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Nature Neuroscience | Article

GABA promotes the competitive selection of dendritic spines by controlling local Ca2+ signaling

Journal name:
Nature Neuroscience
Volume:
16,
Pages:
1409–1416
Year published:
DOI:
doi:10.1038/nn.3496
Received
Accepted
Published online

Abstract

Activity-dependent competition of synapses plays a key role in neural organization and is often promoted by GABA; however, its cellular bases are poorly understood. Excitatory synapses of cortical pyramidal neurons are formed on small protrusions known as dendritic spines, which exhibit structural plasticity. We used two-color uncaging of glutamate and GABA in rat hippocampal CA1 pyramidal neurons and found that spine shrinkage and elimination were markedly promoted by the activation of GABAA receptors shortly before action potentials. GABAergic inhibition suppressed bulk increases in cytosolic Ca2+ concentrations, whereas it preserved the Ca2+ nanodomains generated by NMDA-type receptors, both of which were necessary for spine shrinkage. Unlike spine enlargement, spine shrinkage spread to neighboring spines (<15 μm) and competed with their enlargement, and this process involved the actin-depolymerizing factor ADF/cofilin. Thus, GABAergic inhibition directly suppresses local dendritic Ca2+ transients and strongly promotes the competitive selection of dendritic spines.

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  1. Spine shrinkage and elimination induced with the spike-timing protocol.
    Figure 1: Spine shrinkage and elimination induced with the spike-timing protocol.

    (a,c,e) Pairing of glutamate uncaging and postsynaptic action potentials in the LTP (a) and LTD (c,e) protocols. Scale bars in a also apply to c,e. (b) Spine enlargement induced with the LTP protocol at 1 Hz, 80 times (8 spines, 8 dendrites, 8 slices, 4 rats). (d) The absence of spine shrinkage in the LTD protocol without GABA uncaging (15 spines, 15 dendrites, 8 slices, 5 rats). (f) Time courses of spine shrinkage in the stimulated (s) and neighboring (n1, n2) spines shown in g. (g) Alexa 594 fluorescence images of a dendrite with one spine (arrowhead) to which the LTD protocol was applied with GABA uncaging. GABA uncaging was applied at spike onset as in e, at four points (cyan) around the dendritic shaft for a total duration of 4 ms, each separated by 1 ms. Scale bar, 2 μm. The soma was located to the left. (h) A current trace that was elicited by GABA uncaging at the dendritic shaft shown in g, with it voltage clamped at 0 mV. (i) Spine shrinkage induced with the LTD protocol with GABA uncaging in stimulated (14 spines, 14 dendrites, 14 slices, 10 rats), neighboring (19 spines) and distant spines (9 spines, 5 dendrites, 5 slices, 5 rats). (j) The average spine shrinkage by the LTD protocol in the presence of APV (Supplementary Fig. 3a; 13 spines, 4 dendrites, 4 slices, 2 rats), CGP55845 (22 spines, 5 dendrites, 4 slices, 3 rats), without glutamate uncaging (w/o Glu; 14 spines, 3 dendrites, 3 slices, 2 rats), without the spike (w/o spiking, 33 spines, 6 dendrites, 3 slices, 2 rats) or without GABA uncaging (w/o GABA; 24 spines, 15 dendrites, 8 slices, 5 rats). Data are presented as mean ± s.e.m. ***P < 0.001 versus 0% by Wilcoxon signed-rank test.

  2. Spine shrinkage spread and GABA effects.
    Figure 2: Spine shrinkage spread and GABA effects.

    (a) The average reductions in the spine volumes of stimulated (14 spines, 14 dendrites, 14 slices, 10 rats) and neighboring spines at ≤3 μm (15 spines), 3–10 μm (41 spines), 10–15 μm (17 spines) and ≥15 μm (9 spines) from the stimulated spines. (b) The time courses of current amplitudes evoked by glutamate uncaging in stimulated (9 spines, 9 dendrites, 9 slices, 7 rats), neighboring (<3 μm, 10 spines, 9 dendrites, 9 slices, 7 rats) and distant spines (>15 μm, 8 spines, 8 dendrites, 7 slices, 6 rats). The insets show an example of current traces (two-photon stimulation–induced EPSCs (2pEPSCs)) before and after LTD induction. (c) Time points (blue arrows) of GABA uncaging relative to spike onset. (d) Timing dependence of GABA uncaging on spine shrinkage (Supplementary Fig. 3c,d). (e) A drawing of a dendritic shaft where GABA uncaging was applied 25 μm from the stimulated spine. (f) Spine shrinkage was detected when GABA uncaging was within 5 μm of the stimulated spines (33 spines, 14 dendrites, 14 slices, 10 rats) but not when GABA uncaging was more than 25 μm away from the stimulated spines (21 spines, 6 dendrites, 6 slices, 3 rats). Data are presented as mean ± s.e.m. *P < 0.05, **P < 0.01,***P < 0.001 versus 0% by Wilcoxon signed-rank test.

  3. Pharmacology of spine shrinkage.
    Figure 3: Pharmacology of spine shrinkage.

    (a) LTD protocol with 200 nM muscimol applied using the same pipette as that used for caged glutamate application. bAPs and glutamate uncaging were repeated 80 times at 5 Hz. (b) Spine shrinkage induced with the LTD protocol with 200 nM muscimol. The spine in which glutamate was uncaged is labeled s; neighboring spines, n1–n3. Scale bar, 2 μm. The soma was located to the left. (c) Average time courses of changes in the volumes of stimulated (21 spines, 21 dendrites, 19 slices, 16 rats) and neighboring spines (27 spines). (d) Average reductions in spine volumes of stimulated (21 spines, 21 dendrites, 19 slices, 16 rats) and neighboring spines at ≤3 μm (27 spines), 3–10 μm (56 spines), 10–15 μm (38 spines) and ≥15 μm (21 spines) from the stimulated spines. (e) Average reductions in spine volumes with the LTD protocol with muscimol (0 nM, 24 spines, 15 dendrites, 8 slices, 5 rats; 50 nM, 28 spines, 11 dendrites, 7 slices, 4 rats; 200 nM, 56 spines, 18 dendrites, 17 slices, 14 rats; Supplementary Fig. 5a) and APV (50 μM, 41 spines, 13 dendrites, 6 slices, 4 rats), MK-801 (1 mM, 14 spines, 5 dendrites, 2 slices, 2 rats), MCPG (1 mM, 28 spines, 10 dendrites, 5 slices, 3 rats), CPA (30 μM, 23 spines, 8 dendrites, 4 slices, 2 rats), Y-27632 Rho-kinase inhibitor (20 μM, 21 spines, 8 dendrites, 3 slices, 3 rats), FK506 (1 μM, 28 spines, 10 dendrites, 5 slices, 4 rats) and P-cofilin peptide (0.5 mM, 18 spines, 5 dendrites, 5 slices, 3 rats) (Supplementary Fig. 5b). Wilcoxon signed-rank test versus 0%: **P < 0.01, ***P < 0.001. Steel's test versus muscimol 200 nM: 0 nM, 50 nM muscimol, APV, internal MK-801, FK506 and P-cofilin peptide (P < 0.0001), CPA (P = 0.028), MCPG (P = 0.81) and Y-27632 (P = 0.99). (f) Spine volumes during the LTD protocol in controls and with MCPG and FK506 in the bathing solution or MK-801 and P-cofilin peptide in the pipette. The shrinkage amplitudes were averaged among neighboring spines (2–5 spines) in dendrites within 3 μm of the stimulated spines. Data are presented as mean ± s.e.m.

  4. Competition of the enlargement and shrinkage of neighboring spines.
    Figure 4: Competition of the enlargement and shrinkage of neighboring spines.

    (a) Spike timing protocol for the heterosynaptic competition of spine enlargement and shrinkage with 200 nM muscimol. Glutamate uncaging was applied 5 ms before the spike and 10 ms after the spike, and repetitive pairing was performed 80 times (5 Hz). (b) Fluorescence images of a dendrite subjected to the LTP (e) and LTD protocols (s). Scale bar, 2 μm. The soma was located to the right. (c) Volume changes in the spines shown in b subjected to the LTD protocol (s) and its neighbors (n) or to the LTP protocol (e) and its neighbors distal to spines with LTD stimuli (ne). The duration of glutamate uncaging in the LTP protocol was 3 ms. (d) Volume changes in spines subjected to the LTD protocol (14 spines, 14 dendrites, 14 slices, 13 rats) and their neighbors (34 spines, 14 dendrites, 14 slices, 13 rats) and to the LTP protocol (14 spines, 14 dendrites, 14 dendrites, 13 rats) and its distal neighbors (12 spines, 12 dendrites, 11 slices, 11 rats). The duration of glutamate uncaging in the LTP protocol was 1.8 ms (8 spines, 8 dendrites, 8 slices, 7 rats) or 3 ms (6 spines, 6 dendrites, 6 slices, 6 rats). (e) Peak amplitude changes in glutamate-induced currents (2pEPSCs) in spines subjected to the LTP protocol (5 spines, 5 dendrites, 5 slices, 5 rats), LTD protocol (5 spines, 5 dendrites, 5 slices, 5 rats), neighboring spines (6 spines, 5 dendrites, 5 slices, 5 rats) and distant ones (4 spines, 4 dendrites, 4 slices, 4 rats) ≥15 μm from stimulated ones. Glutamate-induced currents were recorded with voltage clamp at a holding potential of −65 mV. (f) Competition of spine enlargement against the spreading shrinkage induced by neighboring spine stimulation with the LTD protocol (muscimol, 200 nM or 50 nM; Supplementary Fig. 7a,b). Enlargement outcompeted shrinkage when spines were subjected to LTP protocols as in a with glutamate uncagings of 1.2 ms, 1.8 ms or 3 ms with 200 nM muscimol and 0.6 ms or 1.2 ms with 50 nM muscimol (Supplementary Fig. 7a,b). *P < 0.05, **P < 0.01 by Steel's test versus 0 ms. The curves were drawn by eye. (g) The volume changes in spines subjected to the LTP protocol (5 spines, 5 dendrites, 5 slices, 3 rats, 3 ms), the LTD protocol and neighboring spines (23 spines, 5 dendrites, 5 slices, 3 rats) in the presence of dephosphorylated (dp)-cofilin peptide in the patch pipette and 200 nM muscimol. Data are presented as mean ± s.e.m.

  5. The spatial profile of GABA uncaging-mediated inhibition of bAP-induced dendritic Ca2+ transients.
    Figure 5: The spatial profile of GABA uncaging-mediated inhibition of bAP-induced dendritic Ca2+ transients.

    (a) mCherry fluorescence image of a dendrite to which a combination of bAPs and GABA uncaging was applied at the 0-μm location. The soma was located to the bottom. (b,c) GCaMP6s (see Online Methods) images of the dendrite shown in a. The images were normalized by the images before bAP, and the relative increases in the fluorescence were pseudocolor coded. Each image is an average of 6–8 frames. (d) Time courses of fluorescence changes without (black) or with (blue) GABA uncaging at each dendritic location. The traces are averages of 4–13 dendrites. (e) Relative reductions in the peak amplitudes of bAP-induced Ca2+ transient along the dendrite. Kruskal–Wallis test, P < 0.0055. Steel's test, *P < 0.05, **P < 0.01 (4–13 dendrites, 4–10 slices, 2–5 rats). Data are presented as mean ± s.e.m.

  6. Spine Ca2+ signaling and effects of cytosolic EGTA.
    Figure 6: Spine Ca2+ signaling and effects of cytosolic EGTA.

    (a) Effects of GABA uncaging (blue) on peak fluorescence ratios between the low-affinity (KD = 2.3 μM) Ca2+ indicator Fluo5F and Alexa 594 (G/R) for spine [Ca2+]i increases (Supplementary Fig. 8) by a combination of bAP and glutamate uncaging in LTD (22 spines, 6 dendrites, 3 slices, 3 rats), bAP (25 spines, 9 dendrites, 5 slices, 5 rats) and glutamate uncaging (17 spines, 6 dendrites, 5 slices, 4 rats). Two-sided paired t-test versus without GABA uncaging. (b) Effects of muscimol on G/R ratios between Fluo5F and Alexa 594 (Supplementary Fig. 9) by a combination of bAP and glutamate uncaging in LTD at 0 nM (51 spines, 13 dendrites, 13 slices, 8 spines), 50 nM (33 spines, 6 dendrites, 6 slices, 6 rats) and 200 nM (18 spines, 7 dendrites, 7 slices, 2 rats). Two-sided paired t-test versus 0 nM. (c) LTD protocol with 3 mM EGTA in the patch pipette instead of muscimol in the extracellular solution. (d) Spine shrinkage by the LTD protocol with 3 mM EGTA in the patch pipette. Arrow indicates spine in which glutamate was uncaged. Scale bar, 2 μm. (e) Dependence of shrinkage on EGTA concentration (Supplementary Fig. 7c). (f) Dependence of G/R ratio on EGTA concentration (Supplementary Fig. 10). Averaged fluorescence ratios with 0.5 mM EGTA were normalized to 100%. (g) Spine shrinkage dependence on G/R ratio. Data are presented as mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001 versus 0% by Wilcoxon signed-rank test.

  7. Spine Ca2+ signaling and effects of cytosolic BAPTA.
    Figure 7: Spine Ca2+ signaling and effects of cytosolic BAPTA.

    (a) Effect on spine shrinkage of laser irradiation power used for glutamate uncaging, with 6 mW (24 spines, 15 dendrites, 8 slices, 5 rats), 4 mW (28 spines, 6 dendrites, 2 slices, 2 rats) or 3 mW (15 spines, 6 dendrites, 2 slices, 2 rats) (Supplementary Fig. 7e). (b) Effect on fluorescence ratio, for the same spines, of laser power used for glutamate uncaging (30 spines, 5 dendrites, 5 slices, 3 rats), with 6 mW, 4 mW or 3 mW (Supplementary Fig. 9e). P = 0.014 by ANOVA. P = 0.016 and 0.0016 by Scheffe test for 4 mW and 3 mW, respectively, versus 6 mW. (c) Dependence of shrinkage on the BAPTA concentration (Supplementary Fig. 7d), relative to that of EGTA (same as in Fig. 6e). (d) Dependence of peak fluorescence ratio (G/R) on BAPTA concentration (Supplementary Fig. 10), relative to that of EGTA (same as in Fig. 6f). Averaged fluorescence ratios with 0.5 mM EGTA were normalized as 100%. (e) Spine shrinkage dependence on G/R ratio in the presence of BAPTA and EGTA (same as Fig. 6g). Data are presented as mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001 versus 0% by Wilcoxon signed-rank test.

References

  1. Hensch, T.K. Critical period plasticity in local cortical circuits. Nat. Rev. Neurosci. 6, 877–888 (2005).
  2. Nakayama, H. et al. GABAergic inhibition regulates developmental synapse elimination in the cerebellum. Neuron 74, 384–396 (2012).
  3. Matsuzaki, M. et al. Dendritic spine geometry is critical for AMPA receptor expression in hippocampal CA1 pyramidal neurons. Nat. Neurosci. 4, 1086–1092 (2001).
  4. Noguchi, J. et al. In vivo two-photon uncaging of glutamate revealing the structure-function relationships of dendritic spines in the neocortex of adult mice. J. Physiol. (Lond.) 589, 2447–2457 (2011).
  5. Matsuzaki, M., Honkura, N., Ellis-Davies, G. & Kasai, H. Structural basis of long-term potentiation in single dendritic spines. Nature 429, 761–766 (2004).
  6. Harvey, C.D. & Svoboda, K. Locally dynamic synaptic learning rules in pyramidal neuron dendrites. Nature 450, 1195–1200 (2007).
  7. Zhou, Q., Homma, K.J. & Poo, M.M. Shrinkage of dendritic spines associated with long-term depression of hippocampal synapses. Neuron 44, 749–757 (2004).
  8. Holtmaat, A. & Svoboda, K. Experience-dependent structural synaptic plasticity in the mammalian brain. Nat. Rev. Neurosci. 10, 647–658 (2009).
  9. Kasai, H., Fukuda, M., Watanabe, S., Hayashi-Takagi, A. & Noguchi, J. Structural dynamics of dendritic spines in memory and cognition. Trends Neurosci. 33, 121–129 (2010).
  10. Yang, G., Pan, F. & Gan, W. Stably maintained dendritic spines are associated with lifelong memories. Nature 462, 920–924 (2009).
  11. Nowak, L., Bregestovski, P., Ascher, P., Herbet, A. & Prochiantz, A. Magnesium gates glutamate-activated channels in mouse central neurones. Nature 307, 462–465 (1984).
  12. Mori, H. & Mishina, M. Structure and function of the NMDA receptor channel. Neuropharmacology 34, 1219–1237 (1995).
  13. Magee, J.C. & Johnston, D. A synaptically controlled, associative signal for Hebbian plasticity in hippocampal neurons. Science 275, 209–213 (1997).
  14. Markram, H., Lubke, J., Frotscher, M. & Sakmann, B. Regulation of synaptic efficacy by coincidence of postsynaptic APs and EPSPs. Science 275, 213–215 (1997).
  15. Dan, Y. & Poo, M.M. Spike timing-dependent plasticity: from synapse to perception. Physiol. Rev. 86, 1033–1048 (2006).
  16. Hensch, T.K. & Fagiolini, M. Excitatory-inhibitory balance and critical period plasticity in developing visual cortex. Prog. Brain Res. 147, 115–124 (2005).
  17. Mataga, N., Mizuguchi, Y. & Hensch, T.K. Experience-dependent pruning of dendritic spines in visual cortex by tissue plasminogen activator. Neuron 44, 1031–1041 (2004).
  18. Tanaka, J. et al. Protein synthesis and neurotrophin-dependent structural plasticity of single dendritic spines. Science 319, 1683–1687 (2008).
  19. Lee, S.J., Escobedo-Lozoya, Y., Szatmari, E.M. & Yasuda, R. Activation of CaMKII in single dendritic spines during long-term potentiation. Nature 458, 299–304 (2009).
  20. Kanemoto, Y. et al. Spatial distributions of GABA receptors and local inhibition of Ca2+ transients studied with GABA uncaging in the dendrites of CA1 pyramidal neurons. PLoS ONE 6, e22652 (2011).
  21. Steele, P.M. & Mauk, M.D. Inhibitory control of LTP and LTD: stability of synapse strength. J. Neurophysiol. 81, 1559–1566 (1999).
  22. Nishiyama, M., Togashi, K., Aihara, T. & Hong, K. GABAergic activities control spike timing- and frequency-dependent long-term depression at hippocampal excitatory synapses. Front. Synaptic Neurosci. 2, 22 (2010).
  23. Abraham, W.C. & Goddard, G.V. Asymmetric relationships between homosynaptic long-term potentiation and heterosynaptic long-term depression. Nature 305, 717–719 (1983).
  24. Nishiyama, M., Hong, K., Mikoshiba, K., Poo, M.M. & Kato, K. Calcium stores regulate the polarity and input specificity of synaptic modification. Nature 408, 584–588 (2000).
  25. Wang, S.S., Khiroug, L. & Augustine, G.J. Quantification of spread of cerebellar long-term depression with chemical two-photon uncaging of glutamate. Proc. Natl. Acad. Sci. USA 97, 8635–8640 (2000).
  26. Fino, E. et al. RuBi-glutamate: two-photon and visible-light photoactivation of neurons and dendritic spines. Front. Neural Circuits 3, 2 (2009).
  27. Matsuzaki, M., Hayama, T., Kasai, H. & Ellis-Davies, G.C.R. Two-photon uncaging of gamma-aminobutyric acid in intact brain tissue. Nat. Chem. Biol. 6, 255–257 (2010).
  28. Bai, D. et al. Distinct functional and pharmacological properties of tonic and quantal inhibitory postsynaptic currents mediated by gamma-aminobutyric acid(A) receptors in hippocampal neurons. Mol. Pharmacol. 59, 814–824 (2001).
  29. Bernstein, B.W. & Bamburg, J.R. ADF/cofilin: a functional node in cell biology. Trends Cell Biol. 20, 187–195 (2010).
  30. Lai, F.P. et al. Arp2/3 complex interactions and actin network turnover in lamellipodia. EMBO J. 27, 982–992 (2008).
  31. Pontrello, C.G. et al. Cofilin under control of beta-arrestin-2 in NMDA-dependent dendritic spine plasticity, long-term depression (LTD), and learning. Proc. Natl. Acad. Sci. USA 109, E442–E451 (2012).
  32. Lai, C.S., Franke, T.F. & Gan, W.B. Opposite effects of fear conditioning and extinction on dendritic spine remodelling. Nature 483, 87–91 (2012).
  33. Bienenstock, E.L., Cooper, L.N. & Munro, P.W. Theory for the development of neuron selectivity: orientation specificity and binocular interaction in visual cortex. J. Neurosci. 2, 32–48 (1982).
  34. Lisman, J. A mechanism for the Hebb and the anti-Hebb processes underlying learning and memory. Proc. Natl. Acad. Sci. USA 86, 9574–9578 (1989).
  35. Yang, S.N., Tang, Y.G. & Zucker, R.S. Selective induction of LTP and LTD by postsynaptic [Ca2+]i elevation. J. Neurophysiol. 81, 781–787 (1999).
  36. Naraghi, M. & Neher, E. Linearized buffered Ca2+ diffusion in microdomains and its implications for calculation of [Ca2+] at the mouth of a calcium channel. J. Neurosci. 17, 6961–6973 (1997).
  37. Naraghi, M. T-jump study of calcium binding kinetics of calcium chelators. Cell Calcium 22, 255–268 (1997).
  38. Adler, E.M., Augustine, G.J., Duffy, S.N. & Charlton, M.P. Alien intracellular calcium chelators attenuate neurotransmitter release at the squid giant synapse. J. Neurosci. 11, 1496–1507 (1991).
  39. Markram, H., Roth, A. & Helmchen, F. Competitive calcium binding: implications for dendritic calcium signaling. J. Comput. Neurosci. 5, 331–348 (1998).
  40. Wittenberg, G.M. & Wang, S.S. Malleability of spike-timing-dependent plasticity at the CA3–CA1 synapse. J. Neurosci. 26, 6610–6617 (2006).
  41. Abraham, W.C. & Wickens, J.R. Heterosynaptic long-term depression is facilitated by blockade of inhibition in area CA1 of the hippocampus. Brain Res. 546, 336–340 (1991).
  42. Rusnak, F. & Mertz, P. Calcineurin: form and function. Physiol. Rev. 80, 1483–1521 (2000).
  43. Yang, E.J., Lin, E.W. & Hensch, T.K. Critical period for acoustic preference in mice. Proc. Natl. Acad. Sci. USA 109, 17213–17220 (2012).
  44. Paulsen, O. & Moser, E.I. A model of hippocampal memory encoding and retrieval: GABAergic control of synaptic plasticity. Trends Neurosci. 21, 273–278 (1998).
  45. Klausberger, T. & Somogyi, P. Neuronal diversity and temporal dynamics: the unity of hippocampal circuit operations. Science 321, 53–57 (2008).
  46. Oh, W.C., Hill, T.C. & Zito, K. Synapse-specific and size-dependent mechanisms of spine structural plasticity accompanying synaptic weakening. Proc. Natl. Acad. Sci. USA 110, E305–E312 (2013).
  47. Fiete, I.R., Senn, W., Wang, C.Z. & Hahnloser, R.H. Spike-time-dependent plasticity and heterosynaptic competition organize networks to produce long scale-free sequences of neural activity. Neuron 65, 563–576 (2010).
  48. Govindarajan, A., Kelleher, R.J. & Tonegawa, S. A clustered plasticity model of long-term memory engrams. Nat. Rev. Neurosci. 7, 575–583 (2006).
  49. Losonczy, A. & Magee, J.C. Integrative properties of radial oblique dendrites in hippocampal CA1 pyramidal neurons. Neuron 50, 291–307 (2006).
  50. Rust, M.B. et al. Learning, AMPA receptor mobility and synaptic plasticity depend on n-cofilin-mediated actin dynamics. EMBO J. 29, 1889–1902 (2010).
  51. Matsuzaki, M. & Kasai, H. Two-photon uncaging microscopy. Cold Spring Harbor Protocols 2011, pdb prot5620 (2011).
  52. Ellis-Davies, G.C.R. A practical guide to the synthesis of dinitroindolinyl-caged neurotransmitters. Nat. Protoc. 6, 314–326 (2011).
  53. Ellis-Davies, G.C.R., Matsuzaki, M., Paukert, M., Kasai, H. & Bergles, D. 4-Carboxymethoxy-5,7-dinitroindolinyl-Glu: an improved caged glutamate for expeditious ultraviolet and two-photon photolysis in brain slices. J. Neurosci. 27, 6601–6604 (2007).
  54. Rial Verde, E.M., Zayat, L., Etchenique, R. & Yuste, R. Photorelease of GABA with visible light using an inorganic caging group. Front Neural Circuits 2, 2 (2008).
  55. Chen, T.W. et al. Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature 499, 295–300 (2013).

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Author information

  1. These authors contributed equally to this work.

    • Tatsuya Hayama &
    • Jun Noguchi

Affiliations

  1. Laboratory of Structural Physiology, Center for Disease Biology and Integrative Medicine, Faculty of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.

    • Tatsuya Hayama,
    • Jun Noguchi,
    • Satoshi Watanabe,
    • Noriko Takahashi,
    • Akiko Hayashi-Takagi &
    • Haruo Kasai

  2. CREST, Japan Science and Technology Agency, Kawaguchi, Saitama, Japan.

    • Tatsuya Hayama,
    • Jun Noguchi,
    • Satoshi Watanabe,
    • Noriko Takahashi,
    • Akiko Hayashi-Takagi,
    • Masanori Matsuzaki &
    • Haruo Kasai

  3. PRESTO, Japan Science and Technology Agency, Kawaguchi, Saitama, Japan.

    • Akiko Hayashi-Takagi &
    • Masanori Matsuzaki

  4. Department of Neuroscience, Mount Sinai School of Medicine, New York, New York, USA.

    • Graham C R Ellis-Davies

  5. Division of Brain Circuits, National Institute for Basic Biology, The Graduate University of Advanced Studies (Sokendai), Okazaki, Japan.

    • Masanori Matsuzaki

Contributions

T.H., J.N. and H.K. conceived the experiments. T.H., J.N., M.M. and S.W. performed the slice experiments. A.H. and N.T. contributed to the molecular experiments. G.C.R.E.-D. synthesized the caged glutamate compound. H.K., J.N., T.H. and G.C.R.E.-D. wrote the paper.

Competing financial interests

G.C.R.E.-D. has filed a preliminary patent declaration in the USA for the synthesis of dinitroindolinyl-caged neurotransmitters.

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