metallothionein and Abnormalities--Drug-Induced

We have previously demonstrated that ethanol teratogenicity is associated with metallothionein-induced fetal zinc (Zn) deficiency, and that maternal subcutaneous Zn treatment given with ethanol in early pregnancy prevents fetal abnormalities and spatial memory impairments in mice. Here we investigated whether dietary Zn supplementation throughout pregnancy can also prevent ethanol-related dysmorphology.. Pregnant mice were injected with saline or 25% ethanol (0.015 ml/g intraperitoneally at 0 and 4 hours) on gestational day (GD) 8 and fed either a control (35 mg Zn/kg) or a Zn-supplemented diet (200 mg Zn/kg) from GD 0 to 18. Fetuses from the saline, saline + Zn, ethanol and ethanol + Zn groups were assessed for external birth abnormalities on GD 18. In a separate cohort of mice, postnatal growth and survival of offspring from these treatment groups were examined from birth until postnatal day 60.. Fetuses from dams treated with ethanol alone in early pregnancy had a significantly greater incidence of physical abnormalities (26%) compared to those from the saline (10%), saline + Zn (9%), or ethanol + Zn (12%) groups. The incidence of abnormalities in ethanol + Zn-supplemented fetuses was not different from saline-treated fetuses. While ethanol exposure did not affect the number of fetal resorptions or pre- or postnatal weight, there were more stillbirths with ethanol alone, and cumulative postnatal mortality was significantly higher in offspring exposed to ethanol alone (35% deaths) compared to all other treatment groups (13.5 to 20.5% deaths). Mice supplemented with Zn throughout pregnancy had higher plasma Zn concentrations than those in un-supplemented groups.. These findings demonstrate that dietary Zn supplementation throughout pregnancy ameliorates dysmorphology and postnatal mortality caused by ethanol exposure in early pregnancy.

Topics: Abnormalities, Drug-Induced; Alcoholism; Animals; Central Nervous System Depressants; Dietary Supplements; Disease Models, Animal; Embryonic Development; Ethanol; Female; Liver; Male; Metallothionein; Mice; Mice, Inbred C57BL; Pregnancy; Pregnancy Outcome; Pregnancy, Animal; Prenatal Exposure Delayed Effects; Sodium Chloride; Zinc

During pregnancy, exposure to lipopolysaccharide (LPS) can lead to abortion, preterm delivery, and teratogenicity. The mechanisms underlying these effects are unclear. Both LPS and ethanol are potent inducers of liver metallothionein (MT), a key Zn binding protein. The teratogenic effects of ethanol have been linked to MT-induced changes in maternal-fetal Zn homeostasis, leading tofetal deficiency. This study was designed to assess whether the teratogenic effects of LPS are also related to MT induction and changes in Zn homeostasis.. Non-pregnant normal (MT +/+) and MT-null (MT -/-) mice were injected subcutaneously with 0.5 microg/gm LPS and killed over 48 hr. In MT +/+ mice, liver MT concentrations were elevated from 6 hr, and were maximal at 24 hr (30-fold basal), whereas liver Zn levels were also increased from 6 hr. Plasma Zn concentrations decreased by 80% at 6 hr, and were below normal between 6 and 24 hr. In MT -/- mice, plasma Zn levels were increased from basal between 6 and 16 hr. Dams were injected with LPS, saline, or LPS and ZnSO4 (2 microg/gm, MT +/+ only) on Day 8 of gestation (GDS), killed on GD18, and the fetuses examined for malformations.. External abnormalities were most prevalent in fetuses from MT +/+ dams exposed to LPS, where 34% of fetuses in each litterwere affected. MT +/+ dams treated with LPS and ZnSO4, and MT -/- dams treated with LPS had litters in which 5.4 and 4.8% of fetuses were abnormal respectively.. The findings of this study strongly support the hypothesis that LPS teratogenicity is mediated at least in part by MT-induced changes in maternal Zn homeostasis,which compromises fetal Zn supply.

Topics: Abnormalities, Drug-Induced; Animals; Escherichia coli; Female; Lipopolysaccharides; Liver; Male; Metallothionein; Mice; Mice, Inbred C57BL; Mice, Knockout; Pregnancy; Zinc

Since the level of platinum in the environment is destined to increase, because of its use in vehicle catalytic converters, the toxicity of platinum needs further investigation. In this study, the frog embryo teratogenesis assay-Xenopus (FETAX) was used to compare the embryotoxicity and teratogenicity of two common platinum species, (NH4)2PtCl4 and (NH4)2PtCl6. The uptake rates of the two platinum species were studied, and also their effects on the expression of genes encoding metallothionein and heat-shock protein 70, which are known to be induced by several stress factors. In addition, the differential display technique was used to search for genes that were specifically induced by platinum. A gene for the type I collagen alpha-chain and a novel gene were identified.

Topics: Abnormalities, Drug-Induced; Animals; Base Sequence; Blotting, Northern; Body Weights and Measures; Collagen Type I; DNA Primers; DNA, Complementary; Embryo, Nonmammalian; Gene Expression Profiling; Gene Expression Regulation; HSP70 Heat-Shock Proteins; Metallothionein; Molecular Sequence Data; Platinum; Reverse Transcriptase Polymerase Chain Reaction; Sequence Analysis, DNA; Survival Analysis; Xenopus laevis

Butyl benzyl phthalate (BBP) has been shown to be teratogenic. One mechanism contributing to the teratogenicity of several developmental toxicants, is chemical-induced changes in maternal zinc (Zn) metabolism which result in an increased synthesis of maternal liver metallothionein (Mt), and a subsequent reduction in Zn delivery to the conceptus. We investigated the effects of maternal BBP exposure on maternal-fetal Zn metabolism in Wistar rats. In study I, dams were gavaged with BBP (0,250,1000,1500 or 2000 mg/kg) on gestation days (GD) 11 through 13, and killed on GD 20. Maternal toxicity was evident in the three highest dose groups. Embryo/fetal death and small pup weights and lengths were noted in the 2000 mg BBP/kg group. Fetuses in the 1500 and 2000 mg/kg groups were characterized by poor skeletal ossification, and a high frequency of cleft palate. Rib anomalies were observed in the three highest dose groups. Maternal liver Mt concentrations were only slightly elevated in the 1500 and 2000 mg/kg groups. In study II, dams treated as above, were gavaged with 65Zn and killed 18 h later. While the 2000 mg/kg group had high percentages of 65Zn in some maternal tissues, sequestration of 65Zn in maternal liver was not evident. Thus, BBP is not a strong inducer of Mt, and the teratogenicity of BBP does not appear to be due to alterations in maternal and/or embryonic Zn metabolism.

Topics: Abnormalities, Drug-Induced; Animals; Body Weight; Calcification, Physiologic; Eating; Female; Metallothionein; Phthalic Acids; Plasticizers; Pregnancy; Rats; Rats, Wistar; Reproduction; Teratogens; Tissue Distribution; Trace Elements; Zinc; Zinc Radioisotopes

Isotretinoin (ITR), a teratogen in many species, is associated with increased oxidative stress. Metallothionein (MT) is an important tissue antioxidant whose concentrations are induced by zinc. To study the role of supplemental Zn as an inducer of embryonic MT, we injected pregnant CD-1 mice subcutaneously with saline vehicle, or 20 or 40 mg/kg Zn on gestational day (GD) 6.5. After 48 h, embryonic MT concentrations increased in a dose-related manner (r = 0.64, P < 0.05) with Zn treatment. The possible protective role of Zn pretreatment against ITR teratogenicity was investigated in vivo and in vitro. CD-1 mice were pretreated with saline or Zn (20 and 40 mg/kg) on GD 8.5 and 9.5. ITR was administered to both groups of mice via three intragastric intubations of 100 mg ITR/kg at 4 h intervals on GD 10.5. On GD 18.5, Zn pre-treated mice demonstrated decreased ITR-mediated growth retardation, cleft palates and postpartum mortality. A reduction in embryonic MT concentrations was observed in mice exposed to ITR. Mouse embryos cultured on GD 8.5 with an addition of 15 micromol/L Zn for 48 h had a sixfold greater MT concentration (688 microg/g protein) than controls. The Zn pretreatment of cultured embryos prevented malformations and lessened growth retardation caused by 24 h exposure to 17 micromol/L ITR. These results suggest that Zn-mediated induction of MT in mouse embryos could protect against ITR teratogenicity.

Topics: Abnormalities, Drug-Induced; Animals; Culture Techniques; Embryo, Mammalian; Female; Fetus; Gestational Age; Isotretinoin; Metallothionein; Mice; Pregnancy; Yolk Sac; Zinc

Xenopus laevis embryos were analyzed for metallothionein by silver-saturation assay and metallothionein-mRNA by reverse transcriptase/polymerase chain reaction following exposures to the following metal chlorides at levels that caused > 95% malformations and < 7% mortality: Zn2+ (300 microM); Cd2+ (18 microM); Ni2+ (56 microM); Co2+ (1,800 microM); and Cu2+ (5.6 microM). At the beginning of the exposure (stages 8), metallothionein-mRNA and metallothionein levels averaged 2.0 x 10(6) copies/embryo and 19 pmol/embryo, respectively. In control embryos at stages 26, 36, 42, and 46, metallothionein-mRNA content averaged 9, 37, 104, and 97 copies x 10(6)/embryo, and metallothionein content averaged 6, 11, 15, and 18 pmol/embryo. In Zn(2+) -exposed embryos at the same stages, metallothionein-mRNA content averaged 116*, 11,400*, 3,210*, and 14 copies x 10(6)/embryo and metallothionein content averaged 10, 18*, 46*, and 90* pmol/embryo; in Cd(2+)-exposed embryos, metallothionein-mRNA content averaged 22, 7,170*, 1,783*, and 240 copies x 10(6)/embryo and metallothionein content averaged 8, 14, 33*, and 56* pmol/embryo, respectively (*P < 0.05 versus controls). Exposure-response curves (Cd2+, 1-18 microM; Zn2+, 3-300 microM) indicated that Cd2+ was 3- to 5-times more potent than Zn2+, based on metallothionein-mRNA response at stage 36 and metallothionein response at stage 46. In Ni(2+)-, Co(2+)-, or Cu(2+)-exposed embryos, metallothionein-mRNA and metallothionein contents did not differ significantly from controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Abnormalities, Drug-Induced; Animals; Base Sequence; Cadmium; Cobalt; Copper; Embryo, Nonmammalian; Female; Male; Metallothionein; Metals; Molecular Sequence Data; Nickel; RNA, Messenger; Teratogens; Xenopus laevis; Zinc

We tested the hypothesis that tumor necrosis factor-alpha (TNF-alpha) would be teratogenic in mice due in part to its effects on zinc metabolism. In Experiment 1, nonpregnant mice were injected with a single dose of TNF-alpha (40,000 U) or PBS and then received a 65Zn-labeled meal. Mice killed 10 h after TNF-alpha treatment had high liver 65Zn and low plasma 65Zn, compared with controls. In Experiment 2, gestation day 8 (GD 8) mice were injected with PBS or TNF-alpha and then received a 65Zn-labeled meal. Dams killed 10 h after TNF-alpha treatment had higher liver and kidney 65Zn and lower plasma and embryonic 65Zn accumulation than controls. In Experiment 3, TNF-alpha dosing from GD 7-12 was associated with high maternal liver Zn and metallothionein concentrations on GD 13 and a high frequency of exencephaly on GD 18. In Experiment 4, dams fed diets containing 4.5, 12.5 or 50.0 micrograms Zn/g were given PBS or TNF-alpha on GD 7-12. Gross fetal defects were not observed in the PBS-treated litters evaluated on GD 18. In contrast, TNF-alpha-treated litters were characterized by multiple defects, with the incidence and severity being highest in the low Zn diet group. In Experiment 5, embryos cultured in serum from TNF-alpha-treated animals exhibited a high frequency of defects; the developmental toxicity of this serum was ameliorated when it was supplemented with Zn. Thus, the developmental toxicity of TNF-alpha is due in part to its influence on Zn metabolism.

Topics: Abnormalities, Drug-Induced; Animals; Embryo, Mammalian; Embryonic and Fetal Development; Female; Fetus; Kidney; Liver; Metallothionein; Mice; Mice, Inbred C3H; Pregnancy; Tumor Necrosis Factor-alpha; Zinc; Zinc Radioisotopes

We hypothesize that maternal metallothionein (MT) induction by toxic dosages of chemicals may contribute to or cause developmental toxicity by a chain of events leading to a transient but developmentally adverse decrease in Zn availability to the embryo. This hypothesis was tested by evaluating hepatic MT induction, maternal and embryonic Zn status, and developmental toxicity after exposure to urethane, a developmental toxicant, or styrene, which is not a developmental toxicant. Pregnant Sprague-Dawley rats were given 0 or 1 g/kg urethane ip, or 0 or 300 mg/kg styrene in corn oil po, on Gestation Day 11 (sperm positive = Gestation Day 0). These were maternally toxic dosages. As both treatments decreased food consumption, separate pair-fed control groups were also evaluated for effects on MT and Zn status and development. In addition, Gestation Day 11 rat embryos were exposed to urethane in vitro in order to determine whether urethane has the potential to be directly embryotoxic. Urethane treatment induced hepatic MT 14-fold over control; styrene treatment induced MT 2.5-fold. The MT induction by styrene could be attributed to decreased food intake, as a similar level of induction was observed in a pair-fed untreated control group. However, the level of MT induction by urethane was much greater than that produced by decreased food intake alone. Hepatic Zn concentration, particularly in the cytosol, was increased in the presence of increased hepatic MT concentration. Plasma Zn concentration was significantly decreased (approximately 30%) by urethane treatment, but not by styrene or food restriction (pair-feeding). Distribution of 65Zn to the liver of urethane-treated dams was significantly greater (by 30%), while distribution to embryonic tissues was significantly lower (by at least 50%) than in pair-fed or ad lib.-fed controls. Styrene treatment had no effect on 65Zn distribution. Urethane was developmentally toxic, causing an 18% decrease in fetal weight and a significant delay in skeletal ossification, but was not toxic to rat embryos in vitro. Styrene was not developmentally toxic. The changes observed after urethane treatment, namely substantial hepatic MT induction and altered maternal and embryonic Zn status, along with the lack of direct embryotoxicity of urethane in vitro, support the hypothesis that these maternal effects contribute to developmental toxicity. The lack of similar changes in styrene-intoxicated dams provides one explanation for its

Topics: Abnormalities, Drug-Induced; Animals; Embryo, Mammalian; Female; Fetus; Liver; Maternal-Fetal Exchange; Metallothionein; Pregnancy; Pregnancy, Animal; Rats; Styrene; Styrenes; Tissue Distribution; Urethane; Zinc; Zinc Radioisotopes

As an approach toward understanding the mechanisms by which cadmium (Cd) exerts its teratogenic effects, the expression and metal regulation of the metallothionein (MT) genes in midgestation mouse embryos were studied by Northern blot and in situ hybridization. Maternal injection of a teratogenic dosage of Cd (50 mumol Cd/kg body wt) did not induce MT mRNA in day 10 (D10) CD-1 mouse embryos, whereas zinc (Zn) (50 mumol/kg was an effective inducer. In contrast, Cd was about 10-fold more potent than Zn at rapidly inducing MT mRNA in D10 embryos incubated in vitro in medium containing micromolar concentrations of these metals. This suggests that following maternal injection, Cd but not Zn is prevented from reaching the D10 embryo and establishes that the embryonic MT genes are not refractory to metal induction, which might have explained the sensitivity of the embryo to Cd. MT mRNA was detected at high levels only in the extraembryonic membranes of D9 embryos exposed to Cd in vivo. On days 9 and 10, no embryonic cell types contained detectable levels of MT mRNA. This mRNA was detected first at low levels in hepatocytes on D11, soon after formation of liver and these levels increased dramatically by D12. Therefore, Cd teratogenicity was not associated with high levels of cell type-specific expression of the MT genes in Cd-sensitive regions of the embryo (neural tube, limb bud), that might have served to target Cd to these cells. Taken together, the results of this study suggest that Cd teratogenicity reflects damage to maternal or extraembryonic tissues. However, the results cannot exclude the possibility that certain cells in the embryo are exceptionally sensitive to low levels of Cd.

Topics: Abnormalities, Drug-Induced; Animals; Blotting, Northern; Cadmium; Dose-Response Relationship, Drug; Embryo, Mammalian; Female; Gene Expression; Metallothionein; Mice; Nucleic Acid Hybridization; Pregnancy; RNA Probes; RNA, Messenger

The participation of maternal hepatic metallothionein (MT) in the amelioration of cadmium teratogenicity in mice was examined. Pretreatment with bismuth nitrate (subcutaneously) ameliorated the teratogenicity, including exencephaly and abnormalities of the axial skeleton, caused by a single intraperitoneal injection of cadmium sulfate. Pretreatment with bismuth nitrate for 3 days induced MT drastically in maternal liver and kidney. Six and 24 hr after the injection of cadmium sulfate, the accumulation of maternal hepatic cadmium increased and that in the decidua, including embryos, decreased after pretreatment with bismuth nitrate. Mouse embryos on day 7 of gestation were cultured for 48 hr. Exposure to cadmium sulfate in vitro induced unfused brain fold, which corresponds to exencephaly in vivo. From the in vitro experiment, it was suggested that the teratogenicity of cadmium on day 7 of gestation is a direct action against the mouse embryo. In the present experiment it was suggested that pretreatment with bismuth nitrate induced maternal hepatic and renal MT; cadmium was therefore trapped and detoxicated, and consequently embryos were exposed to a lower concentration of cadmium.

Topics: Abnormalities, Drug-Induced; Animals; Bismuth; Cadmium; Dose-Response Relationship, Drug; Embryonic and Fetal Development; Female; Metallothionein; Mice; Mice, Inbred ICR; Pregnancy

The teratogenic effect of cadmium can be diminished by a number of mechanisms including zinc and pretreatment with cadmium and mercury. In this study, the oral administration of alpha-mercapto-beta-(2-furyl)-acrylic acid (MFA) protects against cadmium-induced malformations and embryonic death. This protection is probably mediated by the chelation of the cadmium ion rather than metallothionein (MT) synthesis.

Topics: Abnormalities, Drug-Induced; Acrylates; Animals; Cadmium; Cricetinae; Embryo Implantation; Female; Fetal Resorption; Kidney; Liver; Metallothionein; Pregnancy; Teratogens

The teratogenicity of copper deficiency is well known, but underlying mechanisms have not been delineated. One method of studying the biochemical lesions of copper deficiency is the use of chelating drugs with different chemical characteristics. The teratogenicity of a copper deficient diet and of diets containing either D-penicillamine or triethylenetetramine is quite different, although all three diets result in decreased fetal liver copper levels. Feeding D-penicillamine can result in decreased fetal liver zinc, while feeding triethylenetetramine can result in increased fetal liver zinc. The effect of these three diets on fetal liver copper and zinc molecular localization was determined. Gel filtration showed that fetal liver copper and zinc in controls was localized in 3 fractions with MWs of greater than 50,000 (H), 30,000 (I) and 8-10,000 (L). Independent of dietary treatment, as liver copper diminished, copper was missing first from the L peak, then the I peak and with severe deficiency, from the H peak. Drug induced increases and decreases in fetal liver zinc were reflected in the L peak. These data suggest that the absolute levels of copper in the liver of the term fetus determines the distribution of the element among its binding ligands.

Topics: Abnormalities, Drug-Induced; Animals; Chromatography, Gel; Copper; Ethylenediamines; Female; Liver; Maternal-Fetal Exchange; Metallothionein; Penicillamine; Pregnancy; Rats; Rats, Inbred Strains; Superoxide Dismutase; Trientine; Zinc