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:

L-type calcium channels and GSK-3 regulate the activity of NF-ATc4 in hippocampal neurons

Nature volume 401, pages 703–708 (1999)Cite this article

Abstract

The molecular basis of learning and memory has been the object of several recent advances, which have focused attention on calcium-regulated pathways controlling transcription. One of the molecules implicated by pharmacological, biochemical and genetic approaches is the calcium/calmodulin-regulated phosphatase, calcineurin1,2,3,4,5. In lymphocytes, calcineurin responds to specific calcium signals and regulates expression of several immediate early genes by controlling the nuclear import of the NF-ATc family of transcription factors6,7,8,9. Here we show that NF-ATc4/NF-AT3 (ref. 10) in hippocampal neurons can rapidly translocate from cytoplasm to nucleus and activate NF-AT-dependent transcription in response to electrical activity or potassium depolarization. The calcineurin-mediated translocation is critically dependent on calcium entry through L-type voltage-gated calcium channels. GSK-3 can phosphorylate NF-ATc4, promoting its export from the nucleus and antagonizing NF-ATc4-dependent transcription. Furthermore, we show that induction of the inositol 1,4,5-trisphosphate receptor type 1 is controlled by the calcium/calcineurin/NF-ATc pathway. This provides a new perspective on the function of calcineurin in the central nervous system and indicates that NF-AT-mediated gene expression may be involved in the induction of hippocampal synaptic plasticity and memory formation.

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: Expression of NF-ATc4 and activation of NF-AT-dependent transcription by endogenous NF-AT proteins in the hippocampus.
Figure 2: Activation of NF-AT-dependent transcription by NMDA receptors and L-type Ca2+ channels.
Figure 3: Activity-dependent cytoplasmic-to-nuclear translocation of NF-ATc4 in hippocampal neurons.
Figure 4: GSK-3β blocks NF-AT-dependent transcription and promotes nuclear export of NF-ATc4.
Figure 5: NF-AT-dependent regulation of IP3R1.

Similar content being viewed by others

References

  1. Mulkey, R. M, Endo,S., Shenolikar,S. & Malenka,R. C. Involvement of a calcineurin/inhibitor-1 phosphatase cascade in hippocampal long-term depression. Nature 369, 486–488 (1994).

    Article  ADS  Google Scholar 

  2. Lu,Y. F., Hayashi,Y., Moriwaki,A., Tomizawa,K. & Matsui,H. FK506, a Ca2+/calmodulin-dependent phosphatase inhibitor inhibits the induction of long-term potentiation in the rat hippocampus. Neurosci. Lett. 205, 103–106 (1996).

    Article  CAS  Google Scholar 

  3. Winder,D. G., Mansuy,I. M., Osman,M., Moallem,T. M. & Kandel,E. R. Genetic and pharmacological evidence for a novel, intermediate phase of long-term potentiation suppressed by calcineurin. Cell 92, 25–37 (1998).

    Article  CAS  Google Scholar 

  4. 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 

  5. Klee,C. B., Crouch,T. H. & Krinks,M. H. Calcineurin: a calcium- and calmodulin-binding protein of the nervous system. Proc. Natl Acad. Sci. USA 76, 6270–6273 (1979).

    Article  ADS  CAS  Google Scholar 

  6. Flanagan,W. M., Corthesy,B., Bram,R. J. & Crabtree,G. R. Nuclear association of a T-cell transcription factor blocked by FK-506 and cyclosporin A [see comments]. Nature 352, 803–807 (1991).

    Article  ADS  CAS  Google Scholar 

  7. Liu,J. et al. Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP–FK506 complexes. Cell 66, 807–815 (1991).

    Article  CAS  Google Scholar 

  8. Shaw, J.-P. et al. Identification of a putative regulator of early T cell activation genes. Science 241, 202–205 (1988).

    Article  ADS  Google Scholar 

  9. Clipstone,N. A. & Crabtree,G. R. Identification of calcineurin as a key signalling enzyme in T-lymphocyte activation. Nature 357, 695–697 (1992).

    Article  ADS  CAS  Google Scholar 

  10. Hoey,T., Sun, Y.-L., Williamson,K. & Xu,X. Isolation of two new members of the NF-AT gene family and functional characterization of the NF-AT proteins. Immunity 2, 461–472 (1995).

    Article  CAS  Google Scholar 

  11. Crabtree,G. R. Generic signals and specific outcomes: signaling through Ca2+, calcineurin, and NF-AT. Cell 96, 611–614 (1999).

    Article  CAS  Google Scholar 

  12. Jain,J. et al. The T-cell transcription factor NFATp is a substrate for calcineurin and interacts with Fos and Jun. Nature 365, 352–355 (1993).

    Article  ADS  CAS  Google Scholar 

  13. Beals,C. R., Clipstone,N. A., Ho,S. N. & Crabtree,G. R. Nuclear localization of NF-ATc by a calcineurin-dependent, cyclosporin-sensitive intramolecular interaction. Genes Dev. 11, 824–834 (1997).

    Article  CAS  Google Scholar 

  14. Timmerman,L. A., Clipstone,N. A., Ho,S. N., Northrop,J. P. & Crabtree,G. R. Rapid shuttling of NF-AT in discrimination of Ca2+ signals and immunosuppression. Nature 383, 837–840 (1996).

    Article  ADS  CAS  Google Scholar 

  15. Beals,C. R., Sheridan,C. M., Turck,C. W., Gardner,P. & Crabtree,G. R. Nuclear export of NF-ATc enhanced by glycogen synthase kinase-3. Science 275, 1930–1934 (1997).

    Article  CAS  Google Scholar 

  16. Chow,C. W., Rincon,M., Cavanagh,J., Dickens,M. & Davis,R. J. Nuclear accumulation of NFAT4 opposed by the JNK signal transduction pathway. Science 278, 1638–1641 (1997).

    Article  ADS  CAS  Google Scholar 

  17. Zhu,J. et al. Intramolecular masking of nuclear import signal on NF-AT4 by casein kinase I and MEKKI. Cell 93, 851–861 (1998).

    Article  CAS  Google Scholar 

  18. He,X., Saint-Jeannet,J. P., Woodgett,J. R., Varmus,H. E. & Dawid,I. B. Glycogen synthase kinase-3 and dorsoventral patterning in Xenopus embryos [published erratum appears in Nature 375, 253 (1995)]. Nature 374, 617–622 (1995).

    Article  ADS  CAS  Google Scholar 

  19. Genazzani,A. A., Carafoli,E. & Guerini,D. Calcineurin controls inositol 1,4,5-trisphosphate type I receptor expression in neurons. Proc. Natl Acad. Sci. USA 96, 5797–5801 (1999).

    Article  ADS  CAS  Google Scholar 

  20. Crabtree,G. R. Contingent genetic regulatory events in T lymphocyte activation. Science 243, 355–361 (1989).

    Article  ADS  CAS  Google Scholar 

  21. Zhou,P., Sun,L. J., Dotsch,V., Wagner,G. & Verdine,G. L. Solution structure of the core NFATC1/DNA complex. Cell 92, 687–696 (1998).

    Article  CAS  Google Scholar 

  22. Bito,H., Deisseroth,K. & Tsien,R. W. Ca2+-dependent regulation in neuronal gene expression. Curr. Opin. Neurobiol. 7, 419–429 (1997).

    Article  CAS  Google Scholar 

  23. Ghosh,A. & Greenberg,M. E. Calcium signaling in neurons: molecular mechanisms and cellular consequences. Science 268, 239–247 (1995).

    Article  ADS  CAS  Google Scholar 

  24. Inoue,T., Kato,K., Kohda,K. & Mikoshiba,K. Type 1 inositol 1,4,5-trisphosphate receptor is required for induction of long-term depression in cerebellar Purkinje neurons. J. Neurosci. 18, 5366–5373 (1998).

    Article  CAS  Google Scholar 

  25. Kasono,K. & Hirano,T. Involvement of inositol trisphosphate in cerebellar long-term depression. NeuroReport 6, 569–572 (1995).

    Article  CAS  Google Scholar 

  26. Reyes,M. & Stanton,P. K. Induction of hippocampal long-term depression requires release of Ca2+ from separate presynaptic and postsynaptic intracellular stores. J. Neurosci. 16, 5951–5960 (1996).

    Article  CAS  Google Scholar 

  27. Nguyen,P. V., Abel,T. & Kandel,E. R. Requirement of a critical period of transcription for induction of a late phase of LTP. Science 265, 1104–1107 (1994).

    Article  ADS  CAS  Google Scholar 

  28. Flexner,L. B., Flexner,J. B. & Roberts,R. B. Memory in mice analyzed with antibiotics. Antibiotics are useful to study stages of memory and to indicate molecular events which sustain memory. Science 155, 1377–1383 (1967).

    Article  ADS  CAS  Google Scholar 

  29. Deisseroth,K., Heist,E. K. & Tsien,R. W. Translocaton of calmodulin to the nucleus supports CREB phosphorylation in hippocampal neurons. Nature 392, 198–202 (1998).

    Article  ADS  CAS  Google Scholar 

  30. Bito,H., Deisseroth,K. & Tsien,R. W. CREB phosphorylation and dephosphorylation: a Ca(2+)- and stimulus duration-dependent switch for hippocampal gene expression. Cell 87, 1203–1214 (1996).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank T. Hoey for human NF-ATc4 cDNA, I. Bezprozvanny for the IP3R1 polyclonal antibody, D. Virshup for the casein kinase 1α expression construct, J. Healy for the MEKK expression construct and M. Karin for the JNK-1 expression construct. We also thank D. Wheeler, G. Pitt, J. H. Bayle and S. H. Park for helpful discussions. K.S. is a Stanford graduate fellow and a Howard Hughes Medical Institute predoctoral fellow. G.R.C. is an Investigator of the Howard Hughes Medical Institute. R.W.T is a McKnight Senior Investigator. This work was supported by grants from the NIH and the Mathels Charitable Trust.

Author information

Author notes

  1. Isabella A. Graef, Paul G. Mermelstein, Karl Deisseroth and Richard W. Tsien: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Pathology and Department of Developmental Biology, Howard Hughes Medical Institute,

    Isabella A. Graef, Kryn Stankunas, Joel R. Neilson & Gerald R. Crabtree

  2. Department of Molecular and Cellular Physiology, Beckman Center for Molecular and Genetic Medicine, Stanford University Medical School, 300 Pasteur Drive, Stanford, 94305-5426, California, USA

    Paul G. Mermelstein, Karl Deisseroth & Richard W. Tsien

Authors
  1. Isabella A. Graef
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Paul G. Mermelstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kryn Stankunas
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Joel R. Neilson
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Karl Deisseroth
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Richard W. Tsien
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Gerald R. Crabtree
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gerald R. Crabtree.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Graef, I., Mermelstein, P., Stankunas, K. et al. L-type calcium channels and GSK-3 regulate the activity of NF-ATc4 in hippocampal neurons. Nature 401, 703–708 (1999). https://doi.org/10.1038/44378

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/44378

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