Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Autoradiographic localisation of3H-apomorphine binding sites in rat brain

  • 21 Accesses

  • 13 Citations

Summary

The best experimental conditions for a selective binding of3H-apomorphine to dopamine receptors on cryostat sections were first selected by liquid scintillation quantification of the bound radioactivity. In the corpus striatum, a specific binding occurred with a half-maximal saturation concentration of about 1 nM and a maximal capacity of 180 fmol/mg of slice protein, both values in agreement with previous binding data on either membranes or slices incubated in a physiological medium. Inhibition with domperidone was clearly biphasic, indicating two classes of sites corresponding to the D-2 and D-3 sites as previously defined on membranes.

When3H-apomorphine was used at low concentrations (0.8–1.5 nM), a condition ensuring a preferential labelling of D-2 sites, rather well contrasted autoradiographic pictures were generated. The major dopaminergic projection fields in telencephalon (caudate-putamen, nucleus accumbens, olfactory tubercles) were visualised as well as other catecholaminergic regions such as the superficial gray layer of superior colliculi. Within the striatum, differences in density of these sites were observed in three perpendicular planes and confirmed by a computer densitometric image analysis. Labelling of areas of origin of the cerebral dopaminergic neurons in substantia nigra or ventral tegmental area were also observed.

When a higher concentration of3H-apomorphine (3.5 nM) was used in the presence of domperidone, another, but autoradiographically less distinct subclass of sites (D-3 sites) was demonstrated.

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

References

  1. Baudry M, Martres MP, Schwartz JC (1979)3H-domperidone: a selective ligand for dopamine receptors. Naunyn-Schmiedeberg's Arch Pharmacol 308:231–237

  2. 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 22:290–297

  3. Fuxe K (1965) The distribution of monoamine terminals in the central nervous system. Acta Physiol Scand 64:31–85

  4. Fuxe K, Anderson K, Schwarcz R, Agnati LF, Perez De La Mora M, Hokfelt T, Goldstein M, Ferland L, Possani L, Tapia R (1979) Studies on different types of dopamine nerve terminals in the forebrain and their possible interactions with GABA, glutamate and opioid peptides. In: Poirier LI, Sourkes TL, Bedard PJ (eds) Advances in neurology, vol 24. Raven Press, New York, pp 199–215

  5. Fuxe K, Agnati LF, Andersson K, Calza L, Benfenati F, Zini I, Battistini N, Kohler C, Ogren SO, Hökfelt T (1983) Analysis of transmitter identified neurons by morphometry and quantitative microfluorimetry. Evaluation of the actions of psychoactive drugs, especially sulpiride. In: Ackenheil M, Matussek N (eds) Special aspects of psychopharmacology. Expansion Scientifique Française, pp 13–34

  6. Hamblin MW, Creese I (1982) Phenoxybenzamine treatment differentiates dopaminergic3H-ligand binding sites in bovine caudate membranes. Mol Pharmacol 21:44–51

  7. Höllt V, Schubert P (1978) Demonstration of neuroleptic receptor sites in mouse brain by autoradiography. Brain Res 151:149–153

  8. Howlett DR, Nahorski SR (1980) Quantitative assessment of heter ogenous3H-spiperone binding to rat neostriatum and frontal cortex. Life Sci 26:511–517

  9. König JFR, Klippel RA (1963) The rat brain: a stereotaxic atlas of the forebrain and lower parts of the brain stem. Williams and Wilkins, Baltimore

  10. Kramer EB, Lischka MF, Karobath M, Schönbeck G (1979) Is there a selectivity of neuronal degeneration induced by intrastriatal injection of kainic acid? Brain Res 177:577–582

  11. Kuhar MJ, Yamamura HI (1975) Light autoradiographic localization of cholinergic muscarinic receptors in rat brain by specific binding of a potent antagonist. Nature (London) 253:560–561

  12. Lazareno S, Nahorski SR (1982) Selective labelling of dopamine (D2) receptors in rat striatum by3H-domperidone but not by3H-spiperone. Eur J Pharmacol 81:273–275

  13. Leff SE, Creese I (1983) Dopamine receptors re-explained. Trends Pharmacol Sci nov: 463–467

  14. Leysen JE, Niemegeers CJE, Tollemaere JP, Laduron PM (1978) Serotoninergic component of neuroleptic receptors. Nature 272:168–171

  15. Lindvall O, Björklund A (1974) The glyoxylic acid fluorescence histochemical method: a detailed account of the methodology for the visualization of central catecholamine neurons. Histochemistry 39:97–127

  16. Lindvall O, Björklund A (1978) Organisation of catecholamine neurons in the rat central nervous system. In: Iversen LL, Iversen SD, Snyder SH (eds) Handbook of psychopharmacology, chp 4. Plenum Press, New York, pp 139–232

  17. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

  18. Martres MP, Sokoloff P, Schwartz JC (1984) Dopaminergic binding sites in rat striatal slices and the action of guanyl nucleotides. Naunyn-Schmiedeberg's Arch Pharmacol 325:116–123

  19. Martres MP, Bouthenet ML, Salès N, Sokoloff P, Schwartz JC (1985) Widespread distribution of brain dopamine receptors evidenced with [125I]iodosulpride, a highly selective ligand. Science (in press)

  20. Moore RY, Bloom FE (1978) Central catecholamine neuron systems: anatomy and physiology of the dopamine systems. Ann Rev Neurosci 1:129–169

  21. Murrin CL, Kuhar MJ (1979) Dopamine receptors in the rat frontal cortex: an autoradiographic study. Brain Res 177:285

  22. Nagy JI, Lee T, Seeman P, Fibiger HC (1978) Direct evidence for pre-synaptic and post-synaptic dopamine receptors in brain. Nature (London) 274:278–281

  23. Palacios JM (1984) Receptor autoradiography: the last ten years. J Recept Res 4:633–644

  24. Palacios JM, Niehoff DL, Kuhar MJ (1981)3H-spiperone binding sites in brain: autoradiographic localisation of receptors. Brain Res 213:277–289

  25. Paxinos G, Watson C (1982) The rat brain in stereotaxic coordinates. Academic Press, Sydney, New York, London

  26. Rainbow TC, Bleisch WV, Biegon A, McEwen BS (1982) Quantitative densitometry of neurotransmitter receptors. J Neurosci Methods 5:127–138

  27. Schwartz JC, Sokoloff P, Martres MP, Protais P, Costentin J, Bouthenet ML, Sales N (1983) Distinction of dopamine receptors well recognised by antipsychotics agents: binding, autoradiographic and behavioural studies. In: Carlsson A, Lars J, Nilsson G (eds) Proc Swedish Acad Pharm Sci, Dopamine Receptor Agonists 1; Swedish Pharmaceutical Press, Stockholm, pp 47–59

  28. Schwartz JC, Delandre M, Martres MP, Sokoloff P, Protais P, Vasse M, Costentin J, Laibe P, Mann A, Wermuth CG, Gulat G, Laffite A (1984) Biochemical and behavioral identification of discriminant benzamide derivatives: new tools to differentiate subclasses of dopamine receptors. In: Catecholamines: Neuropharmacology and central nervous system — Theoretical aspects. A. R. Liss, Inc. New York, pp 59–72

  29. Seeman P (1980) Brain dopamine receptors. Pharmacol Rev 32:229–313

  30. Seeman P, Lee T, Chau-Wong M, Tedesco J, Wong K (1976) Dopamine receptors in human and calf brains using3H-apomorphine and an antipsychotic drug. Proc Natl Acad Sci USA 73:4354–4358

  31. Sibley DR, De Lean A, Creese I (1982) Anterior pituitary dopamine receptors. Demonstration of interconvertible high- and lowaffinity states of the D-2 dopamine receptor. J Biol Chem 257:6351–6361

  32. Sokoloff P, Martres MP, Schwartz JC (1980a)3H-apomorphine labels both dopamine postsynaptic receptors and autoreceptors. Nature 288:283–286

  33. Sokoloff P, Martres MP, Schwartz JC (1980b) Three classes of dopamine receptor (D-2, D-3, D-4) identified by binding studies with3H-apomorphine and3H-domperidone. Naunyn-Schmiedeberg's Arch Pharmacol 315:89–102

  34. Sokoloff P, Martres MP, Delandre M, Redouane K, Schwartz JC (1984)3H-domperidone binding sites differ in rat striatum and pituitary. Naunyn-Schmiedeberg's Arch Pharmacol 327:221–227

  35. Strong R, Samorajski T, Gottesfeld Z (1982) Regional mapping of neostriatal neurotransmitter systems as a function of aging. J Neurochem 39 (3):831–836

  36. Takano Y, Kohjimoto Y, Uchimura K, Kamiya HO (1980) Mapping of the distribution of high affinity choline uptake and choline acetyltransferase in the striatum. Brain Res 194:583–587

  37. Westerink BHC, Horn AS (1979) Do neuroleptics prevent the penetration of dopamine agonists into the brain? Eur J Pharmacol 58:39–48

  38. Young WS, Kuhar MJ (1979) A new method for receptors autoradiography: (3H)-opioid receptors in rat brain. Brain Res 179:255–270

Download references

Author information

Correspondence to Jean-Charles Schwartz.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Bouthenet, M., Sales, N. & Schwartz, J. Autoradiographic localisation of3H-apomorphine binding sites in rat brain. Naunyn-Schmiedeberg's Arch. Pharmacol. 330, 1–8 (1985). https://doi.org/10.1007/BF00586702

Download citation

Key words

  • Dopamine receptors
  • 3H-apomorphine
  • Autoradiography
  • Rat brain