[en] There is increasing interest in cultured hepatocytes as a tool for solving toxicological and pharmacological problems while reducing laboratory animal experimentation. In the present study, fetal hepatocytes from the Japanese quail (Coturnix coturnix japonica) were used as an in vitro alternative model for evaluating the effects of PCBs and various pesticide-type chemicals on cell ultrastructure. Major alterations were demonstrated. The most striking effects of toxicants were an increase in the number of cisternae of the rough endoplasmic reticulum (RER), various alterations of mitochondrial morphology, a decreased glycogen content, vacuolization of the cytoplasm, and the appearance of concentric membrane arrays (CMA's), also called myelin-like figures. Other changes were sometimes observed, such as altered cell junctions, an increased lipid content, deformations of the nuclei, or the appearance of crystalline structures. These ultrastructural modifications seem to be dose-dependent. The present in vitro findings are validated by similar observations previously made in vivo on Japanese quail. They confirm the effectiveness of this technique as a biomonitoring tool for the evaluation of environmental quality. Yet the multiplicity of possible toxic effects, even for xenobiotics of a same category, makes it necessary to screen additional indicators of toxicity, such as the detoxifying activity of monooxygenases.
Disciplines :
Environmental sciences & ecology
Author, co-author :
Hugla, J. L.
Goffinet, Gerhard ; Université de Liège - ULiège > Services généraux (Faculté des sciences) > Relations académiques et scientifiques (Sciences)
Kremers, Pierre ; Université de Liège - ULiège > Relations académiques et scientifiques (Sciences)
Dubois, Michel ; Université de Liège - ULiège > Services généraux (Faculté de médecine) > Relations académiques et scientifiques (Médecine)
Lambert, Vincent ; Centre Hospitalier Universitaire de Liège - CHU > Ophtalmologie
Stouvenakers, Nadine ; Université de Liège - ULiège > Didactique des sciences biologiques
Thomé, Jean-Pierre ; Université de Liège - ULiège > Département des sciences et gestion de l'environnement > Ecologie animale et écotoxicologie
Language :
English
Title :
Ultrastructural Modifications in Cultured Fetal Quail Hepatocytes Exposed to Pesticides and Pcbs
Abiola, F., Lorgue, G., Benoit, E., Soyez, D., and Rivière, J. L. (1989). Effects of PCBs on plasma enzymes, testosterone level and hepatic xenobiotic metabolism in the grey partridge, Perdix perdix. Bull. Environ. Contam. Toxicol. 43, 473-480.
Althaus, F. R., Sinclair, J. F., Sinclair, P., and Meyer, U. A. (1979). Drug mediated induction of cytochromes P450 and drug metabolism in cultured hepatocytes maintained in chemically defined medium. J. Biol. Chem. 245, 2148-2153.
Alvares, A. P., Bickers, D. R., and Kappas, A. (1973). Polychlorinated biphenyls: A new type of inducer of cytochrome P-448 in the liver. Proc. Natl. Acad. Sci. USA 70, 1321-1325.
Boehm, K. D., Hood, R. L., and Ilan, J. (1988). Induction of vitellogenin in primary cultures of cockerel hepatocytes. Proc. Natl. Acad. Sci. USA 85, 3450-3454.
Bolline, A., Kremers, P., Kolodjici, C., and Gielen, J. (1987). Long term maintenance of monooxygenase activities in cultured fetal rat hepatocytes. Cell Diff. 21, 239-246.
Borlakoglu, J. T., and Haegele, K. D. (1991). Comparative aspects on the bioaccumulation, metabolism and toxicity with PCBs. Comp. Biochem. Physiol. 100C, 327-338.
Bulterworth, B., Smith-Oliver, T., and Earle, L. (1989). Use of primary cultures of human hepatocytes in toxicological studies. Cancer Res. 49, 1075-1084.
Buyan, P. J., and Page, J. M. (1978). Polychlorinated biphenyls, the effect of structure on the induction of quail hepatic microsomal enzymes. Toxicol. Appl. Pharmacol. 43, 507-518.
Chefurka, W., McLeod, H. L., and Steele, J. E. (1987). The role of lipid fluidity in the interaction of DDT and some analogs with synthetic membranes. Pest. Biochem. Physiol. 28, 93-102.
De Los Reyes, M., and Mora, E. C. (1978). Tissue residues and ultrastructural changes induced by DDT in chickens. Poult. Sci. 58, 1183-1191.
Denizeau, F., and Marion, M. (1984). Cultured rainbow trout and human cells exposed to PCBs in the evaluation of cytotoxicity of aquatic pollutants. Water Res. 18, 247-251.
Donato, T., Gomez-Lechon, M. J., and Castell, J. V. (1990). Drug metabolizing enzymes in rat hepatocytes co-cultured with cell lines. In Vitro Cell. Dev. Biol. 26, 1057-1062.
Dubois, M., Dupond, L., Thome, J. P., and Kremers, P. (1995). Molecular conformation and induction of hepatic CYP 1A1 in rat, avian and human hepatocytes - A screening test based on structure-activity relationships. Comp. Biochem. Physiol., submitted.
Ellenton, J. A., Brownlee, L. J., and Hollebone, B. R. (1985). Aryl hydrocarbon hydroxylase levels in herring gulls embryos from different locations of the Great Lakes. Environ. Toxicol. Chem. 4, 615-621.
Flödström, S., Hemming, H., Warngard, L., and Ahlborg, U. G. (1990). Promotion of altered hepatic foci development in rat liver, cytochrome P450 enzyme induction and inhibition of cell-cell communication by DDT and some structurally related organohalogen pesticides. Carcinogenesis 11, 1413-1417.
Fraslin, J. M., Touquette, L., Douaire, M., Menezo, Y., Guillemot, J.-C., and Mallard, J. (1992). Isolation and long-term maintenance of differentiated adult chicken hepatocytes in primary culture. In Vitro Cell. Dev. Biol. 28A, 615-620.
Hacking, M. A., Budd, J., and Hodson, K. (1977). The ultrastructure of the liver of the rainbow trout: Normal structure and modifications after chronic administration of a polychlorinated biphenyl Aroclor 1254. Can. J. Zool. 56, 477-491.
Hinton, D. E., and Pool, C. R. (1976). Ultrastructure of the liver in channel catfish Ictalurus punctatus (Rafinesque). J. Fish Biol. 8, 209-219.
Hinton, D. E., Glaumann, H., and Trump, B. F. (1978). Studies on the cellular toxicity of polychlorinated biphenyls (PCBs). 1. Effect of PCBs on microsomal enzymes and on synthesis and turnover of microsomal and cytoplasmic lipids of rat liver - A morphological and biochemical study. Virchows Arch. B Cell Pathol. 27, 279-306.
Hoopingarner, R., Samuel, A., and Krause, D. (1972). Polychlorinated biphenyl interaction with tissue culture cells. Environ. Hlth. Persp. 1, 155-158.
Hugla, J. L., Philippart, J. C., Kremers, P., Goffinet, G., and Thomé, J. P. (1995). PCB contamination of the common barbel, Barbus barbus (Pisces, Cyprinidae), in the river Meuse in relation to hepatic monooxygenase activity and ultrastructural liver change. Neth. J. Aquatic Ecol. 29, 121-131.
Jiger, U., and Meyer, U. A. (1983). The substituted pyridins, metyrapone and nicotinamide are inducers of 5-aminolevulinate synthase and cytochrome P450 in hepatocyte culture. Biochem. Pharmacol. 31, 1735-1741.
Jones, G., and Butler, W. H. (1973). Morphological study of the liver lesion induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in rats. J. Pathol. 112, 93-97.
Jonsson, H. T., Walker, M. E., Greene, W. B., Hughson, M. D., and Hennigar, G. R. (1981). Effects of prolonged exposure to dietary DDT and PCB on rat liver morphology. Arch. Environ. Contam. Toxicol. 10, 171-183.
Kimbrough, R. D., Linder, R. E., and Gaines, T. B. (1972). Morphological changes in livers of rats fed with polychlorinated biphenyls. Arch. Environ. Health 25, 354-364.
Klaunig, J. E., Lipsky, M. M., Trump, B. F., and Hinton, D. E. (1979). Biochemical and ultrastructural changes in teleost liver following subacute exposure to PCB. J. Environ. Toxicol. 2, 953-963.
Köhler, A. (1990). Identification of contaminant-induced cellular and subcellular lesions in the liver of flounder (Platichthys flesus L.) caught at differently polluted estuaries. Aquatic Toxicol. 16, 271-294.
Kremers, P., Roelandt, L., Todaro, A., Stouvenakers, N., Louvet, M., Goffinet, G., and Thomé, J. P. (1993). Use of cultured hepatocytes as an alternative method to study the effects of PCBs on living organisms. Toxicol. in Vitro 7, 433-437.
Lemaire, P., Berthaut, J., Lemaire-Gony, S., and Lafaurie, M. (1992). Ultrastructural changes induced by benzo(a)pyrene in sea bass (Dicentrarchus labrax) liver and intestine: Importance of the intoxication route. Environ. Res. 57, 59-72.
Lincer, J. L., and Peakall, D. B. (1970). Metabolic effects of polychlorinated biphenyls in the American Kestrel. Nature 228, 783-784.
Lipsky, M. M., Klaunig, J. E., and Hinton, D. E. (1978). Comparison of acute response to polychlorinated biphenyl in liver of rat and channel catfish: A biochimical and morphological study. J. Toxicol. Environ. Health 4, 107-121.
Nagaoka, S., Kamuro, H., Oda, H., and Yoshida, A. (1991). Effects of polychlorinated biphenyls on cholesterol and ascorbic acid metabolism in primary cultured rat hepatocytes. Biochem. Pharmacol. 41, 1259-1261.
Nigam, S. K., Lakkad, B. C., Karnik, A. B., and Thakore, K. N. (1984). Ultrastructural changes in liver of mice exposed to hexachlorocyclohexane. Int. J. Exp. Biol. 22, 194-204.
Norback, D. H., and Allen, J. R. (1969). Morphogenesis of toxic fat-induced concentric membrane arrays in rat hepatocytes. Lab. Invest. 20, 338-346.
Ohnishi, Y., and Arakawa, T. (1977). Experimental studies on the effects of Kanechlor 400 on cultured conjunctival cells. Fukuoka Acta Med. 68, 109-122.
Ortega, P. (1966). Light and electron microscopy of DDT poisoning in rat liver. Lab. Invest. 15, 657-679.
Porter, T. D., and Coon, M. J. (1991). Cytochrome P-450: Multiplicity of isoforms, substrates and catalytic and regulatory mechanism. J. Biol. Chem. 266, 13469-13472.
Ratanasavan, D., Baffet, G., Latinier, M. F., Rissel, M., and Guillouzo, A. (1988). Use of hepatocyte co-cultures in the assessment of drug toxicity from chronic exposure. Xenobiotica 18, 773-783.
Rivière, J. L., Grolleau, G., and Bach, J. (1985). Hepatic biotransformation in the buzzard (Buteo buteo) and the Japanese quail (Coturnix coturnix): Effect of PCBs. Comp. Biochem. Physiol. 82C, 439-443.
Rodman, L. E., Shedlofsky, S. I., Mannschreck, A., Puttmann, M., Swim, A. T., and Roberstson, L. W. (1991). Differential potency of atropisomers of polychlorinated biphenyls on cytochrome P450 induction and uroporphyrin accumulation in the chick embryo hepatocyte culture. Biochem. Pharmacol. 41, 915-922.
Roelandt, L., Todaro, A., Thomé, J. P., and Kremers, P. (1995). Effect of PCBs (Aroclor 1254) on cytochrome P450 expression and monooxygenase activities in cultured fetal rat hepatocytes. Toxicology 98, 95-103.
Rubin, L. (1983). Toxicological update of dimethyl sulfoxide. Ann. N.Y. Acad. Sci. 411, 6-9.
Schecter, A., Schaffner, F., Tiernan, T., and Taylor, M. (1984). Ultrastructural alterations of liver mitochondria in response to dioxins, furans, PCBs and biphenylenes. In Biological Mechanisms of Dioxin Action (A. Poland and R. D. Kimbrough, Eds.), pp. 177-190, Vol. 18. Branbury Report.
Stadnicki, S., and Allen, S. (1979). Toxicity of 2,2′,5,5′-tetrachlorobiphenyl and its metabolites 2,2′,5,5′-tetrachlorobiphenyl-3,4-oxide and 2,2′,5,5′-tetrachlorobiphenyl 4-ol to cultured cells in vitro. Environ. Contam. Toxicol. 23, 788-796.
Stäubli, W., Hess, R., and Weibel, E. R. (1969). Correlated morphometric and biochemical studies on the liver cell. 2. Effects of phenobarbital on rat hepatocytes. J. Cell Biol. 42, 92-112.
Stotz, I. J., and Greichus, Y. J. (1978). The effects of a polychlorinated biphenyl (Aroclor 1254) on the white pelican: Ultrastructure of hepatocytes. Bull. Environ. Contam. Toxicol. 19, 319-326.
Stouvenakers, N., Hugla, J. L., Goffinet, G., Kremers, P., and Thome, J. P. (1996). Effects of PCBs on liver ultrastructure and monooxygenase activities in Japanese quail. Bull. Environ. Contam. Toxicol. 56, 839-846.
Tarlow, D., Watkins, P. A., Reed, R. E., Miller, R. S., Zwergel, E. E., and Lane, M. D. (1977). Lipogenesis and the synthesis and secretion of very low density lipoprotein by avian cells in nonproliferating monolayer culture. J. Cell Biol. 73, 332-353.
Thomé, J. P., Hugla, J. L., Adam, C., and Goffinet, G. (1993). Incidence d'une contamination chronique par les PCBs sur la morphologie et l'ultrastructure du foie chez le barbeau fluviatile (Barbus barbus). Cah. Ethol. 13, 151-154.
Thomé, J. P., Roelandt, L., Goffinet, G., Stouvenakers, N., and Kremers, P. (1995). Cytotoxic effects of Aroclor 1254 on ultrastructure and biochemical parameters in cultured foetal rat hepatocytes. Toxicology 98, 83-94.
Toftgard, R., Nielsen, O. G., Carlstedt-Duke, J., and Glaumann, H. (1986). Polychlorinated terphenyls: Alterations in liver morphology and induction of cytochrome P-450. Toxicology 41, 131-144.
Turner, J. N., and Collins, D. N. (1985). Alterations in Guinea pig hepatocytes exposed to chlorinated hydrocarbons: Membrane proliferation and excretion. J. Exp. Pathol. 2, 75-92.
Weis, P. (1974). Ultrastructural changes induced by low concentrations of DDT in the livers of the zebrafish and the guppy. Chem. Biol. Interact. 8, 25-30.
Williams, C. S., Jones, B. C., Nchege, O., and Chung, R. A. (1993). Hepatocyte ultrastructure following exposure to Aroclors and pure polychlorinated biphenyls. J. Environ. Pathol. Toxicol. Oncol. 12, 17-33.
Yeoh, G. C. T., Wassemburg, J. A., Edkins, E., and Oliver, I. T. (1979). Synthesis and secretion of albumin and transferritin by foetal rat hepatocyte cultures. Biochim. Biophys. Acta 565, 347-355.