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BDNF-Induced Local Protein Synthesis in Synaptoneurosomes Assessed with Click-iT L-Azidohomoalanine

  • Victor Briz
  • Michel Baudry
Protocol
Part of the Neuromethods book series (NM, volume 143)

Abstract

The stimulation of dendritic protein synthesis by brain-derived neurotrophic factor (BDNF) plays a major role in many forms of synaptic plasticity and memory consolidation. This cellular process has been traditionally studied by metabolic labeling using radioactive isotopes such as [35S]-methionine and [35S]-cysteine. Due to the harmful effects of these compounds to human health, there has been an effort to find nonradioactive alternative methods for the study of protein translation. Here, we describe a simple methodology that uses Click-iT® L-azidohomoalanine, a modified amino acid that can be, after being incorporated into nascent proteins, covalently conjugated to biotin molecules and later detected by western blotting. We show that this methodology can be used not only to sensitively detect global changes in protein synthesis but also to monitor local translation of specific synaptic proteins in synaptoneurosomes and its regulation by BDNF using classical immunoprecipitation techniques.

Keywords

BDNF Dendritic translation Learning and memory Synaptic plasticity Synaptosomes 

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Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  • Victor Briz
    • 1
  • Michel Baudry
    • 2
  1. 1.Department of Molecular Neuropathology, Centro de Biología Molecular Severo OchoaCSIC-Universidad Autónoma de MadridMadridSpain
  2. 2.Graduate College of Biomedical SciencesWestern University of Health SciencesPomonaUSA

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