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Author Huang, Zhenlie ♦ Ichihara, Sahoko ♦ Oikawa, Shinji ♦ Chang, Jie ♦ Zhang, Lingyi ♦ Hu, Shijie ♦ Huang, Hanlin ♦ Ichihara, Gaku
Source United States Department of Energy Office of Scientific and Technical Information
Content type Text
Language English
Subject Keyword APPLIED LIFE SCIENCES ♦ APOPTOSIS ♦ BIOLOGICAL MARKERS ♦ CYTOPLASM ♦ GELS ♦ HIPPOCAMPUS ♦ HUMAN POPULATIONS ♦ INHALATION ♦ MANGANESE ♦ MITOCHONDRIA ♦ MYELIN ♦ PH VALUE ♦ PHOSPHOPROTEINS ♦ PHOSPHORYLATION ♦ RATS ♦ TIME-OF-FLIGHT METHOD
Abstract 1-Bromopropane (1-BP) is neurotoxic in both experimental animals and human. To identify phosphorylated modification on the unrecognized post-translational modifications of proteins and investigate their role in 1-BP-induced neurotoxicity, changes in hippocampal phosphoprotein expression levels were analyzed quantitatively in male F344 rats exposed to 1-BP inhalation at 0, 400, or 1000 ppm for 8 h/day for 1 or 4 weeks. Hippocampal protein extracts were analyzed qualitatively and quantitatively by Pro-Q Diamond gel staining and SYPRO Ruby staining coupled with two-dimensional difference in gel electrophoresis (2D-DIGE), respectively, as well as by matrix-assisted laser-desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) to identify phosphoproteins. Changes in selected proteins were further confirmed by Manganese II (Mn{sup 2+})-Phos-tag SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Bax and cytochrome c protein levels were determined by western blotting. Pro-Q Diamond gel staining combined with 2D-DIGE identified 26 phosphoprotein spots (p < 0.05), and MALDI-TOF/MS identified 18 up-regulated proteins and 8 down-regulated proteins. These proteins are involved in the biological process of response to stimuli, metabolic processes, and apoptosis signaling. Changes in the expression of phosphorylated 14-3-3 θ were further confirmed by Mn{sup 2+}-Phos-tag SDS-PAGE. Western blotting showed overexpression of Bax protein in the mitochondria with down-regulation in the cytoplasm, whereas cytochrome c expression was high in the cytoplasm but low in the mitochondria after 1-BP exposure. Our results suggest that the pathogenesis of 1-BP-induced hippocampal damage involves inhibition of antiapoptosis process. Phosphoproteins identified in this study can potentially serve as biomarkers for 1-BP-induced neurotoxicity. - Highlights: • 1-BP modified hippocampal phosphoproteome in rat and 23 altered proteins were identified. • 1-BP changed phosphorylation of GRP78, 14-3-3 θ, PSMC3, ST13, PURA, GNB2, APOE, PEA15 and ATP5H. • 1-BP-induced hippocampal damage involves inhibition of antiapoptosis process.
ISSN 0041008X
Educational Use Research
Learning Resource Type Article
Publisher Date 2015-01-15
Publisher Place United States
Journal Toxicology and Applied Pharmacology
Volume Number 282
Issue Number 2


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