Gene cloning and expression analysis of AhR and CYP4 from Pinctada martensii after exposed to pyrene

• Published in: Ecotoxicology (London) Vol. 24; no. 7-8; pp. 1574 - 1582
• Main Authors: Du, Junqiao, Liao, Chenghong, Zhou, Hailong, Diao, Xiaoping, Li, Yuhu, Zheng, Pengfei, Wang, Fuqiang
• Format: Journal Article
• Language: English
• Published: New York Springer US 01.10.2015; Springer; Springer Nature B.V
• Subjects: Amino acids; Analysis; Animals; Biomarkers - metabolism; Bivalvia; Cloning; Cloning, Molecular; complementary DNA; Cytochrome; Cytochrome P-450; Cytochrome P-450 CYP4A - genetics; Cytochrome P-450 CYP4A - metabolism; Deoxyribonucleic acid; Detoxification; DNA; DNA sequencing; Earth and Environmental Science; Ecology; Ecotoxicology; Environment; Environmental Management; enzymes; Ethylenediaminetetraacetic acid; Gene Expression - drug effects; Genetic research; gills; Marine; Marine environment; Marine pollution; messenger RNA; metabolism; molecular cloning; Molecular Sequence Data; Mollusks; monitoring; muscles; Nucleotide sequencing; open reading frames; Oysters; Physiological aspects; Pinctada; Pinctada - drug effects; Pinctada - genetics; Pinctada - metabolism; Pinctada martensii; pollutants; Polycyclic aromatic hydrocarbons; Pyrene; Pyrenes - toxicity; Receptors, Aryl Hydrocarbon - genetics; Receptors, Aryl Hydrocarbon - metabolism; RNA; sequence analysis; Sequence Analysis, DNA; Shellfish; Water Pollutants, Chemical - toxicity; Xenobiotics; Pyrene; cDNA cloning; AhR; CYP4; mRNA expression; Pinctada martensii
• ISSN: 0963-9292; 1573-3017
• DOI: 10.1007/s10646-015-1424-x

Pyrene, a typical polycyclic aromatic hydrocarbon, is a common pollutant in the marine environment. Polycyclic aromatic hydrocarbons initiate cellular detoxification in an exposed organism via the activation of the aryl hydrocarbon receptor (AhR). Subsequent metabolism of these xenobiotics is mainly by the cytochrome P450 enzymes of the phase I detoxification system. Full-length complementary DNA sequences from the pearl oyster Pinctada martensii (pm) encoding AhR and cytochrome P4 were cloned. The P. martensii AhR complementary DNA sequence constitutes an open reading frame that encodes for 848 amino acids. Sequence analysis indicated PmAhR showed high similarity with its homologues of other bivalve species. The cytochrome P(CYP)4 complementary DNA sequence of P. martensii constitutes an open reading frame that encodes for 489 amino acids. Quantitative real-time analysis detected both PmAhR and PmCYP4 messenger RNA expressions in the mantle, gill, hepatapancreas and adductor muscle of P. martensii exposed to pyrene. The highest transcript-band intensities of PmAhR and PmCYP4 were observed in the gill. Temporal expression of PmAhR and PmCYP4 messenger RNAs induction was observed in gills and increased between 3 and 5 days post exposure; then returned to control level. These results suggest that messenger RNAs of PmAhR and PmCYP4 in pearl oysters might be useful parameters for monitoring marine environment pyrene pollution.