Control of the establishment of aversive memory by calcineurin and Zif268

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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.

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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.

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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

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.

Correspondence to Isabelle M Mansuy.

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

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