Low Glutamine Synthetase Activity Research Articles

Zinc Deficiency Causes Oxidative Damage to Proteins, Lipids and DNA in Rat Testes

To investigate the effects of zinc deficiency on oxidative damage to testes proteins, lipids and DNA, weanling male rats were allowed free access to low (0.5 µg Zn/g) or adequate (25 µg Zn/g) zinc diets for 14 d. A third group was restricted intake of the adequate Zn diet to the amount consumed by the low Zn diet-fed group. Compared with ad libitum-fed controls, testes from rats fed the low zinc diet had lower glutamine synthetase activity, lower Fe2+-stimulated 2-thiobarbituric acid-reactive substances (TBARS) production, higher protein carbonyl concentrations (P < 0.05), and higher 8-oxo-2′-deoxyguanosine levels (P = 0.06). Glutamine synthetase activity in testes of the food-restricted controls was between the values for the ad libitum controls and zinc-deficient animals. Protein carbonyls were higher in the restricted controls compared with the ad libitum controls, whereas stimulated TBARS production was lower (P < 0.05). Levels of 8-oxo-2′-deoxyguanosine were lower in testes DNA of the restricted controls than in the zinc-deficient group (P < 0.05). Testes iron concentrations were higher in the zinc-deficient and restricted control rats than in ad libitum controls (P < 0.05). The oxidative damage observed may have occurred as a consequence of increased reactive oxygen species generation secondary to tissue iron accumulation and/or reductions in zinc-dependent antioxidant processes.

Comparative Study of the Effects of Salinity on Growth, Gas Exchange, N Accumulation and Stable Isotope Signatures of Forage Oat (Avena sativa L.) Genotypes.

Identifying suitable salt stress-tolerant phenotypes based on their agronomic and physiological traits remains a herculean task in forage-type oat (Avena sativa L.) breeding. This study examined the responses of six forage-type oat cultivars under four levels of saline stress over the vegetative growth cycle. Crop growth, water status-related traits and nitrogen status-related traits were analyzed in different plant parts to evaluate effective approaches for identifying salt tolerance. Plant biomass, height, tiller number and culm thickness changed substantially during salinity, but they were not precise enough for use in estimating genotypic salinity tolerance during long-term stress. Genotypes bearing larger numbers of tillers showed greater sensitivity to salinity due to its effects on biomass loss. Tolerant genotypes exhibited higher relative shoot biomass together with higher water use efficiency. The concentrations of Na+, K+ and their ratio, combined with the δ13C in shoots and roots were effective indicators for estimating tolerant genotypes through better water maintenance. N concentrations of shoots were the most efficient for evaluating genotypic tolerance. Low nitrate reductase (NR) and glutamine synthetase (GS) activity might be key factors limiting N accumulation. Chlorophyll (Chl) content and net photosynthetic rate, as well as stomatal conductance and evaporation, were useful for identifying salinity tolerance physiological mechanisms, but the effectiveness was low for genotypic tolerance testing for forage type oats due to the interaction between genotypes and salinity levels. The selection of high salinity-tolerant genotypes should focus on genotypes with photosynthetic resilience to salt, followed by high N metabolism (higher NR and GS activities) to ensure accumulation of more N in the shoot dry matter.

CHAPERONES AS MOLECULAR TARGET FOR ONCOLOGICAL AND NEUROLOGICAL DISEASES

Brain manganese deposition is led by liver dysfunction and/or portal-systemic shunting in minimal hepatic encephalopathy (MHE). Manganese is toxic and can cause cognitive disorders and extrapyramidal symptoms. Thus, reduction of manganese intake might be considered as a potential treatment strategy for MHE. In this study we aimed to investigate whether low- or no-manganese feed can improve the neuropsychological manifestations in MHE rats. Rats with MHE were established by partially ligating the portal vein and fed a manganese diet (MHE-M, 10 mg per kg feed; n = 24), a no-manganese diet (MHE-N; n = 24) and a half-manganese diet (MHE-H; n = 24) for 2, 4, 6 and 8 weeks, with six rats in each subgroup. Morris water maze (MWM), open-field test and narrow beam test (NBT) were used to evaluate the cognitive and locomotor situations. Fasted blood ammonia, manganese content and glutamine synthetase (GS) activity in basal ganglia and cortex were measured. A significantly longer MWM escape latency, less locomotor activity, longer NBT latency and total time, higher blood ammonia, higher brain manganese content and GS activity were found in MHE-M rats. However, a significantly shorter MWM escape latency, increased locomotor activity, shorter NBT latency and total time, lower blood ammonia, lower brain manganese content and lower GS activity were found in MHE-N rats after no-manganese feed treatment. Partial improvements were found in MHE rats with half-manganese feed treatment. Reduction of manganese intake can significantly improve the cognitive and locomotor situations in MHE rats by reducing brain manganese content and regulating GS activity.

Oral administration of trace element magnesium significantly improving the cognition and locomotion in hepatic encephalopathy rats

The therapeutic effects of iron, zinc and magnesium trace elements, as well as rifaximin were investigated and compared in HE rats. In this study, HE rats were treated with either ferrous sulfate (HE-Fe, 30 mg/kg/day), zinc sulfate (HE-Zn, 30 mg/kg/day), magnesium sulfate (HE-Mg, 50 mg/kg/day) or rifaximin (HE-Rf, 50 mg/kg/day), which was mixed with water and administered orally for 61 days. The Morris water maze (MWM) and open-field tests were used to evaluate cognitive and locomotor function. The blood ammonia levels before and after administration of the glutamine challenge test, manganese concentration and glutamine synthetase (GS) activity were measured. Significantly longer MWM escape latencies, less locomotor activity, higher blood ammonia levels, higher brain manganese concentrations and higher GS activity were observed in HE rats. However, HE-Mg and HE-Rf rats had significantly shorter MWM escape latencies, increased locomotor activity, lower blood ammonia, lower brain manganese concentrations and lower GS activity. Partial improvements were observed in HE-Fe and HE-Zn rats. The results indicated that oral administration of magnesium can significantly improve the cognitive and locomotor functions in HE rats by reducing the brain manganese concentration and regulating GS activity.

Reduction of manganese intake improves neuropsychological manifestations in rats with minimal hepatic encephalopathy

Brain manganese deposition is led by liver dysfunction and/or portal-systemic shunting in minimal hepatic encephalopathy (MHE). Manganese is toxic and can cause cognitive disorders and extrapyramidal symptoms. Thus, reduction of manganese intake might be considered as a potential treatment strategy for MHE. In this study we aimed to investigate whether low- or no-manganese feed can improve the neuropsychological manifestations in MHE rats. Rats with MHE were established by partially ligating the portal vein and fed a manganese diet (MHE-M, 10mg per kg feed; n=24), a no-manganese diet (MHE-N; n=24) and a half-manganese diet (MHE-H; n=24) for 2, 4, 6 and 8weeks, with six rats in each subgroup. Morris water maze (MWM), open-field test and narrow beam test (NBT) were used to evaluate the cognitive and locomotor situations. Fasted blood ammonia, manganese content and glutamine synthetase (GS) activity in basal ganglia and cortex were measured. A significantly longer MWM escape latency, less locomotor activity, longer NBT latency and total time, higher blood ammonia, higher brain manganese content and GS activity were found in MHE-M rats. However, a significantly shorter MWM escape latency, increased locomotor activity, shorter NBT latency and total time, lower blood ammonia, lower brain manganese content and lower GS activity were found in MHE-N rats after no-manganese feed treatment. Partial improvements were found in MHE rats with half-manganese feed treatment. Reduction of manganese intake can significantly improve the cognitive and locomotor situations in MHE rats by reducing brain manganese content and regulating GS activity.

Metabolic regulation of ammonia emission in different senescence phenotypes of Nicotiana tabacum

In order to reveal the character of ammonia emission in senescent tobacco (Nicotiana tabacum), the content of NH4 +, total nitrogen, and soluble protein, and the activities of nitrogen metabolism-related enzymes were measured in leaves of a quick-leaf-senescence phenotype ZY90 and a slow-leaf-senescence phenotype NC89. Compared with NC89, ZY90 had a higher NH4 + accumulation, a lower glutamine synthetase activity, and a significantly higher stomatal ammonia compensation point, and ammonia emission during 40 to 60 d after leaf emergence. During senescence, the quick-leafsenescence phenotype was characterized by nitrogen re-transfer by ammonia emmission, whereas the slow-leafsenescence phenotype by nitrogen re-assimilation. The ammonia emission was primarily regulated by glutamine synthetase activity, apoplastic pH, and NH4 + content.

Type 1 diabetes alters astrocytic properties related with neurotransmitter supply, causing abnormal neuronal activities

Glutamine synthetase (GS), an astrocytic protein in the brain, mediates the process by which glutamate (Glu) is transformed into glutamine (Gln) during Glu and gamma-aminobutyric acid (GABA) de novo synthesis. There are many types of neural complications related with those neurotransmitters in type 1 diabetes (T1D) patients, but there is little information about the change GS. Therefore, we examined changes in GS activity and expression, as well as the amount of Glu, Gln, and GABA in the brain of a T1D animal model. Using primary culture we found that glucose fluctuation caused glial fibrillary acidic protein (GFAP) and GS changes but constant high glucose level didn׳t. In T1D mouse, GS expression increased in the prefrontal cortex (PFC) and hippocampus (HI), but decreased GS activity was only observed in the HI whereas GFAP expression decreased in both regions. Gln increased in both regions, but Glu and GABA were only increased in the HI of T1D animals where GS activity decreased with higher reactive oxygen/nitrogen species. Collectively, low GS activity may be closely related with high levels of Glu and GABA in the HI of T1D brain, and this would result in abnormal neurotransmissions.

Effect of elevated carbon dioxide on nitrogen assimilation and mobilization in wheat and rye genotypes of different ploidy levels

Two wheat genotypes differing in ploidy level viz. PBW 343 (hexaploid) and PDW 274 (tetraploid), and rye genotype WSP 540-2 (diploid) were grown under elevated CO2 (EC: 550 ± 50 ppm) and ambient CO2 (AC: 380 ppm) to study the changes in nitrogen assimilatory pathway enzymes. Elevated CO2 increased nitrate reductase (NR) activity in flag leaves of the three genotypes at milk stage, and activity was highest in rye (diploid). At dough stage, hexaploid and tetraploid genotypes showed higher NR activity in plants grown under EC. Elevated CO2 resulted in higher expression of NR gene (NIA1). Activity of glutamine synthetase (GS) in flag leaves was higher in all the three genotypes under EC at milk stage but the activity declined at dough stage. The expression of GS1 increased in flag leaves of plants grown under EC at both milk and dough stages, while the expression of GS2 declined during the reproductive stages, especially in ears of EC grown plants. Nitrate content decreased in leaf tissues of all the three genotypes by 90 DAS in EC grown plants. This indicated enhanced nitrate assimilation in leaves by NR under EC at reproductive stage. However, lower GS2 expression and lower GS activity during late reproductive phase (dough stage) indicated inhibition of consequent steps.

Calcium-induced proline accumulation contributes to amelioration of NaCl injury and expression of glutamine synthetase in greater duckweed (Spirodela polyrhiza L.)

The calcium-mediated proline accumulation is a critical response under NaCl stress and the function of the induced proline as a glutamine synthetase (GS) protectant in greater duckweed was investigated. The plants were treated with solutions containing 100mM NaCl, 200mM NaCl, 200mM NaCl plus 10mM CaCl2, or 10mM CaCl2 alone for 4 days. At the end of the experiment, the fronds of inoculum treated with 200mM NaCl showed the chlorotic effect, higher glutamate dehydrogenase (NADH-GDH) activity and lower GS activity. At the lower salinity, the activities of GS and NADH-GDH were not altered markedly. A significant accumulation of proline was not found under either low or high salinity. The activity of Δ1-pyrroline-5-carboxylate reductase (P5CR) was enhanced only at 200mM NaCl but remained unchanged at 100mM NaCl. The activity of Δ1-pyrroline-5-carboxylate synthetase (P5CS) did not change under salinity-stressed. Addition of CaCl2 to the salt stressed plants not only lowered NaCl injury but also showed an elevated level of proline contents in response to the salinity treatment. In addition, both GS activity and corresponding polypeptides were expressed close to the level of control. Exogenous proline protects GS2 and the 32kDa protein in photosystem II reaction center (D1) from H2O2-induced redox degradation in the chloroplast lysates of duckweed. The results suggest that calcium-induced proline accumulation may play an important role as a GS protectant under NaCl exposure in S. polyrhiza.

Ammonia emission from rice leaves in relation to photorespiration and genotypic differences in glutamine synthetase activity

Rice (Oryza sativa) plants lose significant amounts of volatile NH(3) from their leaves, but it has not been shown that this is a consequence of photorespiration. Involvement of photorespiration in NH(3) emission and the role of glutamine synthetase (GS) on NH(3) recycling were investigated using two rice cultivars with different GS activities. NH(3) emission (AER), and gross photosynthesis (P(G)), transpiration (Tr) and stomatal conductance (g(S)) were measured on leaves of 'Akenohoshi', a cultivar with high GS activity, and 'Kasalath', a cultivar with low GS activity, under different light intensities (200, 500 and 1000 µmol m(-2) s(-1)), leaf temperatures (27·5, 32·5 and 37·5 °C) and atmospheric O(2) concentrations ([O(2)]: 2, 21 and 40 %, corresponding to 20, 210 and 400 mmol mol(-1)). An increase in [O(2)] increased AER in the two cultivars, accompanied by a decrease in P(G) due to enhanced photorespiration, but did not greatly influence Tr and g(S). There were significant positive correlations between AER and photorespiration in both cultivars. Increasing light intensity increased AER, P(G), Tr and g(S) in both cultivars, whereas increasing leaf temperature increased AER and Tr but slightly decreased P(G) and g(S). 'Kasalath' (low GS activity) showed higher AER than 'Akenohoshi' (high GS activity) at high light intensity, leaf temperature and [O(2)]. Our results demonstrate that photorespiration is strongly involved in NH(3) emission by rice leaves and suggest that differences in AER between cultivars result from their different GS activities, which would result in different capacities for reassimilation of photorespiratory NH(3). The results also suggest that NH(3) emission in rice leaves is not directly controlled by transpiration and stomatal conductance.

Effect of N-(n-butyl) thiophosphoric triamide on urea metabolism and the assimilation of ammonium by Triticum aestivum L.

The use of urea as an N fertilizer has increased to such an extent that it is now the most widely used fertilizer in the world. However, N losses as a result of ammonia volatilization lead to a decrease in its efficiency, therefore different methods have been developed over the years to reduce these losses. One of the most recent involves the use of urea combined with urease inhibitors, such as N-(n-butyl) thiophosphoric triamide (NBPT), in an attempt to delay the hydrolysis of urea in the soil. The aim of this study was to perform an in-depth analysis of the effect that NBPT use has on plant growth and N metabolism. Wheat plants were cultivated in a greenhouse experiment lasting 4 weeks and fertilized with urea and NBPT at different concentrations (0, 0.012, 0.062, 0.125%). Each treatment was replicated six times. A non-fertilized control was also cultivated. Several parameters related with N metabolism were analysed at the end of growth period. NBPT use was found to have visible effects, such as a transitory yellowing of the leaf tips, at the end of the first week of treatment. At a metabolic level, plants treated with the inhibitor were found to have more urea in their tissues and a lower amino acid content, lower glutamine synthetase activity, and lower urease and glutamine synthetase content at the end of the study period, whereas their urease activity seemed to have recovered by this stage.

Impact of the brine from a desalination plant on a shallow seagrass ( Posidonia oceanica) meadow

Although seawater desalination has increased significantly over recent decades, little attention has been paid to the impact of the main by-product (hypersaline water: brine) on ecosystems. In the Mediterranean, potentially the most affected ecosystems are meadows of the endemic seagrass Posidonia oceanica. We studied the effect of brine on a shallow P. oceanica meadow exposed to reverse osmosis brine discharge for more than 6 years. P. oceanica proved to be very sensitive to both eutrophication and high salinities derived from the brine discharge. Affected plants showed high epiphyte load and nitrogen content in the leaves, high frequencies of necrosis marks, low total non-structural carbohydrates and low glutamine synthetase activity, compared to control plants. However, there was no indication of extensive decline of the affected meadow. This is probably due to its very shallow situation, which results in high incident radiation as well as fast dilution and dispersion of the brine plume.

Alterations in glutamine synthetase activity in rat skeletal muscle are associated with advanced age

Objective Glutamine synthetase (GS), a key enzyme in the production of glutamine, is preserved in skeletal muscle during early aging (<24 mo). Because the effects of advanced age on GS are unknown, we investigated the effect of advanced age (>24 mo) on GS activity in skeletal muscle. We hypothesized that advanced age would enhance muscle GS activity. Methods Muscle GS activities were assessed in adult (8 mo), mature adult (15 mo), aged (20–22 mo), advanced age (25–27 mo), or very advanced age (29–32 mo) female Wistar rats. Male Wistar (6–27 mo) were used to investigate the effect of gender on this activity. Results Glutamine synthetase activity remained low and unaltered in rats from 8 to 22 mo of age, as previously demonstrated. In contrast, GS activity was high (∼75% of individual values were higher than the low value mean) in 25-mo to 27-mo-old rats. In very-old-aged rats (29–32 mo), ∼55% of GS activity data points exhibited low values. Changes in GS protein content paralleled those in GS activities. In male rats, GS activity was also high (∼80% of individual values were higher than the mean value of 6-mo to 19-mo-old rats) at the upper limit of life expectancy (27 mo). Conclusion There is enhanced GS activity in old female and male rats suggesting a greater need for glutamine. In some very old rats, low GS activity may be associated with longevity or reflect a limitation in glutamine production due to extremely advanced age per se.

GlnD is essential for NifA activation, NtrB/NtrC-regulated gene expression, and posttranslational regulation of nitrogenase activity in the photosynthetic, nitrogen-fixing bacterium Rhodospirillum rubrum.

GlnD is a bifunctional uridylyltransferase/uridylyl-removing enzyme and is thought to be the primary sensor of nitrogen status in the cell. It plays an important role in nitrogen assimilation and metabolism by reversibly regulating the modification of P(II) proteins, which in turn regulate a variety of other proteins. We report here the characterization of glnD mutants from the photosynthetic, nitrogen-fixing bacterium Rhodospirillum rubrum and the analysis of the roles of GlnD in the regulation of nitrogen fixation. Unlike glnD mutations in Azotobacter vinelandii and some other bacteria, glnD deletion mutations are not lethal in R. rubrum. Such mutants grew well in minimal medium with glutamate as the sole nitrogen source, although they grew slowly with ammonium as the sole nitrogen source (MN medium) and were unable to fix N(2). The slow growth in MN medium is apparently due to low glutamine synthetase activity, because a DeltaglnD strain with an altered glutamine synthetase that cannot be adenylylated can grow well in MN medium. Various mutation and complementation studies were used to show that the critical uridylyltransferase activity of GlnD is localized to the N-terminal region. Mutants with intermediate levels of uridylyltransferase activity are differentially defective in nif gene expression, the posttranslational regulation of nitrogenase, and NtrB/NtrC function, indicating the complexity of the physiological role of GlnD. These results have implications for the interpretation of results obtained with GlnD in many other organisms.

Glutamine metabolism in uricotelic species: variation in skeletal muscle glutamine synthetase, glutaminase, glutamine levels and rates of protein synthesis

High intracellular glutamine levels have been implicated in promoting net protein synthesis and accretion in mammalian skeletal muscle. Little is known regarding glutamine metabolism in uricotelic species but chicken breast muscle exhibits high rates of protein accretion and would be predicted to maintain high glutamine levels. However, chicken breast muscle expresses high glutaminase activity and here we report that chicken breast muscle also expresses low glutamine synthetase activity (0.07±0.01 U/g) when compared to leg muscle (0.50±0.04 U/g). Free glutamine levels were 1.38±0.09 and 9.69±0.12 nmol/mg wet weight in breast and leg muscles of fed chickens, respectively. Glutamine levels were also lower in dove breast muscle (4.82±0.35 nmol/mg wet weight) when compared to leg muscle (16.2±1.0 nmol/mg wet weight) and much lower (1.80±0.46 nmol/mg wet weight) in lizard leg muscle. In fed chickens, rates of fractional protein synthesis were higher in leg than in breast muscle, and starvation (48 h) resulted in a decrease in both glutamine content and rate of protein synthesis in leg muscle. Thus, although tissue-specific glutamine metabolism in uricotelic species differs markedly from that in ureotelic animals, differences in rates of skeletal muscle protein synthesis are associated with corresponding differences in intramuscular glutamine content.

EFFECT OF NITROGEN STRESS ON GROWTH, SOLUBLE METABOLITES, AND GLUTAMINE SYNTHETASE ACTIVITY OF JACK PINE CALLUS CULTURES

Jack pine (Pinus banksiana Lamb.) callus cultures were grown in half-strength unmodified Litvay medium (control) and in media containing either no-nitrogen or nitrate. Almost no growth occurred in absence of nitrogen and in medium containing only nitrate. Glutamine synthetase (GS) activity was drastically reduced in comparison to callus grown in control medium. The pool of amino acids decreased by 66% in medium containing nitrate, and further by 50% in absence of nitrogen. Meanwhile total and reducing sugars sharply increased, and were 14 times higher in N-NO3 − and 22 times higher in nitrogen-free medium. Thirteen amino acids were identified in the control. Changes in the abundance and the relative content of amino acids confirmed that glutamine is the preferred nitrogen transport compound in Pinus. Jack pine cultures seemed to have a low capacity for nitrate uptake and assimilation as shown by the low GS activity and the decrease of glutamine content in nitrate-fed calli.

LEVELS OF NITROGEN ASSIMILATION IN BROMELIADS WITH DIFFERENT GROWTH HABITS

Bromeliads exhibit several growth habits, ranging from terrestrial to epiphytic and associated shoot and root architectures. Accordingly, they are probably exposed to different sources of supply and forms of nitrogen (N), including mineral and organic. Nitrogen uptake and assimilation could reflect this. We compared the activities of nitrate reductase (NR) and glutamine synthetase (GS) as well as NO3 − contents in plantlets of two bromeliad species. Vriesea fosteriana, a C3saxicolous tank-forming type, and Tillandsia pohliana, an atmospheric CAM epiphyte, were grown in vitro with NO3 −, NH4 + or NH4NO3as the sole N sources (4 mM of N). Nitrate alone was a poor N source to support growth of both plant species but it was effective with NH4 +. Consistently, both bromeliads exhibited very low levels of NO3 − assimilation. The tank-forming species presented GS activity 2.5–3.0 fold higher than T. pohliana. The low GS activity found in T. pohliana compared to V. fosteriana may be related to distinct photosynthetic pathways and may also reflect an adaptation to survival in a nutrient-limited habitat.

The transcriptional activator NtrC controls the expression and activity of glutamine synthetase in Herbaspirillum seropedicae

The role of the Ntr system in Herbaspirillum seropedicae was determined via ntrB and ntrC mutants. Three phenotypes were identified in these mutants: Nif −, deficiency in growth using nitrate, and low glutamine synthetase (GS) activity. All phenotypes were restored by the plasmid pKRT1 containing the intact glnA, ntrB and ntrC genes of H. seropedicae. The promoter region of glnA was subcloned into a β-galactosidase fusion vector and the results suggested that NtrC positively regulates the glnA promoter in response to low nitrogen. The H. seropedicae ntrC and ntrB mutant strains showed a deficiency of adenylylation/deadenylylation of GS, indicating that NtrC and NtrB are involved in both transcription and activity control of GS in this organism.

The transcriptional activator NtrC controls the expression and activity of glutamine synthetase in Herbaspirillum seropedicae.

The role of the Ntr system in Herbaspirillum seropedicae was determined via ntrB and ntrC mutants. Three phenotypes were identified in these mutants: Nif(-), deficiency in growth using nitrate, and low glutamine synthetase (GS) activity. All phenotypes were restored by the plasmid pKRT1 containing the intact glnA, ntrB and ntrC genes of H. seropedicae. The promoter region of glnA was subcloned into a beta-galactosidase fusion vector and the results suggested that NtrC positively regulates the glnA promoter in response to low nitrogen. The H. seropedicae ntrC and ntrB mutant strains showed a deficiency of adenylylation/deadenylylation of GS, indicating that NtrC and NtrB are involved in both transcription and activity control of GS in this organism.

Cadmium-induced testes oxidative damage in rats can be influenced by dietary zinc intake

We tested the hypothesis that zinc deficient animals would be characterized by an increased sensitivity to cadmium-induced oxidative damage to the testes. Weanling male rats were given free access to either a control (25 μg Zn/g) or a zinc deficient (0.5 μg Zn/g) diet; or restricted access to the 25 μg Zn/g diet at a level of intake similar to that of rats fed the 0.5 μg Zn/g diet. After 14 days on the diets, animals were injected sc with either saline or CdCl2 (2 mg Cd/kg body weight) solutions, and killed 24 h later. In the zinc-deficient group, testes weight and testes/body weight were higher in the cadmium-injected rats than in the saline-injected rats. The extent of hemorrhages, as indicated by high hemoglobin and testes iron concentrations was higher in the cadmium-treated zinc deficient group than in the cadmium-injected controls. In the zinc-deficient group, cadmium injection was associated with higher levels of lipid peroxidation (33% higher TBARS content) and protein oxidation (17% lower glutamine synthetase activity). Cadmium injection did not influence these parameters in the zinc-adequate groups. Extracellular superoxide dismutase activity was lower in the zinc-deficient group than in the zinc-sufficient groups; there was a trend (P<0.06) for a lower activity in the cadmium- versus the saline-injected rats. These results support the concept that zinc deficiency increases the susceptibility of testes to cadmium-mediated free radical damage.