Molecular Characterization of Cytochrome P450 1A (CYP1A) in Asian Sea bass (Lates calcalifer Bloch) and Its Application as a Biomarker in the Gulf of Thailand
This study focusses on aquatic pollution in Thailand, using induction of hepatic cytochrome P450 1A
(CYP1A) in Asian sea bass as a biomarker to assess exposure to polyaromatic hydrocarbons (PAHs). Polyclonal antibodies (PAb) were raised against Asian sea bass CYP1A in mice. Western blot analyses revealed that these antibodies recognized a CYP1A protein in Asian sea bass treated with benzo[a]pyrene (BaP). The presence of a CYP1A orthologue in Asian sea bass liver was confirmed by isolating a partial cDNA, using a reverse transcriptase polymerase chain reaction (RT-PCR) approach. Quantitative RT-PCR analysis revealed that CYP1A mRNA was 2.5-fold higher in fish injected with BaP. The PAb were next applied on 15 different tropical fish species, caught off the Chonburi from three different stations: (1) Koh Loi, Si Racha, (2) Ao Udom and (3) Laem chabang. Although, species variation in CYP1A protein levels was observed, the data suggest that Si Racha and Ao Udom were more contaminated with PAH-type contaminants compared with Laem chabang. This study confirm studies in temperate waters that CYP1A could be used as early indicators of PAH-type exposure in Thailand, and that PAb against Asian sea bass is suitable for analyses of CYP1A in different tropical fish species.
Celander, M. and L. Förlin (1991). Catalytic activity and immunochemical quantification of hepatic cytochrome P- 450 in b-naphthoflavone and isosafrol treated rainbow trout (Oncorhynchus mykiss). Fish Physiology and Biochemistry,9: 189-197.
Celander, M., Näf, C., Bromann, D. and L. Förlin (1994). Temporal aspects of induction of hepatic cytochrome P450 1A and conjugating enzymes in the viviparous Blenny (Zoarces viviparous) treated with petroleum hydrocarbons. Aquatic Toxicology, 29: 183-196.
Cheevaporn, V. and Beamish, F.W.H. (2007). Cytochrome P450 1A activity in liver and fixed wavelength fluorescence detection of polycyclic aromatic hydrocarbons in the bile of tongue-fish (Cynoglossus acrolepidotus, Bleeker) in relation to petroleum hydrocarbons in the eastern Gulf of Thailand. Journal of Environmental Biology, 28: 701-705.
Chumchuchan, S., Wattayakorn, G., Tuanwechayan, W. and P. Suthannaruk (1998). Petroleum hydrocarbon contamination in coastal environment of the upper Gulf of Thailand: Extent and sources. In: ASEAN-Canada Cooperative Programme on Marine Science. October 26-30, 1998. Langkavvi, Malaysia, 217-224.
Förlin, L. and M. Celander (1993). Induction of cytochrome P4501A in teleosts: Environmental monitoring in Swedish fresh, brackish, and marine waters. Aquatic Toxicology, 26: 41-56.
Miller, H.C., Mills, G.N., Bembo, D.G., Macdonald, J.A. and C.W. Evans (1999). Induction of CYP1A in Trematomus bernachii as an indicator of environmental pollution in Antarctica: Assessment by quantitative RT-PCR. Aquatic Toxicology, 44: 183-193.
Rice, C.D. and D. Schlenk (1995). Immune function and cytochrome P450 1A activity after acute exposure to 3,3’,4, 4’,5-Pentachlorobiphenyl (PCB126) in channel catfish. Journal of Aquatic Animal Health, 7: 195-204.
Sarin, C. and K. Srisuksawad (2000). Distribution of aliphatic and aromatic hydrocarbon in dated sediment cores of The Gulf of Thailand. Regional Training Course (RCA) on Application of Traces to Study Transport Processes and Sedimentation Rates in the Marine Environment. 21 Feb-4 Mar 2000, Thailand. IAEA-SM-354/210P, 563-564.
Śiroká, Z. and J. Drastichová (2004). Biochemical markers of aquatic environment contamination— cytochrome P450— in fish. A review. Acta Veterinaria Brunensis, 73: 123-132.
Schlenk, D., George, S., James, M., Van den Hurk, P., Willett, K., Kullman, S., Celander, M. and E. Gallagher (2008). Biotransformations. In: The Toxicology of Fishes. (Eds.) R. Di Giulio and D. Hinton. Taylor and Francis Publishers, Washington DC. 153-234.
Stegeman, J.J. and M.E. Hahn (1994). Biochemistry and molecular biology of monooxygenases: current perspectives on forms, functions, and regulation of cytochrome P450 in aquatic species. In: Ostrander, G.K., Malins, D. (eds), Aquatic Toxicology: Molecular, Biochemical and Cellular Perspectives. Lewis Publishers, Boca Raton, FL. 87-206.
Stegeman, J.J., Woodin, B.R., Singh, H., Oleksiak, M.F. and M. Celander (1997). Cytochromes P450 (CYP) in tropical fishes: Catalytic activities, expression of multiple CYP proteins and high levels of microsomal P450 in liver of fishes from Bermuda. Comparative Biochemistry and Physiology, 116C: 61-75.
Van der Oost, R., Beyer, J. and N.P.E. Vermeulen (2003). Fish bioaccumulation and biomarkers in environmental risk assessment: A review. Environmental Toxicology and Pharmacology, 13: 57-149.
Vrolijk, N.H., Targett, N.M., Woodin, B.R. and J.J. Stegeman (1994). Toxicological and ecological implications of biotransformation enzymes in the tropical teleost Chaetodon capistratus. Marine Biology, 119: 151-158.