Inhibition Of Cd Uptake Research Articles

Increased zinc supply does not inhibit cadmium accumulation by rice (Oryza sativa L.)

ABSTRACTRice (Oryza sativa L.) grown on cadmium (Cd)-contaminated soils has caused health problems in Asian subsistence rice farmers. For other crops, normal co-contaminant zinc (Zn) inhibits the increased uptake of Cd. We used a multi-chelator-buffered nutrient solution to characterize the interaction of Zn and Cd in uptake-translocation of Cd in “Lemont” rice. The activity of free Zn2+ varied from 10−7.6 to 10−5.2 M, while free Cd2+ held constant at 10−10.7 M. Zinc activity 10−5.6 M and higher was phytotoxic to rice, resulting in severe chlorosis, reduced growth, and increased Cd transport to shoots. In contrast to previous studies with wheat, lettuce, and spinach, free Zn2+ maintained at adequate to sub-phytotoxic levels (10−7.6 to 10−6.1) did not inhibit Cd uptake by rice. The inability of Zn to inhibit Cd uptake by rice is a key factor in Cd risk from zinc-lead mine waste contaminated soil compared with other crops.

A hemicellulose-bound form of silicon inhibits cadmium ion uptake in rice (Oryza sativa) cells.

Silicon (Si) alleviates cadmium (Cd) toxicity in rice (Oryza sativa). However, the chemical mechanisms at the single-cell level are poorly understood. Here, a suspension of rice cells exposed to Cd and/or Si treatments was investigated using a combination of plant cell nutritional, molecular biological, and physical techniques including in situ noninvasive microtest technology (NMT), polymerase chain reaction (PCR), inductively coupled plasma mass spectroscopy (ICP-MS), and atomic force microscopy (AFM) in Kelvin probe mode (KPFM). We found that Si-accumulating cells had a significantly reduced net Cd(2+) influx, compared with that in Si-limited cells. PCR analyses of the expression levels of Cd and Si transporters in rice cells showed that, when the Si concentration in the medium was increased, expression of the Si transporter gene Low silicon rice 1 (Lsi1) was up-regulated, whereas expression of the gene encoding the transporter involved in the transport of Cd, Natural resistance-associated macrophage protein 5 (Nramp5), was down-regulated. ICP-MS results revealed that 64% of the total Si in the cell walls was bound to hemicellulose constituents following the fractionation of the cell walls, and consequently inhibited Cd uptake. Furthermore, AFM in KPFM demonstrated that the heterogeneity of the wall surface potential was higher in cells cultured in the presence of Si than in those cultured in its absence, and was homogenized after the addition of Cd. These results suggest that a hemicellulose-bound form of Si with net negative charges is responsible for inhibition of Cd uptake in rice cells by a mechanism of [Si-hemicellulose matrix]Cd complexation and subsequent co-deposition.

Can the application of rare earth elements improve yield and decrease the uptake of cadmium in ryegrass-dominated pastures?

Rare earth elements (REEs) have been linked to increases and decreases in plant growth and the uptake of biotoxic metals such as Cadmium (Cd). We hypothesised that under the right soil and climatic conditions the application REEs to soil may improve yield of ryegrass-dominated pasture and inhibit Cd uptake in foliage. A soil survey measuring REEs concentrations in soils from across New Zealand showed that REEs were enriched in Volcanic compared with Sedimentary-derived soils, and in soils used for dairying compared with those used for sheep and beef. From a range of REEs applied to ryegrass with and without N, lanthanum (La) showed minor yield improvement under glasshouse conditions, but only for Sedimentary (not Volcanic) soils low in REE concentration and at 40% available soil moisture or less. A similarly mixed yield response occurred in a field trial. Decreases in the Cd concentration of ryegrass (mg kg–1) only occurred in the harvests after application (40% at 20 kg La ha–1 for one soil) and did not translate into an annual effect. We conclude that there was insufficient evidence to recommend the use of La (screened from a range of REEs as the most promising) to increase annual dry matter yield or decrease Cd uptake from ryegrass-dominated pastures.

Exogenous abscisic acid application decreases cadmium accumulation in Arabidopsis plants, which is associated with the inhibition of IRT1-mediated cadmium uptake.

Cadmium (Cd) contamination of agricultural soils is an increasingly serious problem. Measures need to be developed to minimize Cd entering the human food chain from contaminated soils. We report here that, under Cd exposure condition, application with low doses of (0.1–0.5 μM) abscisic acid (ABA) clearly inhibited Cd uptake by roots and decreased Cd level in Arabidopsis wild-type plants (Col-0). Expression of IRT1 in roots was also strongly inhibited by ABA treatment. Decrease in Cd uptake and the inhibition of IRT1 expression were clearly lesser pronounced in an ABA-insensitive double mutant snrk2.2/2.3 than in the Col-0 in response to ABA application. The ABA-decreased Cd uptake was found to correlate with the ABA-inhibited IRT1 expression in the roots of Col-0 plants fed two different levels of iron. Furthermore, the Cd uptake of irt1 mutants was barely affected by ABA application. These results indicated that inhibition of IRT1 expression is involved in the decrease of Cd uptake in response to exogenous ABA application. Interestingly, ABA application increased the iron level in both Col-0 plants and irt1 mutants, suggesting that ABA-increased Fe acquisition does not depend on the IRT1 function, but on the contrary, the ABA-mediated inhibition of IRT1 expression may be due to the elevation of iron level in plants. From our results, we concluded that ABA application might increase iron acquisition, followed by the decrease in Cd uptake by inhibition of IRT1 activity. Thus, for crop production in Cd contaminated soils, developing techniques based on ABA application potentially is a promising approach for reducing Cd accumulation in edible organs in plants.

Effect of potassium deficiency on antioxidant status and cadmium toxicity in rice seedlings.

BackgroundCadmium (Cd) is one of the most toxic heavy metals and inhibits physiological processes of plants. Potassium (K) is an essential macronutrient in plants. K deficiency and Cd stress represent two different abiotic stress conditions that occur in the field simultaneously. In this study, effects of K deficiency on antioxidant status and Cd toxicity in rice seedlings were investigated.ResultsK deficiency significantly decreased K concentration in shoots and roots. However, fresh weight and dry weight of rice seedlings were not affected by K deficiency. The activities of antioxidant enzymes (superoxide dismutase, ascorbate peroxidase, glutathione reductase, and catalase) in K-deficient leaves were higher than respective control leaves. However, K deficiency had no effect on the content of antioxidants (ascorbate and glutathione). Cd toxicity was judged by the decrease in biomass production, chlorosis, and induction of oxidative stress. Based on these criteria, we demonstrated that K deficiency protected rice seedling from Cd stress. Moreover, chlorophyll concentration was higher in K-deficient shoots and roots than their respective control shoots and roots.ConclusionsOur results indicated that K deficiency protects rice seedlings from Cd toxicity. This protective effect of K deficiency is mainly due to enhanced antioxidant enzyme activities but not inhibition of Cd uptake.Electronic supplementary materialThe online version of this article (doi:10.1186/1999-3110-54-2) contains supplementary material, which is available to authorized users.

Iron uptake system mediates nitrate-facilitated cadmium accumulation in tomato (Solanum lycopersicum) plants

Nitrogen (N) management is a promising agronomic strategy to minimize cadmium (Cd) contamination in crops. However, it is unclear how N affects Cd uptake by plants. Wild-type and iron uptake-inefficient tomato (Solanum lycopersicum) mutant (T3238fer) plants were grown in pH-buffered hydroponic culture to investigate the direct effect of N-form on Cd uptake. Wild-type plants fed NO3− accumulated more Cd than plants fed NH4+. Iron uptake and LeIRT1 expression in roots were also greater in plants fed NO3−. However, in mutant T3238fer which loses FER function, LeIRT1 expression in roots was almost completely terminated, and the difference between NO3− and NH4+ treatments vanished. As a result, the N-form had no effect on Cd uptake in this mutant. Furthermore, suppression of LeIRT1 expression by NO synthesis inhibition with either tungstate or L-NAME, also substantially inhibited Cd uptake in roots, and the difference between N-form treatments was diminished. Considering all of these findings, it was concluded that the up-regulation of the Fe uptake system was responsible for NO3− -facilitated Cd accumulation in plants.

Uptake pathways and subcellular fractionation of Cd in the polychaete Nereis diversicolor

Polychaetes have often been utilized as indicator species to investigate the impacts of pollutants, such as heavy metals. The uptake of Cd by the polychaete Nereis diversicolor was determined at varying Ca concentrations and with pre-exposure to Ca ion channel blockers and metabolic inhibitors in simulated sea water over 1week period. The supply of Ca in simulated sea water inhibited Cd uptake and increased Ca concentration in N. diversicolor after 10μM Cd exposure. Pre-exposure to a Ca-channel blocker (Lanthanum) significantly inhibited Cd uptake, suggesting that the uptake of Cd was exerted at a Ca channel. N-ethylmaleimide, which specifically binds to sulfhydryl groups, inhibited Cd uptake at 10μM, implying that the transport of Cd is carrier-mediated by proteins or other SH-containing compounds. Subcellular Cd distribution analysis showed that more than 60% of the total Cd associated with the cytosolic fraction. The presence of higher concentration of Ca in simulated sea water did not impact the proportional subcellular distribution of Cd in N. diversicolor. Nevertheless, the supply of Ca could significantly lower Cd concentration in cytosol and cellular debris. The present study provides evidence that Cd transport by N. diversicolor was mediated mainly through lanthanum- sensitive Ca ion channels and accumulated by SH-containing compounds. These results help to understand the uptake mechanism and subcellular distribution of Cd in polychaetes.

Fe- and Zn-induced inhibition of Cd uptake in human lung cell lines: Speciation studies with H441 and A549 cells

Cadmium transport was studied in human lung cell lines A549 and H441 as a function of inorganic metal speciation. A 2-fold higher equilibrium accumulation was obtained in the A549 cells, but a specific system of transport of high affinity and low capacity was characterized in both these cells. Exposure conditions optimizing [Cd2+] increased Cd uptake, but CdCln2-n species are also taken up. Studies on Cd-Ca reciprocal inhibition do not support a role for Ca pathways in Cd uptake. Nramp2 and Zip8 mRNAs were detected in both cell lines. Fe inhibited Cd uptake in the nitrate medium with an apparent at pH 5.5. NRAMP2 may contribute to Cd uptake but a major role is precluded since the overall process of accumulation was inhibited by acidic conditions. Zn and Mn inhibited Cd uptake with the following observations: (1) Similar apparent Ki values for Zn-induced inhibition in the chloride and nitrate media. (2) Whatever the medium, a higher Mn-induced inhibition at pH 5.5 compared to pH 7.4, with much lower Ki values under acidic conditions. (3) Elimination of the stimulatory effect of HCO3- by Zn but not by Mn. Zn may inhibit a ZIP8-mediated Cd uptake, whereas the Mn-sensitive component of uptake would be related to other transport processes.

Uptake pathways and toxicity of Cd and Zn in the earthworm Eisenia fetida

The uptake of Cd and Zn by the earthworm Eisenia fetida was determined at varying Ca concentrations and with pre-exposure to different metabolic inhibitors in simulated soil solutions over a 48-h period. The presence of Ca in the solution had complex actions on Cd uptake. At a low Cd concentration of 0.1 μM, Ca (0.1–1 mM) slightly but significantly stimulated Cd uptake, whereas it inhibited Cd uptake at a higher Cd level (10 μM). Pre-exposure to a Ca-channel blocker (Lanthanum) inhibited Cd uptake over a relatively wide range of Cd concentrations, but not Zn uptake, suggesting that the uptake of Zn was not exerted at a Ca channel. N-ethylmaleimide, which specifically binds to sulfhydryl groups, inhibited Zn uptake at both 0.1 and 10 μM, implying that the transport of Zn is carrier-mediated by proteins or other SH-containing compounds. The present study provides evidence that the mechanisms of Cd and Zn uptake in earthworms are pharmacologically different, although both metals have similar geochemical and environmental properties. After 24 h pre-exposure to a sublethal concentration (1.0 μM) of Cd, Zn toxicity for E. fetida was significantly reduced with 48-h LC 50 values (with 95% confidence interval) increasing from 145 (105–201) to 316 (212–470) μM Zn. Pre-exposure to Zn (1.0 μM) did not, however, affect Cd toxicity. Pre-exposure to Cd significantly changed the subcellular Zn distribution, with a decreasing fraction of Zn associated with Fraction B (associated with granules and cell membranes), which is believed to be most indicative of toxic pressure and an increased fraction associated with Fraction G (associated with cytosol). This most likely explains the observed Zn tolerance of E. fetida after low level Cd pre-exposure. These results help to understand the uptake mechanism and interactions of Zn and Cd in earthworms.

Cadmium Uptake, Translocation, and Tolerance in AHA1OX Arabidopsis thaliana

Information on cadmium (Cd) uptake and transport is essential to understand better the physiology of Cd tolerance in plants. In this study, Cd uptake, translocation, and tolerance were investigated in AHA1 (Arabidopsis plasma membrane H(+)-ATPase gene) overexpressed plants. Exposed to 10 µM CdCl(2), AHA1OX showed a higher root elongation, accumulated more Cd, and maintained better integrity of nucleus membrane of root tips in comparison to the control plant (WT), suggesting that AHA1OX was more Cd tolerant than WT. To investigate Cd tolerance mechanism of AHA1OX plants, we measured the activity of plasma membrane H(+)-ATPase and the secretion of citrate. Results indicated that treatment with 10 µM of Cd stimulated the activity of plasma membrane H(+)-ATPase and the secretion of citrate, while 30 µM of Cd inhibited them. AHA1OX had higher activity of H(+)-ATPase and secretion of citrate than WT. Addition of citrate enhanced root-to-shoot translocation of Cd significantly. A higher root-to-shoot Cd translocation was observed in AHA1OX than in WT plants. Treatment with low temperature or metabolic inhibitor (carbonyl cyanide m-chlorophenylhydrazone) inhibited Cd uptake and translocation. The study of Cd forms using sequential extraction indicated that Cd was mainly present as a protein-bound form, and AHA1OX had more water-soluble Cd than WT. Taken together, our results suggested that the Cd tolerance of AHA1OX was associated with its root-to-shoot Cd translocation and secretion of citrate, which converts Cd(2+) into less toxic and more easily transportable forms in plant cells.

Effect of cation competition on cadmium uptake from solution by the earthworm Eisenia Fetida

Metal speciation alone is insufficient to predict metal accumulation in aquatic and terrestrial organisms, because competition between cations can play an important role. In the present study, the effects of competing cations (Ca2+, Mg2+, and H+) on Cd uptake by the earthworm Eisenia fetida were quantified through short-term experiments over a large Cd concentration range in aqueous exposure solutions. Free Cd2+ activity was a better predictor for Cd accumulation in the earthworms than was total Cd concentration in solution, thus supporting the free ion activity model. The presence of the other cations at environmentally pertinent concentrations inhibited Cd uptake to different degrees. A competitive equilibrium model based on the biotic ligand model was developed to quantify the effect of the cations. The model parameters are the conditional constants for binding of H+ (log K = 4.97), Ca2+ (log K = 3.00), and Mg2+ (log K = 2.64) to the biological uptake sites of E. fetida. Autovalidation of the model with experimental results showed that 48-h Cd accumulation in the earthworm could be successfully predicted with activities of free Cd2+ and the major cations (Ca2+, Mg2+, and H+) as input.

Silicon‐induced cadmium resistance in rice (Oryza sativa)

AbstractSilicon (Si)‐induced cadmium (Cd) tolerance in rice (Oryza sativa L.) was investigated by analyzing Cd uptake, growth, and physiological parameters. Silicon treatments (0.0, 0.2, or 0.6 mM) were added to 6 d–old seedlings, and Cd treatments (0.0 or 5.0 μM) were added to 20 d–old seedlings. Parameters determined included: maximum net CO2 assimilation (Amax), stomatal conductance (gsmax), and transpiration (Emax) rates at varying intercellular CO2 concentrations (Ci). Also measured were chlorophyll fluorescence, growth, and Cd‐uptake parameters. Results showed a Si‐induced inhibition of Cd uptake. However, 0.2 mM or 0.6 mM Si treatment concentrations did not differentially inhibit Cd uptake or differentially alleviate Cd‐induced growth inhibition, despite a significant increase in tissue Si concentration due to 0.6 mM Si treatment compared to 0.2 mM Si treatment. Additionally, photosynthesis and chlorophyll‐fluorescence analysis showed that treatment with Cd significantly inhibited photosynthetic efficiency. Interestingly, the addition of 0.2 mM Si, more so than the addition of 0.6 mM Si, significantly alleviated the inhibitory effects of Cd toxicity on photosynthesis and chlorophyll‐fluorescence parameters. Our results suggest that 0.2 mM Si could be close to an optimum Si‐dose requirement for the alleviation of toxicity symptoms mediated by moderate (5 μM) Cd exposure.

Cadmium uptake and xylem loading are active processes in the hyperaccumulator Sedum alfredii

Sedum alfredii is a well known cadmium (Cd) hyperaccumulator native to China; however, the mechanism behind its hyperaccumulation of Cd is not fully understood. Through several hydroponic experiments, characteristics of Cd uptake and translocation were investigated in the hyperaccumulating ecotype (HE) of S. alfredii in comparison with its non-hyperaccumulating ecotype (NHE). The results showed that at Cd level of 10 microM measured Cd uptake in HE was 3-4 times higher than the implied Cd uptake calculated from transpiration rate. Furthermore, inhibition of transpiration rate in the HE has no essential effect on Cd accumulation in shoots of the plants. Low temperature treatment (4 degrees C) significantly inhibited Cd uptake and reduced upward translocation of Cd to shoots for 9 times in HE plants, whereas no such effect was observed in NHE. Cadmium concentration was 3-4-fold higher in xylem sap of HE, as compared with that in external uptake solution, whereas opposite results were obtained for NHE. Cadmium concentration in xylem sap of HE was significantly reduced by the addition of metabolic inhibitors, carbonyl cyanide m-chlorophenylhydrazone (CCCP) and 2,4-dinitrophenol (DNP), in the uptake solutions, whereas no such effect was noted in NHE. These results suggest that Cd uptake and translocation is an active process in plants of HE S. alfredii, symplastic pathway rather than apoplastic bypass contributes greatly to root uptake, xylem loading and translocation of Cd to the shoots of HE, in comparison with the NHE plants.

Investigating a potential mechanism of Cd resistance in Chironomus riparius larvae using kinetic analysis of calcium and cadmium uptake

The uptake kinetics of waterborne Ca and Cd, both independently and in combination, were examined in C. riparius larvae, which are extremely Cd tolerant. Larvae exposed to Ca (100–2500 μmol L −1), exhibited classic Michaelis–Menten saturation kinetics for Ca influx, measured using 45Ca as a radio-tracer. The maximum rate of Ca influx ( J max Ca ) was 0.39 μmol g −1 h −1, and the Ca concentration where the carrier reached half saturation ( K M Ca ) was 289 μmol L −1. Cd influx was measured using 109Cd as a radio-tracer in larvae exposed to Cd (0–1400 μmol L −1) while the Ca concentration was set to the K M Ca . This revealed a J max Cd (2.26 μmol g −1 h −1) which was nearly 6-fold higher that of Ca. This unusually high capacity for Cd uptake is in accordance with the huge tissue Cd burdens that chironomid larvae are able to accumulate during high level exposures. The apparent K M Cd (1133 μmol Cd L −1), when recalculated to account for the background Ca level, was still high (567 μmol Cd L −1), suggesting that this organism has a low affinity for Cd relative to most aquatic animals, indeed lower or comparable to its affinity for Ca. In consequence, even well above environmentally relevant Cd exposures, C. riparius does not accumulate Cd at the expense of Ca, thereby avoiding internal hypocalcaemia, in contrast to most other organisms which are much more sensitive to Cd. However, Ca influx was significantly reduced when 1200 μmol Cd L −1 was added to Ca exposures (96–2410 μmol L −1). Michaelis–Menten analysis revealed a similar J max Ca in Cd-exposed and control larvae (i.e. exposed only to Ca), but that the apparent K M Ca was many-fold higher in larvae which were simultaneously exposed to Ca and Cd. Conversely, increasing Ca concentrations (96–2410 μmol L −1) progressively inhibited Cd uptake from a Cd exposure concentration (1200 μmol L −1), providing additional support for a common transport system. These results suggest that the interaction of Cd and Ca in C. riparius is one of simple competitive interaction, and that the unusual Cd transport kinetics (low affinity, high capacity) relative to fairly standard Ca transport kinetics help explain the unusual tolerance that this organism has to acute Cd exposure.

Characterization of cadmium uptake and cytotoxicity in human osteoblast-like MG-63 cells

Since bone mass is maintained constant by the balance between osteoclastic bone resorption and osteoblastic bone formation, alterations in osteoblast proliferation and differentiation may disturb the equilibrium of bone remodeling. Exposure to cadmium (Cd) has been associated with the alteration of bone metabolism and the development of osteoporosis. Because little information is available about the direct effects of Cd on osteoblastic cells, we have characterized in vitro the cellular accumulation and cytotoxicity of Cd in human osteoblastic cells. Incubation of osteoblast-like MG-63 cells with increasing concentrations of Cd in serum-free culture medium reduced cell viability in a time- and concentration-dependent manner, suggesting that Cd accumulates in osteoblasts. Consequently, an uptake time-course could be characterized for the cellular accumulation of 109Cd in serum-free culture medium. In order to characterize the mechanisms of Cd uptake, experiments have been conducted under well-defined metal speciation conditions in chloride and nitrate transport media. The results revealed a preferential uptake of Cd 2+ species. The cellular accumulation and cytotoxicity of Cd increased in the absence of extracellular calcium (Ca), suggesting that Cd may enter the cells in part through Ca channels. However, neither the cellular accumulation nor the cytotoxicity of Cd was modified by voltage-dependent Ca channel (VDCC) modulators or potassium-induced depolarization. Moreover, exposure conditions activating or inhibiting capacitative Ca entry (CCE) failed to modify the cellular accumulation and cytotoxicity of Cd, which excludes the involvement of canonical transient receptor potential (TRPC) channels. The cellular accumulation and cytotoxicity of Cd were reduced by 2-APB, a known inhibitor of the Mg and Ca channel TRPM7 and were increased in the absence of extracellular magnesium (Mg). The inhibition of Cd uptake by Mg and Ca was not additive, suggesting that each ion inhibits the same uptake mechanism. Our results indicate that Cd uptake in human osteoblastic cells occurs, at least in part, through Ca- and Mg-inhibitable transport mechanisms, which may involve channels of the TRPM family. The effect of Cd on bone metabolism may be enhanced under low Ca and/or Mg levels.

BIOKINETICS OF CADMIUM AND ZINC IN A MARINE BACTERIUM: INFLUENCES OF METAL INTERACTION AND PRE-EXPOSURE

The uptake kinetics of Cd and Zn, as influenced by metal interaction and metal pre-exposure, was examined in the gram-positive marine bacterium Bacillus firmus over a wide range of ambient free-Cd and -Zn concentrations. Bacteria were exposed to experimental media with different concentrations of Cd and Zn over a short, 15-min period. Zinc was found to be an effective competitive inhibitor of Cd uptake when the Zn(2+) concentration ([Zn(2+)]) was increased to 10(-8) M, whereas the Cd concentration (ranging from 10(-9) to 10(-6) M) did not affect Zn uptake. Inhibition of Cd uptake was dependent on [Zn(2+)] instead of the [Zn(2+)] to Cd(2+) concentration ratio. Cadmium uptake at different [Zn(2+)] was significantly inhibited by a sulfur ligand (SH) blocker (N-ethylmaleimide) and a Ca-channel blocker (lanthanum), suggesting that competition between Cd and Zn most likely occurred via binding to the same transport sites. Cadmium efflux also was determined in the presence of different [Zn(2+)]. A biphasic depuration of Cd was found when [Zn(2+)] was greater than 10(-8) M, whereas the calculated Cd efflux rate was independent of [Zn(2+)]. We further exposed B. firmus at different Cd or Zn concentrations for 24 h, then determined the metal uptake and efflux kinetics as well as the metallothionein (MT) induction. Both the Cd and Zn cellular concentrations increased with greater exposed metal concentration, but the MT levels and efflux were little affected by the elevated metal concentration. To some extent, however, the Cd uptake was reduced with an elevated intracellular Zn concentration, suggesting that at high Cd concentrations, intracellular Zn can suppress the Cd uptake in B. firmus. These results help to understand the interactions of metals in the marine environments.

Reciprocal inhibition of Cd and Ca uptake in isolated head kidney cells of rainbow trout ( Oncorhynchus mykiss)

Some environmental pollutants, including cadmium (Cd) and zinc (Zn), can act as endocrine disruptors in fish, either in vivo or through a direct action on steroidogenic cells, as has been demonstrated in vitro. We have previously characterized Cd uptake in head kidney (homologue of mammalian adrenal) cells of rainbow trout ( Oncorhynchus mykiss) and have provided evidence for a Cd/Ca interaction. Here, we pursued our investigation of metal competition for uptake. Our results show that inorganic speciation conditions favour Cd uptake with optimal level of accumulation for Cd 2+ compared to chlorocomplexes ( CdCl n 2 - n ). Calcium uptake was studied for the first time in the fish head kidney cells and Ca was found to be less efficiently accumulated compared to Cd. A specific saturable mechanism of transport was characterized for Ca uptake but voltage-gated or La-sensitive cationic channels are unlikely to contribute appreciably. A concentration-dependent reciprocal inhibition was observed between Ca and Cd, whereas, Zn proved to inhibit Cd uptake exclusively. Additive inhibitory effect on Cd uptake was obtained with co-exposure to Ca and Zn. We conclude that Cd, but not Zn, may decrease Ca availability to the head kidney tissue. Also, Zn may partially protect against Cd toxicity but Zn would not protect against Cd-induced perturbation of Ca homeostasis.

Cadmium uptake in isolated adrenocortical cells of rainbow trout and yellow perch

Cadmium uptake was studied in isolated adrenocortical cells of rainbow trout ( Oncorhynchus mykiss) and yellow perch ( Perca flavescens) to test the hypothesis that the greater sensitivity of trout cells to Cd-induced disruption of cortisol secretion observed in previous studies is correlated to higher level of metal accumulation. There was no evidence for interspecies differences in accumulation level, and a specific transport mechanism of similar affinity has been characterized in both fish species. However, inhibition of Cd uptake by calcium was observed in rainbow trout exclusively. The free metal ion Cd 2 + and chlorocomplexes CdCl n 2 − n both contribute to Cd accumulation with different level of contribution between fish species. We conclude that interspecies differences in sensitivity to Cd endocrine disrupting effect are not necessarily related to different levels of metal accumulation but would rather be linked to transport pathways and metal speciation. Cadmium / calcium competition for uptake could be a determinant of the early Cd-induced impaired cortisol secretion in trout but not perch cells.

Roles of calcium and cadmium on Cd-containing intra-and extracellular formation of Ca crystals in tobacco

Growth was severely inhibited when tobacco plants were exposed to toxic levels of cadmium (0.2 mM). However, when this treatment was combined with a high concentration of calcium (30 mM), the Cd-induced damage was strongly alleviated. Under these enhanced conditions, i.e., 30 mM Ca and 0.2 mM Cd, Ca crystals not only were heavily deposited in the leaves but were also actively excreted from the trichomes. The X-ray spectrum from our Energy Dispersive analysis revealed that both intra-and extracellular Ca crystals contained detectable amounts of Cd. Moreover, intracellular Ca deposition in the leaves was stimulated only by a high Ca concentration (30 mM); moderate levels of Ca (3 mM) or a toxic amount of Cd (0.2 mM) alone resulted in crystal deposition that was undetectable under a light microscope. In contrast, extracellular crystal formation on the trichomes was stimulated by toxic Cd treatment but not by high Ca concentrations alone. Finally, Inductively Coupled Plasma Spectroscopy revealed that a high level of Ca (30 mM) suppressed Cd accumulation while also increasing the endogeneous Ca concentration in the leaves. These observations imply that the amelioration of Ca against toxic Cd in tobacco plants is a result of not only the inhibition of Cd uptake, but also the extra-and intracellular sequestration of cadmium via Ca crystallization.

Cytotoxicity of Cadmium and Characteristics of Its Transport in Cardiomyocytes

Cadmium (Cd) is reported to produce cardiotoxicity at doses and exposure conditions that cause no effect in kidney or liver. The purpose of the present investigation was to examine the cytotoxicity of Cd to neonatal rat cardiomyocytes in primary culture and to elucidate the transport characteristics of Cd in these cells at a nontoxic concentration. Cd concentrations of 0.1 μM and higher that are well tolerated by hepatocytes and renal cortical epithelial cells were toxic to the cardiomyocyte. The plot of initial uptake rate of Cd at various concentrations was nonlinear suggesting that, in addition to simple diffusion, other processes may also be involved. These processes required metabolic energy as pretreatment with dinitrophenol or sodium fluoride inhibited 58 and 59% of the Cd uptake, respectively. The uptake of Cd was also affected by the incubation temperature and lowering the temperature from 37 to 4°C reduced Cd uptake over 30 min by 61%. Cd uptake required interaction with membrane sulfhydryl groups; pretreatment withp-chloromercuribenzenesulfonic acid or mercuric chloride reduced Cd uptake by 46 and 58%, respectively. Cd utilized the transport pathways for calcium (Ca), zinc (Zn), and copper (Cu). Coincubation with 1.26 mM Ca competitively inhibited Cd uptake by 77%. In the presence of Ca, 30 μM Zn or Cu further inhibited Cd accumulation competitively by as much as 63 and 32%, respectively. Cd could enter the cardiomyocytes through Ca channels and Ca channel blocker, verapamil, inhibited up to 76% of Cd uptake. From the above results it can be concluded that Cd is highly toxic to the cardiomyocytes. A majority of Cd enters these cells through transport processes that exist for Ca, Zn, and Cu. The transport processes utilized by Cd are temperature sensitive and dependent on metabolic energy. Furthermore, these involve membrane sulfhydryl groups and include Ca channels.