Article ID Journal Published Year Pages File Type
4529513 Aquatic Toxicology 2013 8 Pages PDF
Abstract

•A de novo reference transcriptome assembly has been produced.•RNseq gene expression analyses from whale cells exposed to various Cr(VI) levels was performed.•Genes exhibiting modulated expression were identified and validated using qRT-PCR.•As levels of Cr increase there is a shift in expression patterns from cellular repair to apoptosis.•Results suggest why whale cells are more resistant to Cr then human cells.

Hexavalent chromium Cr(VI) is known to produce cytotoxic effects in humans and is a highly toxic environmental contaminant. Interestingly, it has been shown that free ranging sperm whales (Phyester macrocephalus) may have exceedingly high levels of Cr in their skin. Also, it has been demonstrated that skin cells from whales appear more resistant to both cytotoxicity and clastogenicity upon Cr exposure compared to human cells. However, the molecular genetic mechanisms employed in whale skin cells that might lead to Cr tolerance are unknown.In an effort to understand the underlying mechanisms of Cr(VI) tolerance and to illuminate global gene expression patterns modulated by Cr, we exposed whale skin cells in culture to varying levels of Cr(VI) (i.e., 0.0, 0.5, 1.0 and 5.0 μg/cm2) followed by short read (100 bp) next generation RNA sequencing (RNA-seq). RNA-seq reads from all exposures (≈280 million reads) were pooled to generate a de novo reference transcriptome assembly. The resulting whale reference assembly had 11 K contigs and an N50 of 2954 bp.Using the reads from each dose (0.0, 0.5, 1.0 and 5.0 μg/cm2) we performed RNA-seq based gene expression analysis that identified 35 up-regulated genes and 19 down-regulated genes. The experimental results suggest that low dose exposure to Cr (1.0 μg/cm2) serves to induce up-regulation of oxidative stress response genes, DNA repair genes and cell cycle regulator genes. However, at higher doses (5.0 μg/cm2) the DNA repair genes appeared down-regulated while other genes that were induced suggest the initiation of cytotoxicity.The set of genes identified that show regulatory modulation at different Cr doses provide specific candidates for further studies aimed at determination of how whales exhibit resistance to Cr toxicity and what role(s) reactive oxygen species (ROS) may play in this process.

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Life Sciences Agricultural and Biological Sciences Aquatic Science
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