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

NEUROPHARMACOLOGY

卷:100

Article

Bowling, Heather¹  Bhattacharya, Aditi¹  Zhang, Guoan^2,3  Lebowitz, Joseph Z.¹  Alam, Danyal¹  Smith, Peter T.⁴  Neubert, Thomas A.^2,3  Vogel, Christine⁵  Chao, Moses V.^2,6,7,8  Klann, Eric¹ 

[1] NYU, Ctr Neural Sci, New York, NY 10003 USA

[2] NYU, Sch Med, Kimmel Ctr Biol & Med, Skirball Inst, New York, NY USA

[3] NYU, Sch Med, Dept Biochem & Mol Pharmacol, New York, NY USA

[4] NYU, Dept Chem, New York, NY 10003 USA

[5] NYU, Dept Biol, Ctr Genom & Syst Biol, New York, NY 10003 USA

[6] NYU, Sch Med, Dept Cell Biol, New York, NY 10016 USA

[7] NYU, Sch Med, Dept Neurosci & Physiol, New York, NY USA

[8] NYU, Sch Med, Dept Psychiat, New York, NY USA

关键词: BONLAC;    BONCAT;    SILAC;    de nova proteomics;    BDNF;    hippocampal slice;    synaptic plasticity;    protein synthesis;   

DOI  :  10.1016/j.neuropharm.2015.07.017

来源: Elsevier

【 摘 要 】

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'. (C) 2015 Elsevier Ltd. All rights reserved.

【 授权许可】

Free