Zinc toxicity on neonatal cortical neurons: involvement of glutathione chelation

Several mechanisms have been implicated in pathological neuronal death including zinc neurotoxicity, calcium excitotoxicity and oxidative injury. Glutathione (GSH) serves to provide reducing equivalents for the maintenance of oxidant homeostasis, and also plays roles in intracellular and intercellular signaling in the brain. We investigated the role of GSH homeostasis in the neurotoxic action of zinc using both mixed cortical cultures containing neurons and glia, and cortical neurons prepared from 1-day-old rats. Zinc caused neuronal cell death in a concentration-dependent manner. In parallel, a high concentration of zinc depleted GSH, in a time-dependent manner, preceding the onset of neuronal damage. Depletion of GSH by diethylmaleate injured neurons and exacerbated zinc-induced death. In contrast, replenishment of GSH attenuated zinc neurotoxicity. The thiol-containing compounds N-acetylcysteine and GSH chemically chelated zinc leading to decreases in the influx of zinc, the fall in GSH level and neuronal death. Interestingly, the glycolytic substrate pyruvate, but not lactate, chelated zinc concentration dependently and prevented its toxicity. On the other hand, pyrrolidine dithiocarbamate, serving as a zinc chaperon, enhanced its entry and toxicity. The results suggest that zinc non-enzymatically depleted GSH, an intrinsic factor for neuron survival, leading to activation of the cellular death signal and eventually neuronal death.     

Studies of aluminum neurobehavioral toxicity in the intact mammal

Summary 1. Aluminum (Al) has been implicated in neurotoxic syndromes in several conditions, including Alzheimer's disease (AD). The developmental stage of the mammalian brain most susceptible to Al was determined in rabbits systemically exposed to Al during the prenatal, postnatal, or second month or for 1 month as adults or as aged subjects. Eyeblink reflex classical conditioning showed an Al-induced learning deficit only in the adult and aged rabbits.2. 4-Aminopyridine, which was reported to improve learning in AD subjects, attenuated this Al-induced learning deficit.3. Conditioned eyeblink acquisition is slower in AD subjects than controls, supporting the Al-loaded rabbit as a model of some AD effects.4. To determine if the Al-loaded rabbit modeled the AD cholinergic deficit, acetylcholine (Ach) overflow was measured in rabbit hippocampus using microdialysis. Aluminum pretreatment reduced basal and potassium-stimulated Ach overflow compared to controls.5. Acetylcholine overflow increased as control rabbits acquired the conditioned eyeblink reflex, then subsequently decreased, although conditioned eyeblink performance continued. In contrast, Al-loaded rabbits showed a delay in conditioned eyeblink acquisition and greatly attenuated Ach overflow. The Al-induced attenuation of Ach overflow may contribute to the Al-induced learning deficit.6. Brain Al entry was studied using microdialysis of blood, brain, and lateral ventricle. Aluminum rapidly entered the brain and lateral ventricle. Frontal cortical Al was greater than lateral ventricular Al, suggesting that Al primarily enters the brain through the cerebral microvasculature.7. The brain/blood Al ratio was always significantly less than 1. This ratio was influenced by the Al form administered, brain site and animal species. Thus, there appears to be an active process moving Al out of brain extracellular fluid (ECF).8. Brain and blood dialysate Ach concentrations were not different after cyanide addition to the dialysate, supporting the conclusion that an active process moves Al out of brain ECF.     

Continuous flow rhizostat for the study of aluminum toxicity

Abstract. Wide variations in total aluminum (AL) concentration reported to have a toxic effect on plants arc attributed to imprecise definition of the chemical environment at the root interface. Because AL can complex with natural ligands and form time-dependent metastable species in water, the requirements of a fixed AL speciation in the nutrient medium can only be met by simultaneously holding constant the pH, the AL/OH ratio and the renewal time of the solution. It is also important that the root be constantly perfused by a fresh solution so that the root interface is as chemically close to the bulk solution as possible. The rhizostat presented here uses a process controller to constantly replenish four tanks containing different combinations of nutrients and AL. The solution is pumped from each tank to a tray where it is continuously delivered to seedlings growing on artificial medium. This rhizostat provides an accurate control for AL speciation. Preliminary experiments on white spruce seedlings show that levels of 10 micromolal or less total AL are sufficient to cause toxic effects at pH of 4.5 and without the formation of AL complexes.     

Some aspects of aluminum toxicity in plants

Aluminum toxicity is a major factor in limiting growth in plants in most strongly acid soils. Toxic effects on plant growth have been attributed to several physiological and biochemical pathways, although the precise mechanism is still not fully understood. In general, root elongation is hampered through reduced mitotic activity induced by Al, with subsequent increase in susceptibility to drought. The initial site of uptake is usually the root cap and the mucilaginous secretion covering the epidermal cells. Al ions bind very specifically to the mucilage by exchange adsorption on the polyuronic acid, complexing with the pectic substances and by the formation of polyhydroxy forms, increasing the number of Al atoms per positive charge. Toxicity has been suggested to be initiated at the sites of mucopolysaccharide synthesis. Al is absorbed on all Ca-binding sites on the cell surface. In the intact tissues, most of the Al is bound to the pectic substances of the cell wall and a part to the nucleic acids and cell membrane. Al is also reported to enter the plant by moving into meristematic cells via the cortex, bypassing the endodermal barrier. Being a polyvalent cation, it follows principally the apoplasmic pathway of transport through cortical cells, but may also enter the stele through the plasmalemma. Ultrastructural studies have shown the maximum accumulation to be in the epidermal and cortical cells. The interaction of Al with different systems follows different pathways. The plasma membrane at the outer boundary of the root cell is a potential target and its physical properties can be altered by Al through interaction with membrane-bound ATPase, lipids, carbohydrates and proteins. The Golgi apparatus has been suggested as the primary site of action, followed by damage to the plasmalemma. Aluminum interferes with the uptake, transport and use of several essential elements, including Cu, Zn, Ca, Mg, Mn, K, P and Fe. Excess of Al reduces the uptake of certain elements and increases that of others, the patterns being dependent on the element, the plant part and species involved. A major factor is the pH concentration. At an acid pH, below 5.5, the antagonism between Ca and Al is probably the most important factor affecting Ca uptake by plants. The molecular mechanism of tolerance of Al is as yet not clear. Tolerant plants reduce the absorption by the root or detoxify Al after absorption. Al tolerant plants may be grouped into those with higher Al concentrations in tops and those with less. In the latter, more Al is entrapped in roots. Uptake of Al may be reduced by binding to cell wall or to membrane lipid. Tolerance may be different in different species and seems to be controlled by one or more genes. Absorption of Al in non-metabolic conditions is affected only slightly by temperature. Anaerobic conditions, like the presence of nitrogen and metabolic inhibitors, damage the endodermal membrane barrier, increasing the uptake and enhancing injurious effects. Aluminum also causes morphological damage to plant parts. It affects photosynthesis by lowering chlorophyll content and reducing electron flow. Reduced respiratory activity might be due to reduced metabolic energy requirement. Protein synthesis is decreased probably due to effect on ribosome distribution at endoplasmic reticulum. Aluminum is known to bind to DNA and nuclei. However, its penetrance to DNA of mitotically active centers is slow. On accumulating in roots, it initially inhibits mitotic activity, possibly through affecting the integrated control function of the root meristem. Aluminum toxicity in acid soil is of special importance due to the destruction of components of forest ecosystems under specific conditions. It reduces biomass yield and tree growth and represses litter-degrading microflora. Further information is required on the factors affecting membrane permeability, distribution and accumulation of Al in different plant parts and different species. Al tolerance may be studied with relation to the presence of different ligands, nitrogen metabolism (nitrate reductase and protein accumulation), nitrogen tolerance in relation to pH change and metal ion activities, the role of Ca and P and interference with water relations and litter degradation.     

Rapid bioassay of aluminum toxicity in soil

Fifty-five acid soil horizons from 19 profiles were evaluated for aluminum toxicity using root elongation as a criterion in a two-day petri dish bioassay. The method proved to be simple, efficient, and precise enough to clearly distinguish aluminum toxicity differences among horizons within and between profiles. Although toxicity patterns within profiles differed, it was common for surface horizons to be less toxic even when very acid. The R² for correlations of relative root lengths with pH in H₂O, pH in KCl, soluble and exchangeable aluminum and percent aluminum saturation were only 0.42, 0.45, 0.52, 0.66, and 0.54, respectively, which indicates the need for a bioassay. In a further use of the method, and to demonstrate its efficiency, 243 horizons from 26 profiles were screened. Approximately half of the horizons with a pH of 5.0 or below showed Al toxicity. When used by different operators, with a variety of soil and treatment parameter changes, the two-day bioassay in petri dishes gave consistent rankings of soils by degree of aluminum toxicity. Journal Paper 11690, Purdue University, Agricultural Experiment Station, West Lafayette, Indiana, USA. Contribution from the Agronomy Department.     

Cation amelioration of aluminum toxicity in wheat

Aluminum is a major constituent of most soils and limits crop productivity in many regions. Amelioration is of theoretical as well as practical interest because understanding amelioration may contribute to an understanding of the mechanisms of toxicity. In the experiments reported here 2-day-old wheat (Triticum aestivum L. cv Tyler) seedlings with 15-millimeter roots were transferred to solutions containing 0.4 millimolar CaCl₂ at pH 4.3 variously supplemented with AlCl₃ and additional amounts of a chloride salt. Root lengths, measured after 2 days in the test solutions, were a function of both Al activity and the cation activity of the added salt. Percent inhibition = 100 {Al³⁺}/({Al³⁺} + K_m + α{C}^β) where {Al³⁺} is the activity of Al³⁺ expressed in micromolar, {C} is the activity of the added cation expressed in millimolar, and K_m (= 1.2 micromolar) is the {Al³⁺} required for 50% inhibition in the absence of added salt. For Ca²⁺, Mg²⁺, and Na⁺ the values of α were 2.4, 1.6, and 0.011, respectively, and the values for β were 1.5, 1.5, and 1.8, respectively. With regard to relative ameliorative effectiveness, Ca²⁺ > Mg²⁺ ≈ Sr²⁺ K⁺ ≈ Na⁺. Other cations were tested, but La³⁺, Sc³⁺, Li⁺, Rb⁺, and Cs⁺ were toxic at potentially ameliorative levels. The salt amelioration is not solely attributable to reductions in {Al³⁺} caused by increases in ionic strength. Competition between the cation and Al for external binding sites may account for most of the amelioration.     

Effects of silicon on aluminum toxicity in upland rice plants

Aims

This study aimed to determine the capacity of Si to mitigate Al toxicity in upland rice plants (Oryza sativa L.) by evaluating plant growth and the Si and Al uptake kinetics.

Methods

Plants were grown for 40 days, after which the Si and Al uptake kinetics (Cmin, Km and Imax) were analyzed. Then, the shoots and roots were separated, and the dry matter, root morphology and Si and Al concentration and accumulation in the plant were evaluated.

Results

Aluminum decreased plant growth and the Si uptake capacity by decreasing the root growth and Si transport system efficiency in the upland rice roots (> Km and > Cmin). Silicon mitigated Al toxicity in the upland rice plants by decreasing Al transport to the plant shoots, although it did not reduce the Al uptake rate (Imax). Si treatment increased the growth of upland rice plant shoots grown in the presence of Al without influencing the root growth. The alleviation of Al toxicity by Si is more evident in the susceptible upland rice cultivar Maravilha.

Conclusions

Silicon mitigated Al toxicity in the upland rice plants by decreasing Al transport to the plant shoots but did not reduce the Al uptake rate by roots.

     

微生物铝毒害和耐铝毒机制研究进展

在酸性土壤中,铝毒是限制农作物生产的关键问题之一,铝同样对微生物产生毒害作用。研究微生物的铝毒害和耐铝毒机制可以为植物耐铝毒机制的研究提供一种新视角。目前的研究表明,铝作用于微生物细胞的细胞壁、细胞膜、细胞核和细胞器,影响微生物的物质和能量代谢,抑制微生物的生长和发育。针对这些毒害作用,铝毒耐受微生物通过多途径全方位的机制适应外界的铝毒环境。该文结合作者的研究工作,综述了微生物的铝毒害和耐铝毒机制。     

Analysis of time-dependent recovery from beryllium toxicity following chelation therapy and antioxidant supplementation

Efforts have been made to minimize the toxic effect caused by beryllium. Adult cyclic rats of Sprague Dawley strain were administered a bolus dose of 50mg/kg beryllium nitrate intramuscularly. The chelation therapy with glutathione (GSH), dimercapto propane sulfonic acid (DMPS)+ selenium (Se) and D-Penicillamine (DPA) + Se was given for 3 days followed by a rest of 1,3 and 7 days respectively. The results revealed a significant fall in the blood sugar level, serum alkaline phosphatase activity, serum proteins. A significant rise in the transaminases i.e. aspartate aminotranferase and alanine aminotranferase pattern is indicative of leakage of enzymes from liver resulting in alterations in the cell permeability. A rise in the hepatic lipid peroxidation activity is a direct indication of oxidative damage resulting in free radical generation. Results of the distribution studies by atomic absorption spectrophotometry reveal an increased concentration of beryllium in liver and kidney followed by lung and uterus. The relative ability of 3 chelating agents to act as antagonists for acute beryllium poisoning have been examined in liver, kidney, lungs and uterus. The appreciable change in the beryllium concentration in various organs is duration-dependent during the entire period being highly significant after 7 days rest. From the biochemical assays, and distribution studies it can be assumed that DPA+Se was the most effective therapeutic agent followed by DMPS+Se and GSH. Thus it can be concluded that DPA+Se is a better therapeutic agent as compared to DMPS+Se and GSH.     

Apoplastic binding of aluminum is involved in silicon-induced amelioration of aluminum toxicity in maize

The alleviating effect of silicon (Si) supply on aluminum (Al) toxicity was suggested to be based on ex or in planta mechanisms. In our experiments with the Al-sensitive maize (Zea mays) cultivar Lixis, Si treatment but not Si pretreatment ameliorated Al-induced root injury as revealed by less root-growth inhibition and callose formation. Si treatment did not affect monomeric Al concentrations in the nutrient solution, suggesting an in planta effect of Si on Al resistance. A fractionated analysis of Si and Al in the 1-cm root apices revealed that more than 85% of the root-tip Al was bound in the cell wall. Al contents in the apoplastic sap, the symplastic sap, and the cell wall did not differ between -Si and +Si plants. Si did not affect the Al-induced exudation of organic acid anions and phenols from the root apices. However, Al treatment greatly enhanced Si accumulation in the cell wall fraction, reducing the mobility of apoplastic Al. From our data we conclude that Si treatment leads to the formation of hydroxyaluminumsilicates in the apoplast of the root apex, thus detoxifying Al.     

Redox metal chelation ameliorates radiation-induced bone marrow toxicity in a mouse model.

Since zinc desferrioxamine (Zn-DFO) has been shown to be a very potent protector against injuries induced by redox-active metal ions, we examined its protective effect against radiation-induced toxicity. We found that treatment with Zn-DFO given before TBI increased the survival of mice irradiated with 7.5 and 8.5 Gy. Zn-DFO also protected against radiation-induced myelosuppression and body weight loss, while soluble Il6 levels in serum were normalized in mice pretreated with Zn-DFO. We concluded that administration of Zn-DFO prior to TBI protected BALB/c mice from radiation-induced toxicity, increasing survival rates by up to 75%. The biological effect of Zn-DFO is known to result from its effect on the production of intracellular hydroxyl free radicals mediated by redox-active metal ions, and both metal chelation and zinc delivery appear to be equally likely mechanisms for this outcome. We suggest that radiation-induced toxicity is caused by the deleterious effect of redox-active metal ions, and that compounds which modulate this redox activity may act as radioprotectors.     

Effect of oral aluminum and aluminum citrate on blood level and short-term tissue distribution of aluminum in the rat

Aluminum (Al) absorption seems to be very low, but many factors can enhance it in animals and humans. In the present study, we investigated the acute effect of Na citrate on Al absorption by monitoring Al levels in blood and several tissues. For this purpose, 18 Wistar male rats were divided into 3 groups: control, Al, and Al + Na citrate. After a 14-h fasting period, animals were dosed orally with deionized water, or 2 mmol Al chloride, or 2 mmol Al chloride plus 2 mmol Na citrate. Blood samples were taken before and 1, 2, 4, and 6 h after the gavage. Al concentrations in blood, liver, tibia, kidney, and intestinal wall were determined by ICP-OES. In the Al and Al + citrate groups, Al blood concentrations peaked at 1 h and 2 h with higher levels in the Al + citrate group. Al gavage resulted in an increase in Al level in intestinal wall, but not in the other investigated tissues. Simultaneous gavage of citrate with Al significantly increased its tissue levels in tibia, kidney, and in intestinal wall. Our data show clearly that Al as chloride can be absorbed, but not well retained by the organism tissues. Furthermore, the model used in the present study is appropriate for acute studies to investigate the effect of various compounds on Al absorption in the rat.     

Hydrogen sulfide alleviates aluminum toxicity in barley seedlings

Aims

Aluminum (Al) toxicity is one of the major factors that limit plant growth. Low concentration of hydrogen sulfide (H₂S) has been proven to function in physiological responses to various stresses. The objective of this study is to investigate the possible role of H₂S in Al toxicity in barley (Hordeum vulgare L) seedlings.

Methods

Barley seedlings pre-treated with sodium hydrosulfide (NaHS), a H₂S donor, and subsequently exposed to Al treatment were studied for their effects on root elongation, Al accumulation in seedlings, Al-induced citrate secretion and oxidative stress, and plasma membrane (PM) H⁺-ATPase expression.

Results

Our results showed that H₂S had significant rescue effects on Al-induced inhibition of root elongation which was correlated well with the decrease of Al accumulation in seedlings. Meanwhile, Al-induced citrate secretion was also significantly enhanced by NaHS pretreatment. Al-induced oxidative stress as indicated by lipid peroxidation and reactive oxygen species burst was alleviated by H₂S through the activation of the antioxidant system. Moreover, Al-induced reduction in PM H⁺-ATPase expression was reversed by exogenous NaHS.

Conclusions

Altogether, our results suggest H₂S plays an ameliorative role in protecting plants against Al toxicity by inducing the activities of antioxidant enzymes, increasing citrate secretion and citrate transporter gene expression, and enhancing the expression of PM H⁺-ATPase.     

Molecular physiology of aluminum toxicity and tolerance in plants

Aluminum being the third most abundant metal in the earth’s crust poses a serious threat to crop productivity in acid soils, which comprise almost half of the arable land. This review travels across time and updates research done on aluminum stress in plants. In its phytotoxic forms, aluminum affects root growth by acting in the root apical zone, resulting in growth inhibition in a very short time at micromolar concentrations. The mechanisms of aluminum toxicity in plants may proceed by growth inhibition, callose accumulation, cytoskeletal distortion, disturbance of plasma membrane surface charge, and H⁺-ATPase activity, lipid peroxidation of membranes, production of reactive oxygen species in cytosol and mitochondria, respiratory dysfunction, opening of mitochondrial permeability transition pores, collapsing of inner mitochondrial membrane potential, activation of mitochondrial protease, and induction of nuclear apoptosis, resulting ultimately in programmed cell death. In contrast, the mechanism of tolerance involves the exudation of organic acid anions, complexation of aluminum with organic acids, and subsequent detoxification. Many oxidative stress genes and other metabolically important genes have also been found to be induced under aluminum stress and overexpression analyses have also shown some plants to develop some degree of tolerance. In the future, researchers in the area of aluminum research should investigate more basic mechanisms of aluminum toxicity and discover and study more aluminum-responsive genes that confer resistance against this toxic metal, to ensure food security for ever-increasing human populations in the future.     

Interaction between aluminum toxicity and manganese toxicity in soybean (Glycine max)

Plant and Soil - A greenhouse experiment was conducted on weakly acidic and calcareous soils to evaluate the aging and residual effects of three natural organic Zn chelates...     

Effect of aluminum exposure on pteridine metabolism

Occupational and environmental aluminum (Al) exposure cause serious health problems by interaction with biological systems. Al is one of the most documented metals because its cellular targets are unclear biochemical processes and membranes of organisms. The major aim of the present study was to investigate the alteration of serum and urine aluminum in occupational exposure and to observe whether the metal exposure could cause any changes in pteridine-pathway-related critical compounds such as urinary neopterin and biopterin and blood dihydropteridine reductase (DHPR). In this study, determination of the metal concentrations was carried out in Al-exposed workers (n=23) and healthy volunteers (n=18) by using a tomic absorption spectrometer. DHPR enzyme activity and levels of neopterin and biopterin were detected by spectrophotometric and high-performance liquid chromatographic methods, respectively. It was found that occupational exposure to the metal led to a statistically significant increase in serum Al levels compared to the controls (p<0.05). At the same time, urinary neopterin and biopterin concentrations of the exposed group were higher than nonexposed subjects (both p<0.05). The correlations among Al levels and DHPR activity, magnesium concentration in serum and urine, working years, smoking status, and age were evaluated.