Cadmium accumulation and metallothionein-like proteins in the sea starAsterias rubens

Sea stars were exposed for four months to cadmium at 50 g Cd/L. The cadmium content of stomachs, pyloric caeca, gonads and body wall was determined after 2, 4, 6, 8, and 16 weeks of exposure. The highest accumulation rates were found for body wall and pyloric caeca. The lowest accumulation rate was found for the gonads. In all tissues cadmium accumulation showed saturation kinetics. The body wall contained the largest part of the accumulated cadmium.Sephadex G-75 gel filtration experiments demonstrated the presence of metal-binding, metallothionein (MT)-like proteins with a molecular weight of 10.8 kD in cytosolic fractions of pyloric caeca from unexposed sea stars. The MT-like proteins contained mainly zinc and a small amount of copper. During exposure, the MT-like proteins accumulated cadmium at a relatively high rate, but a steady state was reached after 8 weeks, indicating thatin vivo, the Cd-binding capacity of these proteins was limited. When pyloric caeca cytosol from unexposed sea stars was incubated with cadmiumin vitro, the MT-like proteins could bind larger amounts of cadmium thanin vivo during the Cd exposure. After 16 weeks of Cd exposure, thein vitro Cd-binding capacity had hardly increased.Low-molecular-weight metal-binding proteins were absent in cytosolic fractions of the testes, ovaries and oocytes from Cd-exposed sea stars, even after four months of exposure. The cadmium accumulated in the gonads was bound exclusively to high-molecular-weight proteins, which accumulated cadmium at a relatively low rate, compared to the MT-like proteins in the pyloric caeca. The absence of MT-like proteins in the gonads may explain the low cadmium accumulation rates found for these organs, but as a consequence, the accumulated cadmium is not detoxified.     

Cadmium,zinc, copper and metallothionein levels in human liver

The concentrations of cadmium, zinc, copper and metallothionein in the autopsy samples of liver among the inhabitants of Lód? (Poland) were determined. The cadmium levels were low in the range of 1.5 to 5.8 micrograms/g. The concentration of metallothionein determined by the Hg-method was high (0.160-1.665 mumol Hg/g); it was mainly a Zn-thionein. The percentage of hepatic zinc bound in the MT-fraction increased with the overall content of zinc in the liver. The elevation of zinc in the liver occurs in the proportion required for the saturation of metal-binding ligands of metallothionein. The role of cadmium remains less clear. Our results suggest that the metallothionein level in the liver increase significantly in response to elevated cadmium concentrations. This response, however, is in high excess to the demand which is justified stoichiometrically.     

Cadmium kinetics in freshwater clams. III. Effects of zinc on uptake and distribution of cadmium inAnodonta cygnea

Freshwater clams (Anodonta cygnea) were exposed to Cd²⁺ (25 g/L) or to Cd²⁺ (25 g/L) plus Zn²⁺ (2.5 mg/L). In the presence of zinc, uptake of cadmium in whole clam was halved. In organs such as the gills, mantle, and labial palps a still larger reduction of Cd accumulation occurred. By contrast, accumulation in midgut gland and kidney was hardly affected by the presence of zinc. It is shown that in the gills zinc competes with cadmium for metal binding sites both within the particulate fraction and the high-molecular weight fraction of the cytosol. Zinc probably did not induce an enlarged synthesis of specific metal-binding, metallothionein-like, proteins. In conclusion, zinc exerts antagonistic effects on uptake of cadmium by gills, and accelerates Cd transport from the gills towards the internal organs.     

Maternal zinc deprivation and interleukin-1 influence metallothionein gene expression and zinc metabolism of rats

The influence of maternal dietary zinc intake and recombinant human interleukin-1 alpha (rhIL-1 alpha) administration on metallothionein gene expression and the distribution of 65Zn were investigated. Pregnant rats were fed diets containing 1, 5, 30 or 180 mg Zn/kg diet in an equalized regime from d 13-20 of gestation. Metallothionein gene expression was examined by Northern blot and dot blot hybridization using combined 60-mer oligonucleotides specific for rat metallothionein-1 and -2 genes. Expression was progressively depressed in the fetal livers and livers and kidneys of dams fed diets marginal (5 mg/kg) and deficient (1 mg/kg) in zinc content. Administration of rhIL-1 alpha increased expression in maternal liver, placenta and in fetal liver of dams fed adequate or deficient diets. Kinetics of intravenously administered 65Zn showed that in response to rhIL-1 alpha, there was a higher uptake by the maternal liver and bone marrow with less 65Zn uptake by bone, intestine and plasma activity compared to controls. No change was observed in 65Zn taken up by the placenta or transferred to the fetus. Alteration of metallothionein gene expression could represent, in part, the mechanism whereby altered effects of zinc metabolism and function are mediated during fetal development.     

Cadmium, zinc, and copper in rabbit kidney metallothionein--relation to kidney toxicity

Twenty-two rabbits were given repeated subcutaneous injection of cadmium chloride. The cumulative cadmium dose given ranged from 13 to 214 mumole/kg body weight. Five rabbits served as controls. The treatment resulted in cadmium concentrations in kidney cortex that ranged from 0.3 to 3.2 mmole Cd/kg and a subsequent production of metallothionein. The molar ratio of cadmium, zinc, and copper in metallothionein fractions from kidneys with different concentrations of cadmium was determined. At low concentrations of cadmium in rabbit kidneys, zinc was the dominating metal bound to metallothionein (70-90%). At high concentrations of cadmium in kidneys, cadmium was the dominating metal in metallothionein. Evidence of kidney toxicity, in the form of beta2-microglobulinuria, was seen when cadmium constituted 85% of the metal ions recovered from metallothionein fractions. The remaining 15% was zinc. This indicates that at most six of the seven metal-binding sites in mammalian metallothionein are occupied by cadmium and that the remaining site is occupied by zinc. Our data provide further support for the hypothesis that chronic cadmium nephrotoxicity develops when there is a lack of metal-binding sites available for cadmium in metallothionein.     

Mitochondrial effects of low-level cadmium in rats: Interaction with zinc

Male Sprague-Dawley rats were treated with sodium or cadmium (Cd) 4 acetate (25 g Cd per kg body weight) orally 5 times a week for 6 weeks. A second group of animals was repeatedly injected with zinc sulphate (6 and 12 mg zinc (Zn) per kg ip) with or without Cd gavage. Cadmium treatment alone yielded no obvious toxic effects as evidenced by serum constituents or animal weight gain. Similarly, Zn injection did not affect these criteria. Zinc injection increased metallothionein in liver and kidneys and increased renal Cd. Cytosolic sorbitol dehydrogenase was not influenced by either cadmium, Zn or Cd + Zn exposure. However, individual Cd gavage decreased mitchochondrial cytochrome c oxidase in liver by 50%. This was partly protected by Zn. Hepatic adenosine triphosphatase (ATPase) was not affected by any of the treatment regimens. However, renal ATPase was inhibited by combined Cd + Zn administration. The data suggest subcellular toxic effects due to treatment with low Cd doses as evidenced by the decrease in hepatic cytochrome c oxidase. Simultaneous Zn injection may reduce this effect of Cd in liver. However, the treatment of rats by low level Cd gavage combined with zinc administration impairs the animals' health as shown by weight loss.Abbreviations Cd cadmium - Zn zinc - ATP adenosine triphosphate - ATPase adenosine triphosphatase - MT metallo thionein - NADH nicotinamide adenine dinucleotide, reduced Parts of this study have been presented at the 20th Annual Meeting of the Australasian Society for Clinical and Experimental Pharmacologists (ASCEP), Dec 8–10, 1986, Melbourne.     

Analysis of the renal tissue of a woman chronically exposed to cadmium

Summary In a 61-year-old woman, who had been exposed for 20 years to cadmium in the production of Ni-Cd batteries, nephrectomy of the contracted kidney was performed. The removed kidney was examined histologically and the cadmium concentration was determined in the cortex (44.97 g g^–1) and in the medulla (7.71 g g^–1). The homogenates of the renal cortex and medulla were subjected to gel filtration on Sephadex G-75. In the cortex, as well as the medulla, cadmium was predominantly found in the low-molecular (metallothionein) fraction, but in the cortex, Cd content in this fraction was six times higher than in the medulla. The determination of SH groups and proteins in high- and low-molecular fractions indicates an induction of the metallothionein formation primarily in the renal cortex.     

Fetal effects of cadmium in pregnant rats on normal and zinc deficient diets

Summary This investigation has shown that not only the extent of fetal resorption and malformation but also the types of malformation seen in rats depend upon the strain used and day of gestation. Furthermore, the effects of zinc deficiency and cadmium administration on the fetus can be at least additive, as was seen for malformations. For fetal resorption, zinc deficiency potentiated the action of cadmium.     

Fate and comparative toxicity of metallothioneins with differing Cd/Zn ratios in rat kidney

Metallothioneins with differing Cd/Zn ratios were preparedin vitro from rat liver zinc-thionein by replacing zinc with cadmium and were injected intraperitoneally to female rats. The distribution of cadmium, zinc, and copper in the kidney supernatant fraction was determined using a Sephadex G-75 column. The distribution pattern of cadmium and zinc changed dramatically within 24 hr after the injection. The changes were explained by the degradation and re-synthesis of metallothionein in the kidneys. The necrotic changes of renal tubular lining cells were correlated to the amount of cadmium in the metallothionein but not to the amount of metallothionein (protein).     

Hazardous exposure of ground-living small mammals to cadmium and lead in contaminated terrestrial ecosystems

The dietary exposure to cadmium and lead of two ground-living species of small mammals,i.e., shrewsSorex araneus (Insectivora) and volesMicrotus agrestis (Rodentia), was investigated and related to metal loads in target organs (kidneys and liver). The study was done in two natural areas polluted with cadmium and lead originating from urban and industrial metal sources. The average intake of cadmium by the herbivorous voles varied between 0.1 and 0.4 g/g/day and of lead between 2 and 10 g/g/day. The carnivorous shrews showed a considerably higher metal intake rates,i.e., cadmium 3 to 16 g/g/day and lead 19 to 53 g/g/day, which was largely due to the consumption of contaminated earthworms (Oligochaeta). An average cadmium intake of 15 g/g/day or a lead intake of 20 g/g/day corresponded with critical renal metal loads of 120 g/g for cadmium and 25 g/g for lead, which are indicative of adverse health effects. The renal metal loads in shrews reached the critical level, but they remained far below this level in voles. The results indicate a greater risk of toxic exposure to cadmium and lead in soricine shrews than in microtine rodents.     

Binding of endogenous copper and zinc to cadmium-induced metal-binding proteins in various tissues of Perna viridis

Green lipped mussels, Perna viridis, were exposed to cadmium chloride (CdCl2; 0.52 and 1 g/ml) in water for 4 days. The concentrations of cadmium (Cd), copper (Cu) and zinc (Zn) were measured in the viscera, gill, gonads, mantle, and muscle. There was a significant increase (p<0.05) in Cd concentration in all tissues studied. Results from Sephadex G-75 chromatography indicated that most Cd was bound to a fraction of heat-stable proteins similar to the metal-binding protein (MBP) metallothionein. After exposure to Cd, there was no significant change in Cu concentration in total tissue proteins or in total cytosolic proteins. A significant increase (p<0.05) in Cu, however, was detected in heat-stable proteins bound to the Cd-induced MBP in both viscera and gill. Copper bound to MBP also occurred in the gonad, mantle, and muscle, but to a much lesser extent. These results showed that Cd-induced MBP can also bind endogenous Cu. Zinc concentration in total heat-stable protein was increased only in gill and the muscle. Unlike Cu, a small amount of Zn binding to Cd-induced MBP was detected only in these tissues. Considering that both Cu and Zn exist intracellularly in dynamic equilibrium, the binding of Cu, but not Zn, to MBP may be explained by the kinetic reactivity of the two different metals to protein. The results of this study support the thesis that induction of intracellular MBP may also bind endogenous Cu and Zn.     

Short-term and long-term effects of cadmium on glycogen reserves and liver size in rainbow trout (Salmo gairdneri Richardson)

Salmo gairdneri Richardson were exposed to three test concentrations of 36, 12 and 4 g Cd/L for varying exposure periods of 1–84 days; blood glucose and lactate, liver and muscle glycogen and liver size were examined. The highest cadmium concentration produced mark depletion of glycogen content of both liver and muscle. This response was evident after 1 day post cadmium exposure and persisted on the subsequent second and third day exposure but not on 42 day exposure. Tissue glycogen levels were maintained within control values at lower cadmium concentrations. The liver size increased in the lowest cadmium concentration (4 g/L) after days 56 and 84. The mobilization of certain energy reserves to meet the stress of high cadmium exposure and the enlargement of the liver as a result of acclimation to prolonged cadmium exposure are discussed.     

Effect of short-term ethanol administration on cadmium retention and bioelement metabolism in rats continuously exposed to cadmium

The present study was performed to assess the effect of short-term ethanol administration on cadmium retention and accumulation as well as on bioelement metabolism (zinc, copper, calcium, and magnesium) in rats exposed to an aqueous solution of cadmium chloride for 8 weeks. Intoxication with cadmium led to accumulation of this toxic metal, particularly in the liver and kidney, which was linked to metallothionein synthesis as well as to a disturbance in the metabolism of zinc, copper, and calcium. These effects were dependent on the level of exposure. The administration of ethanol in the final phase of cadmium treatment increased cadmium retention and accumulation in the body with simultaneous elevation in liver and kidney metallothionein concentration. Ethanol alone or with cadmium caused or intensified the cadmium-induced changes in metabolism of zinc and copper. Calcium metabolism disturbed by cadmium was not influenced by ethanol. Neither agents had any effect on magnesium metabolism. We conclude that even short-term ethanol consumption in conditions of exposure to cadmium can increase this heavy metal body burden and lead to more serious disturbances in metabolism of important elements such as zinc and copper. Cadmium- and ethanol-induced changes in the homeostasis of these microelements are probably connected with the ability of both xenobiotics to cause metallothionein induction.     

The protective effect of simultaneous selenium administration on acute cadmium toxicity and metallothionein

The present study was conducted to elucidate the protective action of simultaneous selenium administration against acute cadmium toxicity. The remarkable testicular damages caused by cadmium, that is, hemorrhagic inflammation, atrophy and necrosis, were lessened by simultaneous selenium administration. Cadmium concentration in blood, especially in plasma, increased significantly during the early period after cadmium administration with selenium. Cadmium and selenium in plasma were found in the same fractions of high molecular weight reported by previous workers as the high molecular weight complex containing cadmium and selenium. Cadmium in testis was also noted in the high molecular weight fraction during the early period. However, cadmium in the high molecular weight fraction of plasma and testis were unstable and decreased rapidly by lapse in time. Cadmium concentration in liver was lower than that in the group administered cadmium alone during the increasing phase of plasma cadmium. However, in contrast with the decreased cadmium level in plasma, cadmium in liver and testis increased gradually. Cadmium increased in liver and testis were also found in the metallothionein fraction. In the testis protected from acute cadmium toxicity, the inhibitory effect of glutathione S-transferase activity by cadmium was not detectable and the activity was maintained at the level of the control (saline administered group). Moreover, the increased cadmium in the metallothionein fraction was related to the decrease of cadmium in the high molecular weight fraction of the testis homogenate. In addition, a positive correlation was observed between metallothionein concentration and glutathione S-transferase activity.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationships between heavy metal and metallothionein concentrations in lesser black-backed gulls, Larus fuscus, and Cory's shearwater, Calonectris diomedea

Metallothionein, cadmium, zinc, copper, and mercury concentrations were measured in adult lesser black-backed gulls, Larus fuscus; and metallothionein, cadmium, zinc, and copper concentrations were measured in fledgling Cory's shearwaters, Calonectris diomedea. In gulls, metallothionein was positively correlated with cadmium (kidney r=0.83, liver r=0.46), zinc (kidney r=0.46, liver r=0.37), and copper (kidney r=0.28, liver r=0.34). Mercury levels in lesser black-backed gulls showed no correlations with metallothionein or with any other metal. In shearwaters metallothionein was positively correlated with cadmium in the kidney (r=0.41) but not in liver, zinc in kidney (r=0.43) and liver (r=0.52), and copper in kidney (r=0.55) but not in liver. Cadmium levels were the most important factor determining tissue metallothionein concentrations in adult lesser black-backed gulls demonstrating the role of metallothionein in heavy metal detoxification. In fledgling Cory's shearwaters, the most important factor in determining metallothionein concentrations in kidney was copper concentrations, and in liver, zinc concentrations. During the latter phases of chick growth high levels of zinc are required for feather development, and at this time the binding of cadmium may be masked by the presence of a large amount of zinc- and copper-bound metallothionein. These results illustrate disparate roles of metallothionein, the levels of which will be in a state of flux both seasonally and annually.     

Distribution and binding of intratracheally instilled cadmium in rat lung

Retention, subcellular distribution, and the nature of binding of cadmium in lungs were studied following a single intratracheal instillation of cadmium. The clearance of intratracheally instilled cadmium from the lungs was very rapid during the initial periods of exposure and liver accumulated a maximum amount within 1 day. A low-molecular-weight cadmium-binding protein was isolated and characterized from lung cytosol. Cadmium administration increased metallothionein-like protein content but not sulfhydryls and zinc. The decrease in metallothionein at subsequent days of the exposure indicates the possibility of transport of this protein to some other target organs or the destruction of few cell types involved in providing protection against cadmium toxicity.     

Cadmium Interaction with Essential Metals (Zn, Cu, Fe), Metabolism Metallothionein, and Ceruloplasmin in Pregnant Rats and Fetuses

Cadmium administered to rats per os is accumulated in the duodenal mucosa in the form of metallothionein (MT). Therefore, this toxic metal can influence the efficiency of essential metal absorption, especially their concentration in the maternal organism, which plays an essential role during fetal development. The aim of this study is to elucidate the mechanism of the origin of Zn, Cu, and Fe deficiency in fetal rat livers after maternal exposure to cadmium in drinking water and to investigate the roles of MT and ceruloplasmin (Cp) in this phenomenon. Cadmium was given to pregnant dams exposed for 0–20 days of gestation in drinking water at concentrations of 6.25–100 μg Cd/ml. After cessation of exposure, at the lowest dose, a decrease in Cu and Fe concentrations in the duodenal mucosa was found. Simultaneously, diminution in concentration of two cited metals and Cp activity in serum of dam blood was noted. The lowest dose of cadmium developed a drop tendency in microsomal fetal liver iron. Significant correlations were observed between fetal liver Cu contents and Cp activity in serum of dams and Cu concentrations in serum of dams. Diminished Cp activity in serum of dams is related to reduced availability of Cu and Fe in fetuses. In conclusion, it is suggested that the mechanism of Cu and Fe deficit content in fetuses is based on the diminution of absorption of these metals by dam intestines exposed to cadmium on the reduction of metal concentrations in blood serum and, in consequence, their decreasing availability in fetuses.     

Maternal protein restriction and fetal growth: lack of evidence of a role for homocysteine in fetal programming

The disease-programming effects of a maternal low-protein (MLP) diet in rat pregnancy have been suggested to be attributable of hyperhomocysteinaemia. The aim of the present study was to determine whether MLP feeding impacted upon maternal and day 20 fetal homocysteine concentrations, with ensuing effects upon oxidant/antioxidant status. Sixty-four pregnant rats were fed either MLP diet or control diet before termination of pregnancy at days 4, 10, 18 or 20 gestation (full-term gestation 22 d). Maternal plasma homocysteine concentrations were similar in control and MLP-fed dams at all points in gestation. Fetal plasma homocysteine was similarly unaffected by MLP feeding at day 20 gestation. Activities of superoxide dismutase and glutathione peroxidase were similar in livers of mothers and fetuses in the two groups. Whilst catalase activity was not influenced by diet in maternal liver, MLP exposure increased catalase activity in fetal liver at day 20. Oxidative injury (protein carbonyl concentration) was lower in the livers of MLP-fed animals at day 18 gestation (P<0.05), but significantly greater at day 20. Hepatic expression of methionine synthase was similar in control and MLP-fed dams at all stages of gestation. Expression of DNA methyltransferase 1 in fetal liver was altered by maternal diet in a sex- and gestational age-specific manner. In conclusion, MLP feeding does not impact upon maternal or fetal homocysteine concentrations prior to day 20 gestation in the rat. There was no evidence of increased oxidative injury in fetal tissue that might explain the long-term programming effects of the diet.     

The influence of zinc status on the levels of metallothionein in isolated perfused rat liver.

The levels of cadmium and zinc metallothionein in isolated perfused livers of zinc depleted and repleted rats were investigated. Rats (160-200 g or 90-120 g) were fed either a zinc-deficient or zinc-supplemented diet for 62 or 42 days. The 90 to 120 g rats were repleted with 30 ppm Zn2+ in the drinking water for 1, 4 and 10 days after 35 days of depletion, and another group after depletion for 21 days was repleted for 15 days on a 21% casein diet. At appropriate times livers were cannulated, removed and perfused +/- 25 mug Cd2+ as CdCl2, for 2 hours through the portal vein in a perfusion apparatus. Zinc depletion resulted in significant decreases in growth rate (1.4 versus 5.7 g/day) and liver weights. Repletion with Zn for 10 days increased the weight gain to 7.1 g/day. Depletion decreased the incorporation of Zn and Cd into metallothionein by 57 to 60%, while repletion for 1, 4 and 10 days increased the incorporation to nearly 500% of control levels. But, as repletion progressed, zinc in metallothionein fell to normal levels after 15 days. Cadmium in metallothionein did not decrease between 1 and 10 days of repletion. Zinc deficiency also reduced the zinc content of the liver by 44%, which increased to normal levels after 10 days of repletion. These data suport the conclusion that zinc status can influence the incorporation of cadmium and zinc into metallothionein.     

65zinc metabolism is altered during diabetic pregnancy in rats.

To assess further diabetes-induced alterations in gestational Zn metabolism we examined the uptake/retention and distribution of 65Zn during pregnancy. Control and streptozotocin-induced diabetic rats, with and without insulin treatment, were gavaged with 65Zn on gestational d 12 or 18. Six hours post-intubation, untreated diabetic dams at d 12 of gestation had a lower percentage of retained 65Zn in bone, plasma, erythrocytes, pancreas and spleen than did controls. Retention of 65Zn in amniotic fluid and sacs and in embryos was similar among groups. Untreated diabetic dams at d 18 of gestation had a higher percentage of retained 65Zn in liver, intestine and urine, and most notably a lower percentage in amniotic fluid, placenta, fetus and fetal liver. Lower percentages of 65Zn were found in bone, muscle, plasma, erythrocytes, lung, spleen, heart, pancreas and uterus of diabetic dams compared with controls. HPLC fractionation of samples from maternal livers showed a higher percentage of Zn, Cu and 65Zn associated with the metallothionein peak in samples from untreated diabetic dams compared with other groups, whereas the percentages of Zn, Cu and 65Zn per fraction in fetal liver were similar. The marked decrease in retained 65Zn in products of conception at d 18 suggests either decreased placental transport or altered maternal and/or fetal Zn-binding ligands. Insulin treatment significantly reversed the streptozotocin diabetes-induced derangements in maternal and fetal Zn metabolism.