Korean Journal of Ecology and Environment (생태와환경)

The Korean Society of Limnology (한국수생태학회)
권호 표지

cDNA Cloning and Expression of a Cytochrome P450 1A (CYP1A) from the Pale Chub, Zacco platypus

  • Jeon, Hyoung-Joo (Inland Fisheries Research Institute, National Fisheries Research and Development Institute) ;
  • Park, Young-Chul (Inland Aquaculture Research Center, National Fisheries Research and Development Institute) ;
  • Lee, Wan-Ok (Inland Fisheries Research Institute, National Fisheries Research and Development Institute) ;
  • Lee, Jong-Ha (Inland Fisheries Research Institute, National Fisheries Research and Development Institute) ;
  • Kim, Jin-Hyoung (Inland Fisheries Research Institute, National Fisheries Research and Development Institute)
  • Received : 2011.10.21
  • Accepted : 2011.11.24
  • Published : 2011.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

The pale chub (Zacco platypus) is generally found in Asian countries, such as Korea, Japan, and China. Nevertheless, very little information exists about the genes involved in the metabolism of xenobiotics in this species. This species is useful in monitoring the environmental impact on various pollutants in freshwater as a sentinel fish species. We cloned the full-length cDNA sequence of xenobiotic metabolizing cytochrome P450 1A (CYP1A) gene from Z. platypus and characterized it. Tissue distribution and timedependent induction of CYP1A were studied by real-time RT-PCR. Induction pattern of CYP1A was studied by exposing the fish to an arylhydrocarbon receptor agonist, ${\beta}$-naphthoflavone (BNF). The liver showed the highest level of expression in basal state as well as BNF- treated fish. However, appreciable levels of expression were also recorded in Gill and kidney and the least level of expression was observed in the eye. The results of the time-course study revealed an induction in the liver, brain, and gills after 6 h and 12 h in most of the tissues. This study provides an insight into the xenobiotics metabolizing system of Z. platypus and offers baseline information for further research related to biomarker, stress, and adaptive response of this ecologically important fish species in the freshwater environment.

Keywords

References

  1. Arukwe, A. 2002. Complementary DNA cloning, sequence analysis and differential organ expression of $\beta$-naphthoflavone-inducible cytochrome P4501A in Atlantic salmon (Salmo salar). Comparative Biochemistry and Physiology C 133: 613-624.
  2. Billiard, S.M., N.C. Bols and P.V. Hodson. 2004. In vitro and in vivo comparisons of fishspecific CYP1A induction relative potency factors for selected polycyclic aromatic hydrocarbons. Ecotoxicology Environmental Safety 59: 292-299. https://doi.org/10.1016/j.ecoenv.2004.06.009
  3. Campbell, P.M. and R.H. Devlin. 1996. Expression of CYP1A1 in livers and gonads of Pacific salmon: quantitation of mRNA levels by RT-cPCR. Aquatic Toxicology 34: 47-69. https://doi.org/10.1016/0166-445X(95)00032-Y
  4. Cho, S.H., W.M. Sohn, S.S. Shin, H.J. Song, T.G. Choi, C.M. Oh, Y. Kong and T.S. Kim. 2006. Infection status of pond smelts, Hypomesus olidus, and other freshwater fishes with trematode metacercariae in 6 large lakes. The Korean Journal of Parasitology 44: 243-246. https://doi.org/10.3347/kjp.2006.44.3.243
  5. Chung-Davidson, Y.W., C.B. Rees, H. Wu, S.S. Yun and W. Li. 2004. $\beta$-naphthoflavone induction of CYP1A in brain of juvenile lake trout (Salvelinus namaycush Walbaum). Journal of Experimental Biology 207: 1533-1542. https://doi.org/10.1242/jeb.00919
  6. Cousinou, M., B. Nilsen, J. Lopez-Barea and G. Dorado. 2000. New methods to use fish cytochrome P4501A to assess marine organic pollutants. Science of the Total Environment 247: 213-225. https://doi.org/10.1016/S0048-9697(99)00492-1
  7. Evans, D.H. 1987. The fish gill: Site of action and model for toxic effects of environmental pollutants. Environmental Health Perspective 71: 47-58. https://doi.org/10.1289/ehp.877147
  8. Gao, K., I. Brandt, J.V. Goldstone and M.E. Jonsson. 2011. Cytochrome P450 1A, 1B, and 1C mRNA induction patterns in three-spined stickleback exposed to a transient and a persistent inducer. Comparative Biochemistry and Physiology C 154: 42-55.
  9. Goldstone, H.M. and J.J. Stegeman. 2006. A revised evolutionary history of the CYP1A subfamily: Gene duplication, gene conversion, and positive selection. Journal of Molecular Evolution 62: 708-717. https://doi.org/10.1007/s00239-005-0134-z
  10. Hahn, M.E. 1998. The aryl hydrocarbon receptor: A comparative perspective. Comparative Biochemistry and Physiology C 121: 23-53.
  11. Hahn, M.E. and J.J. Stegeman. 1994. Regulation of cytochrome P450 1A1 in teleosts: sustained induction of CYP1A1 messenger-RNA, protein, and catalytic activity by 2,3,7,8 tetrachlorodibenzofuran in the marine fish Stenotomus chrysops. Toxicology and Applied Pharmacology 127: 187-198. https://doi.org/10.1006/taap.1994.1153
  12. Hankinson, O. 1995. The aryl hydrocarbon receptor complex. Annual Review of Pharmacology and Toxicology 35: 307-340. https://doi.org/10.1146/annurev.pa.35.040195.001515
  13. Hayashi, M., T. Ueda, K. Uyeno, K. Wada, N. Kinae, K. Saotome, N. Tanaka, A. Takai, Y.F. Sasaki, N. Asano, T. Sofuni and Y. Ojima. 1998. Development of genotoxicity assay systems that use aquatic organisms. Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis 399: 125-133. https://doi.org/10.1016/S0027-5107(97)00251-0
  14. Hoffer, A., C.Y. Chang and A. Puga. 1996. Dioxin induces transcription of fos and jun genes by Ah receptor- dependent and -independent pathways. Toxicology and Applied Pharmacology 141: 238-247. https://doi.org/10.1016/S0041-008X(96)80029-9
  15. Hu, M.C., H.J. Hsu, I.C. Guo and B.C. Chung. 2004. Function of Cyp11a1 in animal models. Molecular and Cellular Endocrinology 215: 95-100. https://doi.org/10.1016/j.mce.2003.11.024
  16. Isin, E.M. and F.P. Guengerich. 2007. Complex reactions catalyzed by cytochrome P450 enzymes. Biochimica et Biophysica Acta 1770: 314-329. https://doi.org/10.1016/j.bbagen.2006.07.003
  17. Jonsson, M., A. Abrahamson, B. Brunstrom, I. Brandt, K. Ingebrigtsen and E.H. Jorgensen. 2003. EROD activity in gill filaments of anadromous and marine fish as a biomarker of dioxinlike pollutants. Comparative Biochemistry and Physiology C 136: 235-243.
  18. Jonsson, M.E., B. Brunstrom, K. Ingebrigtsen and I. Brandt. 2004. Cellspecific CYP1A expression and benzo[a]pyrene adduct formation in gills of rainbow trout (Oncorhynchus mykiss) following CYP1A induction in the laboratory and in the field. Environmental Toxicology and Chemistry 23: 874-882. https://doi.org/10.1897/03-211
  19. Jonsson, M.E., R. Orrego, B.R. Woodin, J.V. Goldstone and J.J. Stegeman. 2007. Basal and 3,3′,4,4′,5-pentachlorobiphenyl-induced expression of cytochrome P450 1A, 1B and 1C genes in zebrafish. Toxicology and Applied Pharmacology 221: 29-41. https://doi.org/10.1016/j.taap.2007.02.017
  20. Kim, I.S. 1997. Illustrated encyclopedia of fauna and flora of Korea, Freshwater Fishes. 37, National Textbook Company.
  21. Kim, J.H., D.S. Hwang, K.H. Son, S. Raisuddin, J.S. Ki, J.S. Lee and K.N. Han. 2008a. cDNA cloning and expression of a xenobiotic metabolizing cytochrome P4501A (CYP1A) from the yellow catfish, Pelteobagrus fulvidraco (Siluriformes). Environmental Toxicology 23: 346-353. https://doi.org/10.1002/tox.20333
  22. Kim, J.H., S. Raisuddin, J.S. Ki, J.S. Lee and K.N. Han. 2008b. Molecular cloning and betanaphthoflavone-induced expression of a cytochrome P450 1A (CYP1A) gene from an anadromous river pufferfish, Takifugu obscurus. Marine Pollution Bulletin 57: 433-440. https://doi.org/10.1016/j.marpolbul.2008.01.005
  23. Ku, S.K., J.H. Lee and H.S. Lee. 2004. Immunohistochemical study on the endocrine cells in gut of the stomachless teleost, Zacco platypus (Cyprinidae). Journal of Veterinary Medicine Series C 33: 212-219.
  24. Ledirac, N., C. Delescluse, G. de Sousa, M. Pravaloria, P. Lesca, M. Amichot, J.B. Berge and R. Rahmani. 1997. Carbaryl induces CYP1A1 gene expression in HepG2 and HaCaT cells but is not a ligand of the human hepatic Ah receptor. Toxicology and Applied Pharmacology 144: 177-182. https://doi.org/10.1006/taap.1997.8120
  25. Lee, Y.M., T.D. Williams, S.O. Jung and J.S. Lee. 2005. cDNA cloning and expression of a cytochrome P450 1A (CYP1A) gene from the hermaphroditic fish Rivulus marmoratus. Marine Pollution Bulletin 51: 769-75. https://doi.org/10.1016/j.marpolbul.2005.06.015
  26. Liu, Y., J. Wang, Y. Wei, H. Zhang, Y. Liu and J. Dai. 2008. Molecular characterization of cytochrome P450 1A and 3A and the effects of perfluorooctanoic acid on their mRNA levels in rare minnow (Gobiocypris rarus) gills. Aquatic Toxicology 88: 183-190. https://doi.org/10.1016/j.aquatox.2008.04.008
  27. Livak, K.J. and T.D. Schmittgen. 2001. Analysis of relative gene expression data using real time quantitative PCR and the $2^{-{\Delta\Delta}C_T}$ method. Methods 25: 402-408. https://doi.org/10.1006/meth.2001.1262
  28. Marston, C.P., C. Pereira, J. Ferguson, K. Fischer, O. Hedstrom, W.M. Dashwood and W.M. Baird. 2001. Effect of a complex environmental mixture from coal tar containing polycyclic aromatic hydrocarbons (PAH) on the tumor initiation, PAH-DNA binding and metabolic activation of carcinogenic PAH in mouse epidermis. Carcinogenesis 22: 1077-1086. https://doi.org/10.1093/carcin/22.7.1077
  29. Miller, H., D.G. Bembo, J.A. Macdonald and C.W. Evans. 1999. Induction of cytochrome P4501A (CYP1A) in Trematomus bernacchii as an indicator of environmental pollution in Antarctica: assessment by quantitative RT-PCR. Aquatic Toxicology 44: 183-193.
  30. Miranda, C.L., W.G. Chung, J.L. Wang-Buhler, T. Musafia-Jeknic, W.M. Baird and D.R. Buhler. 2006. Comparative in vitro metabolism of benzo[a]pyrene by recombinant zebrafish CYP1A and liver microsomes from $\beta$-naphthoflavone-treated rainbow trout. Aquatic Toxicology 80: 101-108. https://doi.org/10.1016/j.aquatox.2006.07.018
  31. Monostory, K., K. Jemnitz and L. Vereczkey. 1996. Xenobiotic metabolizing enzymes in fish: diversity, regulation and biomarkers for pollutant exposure. Acta Physica Academiae Scientiarum Hungaricae 84: 369-381.
  32. Nakamura, K. 1952. Environment, food habit, spawning, development, growth and fishes of Zacco platypus in Chikuma River. Bulletin of Freshwater Fisheries Research Laboratory 1: 2-25.
  33. Ortiz-Delgado, J.B. and C. Sarasquete. 2004. Toxicity, histopathological alterations and immunohistochemical CYP1A induction in the early life stages of the seabream, Sparus aurata, following waterborne exposure to B(a)P and TCDD. Journal of Molecular Histology 35: 29-45.
  34. Park, J.H., S.M. Guk, T.Y. Kim, E.H. Shin, A. Lin, J. Y. Park, J.L. Kim, S.T. Hong and J.Y. Chai. 2004. Clonorchis sinensis metacercarial infection in the pond smelt Hypomesus olidus and the minnow Zacco platypus collected from the Soyang and Daechung Lakes. The Korean Journal of Parasitology 42: 41-44. https://doi.org/10.3347/kjp.2004.42.1.41
  35. Rees, C.B., S.D. McCormick and W. Li. 2005. A nonlethal method to estimate CYP1A expression in laboratory and wild Atlantic salmon (Salmo salar). Comparative Biochemistry and Physiology C 141: 217-224.
  36. Rees, C.B., S.D. McCormick, J.P. Vanden Heuvel and W. Li. 2003. Quantitative PCR analysis of CYP1A induction in Atlantic salmon (Salmo salar). Aquatic Toxicology 62: 67-78. https://doi.org/10.1016/S0166-445X(02)00062-0
  37. Reiners, J.J.J., C.L. Jones, N. Hong, R.E. Clift and C. Elferink. 1997. Downregulation of aryl hydrocarbon receptor function and cytochrome P450 1A1 induction by expression of Ha-ras oncogenes. Molecular Carcinogenesis 19: 91-100. https://doi.org/10.1002/(SICI)1098-2744(199707)19:2<91::AID-MC4>3.0.CO;2-O
  38. Sarasquete, C. and H. Segner. 2000. Cytochrome P4501A (CYP1A) in teleostean fishes. A review of immunohistochemical studies. Science of the Total Environment 247: 313-332. https://doi.org/10.1016/S0048-9697(99)00500-8
  39. Smith, G., S.H. Ibbotson, M.M. Comrie, R.S. Dawe, A. Bryden, J. Ferguson and C.R. Wolf. 2006. Regulation of cutaneous drugmetabolizing enzymes and cytoprotective gene expression by topical drugs in human skin in vivo. The British Journal of Dermatology 155: 275-281. https://doi.org/10.1111/j.1365-2133.2006.07317.x
  40. Weber, L.P., S.L. Diamond, S.M. Bandiera and D.M. Janz. 2002. Expression of HSP70 and CYP1A protein in ovary and liver of juvenile rainbow trout exposed to $\beta$-naphthoflavone. Comparative Biochemistry and Physiology C 131: 387-394. https://doi.org/10.1016/S1095-6433(01)00490-1
  41. Williams, D.E., J.J. Lech and D.R. Buhler. 1998. Xenobiotics and xenoestrogens in fish: modulation of cytochrome P450 and carcinogenesis. Mutation Research 399: 179-192. https://doi.org/10.1016/S0027-5107(97)00255-8
  42. Wilson, J.M. and P. Laurent. 2002. Fish gill morphology: inside out. Journal of Experimental Zoology 293: 192-213. https://doi.org/10.1002/jez.10124
  43. Yamazaki, M., Y. Tanizaki and T. Shimokawa. 1996. Silver and other trace elements in a freshwater fish, Carasius auratus langsdorfii, from the Asakawa River in Tokyo, Japan. Environmental Pollution 94: 83-90. https://doi.org/10.1016/S0269-7491(96)00053-X
  44. Yeom, D.H., S.A. Lee, G.S. Kang, J. Seo and S.K. Lee. 2007. Stressor identification and health assessment of fish exposed to wastewater effluents in Miho Stream, South Korea. Chemosphere 67: 2282-2292. https://doi.org/10.1016/j.chemosphere.2006.09.071