Applied Sciences (Apr 2020)
Effects of di-(2-ethylhexyl) phthalate on Transcriptional Expression of Cellular Protection-Related <i>HSP60</i> and <i>HSP67B2</i> Genes in the Mud Crab <i>Macrophthalmus japonicus</i>
Abstract
Di-2-ethylhexyl phthalate (DEHP) has attracted attention as an emerging dominant phthalate contaminant in marine sediments. Macrophthalmus japonicus, an intertidal mud crab, is capable of tolerating variations in water temperature and sudden exposure to toxic substances. To evaluate the potential effects of DEHP toxicity on cellular protection, we characterized the partial open reading frames of the stress-related heat shock protein 60 (HSP60) and small heat shock protein 67B2 (HSP67B2) genes of M. japonicus and further investigated the molecular effects on their expression levels after exposure to DEHP. Putative HSP60 and small HSP67B2 proteins had conserved HSP-family protein sequences with different C-terminus motifs. Phylogenetic analysis indicated that M. japonicus HSP60 (Mj-HSP60) and M. Japonicus HSP67B2 (Mj-HSP67B2) clustered closely with Eriocheir sinensis HSP60 and Penaeus vannamei HSP67B2, respectively. The tissue distribution of Heat shock proteins (HSPs) was the highest in the gonad for Mj-HSP60 and in the hepatopancreas for Mj-HSP67B2. The expression of Mj-HSP60 Messenger Ribonucleic Acid (mRNA) increased significantly at day 1 after exposure to all doses of DEHP, and then decreased in a dose-dependent and exposure time-dependent manner in the gills and hepatopancreas. Mj-HSP67B2 transcripts were significantly upregulated in both tissues at all doses of DEHP and at all exposure times. These results suggest that cellular immune protection could be disrupted by DEHP toxicity through transcriptional changes to HSPs in crustaceans. Small and large HSPs might be differentially involved in responses against environmental stressors and in detoxification in M. japonicus crabs.
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