Skip to main content

Advertisement

Log in

Embryonic turkey liver: activities of biotransformation enzymes and activation of DNA-reactive carcinogens

  • Metabolic Activation/Inactivation
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

Avian embryos are a potential alternative model for chemical toxicity and carcinogenicity research. Because the toxic and carcinogenic effects of some chemicals depend on bioactivation, activities of biotransformation enzymes and formation of DNA adducts in embryonic turkey liver were examined. Biochemical analyses of 22-day in ovo turkey liver post-mitochondrial fractions revealed activities of the biotransformation enzymes 7-ethoxycoumarin de-ethylase (ECOD), 7-ethoxyresorufin de-ethylase (EROD), aldrin epoxidase (ALD), epoxide hydrolase (EH), glutathione S-transferase (GST), and UDP-glucuronyltransferase (GLUT). Following the administration of phenobarbital (24 mg/egg) on day 21, enzyme activities of ECOD, EROD, ALD, EH and GLUT, but not of GST, were increased by two-fold or higher levels by day 22. In contrast, acute administration of 3-methylcholanthrene (5 mg/egg) induced only ECOD and EROD activities. Bioactivation of structurally diverse pro-carcinogens was also examined using 32P-postlabeling for DNA adducts. In ovo exposure of turkey embryos on day 20 of gestation to 2-acetylaminofluorene (AAF), 4,4’-methylenebis(2-chloroaniline) (MOCA), benzo[a]pyrene (BaP), and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) resulted in the formation of DNA adducts in livers collected by day 21. Some of the DNA adducts had 32P-postlabeling chromatographic migration patterns similar to DNA adducts found in livers from Fischer F344 rats exposed to the same pro-carcinogens. We conclude that 21-day embryonic turkey liver is capable of chemical biotransformation and activation of genotoxic carcinogens to form DNA adducts. Thus, turkey embryos could be utilized to investigate potential chemical toxicity and carcinogenicity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Bock KW (1974) Oxidation of barbiturates and the glucuronidation of 1-naphthol in perfused rat liver and microsomes. Arch Pharmacol 283:319–330

    CAS  PubMed  Google Scholar 

  • Brunnemann KD, Enzmann HG, Perrone CE, Iatropoulos M, Williams GM (2002) In ovo carcinogenicity assay (IOCA): evaluation of mannitol, caprolactam and nitrosoproline. Arch Toxicol 76:606–612

    Article  CAS  PubMed  Google Scholar 

  • Bucher JR (1998) Update on national toxicology program (NTP) assays with genetically altered or “transgenic” mice. Environ Health Perspect 106:619–621

    CAS  PubMed  Google Scholar 

  • Burke MD, Prough RA, Mayer RT (1977) Characteristics of a microsomal cytochrome P-448-mediated reaction: ethoxyresorufin o-deethylation. Drug Metab Dispos 5:1–8

    CAS  PubMed  Google Scholar 

  • Carlson J, Abraham R (1985) Nuclear ploidy of neonatal rat livers: effects of two hepatic carcinogens. J Toxicol Environ Health 15:551–559

    CAS  PubMed  Google Scholar 

  • Cresteil T, Beaune P, Kremers P, Celier C, Guengerich FP, Leroux JP (1985) Immunoquantification of epoxide hydrolase and cytochrome P450 isozymes in fetal and adult human liver microsomes. Eur J Biochem 151:345–350

    CAS  PubMed  Google Scholar 

  • Dubois M, Pfohl-Leszkowicz A, Grosse Y, Kremers P (1995) DNA adducts and P450 induction in human, rat and avian liver cells after exposure to polychlorobiphenyls. Mutat Res 345:181–190

    CAS  PubMed  Google Scholar 

  • Endo-Ichikawa Y, Tokunaga KH, Yabusaki Y, Sakaki T, Ohkawa H, Taketani S (1995) Formation of 4,4’-methylene-bis(2-chloroaniline)-DNA adducts in yeast expressing recombinant P450 s. Experientia 51:564–568

    CAS  PubMed  Google Scholar 

  • Enzmann H, Brunnemann KD (1997) The in ovo carcinogenicity assay (ioca): a review of an experimental approach for research on carcinogenesis and carcinogenicity testing. Front Biosci 2:c30–c39

    CAS  PubMed  Google Scholar 

  • Enzmann H, Kaliner G, Watta-Gebert B, Loser E (1992) Foci of altered hepatocytes induced in embryonal turkey liver. Carcinogenesis 13:943–946

    CAS  PubMed  Google Scholar 

  • Enzmann H, Kuhlem C, Kaliner G, Loser E, Bannasch P (1995a) Rapid induction of preneoplastic liver foci in embryonal turkey liver by diethylnitrosamine. Toxicol Pathol 23:560–569

    CAS  PubMed  Google Scholar 

  • Enzmann H, Kühlem C, Loser E, Bannasch P (1995b) Dose dependence of diethylnitrosamine-induced nuclear enlargement in embryonal turkey liver. Carcinogenesis 16:1351–1355

    CAS  PubMed  Google Scholar 

  • Enzmann H, Bomhard E, Iatropoulos M, Ahr HJ, Schlueter G, Williams GM (1998a) Short- and intermediate-term carcinogenicity testing—a review. Part 1: the prototypes mouse skin tumor assay and rat liver focus assay. Food Chem Toxicol 36:979–995

    CAS  PubMed  Google Scholar 

  • Enzmann H, Iatropoulos M, Brunnemann KD, Bomhard E, Ahr HJ, Schlueter G, Williams GM (1998b) Short- and intermediate-term carcinogenicity testing—a review. Part 2: available experimental models. Food Chem Toxicol 36:997–1013

    CAS  PubMed  Google Scholar 

  • Giuliano KA, Lau EP, Fall RR (1980) Simplified liquid chromatographic assay for epoxide hydrolase. J Chromatogr 202:447–452

    CAS  PubMed  Google Scholar 

  • Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferase: the first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139

    CAS  PubMed  Google Scholar 

  • Hamilton JW, Bloom SE (1983) Developmental differences in basal and induced aryl hydrocarbon (benzo[a]pyrene) hydroxylase activity in chick embryo liver and lung in ovo. Biochem Pharmacol 32:2986–2988

    Article  CAS  PubMed  Google Scholar 

  • Hamilton JW, Denison MS, Bloom SE (1983) Development of basal and induced aryl hydrocarbon (benzo[a]pyrene) hydroxylase activity in the chicken embryo in ovo. Proc Natl Acad Sci 80:3372–3376

    CAS  PubMed  Google Scholar 

  • Hamilton JW, Bement WJ, Sinclair PR, Sinclair JF, Wetterhahn KE (1988) Expression of 5-aminolaevulinate synthase and cytochrome P-450 mRNAs in chicken embryo hepatocytes in vivo and in culture. Effect of porphyrinogenic drugs and haem. Biochem J 255:267–275

    CAS  PubMed  Google Scholar 

  • Hamilton JW, Bement WJ, Sinclair PR, Sinclair JF, Alcedo JA, Wetterhahn KE (1992) Inhibition of protein synthesis increases the transcription of the phenobarbital-inducible CYP2H1 and CYP2H2 genes in chick embryo hepatocytes. Arch Biochem Biophys 298:96–104

    CAS  PubMed  Google Scholar 

  • Iatropoulos MJ, Jeffrey AM, Enzmann HG, von Keutz E, Schlueter G, Williams GM (2001) Assessment of chronic toxicity and carcinogenicity in an accelerated cancer bioassay in rats of moxifloxacin, a quinolone antibiotic. Exp Toxicol Pathol 53:345–357

    CAS  PubMed  Google Scholar 

  • Ingram AJ, Grasso P (1985) Nuclear enlargement—an early change produced in mouse epidermis by carcinogenic chemicals applied topically in the presence of a promoter. J Appl Toxicol 5:53–60

    CAS  PubMed  Google Scholar 

  • Ingram AJ, Grasso P (1987) Nuclear enlargement produced in mouse skin by carcinogenic mineral oils. J Appl Toxicol 7:289–295

    CAS  PubMed  Google Scholar 

  • Jeffrey AM, Jennette KW, Blobstein SH, Weinstein IB, Beland FA, Harvey RG, Kasai W, Miura I, Nakanishi K (1976) Benzo[a]pyrene-nucleic acid derivative found in vivo: structure of benzo[a]pyrene tetrahydrodiol epoxide-guanosine adduct. J Am Chem Soc 98:5714–5715

    CAS  PubMed  Google Scholar 

  • Jeffrey AM, Weinstein IB, Jennette KW, Grzeskowiak K, Nakanishi K, Harvey RG, Autrup H, Harris C (1977) Structures of benzo[a]pyrene—nucleic acid adducts formed in human and bovine bronchial explants. Nature 269:348–350

    CAS  PubMed  Google Scholar 

  • Jeffrey AM, Amin SG, Kriminski J, Williams GM, Luo FQ, Zech K (2001) Organ specificity of DNA adduct formation with aromatic amines carcinogenic for the rat nasal mucosa or liver. The Toxicologist 60:153–154

    Google Scholar 

  • Juchau MR, Harris C, Stark KL, Lee QP, Yang HY, Namkung MJ, Fantel AG (1991) Cytochrome P-450 dependent bioactivation of prodysmorphogen in cultured conceptuses. Reprod Toxicol 5:259–263

    Article  CAS  PubMed  Google Scholar 

  • Kitada M, Taneda M, Ohta K, Nagashima K, Itahashi K, Kamataki T (1990) Metabolic activation of aflatoxin B1 and 2-amino-3-methylimidazo[4,5,f]-quinoline by human adult and fetal livers. Cancer Res 50:2641–2645

    CAS  PubMed  Google Scholar 

  • Leaky JEA, Hume R, Burchell B (1987) Development of multiple activities of UDP-glucuronyltransferase in human liver. Biochem J 243:859–861

    PubMed  Google Scholar 

  • Lorr NA, Bloom SE (1987) Ontogeny of the chicken cytochrome P-450 enzyme system. Expression and development of responsiveness to phenobarbital induction. Biochem Pharmacol 36:3059–3067

    CAS  PubMed  Google Scholar 

  • Montandon F, Williams GM (1994) Comparison of DNA reactivity of the polyphenylethylene hormonal agents diethylstilbestrol, tamoxifen and toremifene in rat and hamster liver. Arch Toxicol 68:272–275

    CAS  PubMed  Google Scholar 

  • Montheit DK, Gupta RC (1992) Carcinogen-DNA adducts in cultures of rat and human hepatocytes. Cancer Lett 62:87–93

    CAS  PubMed  Google Scholar 

  • Omiecinski CJ, Aicher L, Swenson L (1994) Developmental expression of human microsomal epoxide hydrolase. J Pharmacol Exp Ther 269:417–423

    CAS  PubMed  Google Scholar 

  • Pacifici GM, Franchi M, Giuliani L, Rane A (1989) Development of glucuronyltransferase and sulfotransferase towards 2-naphthol in human fetus. Dev Pharmacol Ther 14:108–114

    CAS  PubMed  Google Scholar 

  • Pastorelli R, Guanci M, Cerri A, Negri E, La Vecchia C, Fumagalli F, Mezzetti M, Cappelli R, Panigalli T, Fanelli R, Airoldi L (1998) Impact of inherited polymorphisms in glutathione S-transferase M1, microsomal epoxide hydrolase, cytochrome P450 enzymes on DNA, and blood protein adducts of benzo[a]pyrene diolepoxide. Cancer Epidemiol Biomarkers Prev 7:703–709

    PubMed  Google Scholar 

  • Roggeband R, Wolterbeek AP, Rutten AA, Baan RA (1993) Comparative 32P-postlabeling analysis of benzo[a]pyrene—DNA adducts formed in vitro upon activation of benzo[a]pyrene by human, rabbit and rodent liver microsomes. Carcinogenesis 14:1945–1950

    CAS  PubMed  Google Scholar 

  • Shimada T, Yamazaki H, Mimura M, Wakamiya N, Ueng Y-F, Guengerich FP, Inui Y (1996) Characterization of microsomal cytochrome P450 enzymes involved in the oxidation of xenobiotic chemicals in human fetal liver and adult lungs. Drug Metab Dispos 24:515–522

    CAS  PubMed  Google Scholar 

  • Stillwell WG, Turesky RJ, Sinha R, Tannenbaum SR (1999) N-oxidative metabolism of 2-amino-3,8-dimethylimidazo[4,5,f]quinoxaline (MeIQx) in humans: excretion of the N2-glucoronide conjugate of 2-hydroxyamino-MeIQx in urine. Cancer Res 59:5154–5159

    CAS  PubMed  Google Scholar 

  • Storer RD (2000) Current status and use of short/medium term models for carcinogenicity testing of pharmaceuticals—scientific perspective. Toxicol Lett 112–113:557–566

    Google Scholar 

  • Strange RC, Davis BA, Faulder CG, Cotton W, Bain AD, Hopkinson DA, Hume R (1985) The human glutathione S-transferases: developmental aspects of the GST1, GST2, and GST3 loci. Biochem Genet 23:1011–1028

    CAS  PubMed  Google Scholar 

  • Ullrich V, Weber P (1972) The O-dealkylation of 7-ethoxycoumarin by liver microsomes. A direct fluorometric test. Hoppe Seylers. Z Physiol Chem 353:1171–1177

    CAS  PubMed  Google Scholar 

  • van der Laan JW (1998) New perspectives for alternative approaches to carcinogenicity testing: a regulator viewpoint. Toxicol Lett 102–103:561–564

    Google Scholar 

  • Vijg J, van Steeg H (1998) Transgenic assays for mutations and cancer: current status and future perspectives. Mutat Res 400:337–354

    Article  CAS  PubMed  Google Scholar 

  • Walker CH (1998) Avian forms of cytochrome P450. Comp Biochem Physiol 121:65–72

    Article  Google Scholar 

  • Walker MP, Jahnke GD, Snedeker SM, Gladen BC, Lucier GW, DiAugustine RP (1990) 32P postlabeling analysis of the formation and persistence of DNA adducts in mammary glands of parous and nullyparous mice treated with benzo[a]pyrene. Carcinogenesis 11:2009–2015

    PubMed  Google Scholar 

  • Waxman DJ, Sundseth SS, Srivastava PK, Lapenson DP (1992) Gene-specific oligonucleotide probes for alpha, mu, and pi, microsomal rat glutathione S-transferases: analysis of liver transferase expression and its modulation by hepatic enzyme inducers and platinum anticancer drugs. Cancer Res 52:5797–5802

    CAS  PubMed  Google Scholar 

  • Williams GM, Iatropoulos MJ, Wang CX, Jeffrey AM, Thompson S, Pittman B, Palasch M, Gebhardt R (1998) Nonlinearities in 2-acetylaminofluorene exposure responses for genotoxic and epigenetic effects leading to initiation of carcinogenesis in rat liver. Toxicol Sci 45:152–161

    Article  CAS  PubMed  Google Scholar 

  • Wolff T, Deml E, Wanders H (1979) Aldrin epoxidation, a highly sensitive indicator specific for cytochrome P-450-dependent monoxygenase activity. Drug Metab Dispos 7:301–305

    CAS  PubMed  Google Scholar 

  • Yamamoto S, Urano K, Nomura T (2001) Validation of transgenic mice harboring the human prototype c-Ha- ras gene as a bioassay model for rapid carcinogenicity testing. Toxicol Lett 103:473–478

    Article  Google Scholar 

  • Yang H-YL, Namkung MJ, Juchau MR (1994) Expression of functional cytochrome P4501A1 in human embryonic hepatic tissues during organogenesis. Biochem Pharmacol 49:717–726

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Carmen E. Perrone.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Perrone, C.E., Ahr, HJ., Duan, J.D. et al. Embryonic turkey liver: activities of biotransformation enzymes and activation of DNA-reactive carcinogens. Arch Toxicol 78, 589–598 (2004). https://doi.org/10.1007/s00204-004-0580-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-004-0580-1

Keywords

Navigation