Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Involvement of DARPP-32 phosphorylation in the stimulant action of caffeine

Nature volume 418, pages 774–778 (2002)Cite this article

Abstract

Caffeine has been imbibed since ancient times in tea and coffee, and more recently in colas. Caffeine owes its psychostimulant action to a blockade of adenosine A2A receptors1, but little is known about its intracellular mechanism of action. Here we show that the stimulatory effect of caffeine on motor activity in mice was greatly reduced following genetic deletion of DARPP-32 (dopamine- and cyclic AMP-regulated phosphoprotein of relative molecular mass 32,000)2. Results virtually identical to those seen with caffeine were obtained with the selective A2A antagonist SCH 58261. The depressant effect of the A2A receptor agonist, CGS 21680, on motor activity was also greatly attenuated in DARPP-32 knockout mice. In support of a role for DARPP-32 in the action of caffeine, we found that, in striata of intact mice, caffeine increased the state of phosphorylation of DARPP-32 at Thr 75. Caffeine increased Thr 75 phosphorylation through inhibition of PP-2A-catalysed dephosphorylation, rather than through stimulation of cyclin-dependent kinase 5 (Cdk5)-catalysed phosphorylation, of this residue. Together, these studies demonstrate the involvement of DARPP-32 and its phosphorylation/dephosphorylation in the stimulant action of caffeine.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Emulsion autoradiogram illustrating co-expression of adenosine A2A receptor mRNA (silver grains) and DARPP-32 mRNA (dark cells).
Figure 2: DARPP-32 is required for the stimulatory effects of caffeine and the selective adenosine A2A receptor antagonist, SCH 58261, on motor activity.
Figure 3: DARPP-32 is required for the depressant effect of the selective adenosine A2A receptor agonist, CGS 21680, on motor activity.
Figure 4: Effect of caffeine and SCH 58261 on DARPP-32 phosphorylation at Thr 75.
Figure 5: Adenosine A2A receptor regulation of DARPP-32 phosphorylation at Thr 75 by modulation of PP-2A.

Similar content being viewed by others

References

  1. Fredholm, B. B., Battig, K., Holmen, J., Nehlig, A. & Zvartau, E. E. Actions of caffeine in the brain with special reference to factors that contribute to its widespread use. Pharmacol. Rev. 51, 83–133 (1999)

    CAS  PubMed  Google Scholar 

  2. Walaas, S. I., Aswad, D. W. & Greengard, P. DARPP-32, a dopamine- and cAMP-regulated phosphoprotein enriched in dopamine-innervated brain regions. Nature 301, 69–71 (1983)

    Article  CAS  Google Scholar 

  3. Gerfen, C. R. The neostriatal mosaic: multiple levels of compartmental organization in the basal ganglia. Annu. Rev. Neurosci. 15, 285–320 (1992)

    Article  CAS  Google Scholar 

  4. Schiffman, S. N. & Vanderhaeghen, J.-J. Adenosine A2 receptors regulate the gene expression of striatopallidal and striatonigral neurons. J. Neurosci. 13, 1080–1087 (1993)

    Article  Google Scholar 

  5. Fink, J. S. et al. Molecular cloning of the rat A2 adenosine receptor: selective co-expression with D2 dopamine receptor in rat striatum. Mol. Brain Res. 14, 186–195 (1992)

    Article  CAS  Google Scholar 

  6. Kull, B., Svenningsson, P. & Fredholm, B. B. Adenosine A2A receptors are colocalized with and activate Golf in rat striatum. Mol. Pharmacol. 58, 771–777 (2000)

    Article  CAS  Google Scholar 

  7. Fredholm, B. B. Activation of adenylate cyclase from rat striatum and tuberculum olfactorium by adenosine. Med. Biol. 55, 262–267 (1977)

    CAS  PubMed  Google Scholar 

  8. Ledent, C. et al. Aggressiveness, hypoalgesia and increased blood pressure in mice deficient for the adenosine A2a receptor. Nature 388, 674–678 (1997)

    Article  CAS  Google Scholar 

  9. El Yacoubi, M. et al. The stimulant effects of caffeine on locomotor behaviour in mice are mediated through its blockade of adenosine A(2A) receptors. Br. J. Pharmacol. 129, 1465–1473 (2000)

    Article  CAS  Google Scholar 

  10. Ouimet, C. C., Langley-Guillion, K.-C. & Greengard, P. Quantitative immunochemistry of DARPP-32-expressing neurons in the rat caudatoputamen. Brain Res. 808, 8–12 (1998)

    Article  CAS  Google Scholar 

  11. Snyder, S. H., Katims, J. J., Annau, Z., Bruns, R. F. & Daly, J. W. Adenosine receptors and the behavioural actions of methylxanthines. Proc. Natl Acad. Sci. USA 78, 3260–3264 (1981)

    Article  CAS  Google Scholar 

  12. Bibb, J. et al. Phosphorylation of DARPP-32 by Cdk5 modulates dopamine signalling in neurons. Nature 402, 669–671 (1999)

    Article  CAS  Google Scholar 

  13. Nishi, A., Snyder, G. L., Nairn, A. C. & Greengard, P. Role of calcineurin and protein phosphatase-2A in the regulation of DARPP-32 dephosphorylation in neostriatal neurons. J. Neurochem. 72, 2015–2021 (1999)

    Article  CAS  Google Scholar 

  14. Nishi, A. et al. Amplification of dopaminergic signalling by a positive feedback loop. Proc. Natl Acad. Sci. USA 97, 12840–12845 (2000)

    Article  CAS  Google Scholar 

  15. Ongini, E. SCH 58261: a selective A2A adenosine receptor antagonist. Drug Dev. Res. 42, 63–70 (1997)

    Article  CAS  Google Scholar 

  16. Ehrlich, M. E., Kurihara, T. & Greengard, P. Rat DARPP-32: cloning, sequencing, and characterization of the cDNA. J. Mol. Neurosci. 2, 1–10 (1990)

    Article  CAS  Google Scholar 

  17. Le Moine, C. & Bloch, B. D1 and D2 dopamine receptors gene expression in the rat striatum: sensitive cRNA probes demonstrate prominent segregation of D1 and D2 mRNA in distinct neuronal population of the dorsal and ventral striatum. J. Comp. Neurol. 355, 418–427 (1995)

    Article  CAS  Google Scholar 

  18. Svenningsson, P. et al. Dopamine D1 receptor-induced gene transcription is modulated by DARPP-32. J. Neurochem. 75, 248–257 (2000)

    Article  CAS  Google Scholar 

  19. Ögren, S.-Ö., Köhler, C., Fuxe, K. & Ängeby, K. in Dopaminergic Ergot Derivatives and Motor Function (eds Fuxe, K. & Calne, D. B.) 225–266 (Pergamon, Oxford, 1979)

    Google Scholar 

  20. Fienberg, A. A. et al. DARPP-32: regulator of the efficacy of dopaminergic neurotransmission. Science 281, 838–842 (1998)

    Article  CAS  Google Scholar 

  21. Hemmings, H. C. J. & Greengard, P. DARPP-32, a dopamine- and adenosine 3′:5′-monophosphate-regulated phosphoprotein: regional, tissue, and phylogenetic distribution. J. Neurosci. 6, 1469–1481 (1986)

    Article  CAS  Google Scholar 

  22. Liu, F. et al. Regulation of cyclin-dependent kinase 5 and casein kinase 1 by metabotropic glutamate receptors. Proc. Natl Acad. Sci. USA 98, 11062–11068 (2001)

    Article  CAS  Google Scholar 

  23. Rimondini, R., Ferré, S., Giménez-Llort, L., Ögren, S.-O. & Fuxe, K. Differential effects of selective adenosine A1 and A2 receptor agonists on dopamine receptor agonist-induced behavioural responses in rats. Eur. J. Pharmacol. 347, 153–158 (1998)

    Article  CAS  Google Scholar 

  24. Kafka, S. H. & Corbett, R. Selective adenosine A2A receptor/dopamine D2 receptor interactions in animal models of schizophrenia. Eur. J. Pharmacol. 295, 147–154 (1996)

    Article  CAS  Google Scholar 

  25. Svenningsson, P. et al. Regulation of the phosphorylation of the dopamine- and cAMP-regulated phosphoprotein of 32 kDa in vivo by dopamine D1, dopamine D2, and adenosine A2A receptors. Proc. Natl Acad. Sci. USA 97, 1856–1860 (2000)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank R. Rimondini and S.-O. Ögren for help in performing behavioural experiments and A. Nishi for discussions. This work was supported by the Swedish Research Council (G.F. and B.B.F.), the Swedish Society for Medical Research (M.L.), the Foundation Blanceflor Boncompagni-Ludovisi, née Bildt (L.P.) and by funding from the National Institute of Mental Health and the National Institute of Drug Abuse (A.C.N. and P.G.).

Author information

Author notes

  1. Laura Pozzi

    Present address: Department of Neuroscience, “Mario Negri” Institute for Pharmacological Research, Milan, Italy

  2. James A. Bibb

    Present address: Department of Psychiatry, UT Southwestern Medical Center, Dallas, Texas, USA

Authors and Affiliations

  1. Department of Neuroscience, Karolinska Institutet, 17177, Stockholm, Sweden

    Maria Lindskog, Laura Pozzi & Gilberto Fisone

  2. Laboratory of Molecular and Cellular Neuroscience, The Rockefeller University, 10021, New York, New York, USA

    Per Svenningsson, Yong Kim, Allen A. Fienberg, James A. Bibb, Angus C. Nairn & Paul Greengard

  3. Department of Physiology and Pharmacology, Karolinska Institutet, 17177, Stockholm, Sweden

    Bertil B. Fredholm

Authors
  1. Maria Lindskog
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Per Svenningsson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Laura Pozzi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yong Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Allen A. Fienberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. James A. Bibb
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Bertil B. Fredholm
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Angus C. Nairn
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Paul Greengard
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Gilberto Fisone
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gilberto Fisone.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lindskog, M., Svenningsson, P., Pozzi, L. et al. Involvement of DARPP-32 phosphorylation in the stimulant action of caffeine. Nature 418, 774–778 (2002). https://doi.org/10.1038/nature00817

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature00817

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing