Abstract
The form of a neuron's dendritic arbor determines the set of axons with which it may form synaptic contacts, thus establishing connectivity within neural circuits. However, the dynamic relationship between dendrite growth and synaptogenesis is not well understood. To observe both processes simultaneously, we performed long-term imaging of non-spiny dendritic arbors expressing a fluorescent postsynaptic marker protein as they arborized within the optic tectum of live zebrafish larvae. Our results indicate that almost all synapses form initially on newly extended dendritic filopodia. A fraction of these nascent synapses are maintained, which in turn stabilizes the subset of filopodia on which they form. Stabilized filopodia mature into dendritic branches, and successive iterations of this process result in growth and branching of the arbor. These findings support a 'synaptotropic model' in which synapse formation can direct dendrite arborization.
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References
Kaethner, R.J. & Stuermer, C.A. Dynamics of process formation during differentiation of tectal neurons in embryonic zebrafish. J. Neurobiol. 32, 627–639 (1997).
Jontes, J.D., Buchanan, J. & Smith, S.J. Growth cone and dendrite dynamics in zebrafish embryos: early events in synaptogenesis imaged in vivo. Nat. Neurosci. 3, 231–237 (2000).
Wu, G.Y., Zou, D.J., Rajan, I. & Cline, H. Dendritic dynamics in vivo change during neuronal maturation. J. Neurosci. 19, 4472–4483 (1999).
Grueber, W.B., Jan, L.Y. & Jan, Y.N. Different levels of the homeodomain protein cut regulate distinct dendrite branching patterns of Drosophila multidendritic neurons. Cell 112, 805–818 (2003).
Komiyama, T., Johnson, W.A., Luo, L. & Jefferis, G.S. From lineage to wiring specificity. POU domain transcription factors control precise connections of Drosophila olfactory projection neurons. Cell 112, 157–167 (2003).
Furrer, M.P., Kim, S., Wolf, B. & Chiba, A. Robo and Frazzled/DCC mediate dendritic guidance at the CNS midline. Nat. Neurosci. 6, 223–230 (2003).
Sin, W.C., Haas, K., Ruthazer, E.S. & Cline, H.T. Dendrite growth increased by visual activity requires NMDA receptor and Rho GTPases. Nature 419, 475–480 (2002).
Horch, H.W. & Katz, L.C. BDNF release from single cells elicits local dendritic growth in nearby neurons. Nat. Neurosci. 5, 1177–1184 (2002).
Lohmann, C., Myhr, K.L. & Wong, R.O. Transmitter-evoked local calcium release stabilizes developing dendrites. Nature 418, 177–181 (2002).
Cline, H.T. Dendritic arbor development and synaptogenesis. Curr. Opin. Neurobiol. 11, 118–126 (2001).
Scott, E.K. & Luo, L. How do dendrites take their shape? Nat. Neurosci. 4, 359–365 (2001).
Jan, Y.N. & Jan, L.Y. Dendrites. Genes Dev. 15, 2627–2641 (2001).
Wong, R.O. & Ghosh, A. Activity-dependent regulation of dendritic growth and patterning. Nat. Rev. Neurosci. 3, 803–812 (2002).
Garner, C.C., Zhai, R.G., Gundelfinger, E.D. & Ziv, N.E. Molecular mechanisms of CNS synaptogenesis. Trends Neurosci. 25, 243–251 (2002).
McGee, A.W. & Bredt, D.S. Assembly and plasticity of the glutamatergic postsynaptic specialization. Curr. Opin. Neurobiol. 13, 111–118 (2003).
Marrs, G.S., Green, S.H. & Dailey, M.E. Rapid formation and remodeling of postsynaptic densities in developing dendrites. Nat. Neurosci. 4, 1006–1013 (2001).
Cohen-Cory, S. The developing synapse: construction and modulation of synaptic structures and circuits. Science 298, 770–776 (2002).
Washbourne, P., Bennett, J.E. & McAllister, A.K. Rapid recruitment of NMDA receptor transport packets to nascent synapses. Nat. Neurosci. 5, 751–759 (2002).
Wong, W.T. & Wong, R.O. Rapid dendritic movements during synapse formation and rearrangement. Curr. Opin. Neurobiol. 10, 118–124 (2000).
Portera-Cailliau, C., Pan, D.T. & Yuste, R. Activity-regulated dynamic behavior of early dendritic protrusions: evidence for different types of dendritic filopodia. J. Neurosci. 23, 7129–7142 (2003).
Sheng, M. Molecular organization of the postsynaptic specialization. Proc. Natl. Acad. Sci. USA 98, 7058–7061 (2001).
Prange, O. & Murphy, T.H. Modular transport of postsynaptic density-95 clusters and association with stable spine precursors during early development of cortical neurons. J. Neurosci. 21, 9325–9333 (2001).
Rao, A., Kim, E., Sheng, M. & Craig, A.M. Heterogeneity in the molecular composition of excitatory postsynaptic sites during development of hippocampal neurons in culture. J. Neurosci. 18, 1217–1229 (1998).
Ebihara, T., Kawabata, I., Usui, S., Sobue, K. & Okabe, S. Synchronized formation and remodeling of postsynaptic densities: long-term visualization of hippocampal neurons expressing postsynaptic density proteins tagged with green fluorescent protein. J. Neurosci. 23, 2170–2181 (2003).
Gleason, M.R. et al. Translocation of CaM kinase II to synaptic sites in vivo. Nat. Neurosci. 6, 217–218 (2003).
Bresler, T. et al. The dynamics of SAP90/PSD-95 recruitment to new synaptic junctions. Mol. Cell Neurosci. 18, 149–167 (2001).
Douglas, R.H. & Djamgoz, M.B.A. (eds.). The Visual System of Fish (Chapman and Hall, Ltd., London, 1990).
Meek, J. A Golgi-electron microscopic study of goldfish optic tectum. II. Quantitative aspects of synaptic organization. J. Comp. Neurol. 199, 175–190 (1981).
Lendvai, B., Stern, E.A., Chen, B. & Svoboda, K. Experience-dependent plasticity of dendritic spines in the developing rat barrel cortex in vivo. Nature 404, 876–881 (2000).
Grutzendler, J., Kasthuri, N. & Gan, W.B. Long-term dendritic spine stability in the adult cortex. Nature 420, 812–816 (2002).
Matus, A. Actin-based plasticity in dendritic spines. Science 290, 754–758 (2000).
Lichtman, J.W. & Colman, H. Synapse elimination and indelible memory. Neuron 25, 269–278 (2000).
Alsina, B., Vu, T. & Cohen-Cory, S. Visualizing synapse formation in arborizing optic axons in vivo: dynamics and modulation by BDNF. Nat. Neurosci. 4, 1093–1101 (2001).
Friedman, H.V., Bresler, T., Garner, C.C. & Ziv, N.E. Assembly of new individual excitatory synapses: time course and temporal order of synaptic molecule recruitment. Neuron 27, 57–69 (2000).
Fiala, J.C., Feinberg, M., Popov, V. & Harris, K.M. Synaptogenesis via dendritic filopodia in developing hippocampal area CA1. J. Neurosci. 18, 8900–8911 (1998).
Ziv, N.E. & Smith, S.J. Evidence for a role of dendritic filopodia in synaptogenesis and spine formation. Neuron 17, 91–102 (1996).
Colman, H., Nabekura, J. & Lichtman, J.W. Alterations in synaptic strength preceding axon withdrawal. Science 275, 356–361 (1997).
Dunaevsky, A. & Mason, C.A. Spine motility: a means towards an end? Trends Neurosci. 26, 155–160 (2003).
Tashiro, A., Dunaevsky, A., Blazeski, R., Mason, C.A. & Yuste, R. Bidirectional regulation of hippocampal mossy fiber filopodial motility by kainate receptors. A two-step model of synaptogenesis. Neuron 38, 773–784 (2003).
Wong, W.T. & Wong, R.O. Changing specificity of neurotransmitter regulation of rapid dendritic remodeling during synaptogenesis. Nat. Neurosci. 4, 351–352 (2001).
Vaughn, J.E., Barber, R.P. & Sims, T.J. Dendritic development and preferential growth into synaptogenic fields: a quantitative study of Golgi-impregnated spinal motor neurons. Synapse 2, 69–78 (1988).
Vaughn, J.E. Fine structure of synaptogenesis in the vertebrate central nervous system. Synapse 3, 255–285 (1989).
Ackermann, M. & Matus, A. Activity-induced targeting of profilin and stabilization of dendritic spine morphology. Nat. Neurosci. 6, 1194–1200 (2003).
Yoshii, A., Sheng, M.H. & Constantine-Paton, M. Eye opening induces a rapid dendritic localization of PSD-95 in central visual neurons. Proc. Natl. Acad. Sci. USA 100, 1334–1339 (2003).
Yu, X. & Malenka, R.C. beta-catenin is critical for dendritic morphogenesis. Nat. Neurosci. 6, 1169–1177 (2003).
Koster, R.W. & Fraser, S.E. Tracing transgene expression in living zebrafish embryos. Dev. Biol. 233, 329–346 (2001).
Westerfield, M. The Zebrafish Book (Institute of Neuroscience, University of Oregon, Eugene, 1993).
Banker, G. & Goslin, K. (eds.) Culturing Nerve Cells (MIT Press, Cambridge, 1998).
Micheva, K.D., Holz, R.W. & Smith, S.J. Regulation of presynaptic phosphatidylinositol 4,5-biphosphate by neuronal activity. J. Cell Biol. 154, 355–368 (2001).
Hsueh, Y.P., Kim, E. & Sheng, M. Disulfide-linked head-to-head multimerization in the mechanism of ion channel clustering by PSD-95. Neuron 18, 803–814 (1997).
Acknowledgements
We thank B. Barres, L. Luo, W. Talbot and R. Tsien for their comments on the manuscript. GAL4-VP16/UAS expression constructs were graciously provided by S. Fraser. This work was supported by the Wellcome Trust (M.P.M.), Howard Hughes Medical Institute (C.M.N.), the Vincent and Stella Coates Foundation and the National Institute of Neurological Disorders and Stroke (NS043461).
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Supplementary information
Supplementary Fig. 1
Wholemount SV2 labeling of zebrafish tectal neuropil. Due to high synapse density, this technique could not reliably be used to demonstrate specific colocalization of PSD-95:GFP puncta with presynaptic SV2 puncta. Scale bar 5 μm. (JPG 59 kb)
Supplementary Fig. 2
Synaptophysin labelling (red channel) of dissociated hippocampal cells (6 d.i.v.) expressing zebrafish PSD-95:GFP (green channel) shows that 86% of PSD-95:GFP puncta colocalize with a synaptophysin punctum. (n = 456 PSD-95:GFP puncta from 19 cells). Scale bar 10μm. (JPG 39 kb)
Supplementary Fig. 3
Comparison of dendritic branch length over several days between cells expressing the PSD-95:GFP + DsRed construct or simply GFP reveals that neither PSD-95:GFP nor DsRed significantly affects dendrite growth (n = 6 cells each). (JPG 23 kb)
Supplementary Video 1
Timelapse of 3 day post-fertilization tectal cell dendrite expressing actin:GFP and soluble DsRed, shows a large number of highly motile, mostly transient, actin-based protrusions. Frames are at 3 minute intervals, total time 2 hours. (MOV 2371 kb)
Supplementary Video 2
Timelapse series which provided still images for Figure 2a in the text, showing net growth of the arbor and formation of stable PSD-95:GFP puncta. Frames are at 20 minute intervals, total time 20 hours. (MOV 2835 kb)
Supplementary Video 3
Timelapse series which provided still images for Figure 2b in the text, showing highly dynamic filopodia and transient PSD-95:GFP puncta. Frames are at 20 minute intervals, total time 12 hours. (MOV 1189 kb)
Supplementary Video 4
Timelapse series which provided still images for Figure 3 in the text, showing a typical mode of punctum formation on a filopodium. Frames are at 20 minute intervals, total time 6 hours. (MOV 823 kb)
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Niell, C., Meyer, M. & Smith, S. In vivo imaging of synapse formation on a growing dendritic arbor. Nat Neurosci 7, 254–260 (2004). https://doi.org/10.1038/nn1191
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DOI: https://doi.org/10.1038/nn1191
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