectivity of presynaptic autoreceptors modulating dopamine release. Eur J Pharmacol 76: 345–351Google Scholar
  4. Baudry M, Martres MP, Schwartz JC (1979) 3H-domperidone: a selective ligand for dopamine receptors. Naunyn-Schmiedeberg's Arch Pharmacol 308:231–237Google Scholar
  5. Biziere K, Coyle JT (1979) Effects of cortical ablation on the neurotoxicity and receptor binding of kainic acid in striatum. J Neurosci Res 4:383–398Google Scholar
  6. Cheng YC, Prussoff WH (1973) Relationship between the inhibition constant (K i) and the concentration of inhibitor which causes 50% inhibition (I50) of an enzymatic reaction. Biochem Pharmacol 22:3099–3108Google Scholar
  7. Costall B, Naylor RJ (1973) The role of telencephalic dopaminergic systems in the mediation of apomorphine-induced stereotyped behaviour. Eur J Pharmacol 24:8–24Google Scholar
  8. Costall B, Naylor RJ (1979) Behavioural aspects of dopamine agonists and antagonists. In: Horn AS, Korf J, Westerink BHC (eds) The neurobiology of dopamine. Academic Press, London, p 555Google Scholar
  9. Creese I, Sibley DR (1982) Comments on the commentary by Dr. Seeman. Biochem Pharmacol 31:2568–2569Google Scholar
  10. Creese I, Usdin T, Snyder SH (1979) Guanine nucleotides distinguish between two dopamine receptors. Nature 278:577–578Google Scholar
  11. De Lean A, Kilpatrick BF, Caron M (1982) Dopamine receptor of the porcine anterior pituitary gland. Evidence for two affinity states discriminated by both agonists and antagonists. Mol Pharmacol 22:290–297Google Scholar
  12. Ernst A (1967) Mode of action of apomorphine and dexamphetamine on gnawing compulsion in rats. Psychopharmacologia 10:316–323Google Scholar
  13. Gray EG, Whittaker VP (1962) The isolation of nerve endings from brain: an electron microscopic study of cell fragments derived by homogenisation and centrifugation. J Anat 96:79–88Google Scholar
  14. Hamblin MW, Creese I (1982) Phenoxybenzamide treatment differentiates dopaminergic 3H-ligand binding sites in bovine caudate membranes. Mol Pharmacol 21:44–51Google Scholar
  15. Hattori T, Fibiger HC (1982) On the use of lesions of afferents to localize neurotransmitter receptor sites in the striatum. Brain Res 238:245–250Google Scholar
  16. Heidmann T, Changeux JP (1978) Structural and functional properties of the acetylcholine receptor in its purified and membranebound states. Ann Rev Biochem 47:317–357Google Scholar
  17. Janssen PAJ, Niemegeers CJE, Schellekens KHL, Lenaerts FM (1967) Is it possible to predict the clinical effects of neuroleptic drugs (major tranquillizers) from animal data? Arzneimittelforsch 17:841–854Google Scholar
  18. Kebabian JW, Calne DB (1979) Multiple receptors for dopamine. Nature 277:93–96Google Scholar
  19. Kent RS, De Lean A, Lefkowitz RJ (1980) A quantitative analysis of beta-adrenergic receptor interactions: resolution of high and low affinity states of the receptor by computer modeling of ligand binding data. Mol Pharmacol 17:14–23Google Scholar
  20. Köhler C, Ögren SO, Haglund L, Ängeby T (1979) Regional displacement by sulpiride of [3H]spiperone binding in vivo. Biochemical and behavioural evidence for a preferential action on limbic and nigral dopamine receptors. Neurosci Lett 13:51–56Google Scholar
  21. Laville C (1972) Chimie et pharmacologie du sulpiride. Lille Medical, 3ème série XVII:4–13Google Scholar
  22. Lichtfield JT, Wilcoxon F (1949) A simplified method of evaluating dose-effect experiments. J Pharmacol Exp Ther 96:99–113Google Scholar
  23. Ljungberg T, Ungerstedt U (1978) Classification of neuroleptic drugs according to their ability to inhibit apomorphine-induced locomotion and gnawing: evidence for two different mechanisms of action. Psychopharmacology 56:239–247Google Scholar
  24. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  25. Makman MH, Dvorkin B, Klein PN (1982) Sodium ion modulates D2 receptor characteristics of dopamine agonist and antagonist binding sites in striatum and retina. Proc Natl Acad Sci USA 79:4212–4216Google Scholar
  26. McGeer PL, McGeer EG, Sherer U, Singh K (1977) A glutamatergic corticostriatal path? Brain Res 128:369–373Google Scholar
  27. Moore KE, Kelly PE (1978) Biochemical pharmacology of mesolimbic and mesocortical dopaminergic neurons. In: Lipton MA, Di Mascio A, Killam KF (eds) Psychopharmacology: a generation of progress. Raven Press, New York, pp 221–234Google Scholar
  28. Munemura M, Cote TE, Tsuruta K, Eskay RL, Kebabian JW (1980) The dopamine receptor in the intermediate lobe of the rat pituitary gland: pharmacological characterization. Endocrinology 107:1676–1683Google Scholar
  29. Protais P, Costentin J, Schwartz JC (1976) Climbing behaviour induced by apomorphine in mice: a simple test for the study of dopamine receptors in striatum. Psychopharmacology 50:1–6Google Scholar
  30. Protais P, Bonnet JJ, Costentin J, Schwartz JC (1984) Rat climbing behavior elicited by stimulation of cerebral dopamine receptors. Naunyn-Schmiedeberg's Arch Pharmacol 325:93–101Google Scholar
  31. Puech AJ, Simon P, Boissier JR (1978) Benzamides and classical neuroleptics: comparison of their actions using six apomorphine-induced effects. Eur J Pharmacol 50:291–300Google Scholar
  32. Randrup A, Munkvad I (1968) Behavioral stereotypies induced by pharmacological agents. Pharmacopsychiatria 1:18–26Google Scholar
  33. Scatton B (1982) Effect of dopamine agonists and neuroleptic agents on striatal acetylcholine transmission in the rat: evidence against dopamine receptor multiplicity. J Pharmacol Exp Ther 220: 197–202Google Scholar
  34. Schwarcz R, Creese I, Coyle JT, Snyder SH (1978) Dopamine receptors localised on cerebral cortical afferents to rat corpus striatum. Nature 271:766–768Google Scholar
  35. Seeman P (1980) Brain dopamine receptors. Pharmacol Rev 32: 229–313Google Scholar
  36. Seeman P (1982) Nomenclature of central and peripheral dopaminergic sites and receptors. Biochem Pharmacol 31:2563–2568Google Scholar
  37. Sibley DR, De Lean A, Creese I (1982) Anterior pituitary dopamine receptors. Demonstration of interconvertible high and low affinity states of the D-2 dopamine receptor. J Biol Chem 257: 6351–6361Google Scholar
  38. Simon P, Chermat R (1972) Recherche d'une interaction avec les stéréotypies provoquées par l'amphétamine. J Pharmacol 3: 235–238Google Scholar
  39. Snedecor OW, Cochran WG (1967) Multiple regression. In: Statistical methods. The Iowa State University Press, Ames, Iowa, USAGoogle Scholar
  40. Sokoloff P, Martres MP, Schwartz JC (1980a) 3H-apomorphine labels both dopamine post-synaptic receptors and autoreceptors. Nature 288:283–286Google Scholar
  41. Sokoloff P, Martres MP, Schwartz JC (1980b) Three classes of dopamine receptor (D-2, D-3, D-4) identified by binding studies with 3H-apomorphine and 3H-domperidone. Naunyn-Schmiedeberg's Arch Pharmacol 315:89–102Google Scholar
  42. Starke K, Reimann W, Zumstein A, Hertting G (1978) Effect of dopamine receptor agonists and antagonists on release of dopamine in the rabbit caudate nucleus in vitro. Naunyn-Schmiedeberg's Arch Pharmacol 305:27–36Google Scholar
  43. Ungerstedt U (1979) Central dopamine mechanisms and unconditioned behaviour. In: Horn AS, Korf J, Westerink BHC (eds) The neurobiology of dopamine. Academic Press, London, p 557Google Scholar
  44. Ungerstedt U (1980) Behavioral pharmacology reflecting catecholamine neurotransmission. In: Szekeres L (ed) Adrenergic activators and inhibitors, part I. Springer, Berlin Heidelberg New York, pp 449–519Google Scholar
  45. Wallach MB, Hedley LR, Peterson KE, Schulz CH (1980) Antagonism of apomorphine-induced climbing — is it a valid model for neuroleptic activity? Proc West Pharmacol Soc 23:93–98Google Scholar
  46. Westerink BHC (1979) The effects of drugs on dopamine biosynthesis and metabolism in the brain. In: Horn AS, Korf J, Westerink BHC (eds) The neurobiology of dopamine. Academic Press, London, pp 255–291Google Scholar
  47. Zahniser NR, Molinoff PB (1978) Effect of guanine nucleotides on striatal dopamine receptors. Nature 275:453–454Google Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • M. P. Martres
    • 1
  • P. Sokoloff
    • 1
  • M. Delandre
    • 1
  • J. -C. Schwartz
    • 1
  • P. Protais
    • 2
  • J. Costentin
    • 2
  1. 1.Unité de NeurobiologieCentre Paul Broca de l'INSERMParisFrance
  2. 2.Laboratoire de Pharmacodynamie et PhysiologieUER de Médecine et de PharmacieSt. Etienne Du RouvrayFrance

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