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Synaptosomal protein expression in nucleus accumbens after EtOH self‐administration in the posterior VTA
Author(s) -
Witzmann Frank A.,
Hong David,
Rodd Zachary A.,
Simon Jay R.,
Truitt William A.,
Wang Mu
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a477-a
Subject(s) - nucleus accumbens , chemistry , trypsin , gene expression , microbiology and biotechnology , chromatography , biochemistry , gene , biology , enzyme , receptor
Previous microarray data indicates that EtOH administered by self‐infusion into the VTA alters gene expression in the nucleus accumbens (Acb), genes associated with synaptic plasticity, neurite outgrowth, and dendritic spine formation. The objective of this study was to examine the effect of intracranial self‐administration (ICSA) of EtOH in the VTA on protein expression in the Acb. Adult Wistar rats were surgically implanted with guide cannulae aimed at the posterior VTA and self‐administered 150 mg% EtOH for 7 consecutive sessions after which the Acb was dissected and homogenized. Synaptosomes were prepared from pooled samples via discontinuous sucrose gradient centrifugation, and proteins in the enriched samples were subjected to trypsin digestion. Tryptic peptides from each sample were analyzed serially via linear ion‐trap LC‐MS/MS. Proteins were identified via SEQUEST and X!Tandem. Protein quantification was based on total ion chromatograms whereby the integral volume under each selected peptide peak was measured, normalized, and compared for relative abundance. In this study, 1120 proteins were identified and quantified. The identities of significantly altered proteins will be presented in terms of the functional significance of their EtOH‐mediated changes. Supported by NIH Grant R21AA015698‐01.