Abstract
Methods of biochemical monitoring of individual animals for exposure to environmental contaminants are of great potential use. The hepatic metabolism of various alkoxyresorufins, which are highly specific substrates for certain forms of cytochrome(s) P450, is highly induced by a variety of environmental contaminants. Thus, theO-deal-kylation of pentoxy- or benzyloxyresorufin was induced greater than 20-fold in the rat bya-hexachlorocyclohexane, 2,4,5,2′,4′,5′-hexabromobiphenyl, DDT and Aroclor®-1254, while the metabolism of ethoxyresorufin was highly induced by 5,6-benzoflavone, 3,4,5,3′,4′,5′-hexabromobiphenyl and Aroclor®-1254. Additionally, rats exposed to diets containing as little as 12 ppm DDT displayed >five-fold increases in the rate of hepaticO-dealkylation of benzyloxyresorufin. Induction of the hepatic metabolism of these resorufin ethers in 9000xg supernatant fractions taken from rats exposed to potential environmental contaminants may constitute a valuable diagnostic indicator of the presence of a variety of pollutants, including poly cyclic aromatic hydrocarbons, organochlorine pesticides, polyhalogenated biphenyls and 2,3,7,8-tetrachlorodibenzo-p-dioxin. These results suggest the potential applicability of these substrates in detecting chemical contamination in the environment.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Addison RF, Sadler MC, Lubet RA (1987) Absence of hepatic microsomal pentyl- or benzyl-resorufinO-dealkylase induction in rainbow trout(Salmo gairdneri) treated with phenobarbitone. Biochem Pharmacol 36:1183–1184
Blaich G, Gottlicher M, Cikryt P, Metzler M (1988) Induction of P-450 isoenzyme activities in Syrian golden hamster liver compared to rat liver as probed by the rate of 7-alkoxyresorufin-O-dealkylation. Chem-Biol Interact 67:129–138
Bohlen P, Stein S, Dairman W, Udenfriend S (1973) Fluorometric assay of proteins in the nanogram range. Arch Biochem Biophys 155:213–220
Burke MD, Mayer RT (1974) Ethoxyresorufin: Direct fluorimetric assay of a microsomalO-dealkylation which is preferentially inducible by 3-methylcholanthrene. Drug Metab Dispos 2:583–588
— (1983) Differential effects of phenobarbitone and 3-methyl-cholanthrene induction on the hepatic microsomal metabolism and cytochrome P-450-binding of phenoxazone and a homologous series of itsn-alkyl ethers (alkoxyresorufins). Chem-Biol Interact 45:243–258
Burke MD, Thompson S, Elcombe CR, Halpert J, Haaparanta T, Mayer RT (1985) Ethoxy-, pentoxy- and benzyloxyphenoxazones and homologues: A series of substrates to distinguish between different induced cytochromes P-450. Biochem Pharmacol 34:3337–3345
Goldstein A, Aaronow A, Kalman SM (1974) Principles of Drug Action: The Basis of Pharmacology, 2nd Ed. J. Wiley and Sons, New York, pp 360–365.
Gorsline J, Holmes WN, Cronshaw J (1981) The effects of ingested petroleum on the naphthalene-metabolizing properties of liver tissue in seawater-adapted mallard ducks (Anas platyrhynchos). Environ Res 24:337–390
Guengerich FP, Dannan GA, Wright ST, Martin MV, Kaminsky LS (1982a) Purification and characterization of liver microsomal cytochromes P-450: Electrophoretic, spectral, catalytic, and immunochemical properties and inducibility of eight isozymes isolated from rats treated with phenobarbital or ß-naphthoflavone. Biochemistry 21:6019–6030
Guengerich FP, Wang P, Davidson NK (1982b) Estimation of isozymes of microsomal cytochrome P-450 in rats, rabbits, and humans using immunochemical staining coupled with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Biochemistry 21:1698–1706
Kinoshita FK, Frawley JP, DuBois KP (1966) Quantitative measurement of induction of hepatic microsomal enzymes by various dietary levels of DDT and toxaphene in rats. Toxicol Appl Pharmacol 9:505–513
Knight GC, Walker CH (1982) A study of the hepatic microsomal monooxygenase of sea birds and its relationship to organochlorine pollutants. Comp Biochem Physiol 73:211–221
Lubet RA, Mayer RT, Cameron JW, Nims RW, Burke MD, Wolff T, Guengerich FP (1985a) Dealkylation of pentoxyresorufin: A rapid and sensitive assay for measuring induction of cytochrome(s) P-450 by phenobarbital and other xenobiotics in the rat. Arch Biochem Biophys 238:43–48
Lubet RA, Nims, RW, Mayer RT, Cameron JW, Schechtman LM (1985b) Measurement of cytochrome P-450 dependent dealkylation of alkoxyphenoxazones in hepatic S9s and hepatocyte homogenates: Effects of dicumarol. Mutat Res 142:127–131
Lucier GW, Nelson KG, Everson RB, Wong TK, Philpot RM, Tiernan T, Taylor M, Sunahara GI (1987) Placental markers of human exposure to polychlorinated biphenyls and polychlorinated dibenzofurans. Environ Health Perspect 76:79–87
Lucier GW, Rumbaugh RC, McCoy Z, Hass R, Harvan D, Albro P (1986) Ingestion of soil contaminated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) alters hepatic enzyme activities in rats. Fund Appl Tox 6:364–371
McConnell EE, Lucier GW, Rumbaugh RC, Albro PW, Harvan DJ, Hass RJ, Harris MW (1984) Dioxin in soil: Bioavailability after ingestion by rats and guinea pigs. Science 223:1077–1079
Nebert DW, Adesnik M, Coon MJ, Estabrook RW, Gonzalez FJ, Guengerich FP, Gunsalus IC, Johnson EF, Kemper B, Levin W, Phillips IR, Sato R, Waterman MR (1987) The P450 gene superfamily: Recommended nomenclature. DNA 6:1–11
Nims RW, Lubet RA (1983) Cytosol-mediated reduction of resorufin fluorescence: Effects on the ethoxyresorufinO-deethylase (ETR) assay. Biochem Pharmacol 22:175–176
Nims RW, Prough RA, Lubet RA (1984) Cytosol-mediated reduction of resorufin: A method for measuring quinone oxidoreductase. Arch Biochem Biophys 229:459–465
Payne JF (1976) Field evaluation of benzopyrene hydroxylase induction as a monitor for marine petroleum pollution. Science 191:945–946
Safe S (1984) Polychlorinated biphenyls (PCBs) and polybrominated biphenyls (PBBs): Biochemistry, toxicology and mechanisms of action. Crit Rev Toxicol 13:319–395
Sloof W, Van Kreyl CF, Baars AJ (1983) Relative liver weights and xenobiotic-metabolizing enzymes of fish from polluted surface waters in the Netherlands. Aquat Toxicol 4:1–14
Stegeman JJ, Kloepper-Sams PJ, Farrington JW (1986) Monooxygenase induction and chlorobiphenyls in the deep-sea fishCoryphaenoides armatus. Science 231:1287–1289
Tice RR, Armiston BG, Baucher R, Luke CA, Paquette DE (1988) Environmental biomonitoring with feral rodent species. In: Sandhu DH, Marini DE, Mass M, Moore M, Mumford JL (eds) Application of short term testing in analysis of complex environmental mixtures. Plenum Press, New York, pp 175–180
Umbreit TH, Hesse EJ, Gallo MA (1986) Bioavailability of dioxin in soil from a 2,4,5-T manufacturing site. Science 232:497–499
— (1987) Reproductive toxicity in female mice of dioxin-conlaminated soils from a 2,4,5-trichlorophenoxyacetic acid manufacturing site. Arch Environ Contam Toxicol 16:461–466
Whitlock JP (1986) The regulation of cytochrome P-450 gene expression. Ann Rev Pharmacol Toxicol 26:333–369
Wong TK, Domin BA, Bent PE, Blanton TE, Anderson MW, Philpot RM (1986) Correlation of placental microsomal activities with protein detected by antibodies to rabbit cytochrome P-450 isozyme 6 in preparations from humans exposed to polychlorinated biphenyls, quaterphenyls, and dibenzofurans. Cancer Res 46:999–1004
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lubet, R.A., Guengerich, F.P. & Nims, R.W. The induction of alkoxyresorufin metabolism: A potential indicator of environmental contamination. Arch. Environ. Contam. Toxicol. 19, 157–163 (1990). https://doi.org/10.1007/BF01056082
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01056082