Changes in global protein expression in sea turtle cells exposed to common contaminants indicates new biomarkers of chemical exposure

Abstract

Non-targeted protein expression at the cellular level can provide insights into mechanistic effects of contaminants in wildlife, and hence new and potentially more accurate biomarkers of exposure and effect. However, this technique has been relatively unexplored in the realm of in vitro biomarker discovery in threatened wildlife, despite the vulnerability of this group of animals to adverse sublethal effects of contaminant exposure. Here we examined the usefulness of non-targeted protein expression for biomarker discovery in green sea turtles (Chelonia mydas) by investigating differences in the response of primary cells from five different tissue types that were exposed to three contaminants known to accumulate in this species. Cells derived from C. mydas skin, liver, kidney, ovary and small intestine were exposed to 100 μg/L of either polychlorinated biphenyl 153 (PCB153), perfluorononanoic acid (PFNA) or phenanthrene for 24 h. The global protein expression was then quantitatively evaluated using sequential window acquisition of all theoretical mass spectra (SWATH-MS). Comparison of the global protein profiles revealed that, while a majority of proteins were mutually expressed in controls of all tissue types (~90%), the response to exposure in terms of protein expression strength was significantly different between tissue types. Furthermore, a comparison to known markers of chemical exposure in sea turtles from the literature indicated that in vitro response can reflect known in vivo responses. In particular, markers such as heat shock protein (HSP) 60, glutathione S-transferases (GSTs) and superoxide dismutases (SODs), cytochrome P450 and catalase were dysregulated in response to exposure. Furthermore, potential new markers of exposure were discovered such as annexin, an important protein in cell signalling processes. While this methodology proved promising further studies are required to confirm the accuracy of in vitro protein expression as a tool for biomarker discovery in wildlife.