First molecular evidence of the toxicogenetic effects of copper on sea urchin Paracentrotus lividus embryo development.

Bioassays with sea urchin embryos are widely used to define the environmental quality of marine waters. Anomalies during embryogenesis are generally considered as end-points, whereas a toxigenomic approach, despite it is wide use in other species, is yet in its infancy. In the present study we evaluated toxigenic effects induced by copper on the sea urchin Paracentrotus lividus embryo, combining morphological observations with gene expression analysis. Many anthropogenic activities release copper in the marine environment, with harmful effects on aquatic organisms. In the present study P. lidivus embryos were exposed to different concentrations of copper (24, 36, 48 μg/L) and the activation of fifty specific marker genes, involved in different biological processes (stress, skeletogenesis, development/differentiation, detoxification) was investigated at early blastula, late gastrula and pluteus stage. At blastula stage no morphological anomalies were found, with early down-regulation of genes involved in development/differentiation and a moderate up-regulation of some detoxification genes. At gastrula stage a slight increase in developmental anomalies (up to 19% of malformed embryos) was followed by an increased number of targeted genes belonging to the same two classes, relative to the blastula stage. At pluteus stage morphological anomalies increased in a dose dependent manner. All the analyzed genes were strongly up-regulated, stress and skeletogenic genes showing a "late response" and almost all genes were targeted by copper at all the concentrations tested. The present study represents the first molecular report on the potential negative effect of copper on P. lividus embryos in the environment. Gene expression analysis should be considered as a promising tool for future environmental biomonitoring programs.