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Glutamate induces de novo growth of functional spines in developing cortex

Nature volume 474, pages 100–104 (2011)Cite this article

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Abstract

Mature cortical pyramidal neurons receive excitatory inputs onto small protrusions emanating from their dendrites called spines. Spines undergo activity-dependent remodelling, stabilization and pruning during development, and similar structural changes can be triggered by learning and changes in sensory experiences1,2,3,4. However, the biochemical triggers and mechanisms of de novo spine formation in the developing brain and the functional significance of new spines to neuronal connectivity are largely unknown. Here we develop an approach to induce and monitor de novo spine formation in real time using combined two-photon laser-scanning microscopy and two-photon laser uncaging of glutamate. Our data demonstrate that, in mouse cortical layer 2/3 pyramidal neurons, glutamate is sufficient to trigger de novo spine growth from the dendrite shaft in a location-specific manner. We find that glutamate-induced spinogenesis requires opening of NMDARs (N-methyl-d-aspartate-type glutamate receptors) and activation of protein kinase A (PKA) but is independent of calcium–calmodulin-dependent kinase II (CaMKII) and tyrosine kinase receptor B (TrkB) receptors. Furthermore, newly formed spines express glutamate receptors and are rapidly functional such that they transduce presynaptic activity into postsynaptic signals. Together, our data demonstrate that early neural connectivity is shaped by activity in a spatially precise manner and that nascent dendrite spines are rapidly functionally incorporated into cortical circuits.

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Figure 1: De novo spine generation is induced by glutamate uncaging.
Figure 2: New spines grow rapidly and acquire morphology similar to pre-existing spines.
Figure 3: Molecular mechanisms of glutamate-induced spine formation.
Figure 4: Functional characterization of new spines.

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Acknowledgements

We thank members of the Sabatini laboratory for their comments on the manuscript and assistance with data analysis. We are grateful to S. Nazia for technical support and for acting as the blind evaluator. This work was supported by a SFARI grant from the Simons Foundation and the National Institute of Neurological Disorders and Stroke (NS046579).

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Authors and Affiliations

  1. Department of Neurobiology, Howard Hughes Medical Institute, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Hyung-Bae Kwon & Bernardo L. Sabatini

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  1. Hyung-Bae Kwon
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  2. Bernardo L. Sabatini
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Contributions

H.B.K. and B.L.S. designed the experiments and wrote the paper. H.B.K. performed all the experiments, analysed the data (other than spine counting by a blind, third-party observer) and prepared the figures.

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Correspondence to Bernardo L. Sabatini.

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The authors declare no competing financial interests.

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This file contains Supplementary Methods, additional references, Supplementary Figures 1-10 with legends and Supplementary Tables 1-2. (PDF 1866 kb)

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Kwon, HB., Sabatini, B. Glutamate induces de novo growth of functional spines in developing cortex. Nature 474, 100–104 (2011). https://doi.org/10.1038/nature09986

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