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.

  • Article
  • Published:

Control of the establishment of aversive memory by calcineurin and Zif268

Nature Neuroscience volume 11, pages 572–578 (2008)Cite this article

Abstract

Emotional memory is a rapidly acquired and persistent form of memory, and its robustness is in part determined by the initial strength of the memory. Here, we provide new evidence that the protein phosphatase calcineurin (CaN), a potent negative regulator of neuronal signaling that is known to constrain learning and memory, critically regulates the establishment of emotional memory through mechanisms involving the immediate early gene Zif268 (also known as Egr1). We found that CaN is inhibited in the amygdala during the establishment of aversive memory, but Zif268 is activated. Using inducible transgenesis in mice, we further saw that CaN inhibition and Zif268 overexpression during memory establishment strengthen the memory trace and enhance its resistance to extinction. We found that CaN inhibition correlates with increased Zif268 expression and that a common pool of proteins is regulated in the amygdala after CaN inhibition and Zif268 overexpression. Together, these findings reveal a previously unknown mechanism for the control of emotional memory that depends on CaN and Zif268.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: CaN activity is decreased in the amygdala by conditioning in the CTA test.
Figure 2: Zif268 mRNA expression is increased by CTA in the amygdala.
Figure 3: Aversive memory is enhanced by CaN inhibition and weakened by CaN overactivation.
Figure 4: CaN inhibition or overactivation modulate CTA during extinction.
Figure 5: CaN inhibition or overactivation after conditioning does not alter CTA extinction.
Figure 6: Zif268 mRNA expression is increased by CaN inhibition in the amygdala.
Figure 7: Zif268 overexpression mimics the effect of CaN inhibition on CTA.

Similar content being viewed by others

References

  1. Phelps, E.A. & LeDoux, J.E. Contributions of the amygdala to emotion processing: from animal models to human behavior. Neuron 48, 175–187 (2005).

    Article  CAS  Google Scholar 

  2. Dudai, Y. Molecular bases of long-term memories: a question of persistence. Curr. Opin. Neurobiol. 12, 211–216 (2002).

    Article  CAS  Google Scholar 

  3. Eisenberg, M., Kobilo, T., Berman, D.E. & Dudai, Y. Stability of retrieved memory: inverse correlation with trace dominance. Science 301, 1102–1104 (2003).

    Article  CAS  Google Scholar 

  4. Abel, T., Martin, K.C., Bartsch, D. & Kandel, E.R. Memory suppressor genes: inhibitory constraints on the storage of long-term memory. Science 279, 338–341 (1998).

    Article  CAS  Google Scholar 

  5. Han, J.H. et al. Neuronal competition and selection during memory formation. Science 316, 457–460 (2007).

    Article  CAS  Google Scholar 

  6. Mansuy, I.M., Mayford, M., Jacob, B., Kandel, E.R. & Bach, M.E. Restricted and regulated overexpression reveals calcineurin as a key component in the transition from short-term to long-term memory. Cell 92, 39–49 (1998).

    Article  CAS  Google Scholar 

  7. Genoux, D. et al. Protein phosphatase 1 is a molecular constraint on learning and memory. Nature 418, 970–975 (2002).

    Article  CAS  Google Scholar 

  8. Mansuy, I.M. & Shenolikar, S. Protein serine/threonine phosphatases in neuronal plasticity and disorders of learning and memory. Trends Neurosci. 29, 679–686 (2006).

    Article  CAS  Google Scholar 

  9. Rusnak, F. & Mertz, P. Calcineurin: form and function. Physiol. Rev. 80, 1483–1521 (2000).

    Article  CAS  Google Scholar 

  10. Coghlan, V.M. et al. Association of protein kinase A and protein phosphatase 2B with a common anchoring protein. Science 267, 108–111 (1995).

    Article  CAS  Google Scholar 

  11. Yakel, J.L. Calcineurin regulation of synaptic function: from ion channels to transmitter release and gene transcription. Trends Pharmacol. Sci. 18, 124–134 (1997).

    Article  CAS  Google Scholar 

  12. Oliveria, S.F., Dell'Acqua, M.L. & Sather, W.A. AKAP79/150 anchoring of calcineurin controls neuronal L-type Ca2+ channel activity and nuclear signaling. Neuron 55, 261–275 (2007).

    Article  CAS  Google Scholar 

  13. Smith, K.E., Gibson, E.S. & Dell'Acqua, M.L. cAMP-dependent protein kinase postsynaptic localization regulated by NMDA receptor activation through translocation of an A-kinase anchoring protein scaffold protein. J. Neurosci. 26, 2391–2402 (2006).

    Article  CAS  Google Scholar 

  14. Winder, D.G. & Sweatt, J.D. Roles of serine/threonine phosphatases in hippocampal synaptic plasticity. Nat. Rev. Neurosci. 2, 461–474 (2001).

    Article  CAS  Google Scholar 

  15. Kingsbury, T.J., Bambrick, L.L., Roby, C.D. & Krueger, B.K. Calcineurin activity is required for depolarization-induced, CREB-dependent gene transcription in cortical neurons. J. Neurochem. 103, 761–770 (2007).

    Article  CAS  Google Scholar 

  16. Hogan, P.G. & Li, H. Calcineurin. Curr. Biol. 15, R442–R443 (2005).

    Article  CAS  Google Scholar 

  17. Limback-Stokin, K., Korzus, E., Nagaoka-Yasuda, R. & Mayford, M. Nuclear calcium/calmodulin regulates memory consolidation. J. Neurosci. 24, 10858–10867 (2004).

    Article  Google Scholar 

  18. Rescorla, R.A. Inhibition of delay in Pavlovian fear conditioning. J. Comp. Physiol. Psychol. 64, 114–120 (1967).

    Article  CAS  Google Scholar 

  19. Welzl, H., D'Adamo, P. & Lipp, H.P. Conditioned taste aversion as a learning and memory paradigm. Behav. Brain Res. 125, 205–213 (2001).

    Article  CAS  Google Scholar 

  20. Bahar, A., Samuel, A., Hazvi, S. & Dudai, Y. The amygdalar circuit that acquires taste aversion memory differs from the circuit that extinguishes it. Eur. J. Neurosci. 17, 1527–1530 (2003).

    Article  Google Scholar 

  21. Rosenblum, K., Meiri, N. & Dudai, Y. Taste memory: the role of protein synthesis in gustatory cortex. Behav. Neural Biol. 59, 49–56 (1993).

    Article  CAS  Google Scholar 

  22. Dodge, K.L. & Scott, J.D. Calcineurin anchoring and cell signaling. Biochem. Biophys. Res. Commun. 311, 1111–1115 (2003).

    Article  CAS  Google Scholar 

  23. Munton, R.P., Vizi, S. & Mansuy, I.M. The role of protein phosphatase 1 in the modulation of synaptic and structural plasticity. FEBS Lett. 567, 121–128 (2004).

    Article  CAS  Google Scholar 

  24. Fleischmann, A. et al. Impaired long-term memory and NR2A-type NMDA receptor–dependent synaptic plasticity in mice lacking c-Fos in the CNS. J. Neurosci. 23, 9116–9122 (2003).

    Article  CAS  Google Scholar 

  25. Jones, M.W. et al. A requirement for the immediate early gene Zif268 in the expression of late LTP and long-term memories. Nat. Neurosci. 4, 289–296 (2001).

    Article  CAS  Google Scholar 

  26. Malkani, S., Wallace, K.J., Donley, M.P. & Rosen, J.B. An egr-1 (zif268) antisense oligodeoxynucleotide infused into the amygdala disrupts fear conditioning. Learn. Mem. 11, 617–624 (2004).

    Article  Google Scholar 

  27. Malleret, G. et al. Inducible and reversible enhancement of learning, memory and long-term potentiation by genetic inhibition of calcineurin. Cell 104, 675–686 (2001).

    Article  CAS  Google Scholar 

  28. Mansuy, I.M. et al. Inducible and reversible gene expression with the rtTA system for the study of memory. Neuron 21, 257–265 (1998).

    Article  CAS  Google Scholar 

  29. Michalon, A., Koshibu, K., Baumgartel, K., Spirig, D.H. & Mansuy, I.M. Inducible and neuron-specific gene expression in the adult mouse brain with the rtTA2S-M2 system. Genesis 43, 205–212 (2005).

    Article  CAS  Google Scholar 

  30. Fanselow, M.S. Conditioned and unconditional components of post-shock freezing. Pavlov. J. Biol. Sci. 15, 177–182 (1980).

    CAS  PubMed  Google Scholar 

  31. Groth, R.D., Dunbar, R.L. & Mermelstein, P.G. Calcineurin regulation of neuronal plasticity. Biochem. Biophys. Res. Commun. 311, 1159–1171 (2003).

    Article  CAS  Google Scholar 

  32. Antoni, F.A. et al. Calcium control of adenylyl cyclase: the calcineurin connection. Adv. Second Messenger Phosphoprotein Res. 32, 153–172 (1998).

    Article  CAS  Google Scholar 

  33. Chan, G.C., Tonegawa, S. & Storm, D.R. Hippocampal neurons express a calcineurin-activated adenylyl cyclase. J. Neurosci. 25, 9913–9918 (2005).

    Article  CAS  Google Scholar 

  34. Ahi, J., Radulovic, J. & Spiess, J. The role of hippocampal signaling cascades in consolidation of fear memory. Behav. Brain Res. 149, 17–31 (2004).

    Article  CAS  Google Scholar 

  35. Sheng, M. & Kim, M.J. Postsynaptic signaling and plasticity mechanisms. Science 298, 776–780 (2002).

    Article  CAS  Google Scholar 

  36. Dash, P.K., Orsi, S.A. & Moore, A.N. Sequestration of serum response factor in the hippocampus impairs long-term spatial memory. J. Neurochem. 93, 269–278 (2005).

    Article  CAS  Google Scholar 

  37. Matheos, D.P., Kingsbury, T.J., Ahsan, U.S. & Cunningham, K.W. Tcn1p/Crz1p, a calcineurin-dependent transcription factor that differentially regulates gene expression in Saccharomyces cerevisiae. Genes Dev. 11, 3445–3458 (1997).

    Article  CAS  Google Scholar 

  38. Blaeser, F., Ho, N., Prywes, R. & Chatila, T.A. Ca2+-dependent gene expression mediated by MEF2 transcription factors. J. Biol. Chem. 275, 197–209 (2000).

    Article  CAS  Google Scholar 

  39. Shibasaki, F., Price, E.R., Milan, D. & McKeon, F. Role of kinases and the phosphatase calcineurin in the nuclear shuttling of transcription factor NF-AT4. Nature 382, 370–373 (1996).

    Article  CAS  Google Scholar 

  40. Liu, J.O. Endogenous protein inhibitors of calcineurin. Biochem. Biophys. Res. Commun. 311, 1103–1109 (2003).

    Article  CAS  Google Scholar 

  41. Wang, X., Culotta, V.C. & Klee, C.B. Superoxide dismutase protects calcineurin from inactivation. Nature 383, 434–437 (1996).

    Article  CAS  Google Scholar 

  42. Zeng, H. et al. Forebrain-specific calcineurin knockout selectively impairs bidirectional synaptic plasticity and working/episodic-like memory. Cell 107, 617–629 (2001).

    Article  CAS  Google Scholar 

  43. Zhuo, M. et al. A selective role of calcineurin Aα in synaptic depotentiation in hippocampus. Proc. Natl. Acad. Sci. USA 96, 4650–4655 (1999).

    Article  CAS  Google Scholar 

  44. Cannich, A. et al. CB1 cannabinoid receptors modulate kinase and phosphatase activity during extinction of conditioned fear in mice. Learn. Mem. 11, 625–632 (2004).

    Article  Google Scholar 

  45. Lin, C.H. et al. Identification of calcineurin as a key signal in the extinction of fear memory. J. Neurosci. 23, 1574–1579 (2003).

    Article  CAS  Google Scholar 

  46. Lin, C.H., Yeh, S.H., Lu, H.Y. & Gean, P.W. The similarities and diversities of signal pathways leading to consolidation of conditioning and consolidation of extinction of fear memory. J. Neurosci. 23, 8310–8317 (2003).

    Article  CAS  Google Scholar 

  47. Cammarota, M., Bevilaqua, L.R., Kohler, C., Medina, J.H. & Izquierdo, I. Learning twice is different from learning once and from learning more. Neuroscience 132, 273–279 (2005).

    Article  CAS  Google Scholar 

  48. Runyan, J.D., Moore, A.N. & Dash, P.K. A role for prefrontal calcium-sensitive protein phosphatase and kinase activities in working memory. Learn. Mem. 12, 103–110 (2005).

    Article  Google Scholar 

  49. Munton, R.P. et al. Qualitative and quantitative analyses of protein phosphorylation in naive and stimulated mouse synaptosomal preparations. Mol. Cell. Proteomics 6, 283–293 (2007).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank H. Russig, F. Helmchen and B. Kampa for advice with statistical analyses, T. Bliss, S. Laroche and D. Kuhl for discussion, P. Gehrig for mass spectrometry analysis, U. Haditsch and L. Spassova for technical help, T. Rülicke and P. Pelzar for egg microinjection, G. Fischer for help with animal maintenance, and R. Schöb for graphical assistance. Mouse Zif268 cDNA was generously provided by T. Bliss (NMRC, London, UK) and the pBL-G vector by H. Bujard (ZMBH, Heidelberg, Germany). This project was funded by the University of Zürich, the Swiss Federal Institute of Technology, the National Center of Competence in Research “Neural Plasticity and Repair”, the Swiss National Science Foundation, the Human Frontier Science Program, the Slack-Gyr Foundation and the EMBO Young Investigator Program.

Author information

Author notes

  1. David Genoux & Kyoko Koshibu

    Present address: Present addresses: Department of Physiology, Faculty of Medical and Health Sciences, University of Auckland, 85 Park Road, Auckland, New Zealand (D.G.) and The Queensland Brain Institute, University of Queensland, St. Lucia, Brisbane 4072, Australia (K.K.).,

Authors and Affiliations

  1. Medical Faculty of the University of Zürich and Department of Biology, Brain Research Institute, Swiss Federal Institute of Technology, Winterthurerstrasse 190, Zürich, 8057, Switzerland

    Karsten Baumgärtel, David Genoux, Ry Y Tweedie-Cullen, Kyoko Koshibu, Magdalena Livingstone-Zatchej, Céline Mamie & Isabelle M Mansuy

  2. Division of Neuroanatomy and Behavior, Institute of Anatomy, University of Zürich, Winterthurerstrasse 190, Zürich, 8057, Switzerland

    Hans Welzl

Authors
  1. Karsten Baumgärtel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. David Genoux
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hans Welzl
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ry Y Tweedie-Cullen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kyoko Koshibu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Magdalena Livingstone-Zatchej
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Céline Mamie
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Isabelle M Mansuy
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

K.B. was responsible for the project and wrote the manuscript together with I.M.M. He conducted all CTA experiments except those presented in Figure 3a and Supplementary Figure 4, performed all biochemical and molecular experiments, and generated and characterized the Zif268 mutant mice. H.W. provided the expertise for the behavioral experiments and actively contributed to them. D.G. initiated this project and performed the CTA experiments shown in Figure 3a and Supplementary Figure 4. R.Y.T.-C. and M.L.-Z. performed the proteomics screen. K.K. was involved in several CTA experiments and in tissue isolation and developed the PP1 assay used to generate Supplementary Figure 2. C.M. was involved in one CTA experiment and during several sample preparations. I.M.M. is the principal investigator; she generated the CaN and PP1 transgenic lines, conceived the project, provided continual conceptual input and financial support and wrote the manuscript.

Corresponding author

Correspondence to Isabelle M Mansuy.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–9, Supplementary Methods (PDF 1234 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Baumgärtel, K., Genoux, D., Welzl, H. et al. Control of the establishment of aversive memory by calcineurin and Zif268. Nat Neurosci 11, 572–578 (2008). https://doi.org/10.1038/nn.2113

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nn.2113

This article is cited by

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