Abstract
As part of a multi-endpoint systems approach to develop comprehensive methods for assessing endocrine stressors in vertebrates, differential protein profiling was used to investigate expression patterns in the brain of the amphibian model (Xenopus laevis) following in vivo exposure to a suite of T4 synthesis inhibitors. We specifically address the application of Two Dimensional Polyacrylamide Gel Electrophoresis (2D PAGE), Isobaric Tags for Relative and Absolute Quantitation (iTRAQ®) and LC-MS/MS to assess changes in relative protein expression levels. 2D PAGE and iTRAQ proved to be effective complementary techniques for distinguishing protein changes in the developing amphibian brain in response to T4 synthesis inhibition. This information served to evaluate the use of distinctive protein profiles as a potential mechanism to screen chemicals for endocrine activity in anurans. Regulatory pathways associated with proteins expressed as a result of chemical effect are reported. To our knowledge, this is also the first account of the anuran larvae brain proteome characterization using proteomic technologies. Correlation of protein changes to other cellular and organism-level responses will aid in the development of a more rapid and cost-effective, non-mammalian screening assay for thyroid axis-disrupting chemicals.
Original language | English (US) |
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Pages (from-to) | 138-150 |
Number of pages | 13 |
Journal | Comparative Biochemistry and Physiology - Part D: Genomics and Proteomics |
Volume | 5 |
Issue number | 2 |
DOIs | |
State | Published - Jun 2010 |
Bibliographical note
Funding Information:Authors want to thank Benji Hanson (Computer Science Corporation, Duluth, MN) and Thomas McGowan (University of MN, St Paul) for very valuable bioinformatics, statistical, and graphics support. We are also grateful to Drs. Daniel Villeneuve, Michael Hornung, Witold Winnik, David Bencic (USEPA), and Nancy Denslow (University of Florida) for the critical review of this manuscript. Research was supported by ACC CRADA (Endocrine Disrupters; 405SA8 ), and in part by the USEPA-ORD Computational Toxicology Program . The authors recognize the Center for Mass Spectrometry and Proteomics at the University of Minnesota and various supporting agencies, including the National Science Foundation for Major Research Instrumentation grants 9871237 and NSF-DBI-0215759 used to purchase the instruments described in this study.
Keywords
- 2D PAGE
- Amphibian
- Brain
- Endocrine
- LC-MS/MS
- Proteomics
- iTRAQ