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In vivo quantitative proteomics of somatosensory cortical synapses shows which protein levels are modulated by sensory deprivation

Academic Article
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Overview

authors

  • Butko, M. T.
  • Savas, J. N.
  • Friedman, B.
  • Delahunty, C.
  • Ebner, F.
  • Yates III, John
  • Tsien, R. Y.

publication date

  • February 2013

journal

  • Proceedings of the National Academy of Sciences of the United States of America  Journal

abstract

  • Postnatal bilateral whisker trimming was used as a model system to test how synaptic proteomes are altered in barrel cortex by sensory deprivation during synaptogenesis. Using quantitative mass spectrometry, we quantified more than 7,000 synaptic proteins and identified 89 significantly reduced and 161 significantly elevated proteins in sensory-deprived synapses, 22 of which were validated by immunoblotting. More than 95% of quantified proteins, including abundant synaptic proteins such as PSD-95 and gephyrin, exhibited no significant difference under high- and low-activity rearing conditions, suggesting no tissue-wide changes in excitatory or inhibitory synaptic density. In contrast, several proteins that promote mature spine morphology and synaptic strength, such as excitatory glutamate receptors and known accessory factors, were reduced significantly in deprived synapses. Immunohistochemistry revealed that the reduction in SynGAP1, a postsynaptic scaffolding protein, was restricted largely to layer I of barrel cortex in sensory-deprived rats. In addition, protein-degradation machinery such as proteasome subunits, E2 ligases, and E3 ligases, accumulated significantly in deprived synapses, suggesting targeted synaptic protein degradation under sensory deprivation. Importantly, this screen identified synaptic proteins whose levels were affected by sensory deprivation but whose synaptic roles have not yet been characterized in mammalian neurons. These data demonstrate the feasibility of defining synaptic proteomes under different sensory rearing conditions and could be applied to elucidate further molecular mechanisms of sensory development.

subject areas

  • Animals
  • Immunohistochemistry
  • Mice
  • Microscopy, Electron
  • Nerve Tissue Proteins
  • Proteomics
  • Sensory Deprivation
  • Synapses
  • Tandem Mass Spectrometry
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Research

keywords

  • experience-dependent plasticity
  • mass spectrometric proteomics
  • somatosensory cortex
  • synaptic protein dynamics
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Identity

PubMed Central ID

  • PMC3581967

International Standard Serial Number (ISSN)

  • 0027-8424

Digital Object Identifier (DOI)

  • 10.1073/pnas.1300424110

PubMed ID

  • 23382246
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Additional Document Info

start page

  • E726

end page

  • E735

volume

  • 110

issue

  • 8

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