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

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.

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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.

References

  1. 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).

  2. 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).

  3. 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).

  4. 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).

  5. 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).

  6. 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).

  7. 7

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

  8. 8

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

  9. 9

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

  10. 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).

  11. 11

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

  12. 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).

  13. 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).

  14. 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).

  15. 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).

  16. 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).

  17. 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).

  18. 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).

  19. 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).

  20. 20

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

  21. 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).

  22. 22

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

  23. 23

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

  24. 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).

  25. 25

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

  26. 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).

  27. 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).

  28. 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).

  29. 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).

  30. 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).

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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.

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Correspondence to Gerald R. Crabtree.

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