A chemical-toxicological study of animal models exposed to organohalogen environmental contaminants
Abstract
Sensitive effect determination, the understanding of molecular toxicity mechanisms and the
discovery of novel biochemical biomarkers are some of the major challenges in ecotoxicology in
dealing with chemicals in the environment. Among several ‘omics’ tools, proteomic approaches are
used to study the whole proteome of organisms and may provide novel insights into the functional
molecular state of a biological system and for discovery of new sensitive biomarkers indicating
exposure or effects at low toxicant concentrations.
In this study, a proteomic approach has been used in Mytilus galloprovincialis as a screening
of changes in protein expression caused by a mixture of polychlorinated biphenyls (PCBs), in order
to characterize the effects of these environmental contaminants on protein profile and to develop
new molecular biomarkers through identification of more drastically altered proteins.
To achieve this objective, 100 mussels were exposed to PCB 138, 153 and 180 for 3 weeks
under controlled conditions at the concentration of 30μg/l. An equal number of mussels was kept
under the same conditions, but not treated, as control. The edible parts were homogenized and
lyophilized. Extracted proteins were quantified and separated by two-dimensional electrophoresis
(2-DE). It has been made a comparative study of two-dimensional electrophoresis gels obtained
from proteomic analysis and the changes in protein expression were assessed by image analysis.
Image analysis included spot detection, quantification, normalization and matching. On average
more than 1000 spots were resolved and altered expression was qualitatively detected. Stained
protein spots of interest were excised from preparative gels and their tryptic digests were subjected
to protein identification by mass spectrometry. It was used Matrix Science Mascot search engine,
database NCBI and for a homology search the program BLAST. .. [edited by Author]