Fish Physiology and Biochemistry

, Volume 38, Issue 4, pp 1107–1116 | Cite as

Cytochrome P4501A mRNA and protein induction in striped bass (Morone saxatilis)

  • Eric D. H. DurieuxEmail author
  • Richard E. Connon
  • Inge Werner
  • Leandro S. D’Abronzo
  • Patrick S. Fitzgerald
  • Jimmy L. Spearow
  • David J. Ostrach


The striped bass (Morone saxatilis) supports a valuable recreational fishery and is among the most important piscivorous fish of the San Francisco Estuary. This species has suffered a significant decline in numbers over the past decades, and there is indication that contaminants are important contributors. Polycyclic aromatic hydrocarbons (PAHs) and polyhalogenated aromatic hydrocarbons (PHAHs) including PCBs and dioxins are widespread in the estuary, they typically bioaccumulate through trophic levels, reaching highest levels in top predators and are known to affect the fish health and development. The aim of this study was to investigate the dynamics of cytochrome P4501A (Cyp1a) induction simultaneously at different levels of biological organization (RNA transcription and protein synthesis) as a biomarker of exposure to PAHs and PHAHs. We utilized β-naphthoflavone (BNF) as a model PAH to induce Cyp1a responses in juvenile striped bass in both dose–response and time–response assessments and determined Cyp1a mRNA and protein levels. Significant responses were measured in both systems at 10 mg ΒΝF kg−1, a concentration used for time–response studies. Messenger RNA levels peaked at 6 h post-injection, while protein levels increased progressively with time, significantly peaking at 96 h post-injection; both remaining elevated throughout the duration of the test (8 days). Our data suggest that rapid induction of gene transcription following exposure and subsequent cumulative protein synthesis could provide a useful means of identifying temporal variants in exposure to Cyp1a inducers in Morone saxatilis. The potential application of this combined Cyp1a gene and protein biomarker in this species for field studies is discussed.


Cyp1a Biomarker Fish Morone saxatilis β-Naphthoflavone Polycyclic aromatic hydrocarbon 



Funding for this study was provided by the Interagency Ecological Program on Pelagic Organism Decline Investigations, Department of Water Resources, Sacramento, California, contract Numbers 4600004664 to D. J. Ostrach, and 4600004445 to I. Werner, with special thanks to Ted Sommer and Fred Feyrer. We thank Dr. Moshe Tom for providing his modified bacteria, advice and encouragement. This study was part of E. D. H. Durieux’s post-doctorate in ecotoxicology at UC Davis in the laboratory of and mentored by D. J. Ostrach. We thank members of the Aquatic Toxicology Laboratory, and Center for Aquatic Biology and Aquaculture; University of California, Davis, for assistance and use of facilities. The opinions expressed in this paper are the authors and do not represent the opinions or policies of the California EPA or Department of Toxic Substances Control.


  1. Abdelhamid G, Anwar-Mohamed A, Badary OA, Moustafa AA, El-Kadi AO (2010) Transcriptional and posttranscriptional regulation of CYP1A1 by vanadium in human hepatoma HepG2 cells. Cell Biol Toxicol 26:421–434PubMedCrossRefGoogle Scholar
  2. Aluru N, Vijayan MM (2008) Brain transcriptomics in response to [beta]-naphthoflavone treatment in rainbow trout: The role of aryl hydrocarbon receptor signaling. Aquat Toxicol 87:1–12PubMedCrossRefGoogle Scholar
  3. Amara IE, Anwar-Mohamed A, El-Kadi AO (2010) Mercury modulates the CYP1A1 at transcriptional and posttranslational levels in human hepatoma HepG2 cells. Toxicol Lett 199:225–233PubMedCrossRefGoogle Scholar
  4. Anderson S, Sadinski W, Shugart L, Brussard P, Depledge M, Ford T et al (1994) Genetic and molecular ecotoxicology: a research framework. Environ Health Perspect 102:3–8PubMedCrossRefGoogle Scholar
  5. Bailey HC, Alexander C, Digiorgio C, Miller M, Doroshov SI, Hinton DE (1994) The effect of agricultural discharge on striped bass (Morone saxatilis) in California’s Sacramento-San Joaquin drainage. Ecotoxicology 3:123–142CrossRefGoogle Scholar
  6. Barron MG, Carls MG, Heintz R, Rice SD (2004) Evaluation of fish early life-stage toxicity models of chronic embryonic exposures to complex polycyclic aromatic hydrocarbon mixtures. Toxicol Sci 78:60–67PubMedCrossRefGoogle Scholar
  7. Bennett W, Moyle P (1996) Where have all the fishes gone? Interactive factors producing fish declines in the Sacramento-San Joaquin Estuary. In: Holligaugh J (ed) San Francisco Bay: the ecosystem. Pacific Division of the American Association for the Advancement of Science, San Francisco, pp 479–518Google Scholar
  8. Bennett WA, Ostrach DJ, Hinton DE (1995) Larval striped bass condition in a drought-stricken estuary: evaluating pelagic food-web limitation. Ecol Appl 5:680–692CrossRefGoogle Scholar
  9. Beyer A, Biziuk M (2009) Environmental fate and global distribution of polychlorinated biphenyls. Rev Environ Contam Toxicol 201:137–158PubMedCrossRefGoogle Scholar
  10. Billiard SM, Meyer JN, Wassenberg DM, Hodson PV, Di Giulio RT (2008) Nonadditive effects of PAHs on early vertebrate development: mechanisms and implications for risk assessment. Toxicol Sci 105:5–23PubMedCrossRefGoogle Scholar
  11. Burge EJ, Gauthier DT, Van Veld PA (2004) In vitro response of the striped bass natural resistance-associated macrophage protein, Nramp, to LPS and Mycobacterium marinum exposure. Comp Biochem Physiol C Toxicol Pharmacol 138:391–400PubMedCrossRefGoogle Scholar
  12. Chung-Davidson Y-W, Rees CB, Wu H, Yun S–S, Li W (2004) {beta}-naphthoflavone induction of CYP1A in brain of juvenile lake trout (Salvelinus namaycush Walbaum). J Exp Biol 207:1533–1542PubMedCrossRefGoogle Scholar
  13. Connon RE, Beggel S, D’Abronzo LS, Geist JP, Pfeiff J, Loguinov AV et al (2010) Linking molecular biomarkers with higher level condition indicators to identify effects of copper exposures on the endangered delta smelt (Hypomesus transpacificus). Environ Toxicol Chem 30:1–11Google Scholar
  14. Davis JA, Hetzel F, Oram JJ, McKee LJ (2007) Polychlorinated biphenyls (PCBs) in San Francisco Bay. Environ Res 105:67–86PubMedCrossRefGoogle Scholar
  15. Durieux EDH, Farver TB, Fitzgerald PS, Eder KJ, Ostrach DJ (2011) Natural factors to consider when using acetylcholinesterase activity as neurotoxicity biomarker in Young-Of-Year striped bass (Morone saxatilis). Fish Physiol Biochem 37:21–29PubMedCrossRefGoogle Scholar
  16. Fedorenkova A, Vonk JA, Lenders HJR, Ouborg NJ, Breure AM, Hendriks AJ (2010) Ecotoxicogenomics: bridging the gap between genes and populations. Environ Sci Tech 44:4328–4333CrossRefGoogle Scholar
  17. Feyrer F, Nobriga ML, Sommer TR (2007) Multidecadal trends for three declining fish species: habitat patterns and mechanisms in the San Francisco Estuary, California, USA. Can J Fish Aquat Sci 64:723–734CrossRefGoogle Scholar
  18. Geist J, Werner I, Eder KJ, Leutenegger CM (2007) Comparisons of tissue-specific transcription of stress response genes with whole animal endpoints of adverse effect in striped bass (Morone saxatilis) following treatment with copper and esfenvalerate. Aquat Toxicol 85:28–39PubMedCrossRefGoogle Scholar
  19. Gooch JW, Elskus AA, Kloepper-Sams PJ, Hahn ME, Stegeman JJ (1989) Effects of ortho- and non-ortho-substituted polychlorinated biphenyl congeners on the hepatic monooxygenase system in scup (Stenostomus chrysops). Toxicol Appl Pharmacol 98:422–433PubMedCrossRefGoogle Scholar
  20. Gravato C, Santos MA (2002) Juvenile Sea Bass Liver P450, EROD induction, and erythrocytic genotoxic responses to PAH and PAH-like compounds. Ecotoxicol Environ Saf 51:115–127PubMedCrossRefGoogle Scholar
  21. Hahn ME (2001) Dioxin toxicology and the aryl hydrocarbon receptor: insights from fish and other non-traditional models. Mar Biotechnol 3:224–238CrossRefGoogle Scholar
  22. Hahn M, Stegeman JJ (1994) Regulation of cytochrome P4501A1 in Teleosts: sustained induction of CYP1A1 mRNA, protein and catalytic activity by 2, 3, 7, 8-Tetrachlorodibenzofuran in the marine fish Stenotomus chrysops. Toxicol Appl Pharmacol 127:187–198PubMedCrossRefGoogle Scholar
  23. Hicken CE, Linbo TL, Baldwin DH et al (2011) Sublethal exposure to crude oil during embryonic development alters cardiac morphology and reduces aerobic capacity in adult fish. Proc Natl Acad Sci USA 108:7086–7090PubMedCrossRefGoogle Scholar
  24. Hylland K (2006) Polycyclic aromatic hydrocarbon (PAH) ecotoxicology in marine ecosystems. J Toxicol Environ Health A 69:109–123PubMedCrossRefGoogle Scholar
  25. Kloepper-Sams PJ, Stegeman JJ (1989) The temporal relationships between P450E protein content, catalytic activity, and mRNA levels in the teleost Fundulus heteroclitus following treatment with beta-naphthoflavone. Arch Biochem Biophys 268:525–535PubMedCrossRefGoogle Scholar
  26. Kloepper-Sams PJ, Stegeman JJ (1994) Turnover of hepatic microsomal cytochrome P4501A protein and heme in beta-naphthoflavone-induced Fundulus heteroclitus. Mol Mar Biol Biotechnol 3:171–183PubMedGoogle Scholar
  27. Levine SL, Oris JT (1999) CYP1A expression in liver and gill of rainbow trout following waterborne exposure: implications for biomarker determination. Aquat Toxicol 46:279–287CrossRefGoogle Scholar
  28. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  29. Miller KA, Addison RF, Bandiera SM (2004) Hepatic CYP1A levels and EROD activity in English sole: biomonitoring of marine contaminants in Vancouver Harbour. Mar Environ Res 57:37–54PubMedCrossRefGoogle Scholar
  30. Monosson E (1999/2000) Reproductive and developmental effects of PCBs in fish: a summary of laboratory and field studies. Rev Toxicol 3:25–75Google Scholar
  31. Monostory K, Pascussi J-M, Kóbori L, Dvorak Z (2009) Hormonal regulation of CYP1A expression. Drug Metab Rev 41:547–572PubMedCrossRefGoogle Scholar
  32. Nahrgang J, Camus L, Carls MG, Gonzalez P, Jönsson M, Taban IC et al (2010) Biomarker responses in polar cod (Boreogadus saida) exposed to the water soluble fraction of crude oil. Aquat Toxicol 97:234–242PubMedCrossRefGoogle Scholar
  33. Oris JT, Roberts AP (2007) Statistical analysis of cytochrome P4501A biomarker measurements in fish. Environ Toxicol Chem 26:1742–1750PubMedCrossRefGoogle Scholar
  34. Oros DR, Ross JR, Spies RB, Mumley T (2007) Polycyclic aromatic hydrocarbon (PAH) contamination in San Francisco Bay: a 10-year retrospective of monitoring in an urbanized estuary. Environ Res 105:101–118PubMedCrossRefGoogle Scholar
  35. Ortiz-Delgado JB, Behrens A, Segner H, Sarasquete C (2008) Tissue-specific induction of EROD activity and CYP1A protein in Sparus aurata exposed to B(a)P and TCDD. Ecotoxicol Environ Saf 69:80–88PubMedCrossRefGoogle Scholar
  36. Ostrach DJ, Low-Marchelli JM, Eder KJ, Whiteman SJ, Zinkl JG (2008) Maternal transfer of xenobiotics and effects on larval striped bass in the San Francisco Estuary. Proc Natl Acad Sci USA 105:19354–19359PubMedCrossRefGoogle Scholar
  37. Ostrach DJ, Groff JM, Weber PK, et al. (2009) The role of contaminants, within the context of multiple stressors, in the collapse of the striped bass population in the San Francisco estuary and its watershed). Final year 2 report for DWR agreement no. 4600004664. pp 186 (
  38. Picha ME, Turano MJ, Tipsmark CK, Borski RJ (2008) Regulation of endocrine and paracrine sources of Igfs and Gh receptor during compensatory growth in hybrid striped bass (Morone chrysops x Morone saxatilis). J Endocrinol 199:81–94PubMedCrossRefGoogle Scholar
  39. Pinkney AE, Harshbarger JC, Karouna-Renier NK, Jenko K, Balk L, Skarphéðinsdóttir H, Liewenborg B, Rutter MA (2011) Tumor prevalence and biomarkers of genotoxicity in brown bullhead (Ameiurus nebulosus) in Chesapeake Bay tributaries. Sci Tot Environ 410:248–257CrossRefGoogle Scholar
  40. R Development Core Team (2008) R: A language and environment for statistical computing. R Foundation for Statistical Computing, ViennaGoogle Scholar
  41. Rees CB, McCormick SD, Vanden Heuvel JP, Li W (2003) Quantitative PCR analysis of CYP1A induction in Atlantic salmon (Salmo salar). Aquat Toxicol 62:67–78CrossRefGoogle Scholar
  42. Reynaud S, Deschaux P (2006) The effects of polycyclic aromatic hydrocarbons on the immune system of fish: A review. Aquat Toxicol 77:229–238PubMedCrossRefGoogle Scholar
  43. Sarkar A, Ray D, Shrivastava A, Sarker S (2006) Molecular biomarkers: their significance and application in marine pollution monitoring. Ecotoxicology 15:333–340PubMedCrossRefGoogle Scholar
  44. SFEI (2008) The pulse of the estuary: monitoring and managing water quality in the San Francisco Estuary. San Francisco Estuary Institute Annual report, 92 ppGoogle Scholar
  45. Sommer T, Armor C, Baxter R, Breuer R, Brown L, Chotkowski M et al (2007) The collapse of pelagic fishes in the upper San Francisco estuary. Fisheries 32:270–277CrossRefGoogle Scholar
  46. Spearow JL, Kota RS, Ostrach DJ (2011) Environmental contaminant effects on juvenile striped bass in the San Francisco Estuary, California, USA. Environ Toxicol Chem 30:393–402PubMedCrossRefGoogle Scholar
  47. Stegeman JJ, Hahn ME (1994) Biochemistry and molecular biology of monooxygenases: current perspectives on forms, functions, and regulation of cytochrome P450 in aquatic species. In: Malins DC, Ostrander GK (eds) Aquatic toxicology: molecular, biochemical, and cellular perspectives. Lewis Publishers, Boca Raton, pp 87–206Google Scholar
  48. Stevens DE, Kohlhorst DW, Miller LW, Kelley DW (1985) The decline of striped bass in the Sacramento-San Joaquin Estuary, California. Transac Am Fish Soc 114:12–30CrossRefGoogle Scholar
  49. Strange CJ (2008) Troubling waters. Bioscience 58:1008–1013CrossRefGoogle Scholar
  50. Sundberg H, Ishaq R, Tjarnlund U, Kerman G, Grunder K, Bandh C, Broman D, Balk L (2006) Contribution of commonly analyzed polycyclic aromatic hydrocarbons (PAHs) to potential toxicity in early life stages of rainbow trout (Oncorhynchus mykiss). Can J Fish Aquat Sci 63:1320–1333CrossRefGoogle Scholar
  51. Tanguay RL, Andreasen EA, Walker MK, Peterson RE (2005) Dioxin toxicity and aryl hydrocarbon receptor signaling in fish. In: Dioxins and health. Wiley, New York, pp 603–628Google Scholar
  52. Tom M, Auslander M (2005) Transcript and protein environmental biomarkers in fish: a review. Chemosphere 59:155–162PubMedCrossRefGoogle Scholar
  53. Tom M, Myers CR, Waterman MR (2002) Evaluating molar CYP1A level in fish hepatic microsomes by competitive ELISA using recombinant membrane-free CYP1A standard protein. Aquat Toxicol 59:101–114PubMedCrossRefGoogle Scholar
  54. van der Oost R, Beyer J, Vermeulen NPE (2003) Fish bioaccumulation and biomarkers in environmental risk assessment: a review. Environ Toxicol Pharmacol 13:57–149PubMedCrossRefGoogle Scholar
  55. Viarengo A, Bettella E, Fabbri R, Burlando B, Lafaurie M (1997) Heavy metal inhibition of EROD activity in liver microsomes from the bass Dicentrarchus labrax exposed to organic xenobiotics: Role of GSH in the reduction of heavy metal effects. Mar Environ Res 44:1–11CrossRefGoogle Scholar
  56. Washburn BS, Vines CA, Baden DG, Hinton DE, Walsh PJ (1996) Differential effects of brevetoxin and [beta]-naphthoflavone on xenobiotic metabolizing enzymes in striped bass (Morone saxatilis). Aquat Toxicol 35:1–10CrossRefGoogle Scholar
  57. Willett KL, McDonald SJ, Steinberg MA, Beatty KB, Kennicutt MC, Safe SH (1997) Biomarker sensitivity for polynuclear aromatic hydrocarbon contamination in two marine fish species collected in Galveston Bay, Texas. Environ Toxicol Chem 16:1472–1479Google Scholar
  58. Willett KL, Wassenberg D, Lienesch L, Reichert W, Di Giulio RT (2001) In vivo and in vitro inhibition of CYP1A-dependent activity in Fundulus heteroclitus by the polynuclear aromatic hydrocarbon Fluoranthene. Toxicol Appl Pharmacol 177:264–271PubMedCrossRefGoogle Scholar
  59. Wills LP, Jung D, Koehrn K, Zhu S, Willett KL, Hinton DE, Di Giulio RT (2010) Comparative chronic liver toxicity of benzo[a]pyrene in two populations of the atlantic killifish (Fundulus heteroclitus) with different exposure histories. Environ Health Perspect 118:1376–1381PubMedCrossRefGoogle Scholar
  60. Wirgin I, Konkle B, Pedersen M, Grunwald C, Williams J, Courtenay S (1996) A comparison of cytochrome P4501A (CYP1A) mRNA inducibility in four species of Atlantic coast anadromous fishes. Estuaries Coasts 19:913–922CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Eric D. H. Durieux
    • 1
    • 3
    Email author
  • Richard E. Connon
    • 2
  • Inge Werner
    • 2
    • 4
  • Leandro S. D’Abronzo
    • 2
    • 5
  • Patrick S. Fitzgerald
    • 1
  • Jimmy L. Spearow
    • 1
    • 6
  • David J. Ostrach
    • 1
    • 7
  1. 1.Pathobiology, Conservation and Population Biology Laboratory, John Muir Institute of the Environment, Center for Watershed SciencesUniversity of CaliforniaDavisUSA
  2. 2.Aquatic Toxicology Laboratory, School of Veterinary Medicine, Department of Anatomy, Physiology and Cell BiologyUniversity of CaliforniaDavisUSA
  3. 3.UMR 6134 Sciences Pour l’EnvironnementUniversité de Corse Pasquale PaoliCorteFrance
  4. 4.Swiss Centre for Applied Ecotoxicology, Eawag/EPFLDübendorfSwitzerland
  5. 5.University of California Medical CenterMatherUSA
  6. 6.Department of Toxic Substances ControlSacramentoUSA
  7. 7.Ostrach ConsultingWoodlandUSA

Personalised recommendations