HA4 Ecotoxicogenomics
204 Oregon Ballroom
8:00 AM - 12:00 PM, Thursday
() Effects of endocrine-disrupting chemicals on hepatic protein expression profile in wild cormorants: an ecotoxicoproteomics approach.
Iwata, H1, Nakayama, K1, Kim, E-Y2, Tanabe, S1, Yamagata, A3, Oofusa, K3, 1 Center for Marine Environmental Studies, Ehime University, Matsuyama, Ehime, Japan2 Ehime Prefectural Institute of Public Health and Environmental Sciences, Matsuyama, Ehime, Japan3 ProPhoenix Company Ltd., Higashihiroshima, Hiroshima, Japan
ABSTRACT- The present study examines whether a proteomics approach can be used for the early recognition of toxic effects of endocrine-disrupting chemicals (EDCs) in common cormorant (Phalacrocorax carbo) population from the Lake Biwa, Japan. A variety of EDCs including polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), polychlorinated biphenyls (PCBs), DDTs, chlordanes, HCHs, butyltins, nonylphenol and bisphenol A were analyzed in the cormorant livers. Lysates were prepared from liver samples representing a diverse range of EDC concentrations, and used for two-dimensional gel electrophoresis (2D-GE). Abundance of protein spots stained in the 2D gels were quantified by image analysis. Spearman's rank correlation analysis revealed that 2,3,7,8-TCDD toxic equivalents (TEQs) from PCDDs, PCDFs and coplanar PCBs were positively correlated with abundance of two protein spots, indicating the induction of these proteins by TEQs at translational level. Both spots were excised from the stained gels, and sequenced by nanoflow electrospray ionisation (ESI) in a quadrupole time-of-flight (Q-TOF) mass spectrometer. Peptide sequence tags were used to search the NCBInr protein sequence database by Mascot. The database retrieval succeeded in assigning both proteins as homologues of other species; histidine triad nucleotide-binding protein 1 (protein kinase C inhibitor 1) and mitochondrial F1-ATPase gamma subunit. The amino acid sequences of two spots deduced by ESI-Q-TOF mass spectrometric analysis were further confirmed by sequencing cDNAs isolated from cormorant liver using a set of specific primers. In addition to these spots, densities from three spots were positively correlated with butyltin levels, and one spot with nonylphenol. The sequence analyses of these four spots by ESI-Q-TOF are currently being investigated. The present study demonstrates that the combination of 2D-GE and ESI-Q-TOF mass spectrometry may be a useful tool to identify the effects of chemical exposure on protein expression profiles in wild species.
Key words: Cormorant, Endocrine-disrupting chemicals, Two-dimensional gel electrophoresis, ESI-Q-TOF mass spectrometry