Gene cloning and expression analysis of AhR and CYP4 from Pinctada martensii after exposed to pyrene
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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.
KeywordsPyrene cDNA cloning AhR CYP4 Pinctada martensii mRNA expression
This study was supported by National Natural Sciences Foundation of China (31160126). The authors would like to acknowledge all other members at Dr. Xiaoping Diao Laboratory for their help with sampling and taking care of the pearl oysters.
Conflict of interest
The authors declare that there are no conflicts of interest for their work.
- Bo J, Wu SJ, Li YH, Ren HL, Fan DQ, Chen FY, Wang KJ (2010) The effects of Benzo[a]Pyrene (BaP) exposure on the CYP1A1 mRNA and AhR2 mRNA expression of Red Seabream (Pagrus major). Acta Scientiarum Naturalium Universities Sunyatseni 49:93–97Google Scholar
- Butler RA, Kelley ML, Powell WH, Hahn ME, Van Beneden RJ (2001) An aryl hydrocarbon receptor (AHR) homologue from the soft-shell clam, Mya arenaria: evidence that invertebrate AHR homologues lack 2,3,7,8-tetrachlorodibenzo-p-dioxin and beta-naphthoflavone binding. Gene 278:223–234CrossRefGoogle Scholar
- Butler RA, Kelley ML, Olberding KE, Gardner GR, Van Beneden RJ (2004) Aryl hydrocarbon receptor (AhR)-independent effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on softshell clam (Mya arenaria) reproductive tissue. Comp Biochem Physiol C 138:375–381Google Scholar
- Dong JY, Wang SG, Shang Z (2009) Water environmental health risk assessment of polycyclic aromatic hydrocarbons in the Lanzhou reach of the Yellow River. J Agro-Environ Sci 28:1892–1897Google Scholar
- Feng CL, Lei BL, Wang ZJ (2009) Preliminary ecological risk assessment of polycyclic aromatic hydrocarbons in main rivers of China. China Environ Sci 29:583–588Google Scholar
- Hahn ME (1998) The aryl hydrocarbon receptor: a comparative perspective. Comp Biochem Physiol C 121:23–53Google Scholar
- Jain S, Dolwick KM, Schmidt JV, Bradfield CA (1994) Potent transactivation domains of the Ah receptor and the Ah receptor nuclear translocator map to their carboxyl termin. J Biol Chem 269:31518–31524Google Scholar
- Kann S, Huang M-y, Estes C, Reichard JF, Sartor MA, Xia Y, Puga A (2005) Arsenite-induced aryl hydrocarbon receptor nuclear translocation results in additive induction of phase I genes and synergistic induction of phase II genes. Mol Pharmacol 68:336–346Google Scholar
- Kim E-Y, Iwata H, Suda T, Tanabe S, Amano M, Miyazaki N, Petrov EA (2005) Aryl hydrocarbon receptor (AHR) and AHR nuclear translocator (ARNT) expression in Baikal seal (Pusa sibirica) and association with 2,3,7,8-TCDD toxic equivalents and CYP1 expression levels. Comp Biochem Physiol C 141:281–291CrossRefGoogle Scholar
- Liu N, Pan L, Miao J, Xu C, Zhang L (2010) Molecular cloning and sequence analysis and the response of a aryl hydrocarbon receptor homologue gene in the clam Ruditapes philippinarum exposed to benzo(a)pyrene. Comp Biochem Physiol C 152:279–287Google Scholar
- Nelson DR (1998) Metazoan cytochrome P450 evolution. Comp Biochem Physiol C 121:15–22Google Scholar
- Rewitz KF, Styrishave B, Løbner-Olesen A, Andersen O (2006) Marine invertebrate cytochrome P450: emerging insights from vertebrate and insects analogies. Comp Biochem Physiol C 143:363–381Google Scholar
- Whitelaw ML, Göttlicher M, Gustafsson J, Poellinger L (1993) Definition of a novel ligand binding domain of a nuclear bHLH receptor: co-localization of ligand and hsp90 binding activities within the regulable inactivation domain of the dioxin receptor. EMBO J 12:4169–4179Google Scholar
- Wu YY, Wu QH, Luo H, Zhang HJ, Wu YH, Zhang RZ, Xu ZC (2013) Ecological risk assessment for polycyclic aromatic hydrocarbons in river sediments. Acta Scientiae Circumstantiae 33:544–556Google Scholar
- Zhou C, Li C, Zhang W, Jia X (2010) CYP4 gene cloning and expression level analysis of Perna viridis. J Trop Oceanogr 29:82–88Google Scholar