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Title: Hippocampal phosphoproteomics of F344 rats exposed to 1-bromopropane

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 cytochromemore » 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.« less

Authors:
 [1];  [2];  [3];  [4];  [5];  [2];  [5];  [2];  [1];  [1];  [5];  [2]
  1. Guangdong Provincial Key Laboratory of Occupational Disease Prevention and Treatment, Guangdong Province Hospital for Occupational Disease Prevention and Treatment, Guangzhou 510-300 (China)
  2. (Japan)
  3. Graduate School of Regional Innovation Studies, Mie University, Tsu 514-8507 (Japan)
  4. Department of Environmental and Molecular Medicine, Mie University Graduate School of Medicine, Mie 514-8507 (Japan)
  5. Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Nagoya 466-8550 (Japan)
Publication Date:
OSTI Identifier:
22465678
Resource Type:
Journal Article
Resource Relation:
Journal Name: Toxicology and Applied Pharmacology; Journal Volume: 282; Journal Issue: 2; Other Information: Copyright (c) 2014 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; APOPTOSIS; BIOLOGICAL MARKERS; CYTOPLASM; GELS; HIPPOCAMPUS; HUMAN POPULATIONS; INHALATION; MANGANESE; MITOCHONDRIA; MYELIN; PH VALUE; PHOSPHOPROTEINS; PHOSPHORYLATION; RATS; TIME-OF-FLIGHT METHOD

Citation Formats

Huang, Zhenlie, Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Ichihara, Sahoko, Oikawa, Shinji, Chang, Jie, Graduate School of Regional Innovation Studies, Mie University, Tsu 514-8507, Zhang, Lingyi, Department of Occupational and Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda 278-8510, Hu, Shijie, Huang, Hanlin, E-mail: huanghl@gdoh.org, Ichihara, Gaku, E-mail: gak@rs.tus.ac.jp, and Department of Occupational and Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda 278-8510. Hippocampal phosphoproteomics of F344 rats exposed to 1-bromopropane. United States: N. p., 2015. Web. doi:10.1016/J.TAAP.2014.10.016.
Huang, Zhenlie, Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Ichihara, Sahoko, Oikawa, Shinji, Chang, Jie, Graduate School of Regional Innovation Studies, Mie University, Tsu 514-8507, Zhang, Lingyi, Department of Occupational and Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda 278-8510, Hu, Shijie, Huang, Hanlin, E-mail: huanghl@gdoh.org, Ichihara, Gaku, E-mail: gak@rs.tus.ac.jp, & Department of Occupational and Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda 278-8510. Hippocampal phosphoproteomics of F344 rats exposed to 1-bromopropane. United States. doi:10.1016/J.TAAP.2014.10.016.
Huang, Zhenlie, Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Ichihara, Sahoko, Oikawa, Shinji, Chang, Jie, Graduate School of Regional Innovation Studies, Mie University, Tsu 514-8507, Zhang, Lingyi, Department of Occupational and Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda 278-8510, Hu, Shijie, Huang, Hanlin, E-mail: huanghl@gdoh.org, Ichihara, Gaku, E-mail: gak@rs.tus.ac.jp, and Department of Occupational and Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda 278-8510. Thu . "Hippocampal phosphoproteomics of F344 rats exposed to 1-bromopropane". United States. doi:10.1016/J.TAAP.2014.10.016.
@article{osti_22465678,
title = {Hippocampal phosphoproteomics of F344 rats exposed to 1-bromopropane},
author = {Huang, Zhenlie and Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Nagoya 466-8550 and Ichihara, Sahoko and Oikawa, Shinji and Chang, Jie and Graduate School of Regional Innovation Studies, Mie University, Tsu 514-8507 and Zhang, Lingyi and Department of Occupational and Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda 278-8510 and Hu, Shijie and Huang, Hanlin, E-mail: huanghl@gdoh.org and Ichihara, Gaku, E-mail: gak@rs.tus.ac.jp and Department of Occupational and Environmental Health, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda 278-8510},
abstractNote = {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.},
doi = {10.1016/J.TAAP.2014.10.016},
journal = {Toxicology and Applied Pharmacology},
number = 2,
volume = 282,
place = {United States},
year = {Thu Jan 15 00:00:00 EST 2015},
month = {Thu Jan 15 00:00:00 EST 2015}
}