title: BONLAC: A combinatorial proteomic technique to measure stimulus-induced translational profiles in brain slices.
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'.
authors.affiliation: Center for Neural Science, New York University, New York, NY, USA.
authors.forename: Heather
authors.initials: H
authors.lastname: Bowling
authors.affiliation: Center for Neural Science, New York University, New York, NY, USA.
authors.forename: Aditi
authors.initials: A
authors.lastname: Bhattacharya
authors.affiliation: Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, USA; Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, USA.
authors.forename: Guoan
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Center for Neural Science, New York University, New York, NY, USA.
authors.forename: Joseph Z
authors.initials: JZ
authors.lastname: Lebowitz
authors.affiliation: Center for Neural Science, New York University, New York, NY, USA.
authors.forename: Danyal
authors.initials: D
authors.lastname: Alam
authors.affiliation: Department of Chemistry, New York University, USA.
authors.forename: Peter T
authors.initials: PT
authors.lastname: Smith
authors.affiliation: Department of Chemistry, New York University, USA.
authors.forename: Kent
authors.initials: K
authors.lastname: Kirshenbaum
authors.affiliation: Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, USA; Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, USA.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Neubert
authors.affiliation: Department of Biology, Center for Genomics and Systems Biology, New York University, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Vogel
authors.affiliation: Department of Cell Biology, New York University School of Medicine, USA; Department of Neuroscience and Physiology, New York University School of Medicine, USA; Department of Psychiatry, New York University School of Medicine, USA; Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, USA.
authors.forename: Moses V
authors.initials: MV
authors.lastname: Chao
authors.affiliation: Center for Neural Science, New York University, New York, NY, USA. Electronic address: eklann@cns.nyu.edu.
authors.forename: Eric
authors.initials: E
authors.lastname: Klann
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000096:biosynthesis*; D002478:Cells, Cultured; D006624:Hippocampus / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000378:metabolism*; D014176:Protein Biosynthesis; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results
chemical_list: D019208:Brain-Derived Neurotrophic Factor
keywords: BDNF; BONCAT; BONLAC; SILAC; de novo proteomics; hippocampal slice; protein synthesis; synaptic plasticity