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Binding of adenosine receptor ligands to brain of adenosine receptor knock-out mice: evidence that CGS 21680 binds to A1 receptors in hippocampus

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Abstract

The adenosine receptor agonist 2-[p-(2-carboxyethyl)phenylethylamino]-5′-N-ethylcarboxamidoadenosine (CGS 21680) is generally considered to be a selective adenosine A2A receptor ligand. However, the compound has previously been shown to exhibit binding characteristics that are not compatible with adenosine A2A receptor binding, at least in brain regions other than the striatum. We have examined binding of [3H]CGS 21680 and of antagonist radioligands with high selectivity for adenosine A1 or A2A receptors to hippocampus and striatum of mice lacking either adenosine A1 (A1R(−/−)) or A2A (A2AR(−/−)) receptors. Both receptor autoradiography and membrane binding techniques were used for this purpose and gave similar results. There were no significant changes in the binding of the A1 receptor antagonist [3H]DPCPX in mice lacking A2A receptors, or in the binding of the A2A receptor antagonists [3H]SCH 58261 and [3H]ZM 241385 in mice lacking A1 receptors. Furthermore, [3H]CGS 21680 binding in striatum was abolished in the A2AR(−/−), and essentially unaffected in striatum from mice lacking A1 receptors. In hippocampus, however, binding of [3H]CGS 21680 remained in the A2AR(−/−), whereas binding was virtually abolished in the A1R(−/−). There were no adaptive alterations in A2A receptor expression in this region in A1R(−/−) mice. Thus, most of the [3H]CGS 21680 binding in hippocampus is dependent on the presence of adenosine A1 receptors, but not on A2A receptors, indicating a novel binding site or novel binding mode.

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Abbreviations

CGS 21680:

2-[p-(2-Carboxyethyl)phenylethylamino]-5′-N-ethylcarboxamidoadenosine

ZM 241385:

4-(2-[7-Amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol

SCH 58261:

5-Amino-7-(2-phenylethyl)-2-(2-furyl)-pyrazolo[4,3-e]-1,2,4-triazolo[1,5-c]pyrimidine

DPCPX:

1,3-Dipropyl-8-cyclopentylxanthine

CGS 15943:

9-Chloro-2-(2-furanyl)-5,6-dihydro-[1,2,4]-triazolo-[1,5]quinazolin-5-imine monomethanesulfonate

NECA:

5′-N-ethylcarboxamidoadenosine

A1R(−/−):

Adenosine A1 receptor knock-out

A1R(+/−):

Adenosine A1 receptor heterozygote

A1R(+/+):

Adenosine A1 receptor wild-type

A2AR(−/−):

Adenosine A2A receptor knock-out

A2AR(+/−):

Adenosine A2A receptor heterozygote

A2AR(+/+):

Adenosine A2A receptor wild-type

RT-PCR:

Reverse transcriptase-polymerase chain reaction

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Acknowledgements

The present studies were supported by grants from the Swedish Science Research Council (proj no. 2553), the Bank of Sweden Tercentenary Fund, the Swedish Foundation for Strategic Research, Fundação para a Ciência e Tecnologia (POCTI//36319/99), and the European Commission (project no. QLK1-CT-2000-00069). The authors also want to thank Eva Irenius for helping out with genotyping and RT-PCR work, Professor Brun Ulfhake for kindly letting us use the ABI Prism 7000 Sequence Detector System, and Janet Holmén for help with the English language.

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Authors and Affiliations

  1. Department of Physiology and Pharmacology, Karolinska Institutet, Nanna Svartz väg 2, 171 77, Stockholm, Sweden

    Linda Halldner, Elisabetta Daré, Karin Lindström & Bertil B. Fredholm

  2. Laboratory of Neurosciences, Faculty of Medicine, 1649-028, Lisboa, Portugal

    Luisa V. Lopes

  3. Department of Neuroscience, Karolinska Institutet, 171 77, Stockholm, Sweden

    Björn Johansson

  4. IRIBHN, ULB, Campus Erasme, 808 route de Lennik, 1070, Brussels, Belgium

    Catherine Ledent

  5. Center for Neuroscience of Coimbra, Institute of Biochemistry, Faculty of Medicine, University of Coimbra, Coimbra, Portugal

    Rodrigo A. Cunha

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  1. Linda Halldner
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Correspondence to Linda Halldner.

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Halldner, L., Lopes, L.V., Daré, E. et al. Binding of adenosine receptor ligands to brain of adenosine receptor knock-out mice: evidence that CGS 21680 binds to A1 receptors in hippocampus. Naunyn-Schmiedeberg's Arch Pharmacol 370, 270–278 (2004). https://doi.org/10.1007/s00210-004-0970-1

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