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Originally published In Press as doi:10.1074/mcp.M500415-MCP200 on February 23, 2006.
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Molecular & Cellular Proteomics 5:882-894, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

Differential Expression Profiling of the Hepatic Proteome in a Rat Model of Dioxin Resistance

CORRELATION WITH GENOMIC AND TRANSCRIPTOMIC ANALYSES*,S

Roberta Pastorelli{ddagger},§, Donatella Carpi{ddagger}, Roberta Campagna{ddagger}, Luisa Airoldi{ddagger}, Raimo Pohjanvirta,||,**, Matti Viluksela**, Helen Hakansson{ddagger}{ddagger}, Paul C. Boutros§§, Ivy D. Moffat§§, Allan B. Okey§§ and Roberto Fanelli{ddagger}

From the {ddagger} Department of Environmental Health Sciences, Istituto di Ricerche Farmacologiche "Mario Negri," 20157 Milan, Italy, Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, University of Helsinki, FIN-00014 Helsinki, Finland, || National Veterinary and Food Research Institute and ** Department of Environmental Health, National Public Health Institute, FIN-70701 Kuopio, Finland, {ddagger}{ddagger} Institute of Environmental Medicine, Karolinska Institutet, SE-17177 Stockholm, Sweden, and §§ Department of Pharmacology, University of Toronto, Toronto M5S 1A8, Canada

One characteristic feature of acute 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) toxicity is dramatic interspecies and interstrain variability in sensitivity. This complicates dioxin risk assessment for humans. However, this variability also provides a means of characterizing mechanisms of dioxin toxicity. Long-Evans (Turku/AB) rats are orders of magnitude more susceptible to TCDD lethality than Han/Wistar (Kuopio) rats, and this difference constitutes a very useful model for identifying mechanisms of dioxin toxicity. We adopted a proteomic approach to identify the differential effects of TCDD exposure on liver protein expression in Han/Wistar rats as compared with Long-Evans rats. This allows determination of which, if any, protein markers are indicative of differences in dioxin susceptibility and/or responsible for conferring resistance. Differential protein expression in total liver protein was assessed using two-dimensional gel electrophoresis, computerized gel image analysis, in-gel digestion, and mass spectrometry. We observed significant changes in the abundance of several proteins, which fall into three general classes: (i) TCDD-independent and exclusively strain-specific (e.g. isoforms of the protein-disulfide isomerase A3, regucalcin, and agmatine ureohydrolase); (ii) strain-independent and only dependent on TCDD exposure (e.g. aldehyde dehydrogenase 3A1 and rat selenium-binding protein 2); (iii) dependent on both TCDD exposure and strain (e.g. oxidative stress-related proteins, apoptosis-inducing factor, and MAWD-binding protein). By integrating transcriptomic (microarray) data and genomic data (computational search of regulatory elements), we found that protein expression levels were mainly controlled at the level of transcription. These results reveal, for the first time, a subset of hepatic proteins that are differentially regulated in response to TCDD in a strain-specific manner. Some of these differential responses may play a role in establishing the major differences in TCDD response between these two strains of rats. As such, our work is expected to lead to new insights into the mechanism of TCDD toxicity and resistance.

§ To whom correspondence should be addressed: Laboratory of Molecular Toxicology, Dept. of Environmental Health Sciences, Istituto di Ricerche Farmacologiche "Mario Negri," Via Eritrea 62, 20157 Milan, Italy. Tel.: 39-0239014456; Fax: 39-023546277; E-mail: rpastorelli{at}marionegri.it


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