Dynamic control of caMKII translocation and localization in hippocampal neurons by NMDA receptor stimulation

Science

Volumn 284, Issue 5411, 1999, Pages 162-166

  Shen, Kang ^a   Meyer, Tobias ^a  

^a NONE

Author keywords

[No Author keywords available]

Indexed keywords

F ACTIN; GREEN FLUORESCENT PROTEIN; PROTEIN KINASE (CALCIUM,CALMODULIN) II;

ANIMAL CELL; ARTICLE; AUTOPHOSPHORYLATION; BINDING AFFINITY; CELLULAR DISTRIBUTION; CONTROLLED STUDY; ENZYME ACTIVATION; ENZYME PHOSPHORYLATION; GLIA CELL; HIPPOCAMPUS; NERVE CELL PLASTICITY; NONHUMAN; PRIORITY JOURNAL; PROTEIN PROTEIN INTERACTION;

ACTINS; ANIMALS; CA(2+)-CALMODULIN DEPENDENT PROTEIN KINASE; CALCIUM; CELLS, CULTURED; CYTOSOL; DENDRITES; ELECTRIC STIMULATION; GLUTAMIC ACID; GREEN FLUORESCENT PROTEINS; HIPPOCAMPUS; ISOENZYMES; LUMINESCENT PROTEINS; MICROSCOPY, FLUORESCENCE; NERVE TISSUE PROTEINS; NEURONS; PHOSPHORYLATION; RATS; RECEPTORS, N-METHYL-D-ASPARTATE; SYNAPSES; TUMOR CELLS, CULTURED;

EID: 0033515884     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.284.5411.162     Document Type: Article

References (41)

1. R. Malinow, H. Schulman, R. W. Tsien, Nature 335, 820 (1988); D. L. Pettit, S. Perlman, R. Malinow, Science 266, 1881 (1994); S. Clazewski, C.-M. Chen, A. Silva, K. Fox, ibid. 272, 421 (1996); J. A. Gordon et al., Neuron 17, 491 (1996); M. Mayford et al., Science 274, 1678 (1996); M. Mayford et al., Cell 81, 891 (1995); A. J. Silva, S. Tonegawa, C. F. Stevens, Y. Wang, Science 257, 201 (1992).
2. R. Malinow, H. Schulman, R. W. Tsien, Nature 335, 820 (1988); D. L. Pettit, S. Perlman, R. Malinow, Science 266, 1881 (1994); S. Clazewski, C.-M. Chen, A. Silva, K. Fox, ibid. 272, 421 (1996); J. A. Gordon et al., Neuron 17, 491 (1996); M. Mayford et al., Science 274, 1678 (1996); M. Mayford et al., Cell 81, 891 (1995); A. J. Silva, S. Tonegawa, C. F. Stevens, Y. Wang, Science 257, 201 (1992).
3. R. Malinow, H. Schulman, R. W. Tsien, Nature 335, 820 (1988); D. L. Pettit, S. Perlman, R. Malinow, Science 266, 1881 (1994); S. Clazewski, C.-M. Chen, A. Silva, K. Fox, ibid. 272, 421 (1996); J. A. Gordon et al., Neuron 17, 491 (1996); M. Mayford et al., Science 274, 1678 (1996); M. Mayford et al., Cell 81, 891 (1995); A. J. Silva, S. Tonegawa, C. F. Stevens, Y. Wang, Science 257, 201 (1992).
4. R. Malinow, H. Schulman, R. W. Tsien, Nature 335, 820 (1988); D. L. Pettit, S. Perlman, R. Malinow, Science 266, 1881 (1994); S. Clazewski, C.-M. Chen, A. Silva, K. Fox, ibid. 272, 421 (1996); J. A. Gordon et al., Neuron 17, 491 (1996); M. Mayford et al., Science 274, 1678 (1996); M. Mayford et al., Cell 81, 891 (1995); A. J. Silva, S. Tonegawa, C. F. Stevens, Y. Wang, Science 257, 201 (1992).
5. R. Malinow, H. Schulman, R. W. Tsien, Nature 335, 820 (1988); D. L. Pettit, S. Perlman, R. Malinow, Science 266, 1881 (1994); S. Clazewski, C.-M. Chen, A. Silva, K. Fox, ibid. 272, 421 (1996); J. A. Gordon et al., Neuron 17, 491 (1996); M. Mayford et al., Science 274, 1678 (1996); M. Mayford et al., Cell 81, 891 (1995); A. J. Silva, S. Tonegawa, C. F. Stevens, Y. Wang, Science 257, 201 (1992).
6. R. Malinow, H. Schulman, R. W. Tsien, Nature 335, 820 (1988); D. L. Pettit, S. Perlman, R. Malinow, Science 266, 1881 (1994); S. Clazewski, C.-M. Chen, A. Silva, K. Fox, ibid. 272, 421 (1996); J. A. Gordon et al., Neuron 17, 491 (1996); M. Mayford et al., Science 274, 1678 (1996); M. Mayford et al., Cell 81, 891 (1995); A. J. Silva, S. Tonegawa, C. F. Stevens, Y. Wang, Science 257, 201 (1992).
7. R. Malinow, H. Schulman, R. W. Tsien, Nature 335, 820 (1988); D. L. Pettit, S. Perlman, R. Malinow, Science 266, 1881 (1994); S. Clazewski, C.-M. Chen, A. Silva, K. Fox, ibid. 272, 421 (1996); J. A. Gordon et al., Neuron 17, 491 (1996); M. Mayford et al., Science 274, 1678 (1996); M. Mayford et al., Cell 81, 891 (1995); A. J. Silva, S. Tonegawa, C. F. Stevens, Y. Wang, Science 257, 201 (1992).
8. C.-Y. Wu and H. T. Cline, Science 279, 222 (1998).
9. CA3-CA1 hippocampat cultures (postnatal day 2 to 4) were prepared as previously described (8). Neurons, 6 to 14 days in culture, were transfected by microelectroporation of the respective mRNA or of mixtures of different mRNAs (22). For some of the mutants, DNA constructs were used instead of the mRNA. Three electrical pulses (20 V/mm, 20 s apart) were applied to the neurons immediately after mRNA or DNA application. Localization and translocation studies were done 8 to 16 hours after transfection.
10. Supplementary figures can be found at the Science Web site (www.sdencemag.org/feature/data/986549.shl)
11. K. Shen and T. Meyer, data not shown.
12. S. G. Miller and M. B. Kennedy, J. Biol. Chem. 260, 9039 (1985); S. M. Shields et al., J. Neurochem. 43, 1599 (1984); J. R. Goldenring, ibid. 42, 1077 (1984); B. R. Hu and T. Wieloch, ibid. 64, 277 (1995); T. Suzuki et al., ibid. 63, 1529 (1994); S. Strack et al., J. Biol. Chem. 272, 13467 (1997); Y. Yoshimura and T Yamauchi, ibid., p. 26354.
13. S. G. Miller and M. B. Kennedy, J. Biol. Chem. 260, 9039 (1985); S. M. Shields et al., J. Neurochem. 43, 1599 (1984); J. R. Goldenring, ibid. 42, 1077 (1984); B. R. Hu and T. Wieloch, ibid. 64, 277 (1995); T. Suzuki et al., ibid. 63, 1529 (1994); S. Strack et al., J. Biol. Chem. 272, 13467 (1997); Y. Yoshimura and T Yamauchi, ibid., p. 26354.
14. S. G. Miller and M. B. Kennedy, J. Biol. Chem. 260, 9039 (1985); S. M. Shields et al., J. Neurochem. 43, 1599 (1984); J. R. Goldenring, ibid. 42, 1077 (1984); B. R. Hu and T. Wieloch, ibid. 64, 277 (1995); T. Suzuki et al., ibid. 63, 1529 (1994); S. Strack et al., J. Biol. Chem. 272, 13467 (1997); Y. Yoshimura and T Yamauchi, ibid., p. 26354.
15. S. G. Miller and M. B. Kennedy, J. Biol. Chem. 260, 9039 (1985); S. M. Shields et al., J. Neurochem. 43, 1599 (1984); J. R. Goldenring, ibid. 42, 1077 (1984); B. R. Hu and T. Wieloch, ibid. 64, 277 (1995); T. Suzuki et al., ibid. 63, 1529 (1994); S. Strack et al., J. Biol. Chem. 272, 13467 (1997); Y. Yoshimura and T Yamauchi, ibid., p. 26354.
16. S. G. Miller and M. B. Kennedy, J. Biol. Chem. 260, 9039 (1985); S. M. Shields et al., J. Neurochem. 43, 1599 (1984); J. R. Goldenring, ibid. 42, 1077 (1984); B. R. Hu and T. Wieloch, ibid. 64, 277 (1995); T. Suzuki et al., ibid. 63, 1529 (1994); S. Strack et al., J. Biol. Chem. 272, 13467 (1997); Y. Yoshimura and T Yamauchi, ibid., p. 26354.
17. S. G. Miller and M. B. Kennedy, J. Biol. Chem. 260, 9039 (1985); S. M. Shields et al., J. Neurochem. 43, 1599 (1984); J. R. Goldenring, ibid. 42, 1077 (1984); B. R. Hu and T. Wieloch, ibid. 64, 277 (1995); T. Suzuki et al., ibid. 63, 1529 (1994); S. Strack et al., J. Biol. Chem. 272, 13467 (1997); Y. Yoshimura and T Yamauchi, ibid., p. 26354.
18. S. G. Miller and M. B. Kennedy, J. Biol. Chem. 260, 9039 (1985); S. M. Shields et al., J. Neurochem. 43, 1599 (1984); J. R. Goldenring, ibid. 42, 1077 (1984); B. R. Hu and T. Wieloch, ibid. 64, 277 (1995); T. Suzuki et al., ibid. 63, 1529 (1994); S. Strack et al., J. Biol. Chem. 272, 13467 (1997); Y. Yoshimura and T Yamauchi, ibid., p. 26354.
19. 7. Fluorescent images were taken at an excitation wavelength of 488 nm, using a confocal microscope (LSM410, Zeiss Inc.). For the immunocolocalization experiments, neurons were fixed with methanol for 10 min at -20°C and blocked with bovine serum albumin (10 mg/ml). Monoclonal antibody to PSD-95 (1:200) was used (Affinity Bioreagents), followed by Cy5-labeled secondary antibodies. Cy5 distribution was monitored by excitation at 568 nm.
20. K. Shen et al., Neuron 21, 593 (1998).
21. For photobleaching experiments, the microscope was zoomed onto a small region of the neuron at maximal laser power. Recovery of the fluorescence was then measured at lower laser power by sequential imaging of the entire neuron.
22. AV. Each of the traces shown was calculated from an average of such intensity recordings. The same type of analysis was also used to measure the time course of dissociation of β-CaMKII from F-actin after glutamate addition. The average intensity was measured in small cortical regions instead of the PSD.
23. V. Ramakrishnapillai et al., J. Biol. Chem. 271, 31670 (1996).
24. E. McGlade-McCulloh et al., Nature 362, 640 (1993); A. Barria, D. Muller, V. Derkach, L. C. Griffith, T. R. Soderling, Science 276, 2042 (1997).
25. E. McGlade-McCulloh et al., Nature 362, 640 (1993); A. Barria, D. Muller, V. Derkach, L. C. Griffith, T. R. Soderling, Science 276, 2042 (1997).
26. H. Chen et al., Neuron 20, 895 (1998).
27. S. Strack and R. J. Colbran, J. Biol. Chem. 273, 20689 (1998); F. Gardoni et al., J. Neurochem. 71, 1733 (1998).
28. S. Strack and R. J. Colbran, J. Biol. Chem. 273, 20689 (1998); F. Gardoni et al., J. Neurochem. 71, 1733 (1998).
29. K. Fukunaga, T. R. Soderling, E. Miyamoto, J. Biol. Chem. 267, 22527 (1992).
30. C. M. Schworer, R. J. Colbran, T. R. Soderling, ibid. 261, 8581 (1986); L. L. Lou, S. J. Lloyd, H. Schulman, Proc Natl. Acad. Sci. U.S.A. 83, 9497 (1986).
31. C. M. Schworer, R. J. Colbran, T. R. Soderling, ibid. 261, 8581 (1986); L. L. Lou, S. J. Lloyd, H. Schulman, Proc Natl. Acad. Sci. U.S.A. 83, 9497 (1986).
32. T. Meyer, P. I. Hanson, L. Stryer, H. Schulman, Science 256, 1199 (1992).
33. GFP-tagged α-and β-CaMKII isoforms were characterized previously (8). Point mutations of α-and β-CaMKII were created using polymerase chain reactions in the in vitro translation vector pSHIRO3 (22).
34. P. Hanson et al., Neuron 12, 943 (1994).
35. Y. L. Fong et al., J. Biol. Chem. 264, 16759 (1989); P. Hanson et al., Neuron 3, 59 (1989).
36. Y. L. Fong et al., J. Biol. Chem. 264, 16759 (1989); P. Hanson et al., Neuron 3, 59 (1989).
37. S. Mukherji and T. R. Soderling, J. Biol. Chem. 269, 13744 (1994).
38. M. F. Teruel and T. Meyer [Biophys. J. 73, 1785 (1997)] described the microporation technique: H. Yokoe and T. Meyer [Nature Biotechnol. 14, 1252 (1996)] described the RNA transfection technique; M. F. Teruel et al. (in preparation) described the efficiency of RNA cotransfection in neurons.
39. M. F. Teruel and T. Meyer [Biophys. J. 73, 1785 (1997)] described the microporation technique: H. Yokoe and T. Meyer [Nature Biotechnol. 14, 1252 (1996)] described the RNA transfection technique; M. F. Teruel et al. (in preparation) described the efficiency of RNA cotransfection in neurons.
40. M. F. Teruel and T. Meyer [Biophys. J. 73, 1785 (1997)] described the microporation technique: H. Yokoe and T. Meyer [Nature Biotechnol. 14, 1252 (1996)] described the RNA transfection technique; M. F. Teruel et al. (in preparation) described the efficiency of RNA cotransfection in neurons.
41. We thank B. Wang for support with the neuronal culturing, K. Subramanian for molecular biology support, and M. Teruel for developing the neuronal RNA transfection technique. We also thank E. Oancea, W. Chen, and A. Means and M. Teruel for stimulating discussions. Supported by NIH grant GM-48113.