Cadmium, zinc and copper in horse kidney metallothionein

Cadium, zinc, and copper were determined in kidney cortex samples and in protein fractions obtained from 20 normal Swedish horses. Cadmium concentrations in kidneys ranged from 0.01 to 1.46 mmole/kg. Zinc concentrations in kidneys increased with increasing cadmium concentrations. At low concentrations of cadmium the increase of zinc was almost equimolar with the increase in cadmium, but at higher concentrations of cadmium the increase of zinc was less pronounced. Copper concentrations ranged from 0.07 to 0.22 mmole Cu/kg and were not related to the cadmium concentration. The increase of zinc in kidney cortex took place in the metallothionein (MT) fractions, where also almost all cadmium was recovered. The molar ratio of zinc and cadmium in MT was close to one at low concentrations (below 0.1 mmole/kg) of cadmium but the ratio decreased with increasing cadmium concentrations in the kidney. This change in molar ratio of zinc and cadmium in MT explains the leveling off in zinc increase at cadmium concentrations in kidney exceeding 0.6 mmole/kg.     

Cadmium,zinc, and copper in horse liver and in horse liver metallothionein: Comparisons with kidney cortex

Cadmium, zinc, and copper were determined in liver and in kidney cortex samples obtained from 33 normal Swedish horses. Cadmium concentrations in liver ranged from 0.002 to 0.165 mmole/kg and in kidney from 0.01 to 2.15 mmole/kg. There was a significant correlation between liver and kidney concentrations of cadmium. The average kidney concentration of cadmium was about 15 times that of liver. Zinc concentrations increased with increasing cadmium concentrations in both liver and kidney. The relative increase of zinc with cadmium was more pronounced in liver than in kidney. However, the absolute increase of zinc was larger in kidney due to the much higher concentration of cadmium in kidney compared to liver. Any significant correlation between copper and cadmium, or copper and zinc, could not be revealed. Sephadex gel filtration was performed on supernatants from homogenates of kidney and liver from 19 of the horses. In both organs the major part of cadmium was recovered in protein fractions corresponding to metallothionein (MT), in which the increase of zinc also took place. The molar ratio between zinc and cadmium was higher in MT fractions obtained from liver than in MT fractions obtained from kidney.     

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.     

Cadmium/zinc relationships in kidney cortex and metallothionein of horse and red deer: histopathological observations on horse kidneys

Cadmium and zinc were determined in kidney cortex of 63 horses and 51 red deer (Cervus elaphus). Cadmium and zinc were also determined in protein fractions obtained by Sephadex chromatography of kidney cortex from 10 horses and 4 red deer. Histopathological parameters in kidney cortex of horses were compared to cadmium content. The metal contents (on wet weight basis) in kidney cortex of the horses were 0.31 +/- 0.22 mmole Cd/kg (range 0.03-1.21) and 0.63 +/- 0.17 mmole Zn/kg (range 0.36-1.23). The Zn content increased with the Cd content, the Zn increase being less at higher concentrations. No significant increase of the Cd content with age of the horses (range 2-19 years) was found. The metal contents in kidney cortex of the red deer were 0.030 +/- 0.031 mmole Cd/kg (range 0.002-0.13) and 0.51 +/- 0.37 mmole Zn/kg (range 0.30-2.82). Here the Zn content did not increase significantly with Cd content, and in this case a positive linear relation of Cd content with age of the deer (range 0-11 years) was observed. The molar Zn/Cd ratio in the metallothionein fractions of horse kidney cortex was less variable at higher Cd contents. All Cd and the increase of Zn above an average basal level of 0.36 mmole Zn/kg was recovered in the metallothionein fractions. No gross histopathological changes could be observed in kidney cortex of the horses. Nevertheless a slight increase of the thickness of the basement membrane of Bowman's Capsule and also of the diameter of Bowman's Capsule in relation to Cd content up to 0.3 mmole/kg could be observed.     

Native and induced rat kidney metallothioneins and their relation to cadmium toxicity

The concentrations of zinc, copper, iron, manganese, magnesium, and phosphorus in the kidneys were determined for control female rats of five different ages (4, 8, 12, 16, and 20 weeks old). The concentration of copper increased with age and it was attributed to three metallothionein peaks on a SW 3000 column. Parenteral administration of Cd-thionein caused swelling of kidneys to different degrees, depending on the age of the rats; the youngest rats were affected most. The concentrations of cadmium and the six metals in the kidneys at 12 hr and 7 days after injection were determined and compared with controls. The distribution among protein fractions of cadmium, zinc, and copper, which are related to metallothionein, were examined by gel permeation chromatography. The elution profiles of the three metals changed not only with the ages of the rats but also with time after injection. The changes were correlated with the copper content in the metallothioneins and with swelling of the kidneys. The changes also correlated with native metallothionein levels.     

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).     

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.     

Copper content in cadmium-exposed animal kidney metallothioneins

Cadmium chloride was injected intraperitoneally into five animals (mouse, hamster, rat, guinea-pig, and rabbit) with the same amount of cadmium per body weight. The concentrations of three metals (Cd, Zn, and Cu) in the 105,000 g supernatant and in the heat-treated supernatant of livers and kidneys of the cadmium-exposed animals were determined by atomic absorption analyses. The supernatants of both organs were applied to a Sephadex G-75 column and the distribution profiles of the three metals were analyzed to show the metal contents in the metallothionein fraction. The results indicated that the amount of retained cadmium and the increased amount of zinc in both organs were different from animal to animal and that the metals were bound to metallothionein. The most significant result was that the five animals were divided into two groups according to changes of increased copper in the renal metallothionein of cadmium-exposed animals. One group had a dramatic increase in copper content while the other group had little or no increase in copper content for the renal metallothionein after exposure to cadmium. Rat and guinea-pig belonged to the former group and mouse, hamster, and rabbit belonged to the latter group.     

Induction and metal speciation of metallothionein in wood mice (Apodemus sylvaticus) along a metal pollution gradient

We investigated the binding of Cd, Cu, and Zn to metallothionein (MT) and other metal-binding proteins in free-living wood mice (Apodemus sylvaticus L.) captured in four areas along a metal pollution gradient. We measured total and cytosolic Cd, Cu, and Zn concentrations in mouse liver and kidney by means of inductively coupled plasma-mass spectrometry (ICP-MS). Total (Cu, Cd, Zn)-MT levels were determined in the same tissues by means of the cadmium thiomolybdate saturation assay. Metal speciation of metalloproteins was studied by means of size-exclusion high-performance liquid chromatography-ICP-MS. Liver and kidney of wood mice from the site adjacent to the pollution source showed the highest Cd and Zn concentrations (total and cytosolic) and (Cu, Cd, Zn)-MT levels compared to the other sites farther away from the pollution source. No or only small site differences in tissue Cu concentrations were observed. Almost all the variation (85-95%) in hepatic and renal (Cu, Cd, Zn)-MT levels was explained by the total or cytosolic hepatic Zn and Cd concentration or the renal Cd concentration, respectively. An analysis of the cytosolic metal speciation showed that the Cd-MT, Cu-MT, and Zn-MT fractions in liver and kidney increased significantly with increasing cytosolic metal concentrations. Metals associated with the other cytosolic protein fractions did not increase with increasing exposure. These results illustrate the important role of MT in metal homeostasis and detoxification processes. We conclude that MT is a useful biomarker for environmental metal contamination in free-living wood mice.     

Effects of maternal exposure to zinc and cadmium on metallothionein in fetal rat liver

Experiments were made to observe the effects of the maternal administration of cadmium and zinc and their influence on the metal-binding capacity of metallothionein in the rat fetus. Metallothionein was detected in the fetal liver as early as day 16 of gestation and may have appeared earlier. The zinc concentration in the metallothionein fraction was low on day 16 but increased as gestation progressed. The metal-binding capacity was fairly high on day 16 but increased at a slower rate than the zinc concentration. Following the intraperitoneal administration of zinc (100 moles Zn/kg) or cadmium (5 moles Cd/kg) once a day on days 14, 15 and 16 of gestation to the dam, the concentration of zinc in the metallothionein fraction of the maternal liver was far greater than that in the fetal liver. In contrast, cadmium was found only in the metallothionein fraction of the maternal liver; it was not found in the fetal liver, which suggests an effective placental barrier to cadmium. The metalbinding capacity in the fetal liver was induced by the maternal administration of zinc, but not cadmium.     

Metallothionein gene expression in rats: tissue-specific regulation by dietary copper and zinc

Regulation of metallothionein gene expression by dietary zinc and copper was examined in rat liver, kidney, intestine and brain using a 3 X 3 factorial design. Purified diets containing 5, 30 and 180 mg Zn/kg and 1, 6 and 36 mg Cu/kg were fed for 2 wk. Serum concentrations of copper and zinc were lower at the lowest intakes of either metal than at normal or supplemental levels. Kidney metallothionein levels were proportional to dietary zinc, being 50% less in the 5 mg Zn/kg group than in those fed the highest zinc intake. Metallothionein mRNA was measured by dot blot hybridization to a 32P-labeled oligonucleotide DNA probe representing the terminal 5' sequence of the metallothionein gene. In kidney the number of metallothionein mRNA molecules per cell increased four- to five-fold (from 4 to 29 molecules per cell) with increasing dietary zinc. A less pronounced effect on metallothionein mRNA was observed in response to dietary copper. At the lowest copper intake level and highest intake of zinc intestinal metallothionein mRNA was sevenfold greater than in any other group. Liver and brain did not respond appreciably to the dietary levels of copper and zinc that were fed. Chromatography showed that copper and zinc content of renal metallothionein was directly related to the dietary levels fed. In kidney, both metallothionein-1 and -2 genes were expressed.     

Copper, zinc, and cadmium concentrations in Peromyscus maniculatus sampled near an abandoned copper mine

Concentrations of zinc, copper, and cadmium were determined in soil and liver, kidney, bone and stomach contents of deer mice (Peromyscus maniculatus) from two sites near an abandoned mine and one control site, on Vancouver Island, British Columbia, Canada. Soil concentrations of copper were significantly elevated at the mine and off site vs the reference site. In contrast, there was no difference in soil cadmium and zinc concentrations between the mine and reference site. Concentrations of copper, cadmium and zinc in livers of mice from the mine site were significantly elevated relative to the reference and off site locations. Cadmium kidney concentrations tended to be greater in mice from the mine versus the off site and reference site. No differences in bone cadmium, copper and zinc and, kidney copper and zinc concentrations were noted among mice from the three locations. Diet of mice from mine and off sites contained significantly greater copper concentrations than the reference population; no differences in cadmium or zinc diet concentrations in mice from the three sites were noted. Comparison of ratios of metal concentrations in diet:soil and concentrations in liver:soil suggest that for zinc and copper, soil and diet are of equal importance as a source of metal contamination to these mice. In contrast, cadmium diet:soil and cadmium liver:soil ratios were much greater than one indicative of bioconcentration of cadmium from soil to diet and from soil to liver. For assessing routes of metal exposure, in this case for deer mice inhabiting an abandoned mine site, for copper and zinc, soil will most likely be indicative of exposure conditions. In contrast, concentrations of cadmium in diet will be more representative of amounts that the animal is potentially ingesting. Of further importance is that relative to reference sites, mice inhabiting an abandoned copper mine site have significantly elevated tissue levels of copper. This is turn will provide a route of metal exposure to canivorous birds such as owls and hawks. The toxicological significance of this exposure to birds of prey has yet to be assessed adequately.     

Accumulation and Soluble Binding of Cadmium, Copper, and Zinc in the Polychaete Hediste diversicolor from Coastal Sites with Different Trace Metal Bioavailabilities

Bioaccumulation of cadmium, copper, and zinc was examined in common ragworms Hediste diversicolor from control (Bay of Somme, Blackwater) and metal-rich (Seine estuary, Boulogne harbor, Restronguet Creek) sites in France and the United Kingdom. The degree of exposure in the field was assessed by considering both total concentrations in superficial sediment and the quantities of metals which may be released in vitro at different pH levels. Among the three contaminated sites, release of the three metals was not detectable in Boulogne harbor, in correlation with limited enhancement of the metal concentrations in the common ragworms from this site. Even at those sites where zinc could be released in vitro from the sediment, zinc concentrations were not enhanced in common ragworms, in agreement with previous findings indicating that the body content of this metal is regulated in H. diversicolor. At all the studied sites, bioaccumulated zinc was mainly in cytosolic form. The distribution of cadmium and copper varied according to the origin of the common ragworms, the insoluble fraction increasing with the degree of contamination (cadmium in the Restronguet Creek, copper in the Seine estuary, and even more in Restronguet Creek). In the cytosolic fraction, metals were partly linked to cytosolic heat-stable thiolic compounds (CHSTC) with molecular masses (5–6 kDa and about 12 kDa) consistent with metallothionein-like proteins (MTLP). Metal-binding to MTLP varied with the degree of contamination and with the metal studied. In contrast to many invertebrates, the presence of metal-binding CHSTC (MM about 2 kDa) other than MTLP seems to be a peculiar feature of H. diversicolor.     

Dietary zinc reduces uptake but not metallothionein binding and elimination of cadmium in the springtail,Orchesella cincta

In metal-polluted environments, high concentrations of metals such as zinc, cadmium, copper, and lead are present. Many studies have been conducted to determine uptake and binding of the nonessential, toxic metal cadmium. Effects of essential metals on this uptake and binding remain unclear. The aim of the present study was to determine the effect of zinc on cadmium kinetics and metallothionein induction in the springtail Orchesella cincta. Exposure experiments were conducted with cadmium only, zinc only, and cadmium + zinc. Metallothionein was isolated to determine zinc binding to or induction of this cadmium-binding protein. In animals exposed to cadmium only and to cadmium + zinc, metallothionein was induced. In animals exposed to zinc only, no metallothionein was induced, and zinc did not bind to the metallothionein induced by cadmium. Cadmium uptake rate was significantly lower in animals exposed to cadmium + zinc than in animals exposed to cadmium only. No significant pattern of uptake and elimination of zinc was observed. Zinc concentrations in the animals were not significantly different for the different exposures, demonstrating that this element is regulated by O. cincta. These results suggest a protective role of zinc in the uptake of cadmium but no interference of zinc with the internal cadmium distribution and elimination.     

Disturbances in the metabolism of endogenous metals (Zn and Cu) in nickel-exposed rats

The effect of nickel chloride on tissue concentrations of zinc and copper was studied in female rats. Animals were subjected to repeated exposure to 5 mg Ni/kg. Whole-body retention of zinc increased about 30%, and retention of copper did not change. Increase of the zinc content was observed in liver, but in kidneys, lungs, and brain zinc content decreased. It was also noted that the level of metallothionein was higher in nickel-exposed rat liver than in control rat liver. Concentration of metallothionein was correlated with the metal contents (Ni, Cu, Zn).     

Body mass and caloric value of the ground beetle (Pterostichus oblongopunctatus) (Coleoptera, Carabidae) along a gradient of heavy metal pollution

At five sites located along a metal-pollution gradient in southern Poland, we collected, during the spring and summer of 2000, more than 1,200 individuals of the ground beetle (Pterostichus oblongopunctatus) to examine the relationship between pollution level and body mass. Animals from one additional sampling in May 2001 were used to measure body caloric value to verify whether metal pollution has an effect on the energy content of the body. The study sites were located in an area with a history of zinc and lead mining and smelting dating back to medieval times. Metal concentrations in the humus layer ranged from 200 to 9,600 mg/kg of zinc, 120 to 1,600 mg/kg of lead, and 3 to 82 mg/kg of cadmium. We found a significant increase in body mass with increasing pollution level. The beetles from all sites collected near the end of the season were lighter. However, no statistically significant trend in body caloric value was detected. We suggest that the high metal tolerance of the species, combined with altered interspecies competition at the polluted sites, is responsible for the positive correlation between soil metal concentration and body mass.     

Metallothionein induced by cadmium or zinc inhibits lipid peroxidation in rats exposed to dimethylnitrosamine

Dimethylnitrosamine (N-nitrosodimethyamine) is a potent inducer of microsomal lipid peroxidation in the liver and kidneys. The results of this study show that prior exposure of adult rats to cadmium or zinc antagonises the effect of dimethylnitrosamine on lipid peroxidation by inducing increased metallothionein synthesis. Increased concentration of reduced glutathione (GSH) in the liver and kidney of rats pre-treated with cadmium or zinc and exposed to dimethylnitrosamine suggests that metallothionein inhibits oxidative stress. The presented results have been corroborated by findings that exposure of adult rats to cadmium or zinc may activate genes for metallothionein and glutathione-S-transferase, the protective proteins against oxidative stress.     

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.     

Monitoring of Heavy Metal Contamination Using Tissues of Two Ardeids Chicks, Korea

This study presents concentrations of iron, zinc, manganese, lead and cadmium in livers and kidneys of Little Egret Egretta garzetta (n = 10) and Black-crowned Night-Heron Nycticorax nycticorax (n = 10) chicks from Pyeongtaek, Gyeonggi-do, Korea. Essential element concentrations such as iron, zinc and manganese did not differ between both species chicks in livers and kidneys, but lead and cadmium concentrations were significantly greater in both tissues of Little Egret chicks (lead 2.78 ± 4.06 μg/g wet weight, cadmium 10.3 ± 12.8 kg/g wet weight) than in those of Black-crowned Night-Heron chicks (lead 0.92 ± 0.73 μg/g wet weight, cadmium 1.00 ± 1.00 kg/g wet weight). Lead and cadmium concentrations in livers of Black-crowned Night-Heron chicks were highly related to sediment and/or prey concentrations of their foraging sites in Korean studies. It shows that lead and cadmium concentrations in livers of heron chicks can reflect those of surrounding environment of breeding sites. In this study, cadmium concentrations were higher in kidneys than in livers and it’s not recent high-level exposure but chronic background exposure to cadmium contamination around breeding site. Therefore, we suggest that cadmium concentrations in livers and kidneys can be used as a bioindicator of acute and/or chronic local contamination.