BONLAC: A combinatorial proteomic technique to measure stimulus-induced translational profiles in brain slices

Heather Bowling, Aditi Bhattacharya, Guoan Zhang, Joseph Z. Lebowitz, Danyal Alam, Peter T. Smith, Kent Kirshenbaum, Thomas A. Neubert, Christine Vogel, Moses V. Chao, Eric Klann

Research output: Contribution to journalArticle

Abstract

Stimulus-triggered protein synthesis is critical for brain health and function. However, due to technical hurdles, de novo neuronal translation is predominantly studied in cultured cells, whereas electrophysiological and circuit analyses often are performed in brain slices. The different properties of these two experimental systems create an information gap about stimulus-induced alterations in the expression of new proteins in mature circuits. To address this, we adapted two existing techniques, BONCAT and SILAC, to a combined proteomic technique, BONLAC, for use in acute adult hippocampal slices. Using BDNF-induced protein synthesis as a proof of concept, we found alterations in expression of proteins involved in neurotransmission, trafficking, and cation binding that differed from those found in a similar screen in cultured neurons. Our results indicate important differences between cultured neurons and slices, and suggest that BONLAC could be used to dissect proteomic changes underlying synaptic events in adult circuits. This article is part of the Special Issue entitled 'Synaptopathy - from Biology to Therapy'.

Original languageEnglish (US)
Pages (from-to)76-89
Number of pages14
JournalNeuropharmacology
Volume100
DOIs
StatePublished - Jan 1 2016

Keywords

  • BDNF
  • BONCAT
  • BONLAC
  • SILAC
  • de novo proteomics
  • hippocampal slice
  • protein synthesis
  • synaptic plasticity

ASJC Scopus subject areas

  • Pharmacology
  • Cellular and Molecular Neuroscience

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