RESUMO
Exposure to environmental dioxins via mother's milk may be one causative factor of mineralization defects in children's teeth. A prerequisite for the completion of enamel mineralization is the removal of enamel matrix. To test the hypothesis that dioxins interfere with enamel maturation, we administered lactating Han/Wistar rats a single dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; 50 or 1000 micro g/kg) on the day after delivery and analyzed tissue sections of the pup heads at post-natal days (Pn) 9 and 22. By Pn22, the first and second molars of the exposed pups, but not controls, showed retention of enamel matrix. Predentin was thicker than normal. Immunostaining for the aryl hydrocarbon/dioxin receptor (AhR) and cytochrome P4501A1 (CYP1A1) in ameloblasts and odontoblasts was reduced, suggesting that TCDD interferes with tooth mineralization via AhR. Extinction of AhR may lead to abolition of CYP1A1 expression as a sign of impaired dental cell function.
Assuntos
Esmalte Dentário/efeitos dos fármacos , Dentina/efeitos dos fármacos , Poluentes Ambientais/efeitos adversos , Lactação , Dibenzodioxinas Policloradas/efeitos adversos , Calcificação de Dente/efeitos dos fármacos , Ameloblastos/efeitos dos fármacos , Amelogênese/efeitos dos fármacos , Animais , Distribuição de Qui-Quadrado , Citocromo P-450 CYP1A1/análise , Feminino , Leite , Odontoblastos/efeitos dos fármacos , Ratos , Ratos Endogâmicos , Ratos Wistar , Receptores de Hidrocarboneto Arílico/análiseRESUMO
Although the average cadmium intake in Finland is about 10 microg day(-1), some risk groups can be identified. This study assessed cadmium intake from the consumption of moose meat, liver and kidneys by moose hunters. Consumption data from a postal questionnaire were combined with a representative database on moose cadmium concentrations. Cadmium intakes were calculated as point estimates for all respondents (n = 711), for those consuming moose meat, liver and/or kidneys, and for the highest decile of those. Probabilistic modelling using the Monte Carlo technique was used to simulate the distribution of dietary cadmium exposure. Of the respondents, 69% consumed moose liver and only 23% moose kidneys. The consumption of moose liver or kidneys significantly increased cadmium intake, whereas moose meat (median consumption 17 kg year(-1) person(-1)) contributed only slightly (0.16 microg day(-1) person(-1)) to the daily total cadmium intake. In the simulation, 10% of the moose hunters had an intake of > 8.76 microg day(-1) (14.6% of PTWI for a 60-kg person) from moose. Point estimates provided only a partial understanding of the potential exposure. Simulated distributions of intake were more useful in characterizing exposure. The study revealed that heavy users of moose organs have a relatively narrow safety margin from the levels of cadmium probably causing adverse health effects.
Assuntos
Cádmio/administração & dosagem , Cervos/metabolismo , Contaminação de Alimentos/análise , Carne/análise , Animais , Cádmio/análise , Dieta , Finlândia , Análise de Alimentos/métodos , Humanos , Rim/química , Fígado/químicaRESUMO
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and related halogenated aromatic hydrocarbons are environmental toxicants that act via the AH receptor (AHR). In vitro studies have demonstrated that some indole derivatives present in cruciferous vegetables also bind to the AHR. One of the highest AHR binding affinities is exhibited by indolo[3,2-b]carbazole (ICZ). Since exposure to these dietary indoles is quantitatively far larger than that to halogenated aromatic compounds, their potential toxic risks have raised concern. In the present study, we compared the effects of ICZ with those of a single dose of 20 microg/kg TCDD in the most TCDD-sensitive rat strain (Long-Evans [Turku AB]) (L-E). Whereas TCDD elicited the expected toxicity syndrome, ICZ, either as a single subcutaneous dose (63.5, 127 or 508 microg/kg) or with repeated sc dosing (508 microg/kg for 5 days) failed to reproduce any toxic impacts of TCDD. Furthermore, a simultaneous ICZ treatment (63.5 or 127 microg/kg for 10 days) did not interfere with TCDD (20 microg/kg; single exposure) action. A moderate hepatic induction of CYP1A1 could be triggered by repeated intragastric administration of ICZ (127 microg/kg for 4 days, the last treatment 2.5 h prior to termination). In control experiments in a reconstituted yeast system, ICZ potently and dose-dependently activated L-E rat AHR function demonstrating that it represents a bona fide high-affinity ligand for the rat receptor in vivo. Thus, the present study does not support the view that dietary exposure to ICZ would present a hazard of AHR-mediated adverse health effects to humans.
Assuntos
Carbazóis/toxicidade , Indóis/toxicidade , Dibenzodioxinas Policloradas/toxicidade , Animais , Peso Corporal/efeitos dos fármacos , Comportamento Alimentar/efeitos dos fármacos , Ratos , Ratos Long-Evans , Receptores de Hidrocarboneto Arílico/metabolismoRESUMO
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) brings about a wide spectrum of toxic and biochemical changes, most of which are mediated by the AH receptor (AHR). Recent cloning of the AHR from the two most TCDD-resistant laboratory animals, Han/Wistar (Kuopio) rats and hamsters, suggested a critical role for the C-terminal transactivation domain structure in TCDD sensitivity. Here we cloned the AHR from the most TCDD-susceptible species, guinea pig. The N-terminus of its AHR was highly similar to that in the resistant animals. However, the C-terminal Q-rich subdomain was only about half the size of this subunit in the hamster AHR. There was a distinct correlation across published mammalian species between the number of glutamine residues in the Q-rich subdomain and sensitivity to the acute lethality of TCDD. The closest homolog of the Guinea pig receptor turned out to be the human AHR, which may be relevant for dioxin risk assessment.
Assuntos
Dibenzodioxinas Policloradas/toxicidade , Receptores de Hidrocarboneto Arílico/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Western Blotting , Resistência a Medicamentos/genética , Cobaias , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Especificidade da Espécie , Relação Estrutura-AtividadeRESUMO
Most toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) are mediated by the aryl hydrocarbon receptor (AHR). A single, acute dose of TCDD can alter its own receptor levels thus complicating evaluation of dose-response relationships for AHR-mediated events. Since environmental exposure to dioxins is typically of a repeated low-dose nature, we examined the effect of such exposure on AHR expression. Three rat strains differing greatly in their sensitivity to acute TCDD lethality, Long-Evans (Turku AB) (L-E) (LD50 approximately 10 microg/kg); Sprague Dawley (SD) (LD50 approximately 50 microg/kg); and Han/Wistar (Kuopio) (H/W) (LD50 > 9600 microg/kg), were administered TCDD intragastrically, biweekly for 22 weeks producing doses equivalent to 0, 10, 30, and 100 ng/kg/day. Changes in hepatic AHR levels were quantitated at the protein level by radioligand binding and immunoblotting and at the mRNA level by RT-PCR. Cytosolic AHR protein was elevated at 10 or 30 ng/kg/day TCDD in SD and L-E rats; AHR mRNA was also elevated at these doses, suggesting a pretranslational mechanism. There was no apparent relationship between TCDD-induced AHR regulation and strain sensitivity to TCDD. Overall, "subchronic" TCDD did not greatly perturb AHR expression. The maintenance of relatively constant receptor levels in the face of persistent agonist stimulation is in contrast to the sustained depletion of AHR by TCDD observed in cell culture and to the fluctuations in AHR observed hours to days following acute TCDD exposure in vivo. Changes in AHR levels may affect dose-response relationships; the effect of TCDD on its own receptor at environmentally relevant dosing schemes is therefore important to risk assessment.
Assuntos
Dibenzodioxinas Policloradas/toxicidade , Receptores de Hidrocarboneto Arílico/metabolismo , Teratogênicos/toxicidade , Animais , Peso Corporal/efeitos dos fármacos , Dioxinas/toxicidade , Resistência a Medicamentos/fisiologia , Feminino , Immunoblotting , Fígado/efeitos dos fármacos , Fígado/fisiologia , Masculino , Modelos Animais , Tamanho do Órgão/efeitos dos fármacos , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Ensaio Radioligante , Ratos , Ratos Long-Evans , Ratos Sprague-Dawley , Ratos Wistar , Receptores de Hidrocarboneto Arílico/efeitos dos fármacos , Receptores de Hidrocarboneto Arílico/genética , Timo/efeitos dos fármacos , Timo/fisiologiaRESUMO
Changes in the perinatal testosterone surge have been related to demasculinization of the central nervous system and androgen-dependent growth of the reproductive organs in male mammals. Earlier reports suggest that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) interferes with androgen production, but the perinatal effects have remained elusive. In the present study we explored in utero-effects of TCDD (0.05, 0.1, 0.5, and 1.0 microg/kg), introduced on day 13.5 of pregnancy, on prenatal (day 19.5 post-conception [p.c.]) testosterone (T) surge and pituitary luteinizing hormone (LH) production in TCDD-resistant Han/Wistar (H/W) and TCDD-sensitive Long-Evans (L-E) rats. To elucidate estrogenic effects on T and LH production, Sprague-Dawley (S-D) fetuses with previously known DES-sensitivity were exposed in utero to diethylstilbestrol (DES, 100-300 microg/kg) on days 13.5, 15.5, and 17.5 p.c. For comparison, H/W fetuses that responded to TCDD treatments were exposed to DES at concentration of 100 microg/kg. It was found that TCDD has a stimulatory effect on testicular T synthesis in the H/W fetuses and that their circulating T concentrations increased significantly. The effect was not seen in the inbred L-E fetuses, which throughout the study showed considerably low testicular T levels. Pituitary LH concentrations also increased in the H/W fetuses exposed to TCDD. Effects of TCDD (1.0 microg/kg) in the H/W fetuses could be confirmed in vitro by human chorionic gonadotropin (hCG) stimulation assay showing the highest response rate in the TCDD exposed testes. Stimulation of cyclic AMP (adenosine-3', 5'-cyclic monophosphate[cAMP]) production was not considerably altered by in utero TCDD exposure. A significant depression in testicular and plasma T content was seen in the DES-exposed S-D and H/W fetuses, but pituitary LH levels did not alter considerably. In the presence of hCG, DES-exposed testes showed lower in vitro T and cAMP production rates compared to the untreated testes. TCDD (1.0 microg/kg) increased and DES decreased the male body weight gain, but the changes were not sex-dependent. It is concluded that TCDD may increase the amplitude of the prenatal testosterone surge in male rats by stimulating pituitary LH production and enhancing the sensitivity of the fetal testis to LH. DES, on the contrary, apparently impairs testicular steroidogenesis and pituitary function.
Assuntos
Dietilestilbestrol/toxicidade , Feto/efeitos dos fármacos , Feto/metabolismo , Hormônio Luteinizante/metabolismo , Dibenzodioxinas Policloradas/toxicidade , Testosterona/metabolismo , Animais , Peso Corporal/efeitos dos fármacos , AMP Cíclico/biossíntese , Feminino , Técnicas In Vitro , Masculino , Troca Materno-Fetal , Hipófise/efeitos dos fármacos , Hipófise/metabolismo , Gravidez , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Diferenciação Sexual/efeitos dos fármacos , Testículo/efeitos dos fármacos , Testículo/metabolismoRESUMO
The interference with tooth development by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) was studied in dioxin-resistant Han/Wistar rats. Lactating dams were given a single dose of 50 or 1000 microg TCDD/kg body wt 1 day after delivery and the pup heads were analyzed radiographically or histologically at postnatal days 9 and/or 22. Of 19 animals studied histologically, 10 lacked one or more third molars, which were at the bud stage at the start of the experiment. A higher proportion of pups exposed to the higher dose (9/13) lacked third molars than those exposed to the lower dose (1/6) (27/52 and 2/24 teeth missing, respectively). Missing upper third molars (19/38) were more frequent than were lower (10/38). The development of the third molars present was retarded. The root tips of the more advanced first and second molars were prematurely closed and root formation was arrested, but eruption was not affected. Dentinogenesis of the continuously erupting lower incisor teeth was preeruptively arrested because of pulpal cell death. All the teeth of the control rat pups developed normally. In contrast to the control pups, none of the 11 experimental pups examined radiographically (6 exposed to the higher dose and 5 to the lower) showed mineralization of their third molar cusps. The results show that the effects of TCDD on rat tooth development depend on not only the dose but also the tooth type and developmental stage. Inasmuch as early tooth development is under the control of inductive interactions between the epithelium and the mesenchyme, the interference by TCDD with tooth morphogenesis with the consequent arrest of development is likely to involve epithelial-mesenchymal signaling.
Assuntos
Poluentes Ambientais/toxicidade , Lactação , Dente Molar/crescimento & desenvolvimento , Dibenzodioxinas Policloradas/toxicidade , Animais , Feminino , Incisivo/diagnóstico por imagem , Incisivo/crescimento & desenvolvimento , Masculino , Dente Molar/diagnóstico por imagem , Dente Molar/patologia , Dibenzodioxinas Policloradas/administração & dosagem , Radiografia , Ratos , Ratos Wistar , Anormalidades Dentárias/induzido quimicamente , Anormalidades Dentárias/patologiaRESUMO
The aryl hydrocarbon receptor (AHR) mediates toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and regulates expression of several genes such as CYP1A1. Little is known about what regulates expression of the AHR itself. We tested the ability of TCDD to alter in vivo expression of its own receptor in rat strains that are susceptible to TCDD lethality [Long-Evans (Turku AB) (L-E) and Sprague Dawley (SD)] and in a rat strain that is remarkably resistant to TCDD lethality [Han/Wistar (Kuopio) (H/W)]. Rats were administered a single, intragastric dose of 5 or 50 microg/kg of TCDD. Hepatic cytosol, nuclear extract, and RNA were prepared at 1, 4, and 10 days after TCDD exposure. AHR expression was assessed at three levels: ligand binding function, immunoreactive protein and mRNA. TCDD at 5 microg/kg produced a 2- to 3-fold increase in cytosolic AHR in all strains; 50 microg/kg produced depletion at day 1 followed by recovery in SD and H/W but not L-E rats. Both the increase in AHR above basal levels and the recovery from initial depletion were accompanied by elevations in steady-state AHR mRNA, suggesting a pre-translational mechanism for AHR regulation by its own ligand. This up-regulation in vivo is in contrast to the sustained depletion of AHR caused by TCDD in cell culture. There was no clear relationship between AHR regulation and strain sensitivity; thus, the large inherent strain differences in susceptibility to TCDD lethality probably are not explained by differential regulation of AHR by TCDD.
Assuntos
Dibenzodioxinas Policloradas/farmacologia , Receptores de Hidrocarboneto Arílico/biossíntese , Teratogênicos/farmacologia , Análise de Variância , Animais , Peso Corporal/efeitos dos fármacos , Dioxinas/farmacologia , Feminino , Immunoblotting , Fígado/efeitos dos fármacos , Fígado/fisiologia , Modelos Animais , Tamanho do Órgão/efeitos dos fármacos , RNA Mensageiro/biossíntese , RNA Mensageiro/efeitos dos fármacos , Ensaio Radioligante , Ratos , Ratos Long-Evans , Ratos Sprague-Dawley , Receptores de Hidrocarboneto Arílico/efeitos dos fármacos , Receptores de Hidrocarboneto Arílico/genética , Regulação para Cima/efeitos dos fármacosRESUMO
Hamsters and Han/Wistar (Kuopio; H/W) rats show peculiarly selective responsiveness to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). They are extremely resistant to its acute lethality but sensitive to, e.g. , enzyme induction. The biological effects of TCDD are mediated by the AH receptor (AHR). Recent studies on H/W rat AHR discovered a remodelled transactivation domain which appears to be critical for the TCDD resistance of these animals. Here, molecular cloning and sequencing of hamster AHR reveals another type of restructured transactivation domain. In hamsters, the functionally pivotal Q-rich region is substantially expanded and enriched in glutamine compared with all other AHRs cloned to date. By contrast, the amino-terminal end is highly conserved, which is in agreement with the H/W rat AHR. Because of the additional material in the transactivation domain, hamster AHR protein is larger than that in rats or mice, but the pattern of AHR mRNA expression in tissues is similar.
Assuntos
Resistência a Medicamentos , Dibenzodioxinas Policloradas/toxicidade , Receptores de Hidrocarboneto Arílico/química , Receptores de Hidrocarboneto Arílico/genética , Ativação Transcricional , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Cricetinae , Feminino , Perfilação da Expressão Gênica , Humanos , Masculino , Mesocricetus , Dados de Sequência Molecular , Estrutura Terciária de Proteína , RNA Mensageiro/análise , RNA Mensageiro/genética , Receptores de Hidrocarboneto Arílico/metabolismo , Alinhamento de Sequência , Especificidade da EspécieRESUMO
Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on food selection were studied in TCDD-resistant Han/Wistar and TCDD-sensitive Long-Evans rats and their crosses. The rats were offered a selection diet consisting of chocolate, cheese, and chow, and TCDD was given at the same time or 4 or 16 days later. TCDD persistently reduced the chocolate intake. When the selection diet was started at the time of or less than 11 h after TCDD exposure, TCDD almost completely prevented the intake of chocolate and also cheese in all strains already on the first day, while controls started to consume large amounts of both foods. This may be due to conditioned taste aversion. The effect on food selection with familiar foods seemed to reduce fat intake, while protein and carbohydrate intakes were more variable. There were no major strain differences in the chocolate intake inhibition despite a 1000-fold sensitivity difference in TCDD lethality.
Assuntos
Comportamento Alimentar/efeitos dos fármacos , Preferências Alimentares/efeitos dos fármacos , Dibenzodioxinas Policloradas/toxicidade , Animais , Carboidratos da Dieta/administração & dosagem , Gorduras na Dieta/administração & dosagem , Proteínas Alimentares/administração & dosagem , Ingestão de Energia , Feminino , Masculino , Ratos , Ratos Long-Evans , Ratos WistarRESUMO
Risk assessment of dioxins is currently based on induction of liver tumors in rats. The toxicity of dioxins is characterized by large sensitivity differences among animal species and even strains of the same species, which complicates the risk assessment. The significance of these differences in dioxin-induced carcinogenicity is not known. We therefore studied the liver tumor-promoting activity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in the sensitive Long-Evans (L-E) and the resistant Han/Wistar (H/W) rats differing >1000-fold in their sensitivity to the acute lethaity of TCDD. Female rats were partially hepatectomized, initiated with nitrosodiethylamine, and treated with TCDD for 20 weeks. Altered hepatic foci (AHF) were stereologically quantitated using glutathione S-transferase P as a marker. AHF were significantly (P < 0.001) and dose dependently increased in L-E rats at 10 and 100 ng/kg/day, but in H/W rats only at 1000 ng/kg/day and above, indicating a remarkable (approximately 100-fold) sensitivity difference between L-E and H/W rats. The same sensitivity difference but 10-fold less foci were observed between nonhepatectomized/noninitiated L-E and H/W rats. Induction of AHF was related to hepatotoxicity but not to cytochrome P4501A1 activity in the liver. Liver TCDD concentrations were similar in both strains. H/W rats are exceptionally resistant to induction of AHF by TCDD, and the resistance is associated with an altered transactivation domain of the aryl hydrocarbon receptor. Genetic differences may account for significant interindividual/intraspecies sensitivity differences in dioxin-induced carcinogenesis. Understanding the role of transactivation domain of the aryl hydrocarbon receptor in carcinogenesis is therefore likely to improve dioxin risk assessment.
Assuntos
Carcinógenos , Resistencia a Medicamentos Antineoplásicos , Neoplasias Hepáticas/induzido quimicamente , Dibenzodioxinas Policloradas/farmacologia , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Peso Corporal/efeitos dos fármacos , Citocromo P-450 CYP1A1/metabolismo , Dietilnitrosamina/farmacologia , Relação Dose-Resposta a Droga , Eritrócitos/metabolismo , Feminino , Glutationa Transferase/metabolismo , Fígado/efeitos dos fármacos , Fígado/patologia , Neoplasias Hepáticas/patologia , Testes para Micronúcleos , Tamanho do Órgão/efeitos dos fármacos , Estrutura Terciária de Proteína , Ratos , Ratos Long-Evans , Ratos Wistar , Receptores de Hidrocarboneto Arílico/química , Receptores de Hidrocarboneto Arílico/genética , Ativação Transcricional , gama-Glutamiltransferase/sangueRESUMO
Reduced gluconeogenesis due to decreased activity of key gluconeogenic enzymes in liver, together with feed refusal, has been suggested to play an important role in 2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD)-induced lethality in rats. This study was carried out to further analyse the toxicological significance of reduced gluconeogenesis by comparing dose-responses and time-courses of effects of TCDD on the activity of phosphoenolpyruvate carboxykinase (PEPCK) in liver, liver glycogen concentration as well as plasma concentrations of glucose and amino acids in both genders of TCDD-sensitive Long-Evans (L-E) rats and TCDD-resistant Han/Wistar (H/W) rats. A dose-dependent decrease in PEPCK activity was observed in H/W rats, but in L-E rats the activity was not decreased. However, TCDD impaired the strong increase in liver PEPCK activity observed in pair-fed controls of the L-E strain. Liver glycogen concentrations were severely decreased in L-E rats and moderately in H/W rats. This effect seems to be secondary to reduced feed intake, since a similar decrease was seen in pair-fed controls. Decreases in plasma glucose concentrations were also more profound in L-E rats than in H/W rats, but pair-fed controls were generally less affected. Circulating concentrations of amino acids were markedly increased in TCDD-treated L-E rats, which is likely to reflect increased mobilization of amino acids and their decreased metabolism in liver. Reduction of liver PEPCK activity cannot account for the sensitivity difference of these two strains of rats in terms of mortality. Nevertheless, the response of both strains of TCDD-treated rats regarding gluconeogenesis is different from that seen in pair-fed controls and suggesting that impairment of this pathway contributes to the development of the wasting syndrome.
Assuntos
Aminoácidos/sangue , Glicemia/metabolismo , Poluentes Ambientais/toxicidade , Fígado/enzimologia , Fosfoenolpiruvato Carboxiquinase (GTP)/metabolismo , Dibenzodioxinas Policloradas/toxicidade , Animais , Peso Corporal/efeitos dos fármacos , Citocromo P-450 CYP1A1/metabolismo , Resistência a Medicamentos , Ativação Enzimática/efeitos dos fármacos , Feminino , Gluconeogênese/efeitos dos fármacos , Homeostase/efeitos dos fármacos , Fígado/anatomia & histologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Glicogênio Hepático/metabolismo , Masculino , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Long-Evans , Ratos Wistar , Ureia/sangueRESUMO
Interactions of diet and diet-induced obesity, and the characteristic wasting syndrome caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were studied in TCDD-resistant Han/Wistar and TCDD-sensitive Long-Evans rats. The rats were made obese by feeding them either a high-energy diet (consisting of chocolate, cheese, and chow) or force feeding. TCDD reduced body weight in a parallel manner in lean and obese rats. The high-energy diet diminished the body weight loss and increased the survival time in L-E rats after a lethal dose of TCDD, while energy supplement with high-fat/low-protein food had an opposite effect. In conclusion, diet-induced obesity and TCDD had additive effects on body weight. Dietary manipulations were able to modify the weight loss and survival time after TCDD. Fat seems to have a negative impact, while carbohydrate or protein may have a positive impact in this respect. The results are in agreement with a view that TCDD-exposed rats have a negative fat balance favoring fat loss.
Assuntos
Anorexia/induzido quimicamente , Dieta , Obesidade/patologia , Dibenzodioxinas Policloradas/toxicidade , Síndrome de Emaciação/induzido quimicamente , Animais , Anorexia/psicologia , Análise Química do Sangue , Metabolismo Energético/efeitos dos fármacos , Intubação Gastrointestinal , Ratos , Ratos Long-Evans , Ratos Wistar , Especificidade da Espécie , Análise de Sobrevida , Redução de Peso/efeitos dos fármacosRESUMO
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD),12 the most toxic congener of dioxins, exhibits wide sensitivity differences between a sensitive Long-Evans (L-E) rat and a resistant Han/Wistar (H/W) rat. The sensitivity is determined probably by two autosomal genes and it is highly end point dependent. The difference is more than 1000-fold for acute toxicity and negligible for CYP1A1 induction. The rat strains were recently shown to have differences in the size of AH receptor (AHR), which mediates most effects of TCDD. In the present study, the rat strains were crossed and the resistant alleles of genes determining TCDD sensitivity were segregated to new rat lines. Selection was based on AHR phenotype determined by Western blot and resistance to TCDD lethality. Two genes determining resistance were found: the Ahr and a novel gene designated "B." In homozygous rats, the H/W type Ahrhw allele prevented TCDD lethality up to 2000 microg/kg or more, and the H/W type "Bhw" allele also increased resistance to TCDD lethality but to a lesser extent. Heterozygous rats were only slightly more resistant to acute lethality than the respective sensitive homozygous rats. CYP1A1 induction was similar irrespective of the Ahr and "B" genotypes, but a substantial increase in serum bilirubin seen after low doses in sensitive rats occurred only after large doses in "Bhw/hw" and not at all in Ahrhw/hw rats. In conclusion, the Ahrhw allele is a major determinant of the exceptional resistance of H/W rats to TCDD lethality. There is also an additional gene, whose function remains to be characterized, conferring limited resistance to TCDD toxicity. These two H/W rat-derived alleles are separately expressed in the new rat lines created.
Assuntos
Alelos , Dibenzodioxinas Policloradas/toxicidade , Receptores de Hidrocarboneto Arílico/genética , Receptores de Hidrocarboneto Arílico/fisiologia , Animais , Citocromo P-450 CYP1A1/metabolismo , Resistência a Medicamentos/genética , Feminino , Genes Dominantes , Masculino , Ratos , Ratos Long-Evans , Ratos WistarRESUMO
Long-Evans rats (strain Turku AB; L-E) are at least 1000-fold more sensitive (LD50 about 10 microg/kg) to the acute lethal effects of 2, 3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) than are Han/Wistar (Kuopio; H/W) rats (LD50 > 9600 microg/kg). The AH receptor (AHR) is believed to mediate the toxic effects of TCDD and related halogenated aromatic hydrocarbons. We compared the AHRs of L-E and H/W rats to determine if there were any structural or functional receptor differences that might be related to the dramatic difference in the sensitivity of these two strains to the lethal effects of TCDD. Cytosols from liver and lung of the sensitive L-E rats contained about twofold higher levels of specific binding sites for [3H]TCDD than occurred in H/W rats; the Kd for binding of [3H]TCDD to AHR in hepatic cytosols was similar between the two strains. Addition of the oxyanions, molybdate or tungstate (20 mM), had little effect upon ligand binding to AHR in hepatic cytosols from L-E rats whereas in cytosols from H/W rats these agents substantially diminished or totally abolished TCDD binding. The AHR in H/W cytosols also lost ligand-binding function when NaCl (20 to 400 mM) was added to the buffer whereas, in cytosols from L-E rats, the addition of 400 mM NaCl caused the receptor complex to shift from 9S to 6S during velocity sedimentation but did not destroy ligand binding function. AHR from hepatic cytosol of both the L-E and H/W rats could be transformed to the DNA-binding state in the presence of TCDD or other dioxin congeners as assessed by gel mobility shift assays. The most dramatic difference in AHR properties between L-E and H/W rats is molecular mass. Immunoblotting of cytosolic proteins revealed that the AHR in L-E rats has an apparent mass of approximately 106 kDa, similar to the mass of the receptor previously reported in several other common laboratory rat strains. In contrast, the mass of the AHR in H/W rats is approximately 98 kDa, significantly smaller than the mass of receptor reported in any other rat strains. F1 offspring of a cross between L-E and H/W rats expressed both the 106- and the 98-kDa protein. There was no apparent difference in the mass of the AHR nuclear translocator protein (ARNT) between the two strains, but the hepatic concentration of ARNT was about three times as high in L-E as in H/W rats. It will be interesting to find out how the altered structure of the AHR in H/W rats is related to their remarkable resistance to the lethal effects of TCDD.
Assuntos
Proteínas de Ligação a DNA , Dibenzodioxinas Policloradas/toxicidade , Receptores de Hidrocarboneto Arílico/química , Animais , Translocador Nuclear Receptor Aril Hidrocarboneto , Citocromo P-450 CYP1A1/biossíntese , DNA/metabolismo , Resistência a Medicamentos , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Peso Molecular , Ratos , Ratos Long-Evans , Ratos Sprague-Dawley , Ratos Wistar , Receptores de Hidrocarboneto Arílico/metabolismo , Cloreto de Sódio/farmacologia , Especificidade da Espécie , Fatores de Transcrição/químicaRESUMO
Polychlorinated dioxins (PCDD) are widespread environmental contaminants. The most potent and the general model compound for dioxins is 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Our laboratory has developed a new model for studies of dioxin toxicity based on totally disparate sensitivity to the lethal action of TCDD between Long-Evans (L-E, Turku AB; LD50 ca. 10 micrograms/kg) and Han/Wistar (H/W, Kuopio; LD50 over 10,000 micrograms/kg) rat strains. We have shown that body weight regulation is differentially regulated by TCDD in these rat strains: body weight gain is permanently reduced in the sensitive L-E but not in the resistant H/W strain. In concert with reduced body weight, TCDD increased brain TRP concentration, 5-HT synthesis and its metabolism to 5-HIAA at lethal doses in TCDD-susceptible L-E rats, and almost not at all in resistant H/W rats in which lethal dose levels were not reached. Further studies showed that TCDD indirectly increases free TRP concentration in the circulation in TCDD-susceptible L-E rats. Blood free fatty acids seem to be involved in the latter phenomenon. It is not likely that the enhanced serotonergic tone in the CNS is a causative factor in TCDD-induced anorexia. However, the present results may open up an interesting avenue to better understand physiology of TRP and the complex regulation of energy balance.
Assuntos
Encéfalo/metabolismo , Dioxinas/farmacologia , Dibenzodioxinas Policloradas/toxicidade , Triptofano/metabolismo , Animais , Animais de Laboratório , Peso Corporal/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Homeostase/efeitos dos fármacos , Ácido Hidroxi-Indolacético/metabolismo , Ratos , Ratos Long-Evans , Ratos Wistar , Serotonina/metabolismoRESUMO
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is the most potent dioxin. There are exceptionally wide inter- and intraspecies differences in sensitivity to TCDD toxicity with Han/Wistar (H/W) (Kuopio) rats being the most resistant mammals tested. A peculiar feature of H/W rats is that despite their unresponsiveness to the acute lethality of TCDD, their sensitivity to other biological impacts of TCDD (e.g., CYP1A1 induction) is preserved. The biological effects of TCDD are mediated by the aryl hydrocarbon receptor (AhR). We recently found that the AhR of H/W rats (about 98 kDa) is smaller than the receptor in other rat strains (106 kDa). In the present study, molecular cloning and sequencing of the H/W rat AhR revealed that the reason for its smaller size is a deletion/insertion-type change at the 3' end of exon 10 in the receptor cDNA. This change emanates from a single point mutation at the first nucleotide of intron 10, resulting in altered mRNA splicing. At the protein level, the mutation leads to a total loss of either 43 or 38 amino acids (with altered sequence for the last seven amino acids in the latter case) toward the carboxyl-terminal end in the trans-activation domain of the AhR. H/W rats also harbor a point mutation in exon 10 that will cause a Val-to-Ala substitution in codon 497, but this occurs in a variable region of the AhR. These findings suggest that there is a relatively small region in the AhR trans-activation domain that may be capable of providing selectivity to its function.
Assuntos
Íntrons/genética , Mutação Puntual/genética , Receptores de Hidrocarboneto Arílico/genética , Animais , Sequência de Bases , Feminino , Masculino , Dados de Sequência Molecular , Dibenzodioxinas Policloradas/farmacologia , Reação em Cadeia da Polimerase , Conformação Proteica , Ratos , Ratos Sprague-Dawley , Receptores de Hidrocarboneto Arílico/química , Alinhamento de SequênciaRESUMO
We have previously shown that dioxins at prevailing levels in mothers' milk may cause mineralization defects in the developing teeth of their children. Developmental dental defects have also been reported in rhesus macaques and rats experimentally exposed to dioxin. The most toxic dioxin congener, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), is a potent modulator of epithelial cell growth and differentiation. To clarify whether epidermal growth factor receptor (EGFR), implicated in the mediation of the developmental toxicity of TCDD, is involved in dental toxicity, we cultured embryonic molar teeth from EGFR-deficient mice with TCDD, epidermal growth factor (EGF), and both agents in combination. In teeth of the normal embryos, TCDD caused depolarization of odontoblasts and ameloblasts. Consequently, the dentin matrix failed to undergo mineralization, the enamel matrix was not deposited, and cuspal morphology was disrupted. In teeth of the null mutant embryos, only the cuspal contour was mildly modified. EGF alone retarded the molar tooth development of normal embryos, but not that of EGFR-deficient embryos. When coadministered with TCDD, EGF for the most part prevented the adverse effects of TCDD on teeth of the normal embryos. These results show that the interference of TCDD with mouse molar tooth development in vitro involves EGFR signaling. Thus, EGFR may also play a role in the developmental defects that dioxins cause in human teeth. Because EGFR is widely expressed in developing organs, EGFR signaling may even be of general relevance in the mediation of the developmental toxicity of TCDD.
Assuntos
Receptores ErbB/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Odontogênese/efeitos dos fármacos , Dibenzodioxinas Policloradas/toxicidade , Germe de Dente/efeitos dos fármacos , Animais , Criança , Desenvolvimento Embrionário e Fetal , Receptores ErbB/deficiência , Receptores ErbB/genética , Homozigoto , Humanos , Mandíbula , Camundongos , Camundongos Knockout , Morfogênese/efeitos dos fármacos , Técnicas de Cultura de Órgãos , Ratos , Valores de Referência , Pele/efeitos dos fármacos , Pele/embriologia , Pele/patologia , Glândula Submandibular/efeitos dos fármacos , Glândula Submandibular/embriologia , Glândula Submandibular/patologia , Germe de Dente/patologia , Germe de Dente/fisiologia , Vibrissas/efeitos dos fármacos , Vibrissas/embriologia , Vibrissas/patologiaRESUMO
We compared the effects of 2,3,7,8-tetra (TCDD), 1,2,3,7,8-penta (PeCDD), 1,2,3,4,7,8-hexa (HxCDD) and 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD) on brain serotonin metabolism, plasma tryptophan and liver tryptophan pyrrolase activity in two rat strains, TCDD-sensitive Long-Evans (Turku AB; L-E) and TCDD-resistant Han/Wistar (Kuopio; H/W). Previously it was shown that L-E rats exhibit the expected rank order of potency for CDDs in terms of acute toxicity with TCDD being the most potent, followed by PeCDD, HxCDD and HpCDD. In contrast, to H/W rats HxCDD was the most toxic and TCDD the least toxic of these congeners. In the present study, the CDDs decreased body weight in L-E rats in the following order of potency: TCDD > PeCDD > HxCDD > HpCDD. The same rank order was recorded for elevations in brain tryptophan and plasma free tryptophan concentrations as well as for inhibition of the main hepatic tryptophan metabolizing enzyme, tryptophan pyrrolase. By contrast, in H/W rats HxCDD was the most effective congener in producing loss of body weight, followed by HpCDD, PeCDD and TCDD. This was also true of changes in tryptophan homeostasis. These findings imply that in TCDD-susceptible L-E and TCDD-resistant H/W rats the potency of dioxin congeners in inducing acute toxicity highly correlates with their ability to disrupt tryptophan homeostasis. However, there may not be a direct causal relationship between body weight loss and altered tryptophan homeostasis, because the magnitudes of these two phenomena were not consistently parallel across the dioxin congeners tested.
Assuntos
Dibenzodioxinas Policloradas/toxicidade , Triptofano/metabolismo , Redução de Peso/efeitos dos fármacos , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Resistência a Medicamentos , Feminino , Homeostase/efeitos dos fármacos , Dibenzodioxinas Policloradas/análogos & derivados , Ratos , Ratos Long-Evans , Ratos Wistar , Especificidade da Espécie , Testes de ToxicidadeRESUMO
We have previously shown that the prototype for halogenated aromatic hydrocarbons, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), diminishes serum melatonin concentration at the same dose in both the most TCDD-susceptible (Long-Evans, Turku AB; L-E) and the most TCDD-resistant (Han/Wistar, Kuopio; H/W) rat strain. The change developed within 24 h and persisted for at least 28 days after TCDD exposure; was independent of the time of day and was not associated with any morphological damage to the pineal gland. In the present study, we investigated the mechanism of this endocrine effect. Despite a 40-50% decrease in circulating melatonin levels, the pineal content of melatonin, serotonin and 5-hydroxyindole acetic acid remained unaltered and the rate-limiting enzyme of pineal melatonin biosynthesis, N-acetyltransferase, displayed only a relatively minor suppression in activity (30%) in TCDD-treated L-E rats. Likewise, TSDD did not influence the ability of pineal glands from L-E rats to synthesize and secrete melatonin in ex vivo or in vitro experiments. TCDD accelerated the disappearance of exogenous melatonin from the serum in both rat strains. This enhancement probably did not originate in the liver, because liver perfusion studies revealed that even control rat livers were capable of total melatonin clearance in spite of the fact that the melatonin concentration far exceeded physiological levels. Urine excretion of the normal main metabolite of melatonin, 6-hydroxymelatoninsulfate, was reduced by TCDD treatment in both strains. This was accompanied by an altered HPLC pattern of metabolites, especially in H/W rats. We conclude that TCDD decreases serum melatonin levels in rats by enhancing the peripheral, evidently extrahepatic, metabolism of the hormone.