title: High Resolution Quantitative Synaptic Proteome Profiling of Mouse Brain Regions After Auditory Discrimination Learning.
abstract: The molecular synaptic mechanisms underlying auditory learning and memory remain largely unknown. Here, the workflow of a proteomic study on auditory discrimination learning in mice is described. In this learning paradigm, mice are trained in a shuttle box Go/NoGo-task to discriminate between rising and falling frequency-modulated tones in order to avoid a mild electric foot-shock. The protocol involves the enrichment of synaptosomes from four brain areas, namely the auditory cortex, frontal cortex, hippocampus, and striatum, at different stages of training. Synaptic protein expression patterns obtained from trained mice are compared to naïve controls using a proteomic approach. To achieve sufficient analytical depth, samples are fractionated in three different ways prior to mass spectrometry, namely 1D SDS-PAGE/in-gel digestion, in-solution digestion and phospho-peptide enrichment. High-resolution proteomic analysis on a mass spectrometer and label-free quantification are used to examine synaptic protein profiles in phospho-peptide-depleted and phospho-peptide-enriched fractions of synaptosomal protein samples. A commercial software package is utilized to reveal proteins and phospho-peptides with significantly regulated relative synaptic abundance levels (trained/naïve controls). Common and differential regulation modes for the synaptic proteome in the investigated brain regions of mice after training were observed. Subsequently, meta-analyses utilizing several databases are employed to identify underlying cellular functions and biological pathways.
authors.affiliation: Leibniz Institute for Neurobiology (LIN).
authors.forename: Angela
authors.initials: A
authors.lastname: Kolodziej
authors.affiliation: Leibniz Institute for Neurobiology (LIN).
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Smalla
authors.affiliation: Institute of Experimental Internal Medicine, Medical School, Otto von Guericke University Magdeburg.
authors.forename: Sandra
authors.initials: S
authors.lastname: Richter
authors.affiliation: Leibniz Institute for Neurobiology (LIN).
authors.forename: Alexander
authors.initials: A
authors.lastname: Engler
authors.affiliation: Leibniz Institute for Neurobiology (LIN); Institute of Pharmacology and Toxicology, Medical School, Otto von Guericke University.
authors.forename: Rainer
authors.initials: R
authors.lastname: Pielot
authors.affiliation: Institute of Pharmacology and Toxicology, Medical School, Otto von Guericke University.
authors.forename: Daniela C
authors.initials: DC
authors.lastname: Dieterich
authors.affiliation: Leibniz Institute for Neurobiology (LIN).
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Tischmeyer
authors.affiliation: Institute of Experimental Internal Medicine, Medical School, Otto von Guericke University Magdeburg.
authors.forename: Michael
authors.initials: M
authors.lastname: Naumann
authors.affiliation: Institute of Experimental Internal Medicine, Medical School, Otto von Guericke University Magdeburg; kaehne@med.ovgu.de.
authors.forename: Thilo
authors.initials: T
authors.lastname: Kähne
mesh_terms: D000818:Animals; D001307:Auditory Perception; D001921:Brain; D004193:Discrimination Learning / Q000502:physiology*; D051379:Mice; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D015398:Signal Transduction
chemical_list: D020543:Proteome