Increase In Glutamine Synthetase Activity Research Articles

Modulation of Reserve Mobilization by Sucrose, Glutamine, and Abscisic Acid During Seedling Establishment in Sunflower

We carried out in vitro feeding experiments using sunflower as a model to differentiate the modulatory effects of metabolites (sucrose and glutamine) and hormones (gibberellic acid and abscisic acid) on reserve mobilization, metabolite partitioning, and key enzyme activities. Exogenous sucrose negatively not only modulated the mobilization of carbon reserves (oils and starch), but it also delayed the degradation of nitrogen reserves (storage proteins) in the cotyledons. Similarly, exogenous glutamine negatively not only modulated storage protein hydrolysis, but it also retarded oil and starch degradation. Different from the metabolites, exogenous abscisic acid affected only the mobilization of oils and storage proteins. Sucrose and glutamine caused non-reducing sugar accumulation in the cotyledons and axis, but abscisic acid did not change the content of these compounds in both seedling parts. Curiously, glutamine failed to cause amino acid accumulation in the cotyledons and abscisic acid increased the amino acid content in both cotyledons and axis. Gibberellic acid did not stimulate reserve mobilization and metabolite consumption. Although the mobilization of oils, storage proteins, and starch has been delayed by sucrose and glutamine, these metabolites augmented the activity of isocitrate lyase, acid proteases, and amylases. Only abscisic acid reduced amylase activity and increased glutamine synthetase activity. Accordingly, sucrose and glutamine exert a “crossed effect” on reserve mobilization, that is, sucrose delays storage protein hydrolysis and glutamine retards oil and starch degradation. These effects may be mediated by non-reducing sugars and they are, at least in part, different from those exerted by abscisic acid.

Effect of Liver Damage and Hyperbaric Oxygenation on Glutamine Synthetase of Hepatocytes.

Activity of glutamine synthetase in the hepatocytes of healthy animals and animals with chronic CCl4-induced hepatitis was studied on white mature female rats after liver resection (15-20% of organ weight) and hyperbaric oxygenation (3 atm, 50 min, 3 times). Surgically operated left and non-operated middle lobes of the liver were analyzed on day 3 after liver resection and exposure to hyperbaric oxygenation. On day 65 of CCl4 poisoning, activity of glutamine synthetase decreased in both lobes and did not recover on day 3 after toxin cessation. Liver resection under conditions of CCl4-induced hepatitis restored reduced activity of glutamine synthetase in both liver lobes to the normal level. In healthy rats, the increase in glutamine synthetase activity after liver resection was found only in the middle lobe of the liver. Hyperbaric oxygenation enhanced the stimulatory effect of liver resection on glutamine synthetase activity in hepatocytes during chronic CCl4-induced hepatitis. In healthy animals with liver resection, activity of glutamine synthetase did not change after hyperbaric oxygenation, while normally oxygenation inhibited glutamine synthetase activity.

L-NAME combats excitotoxicity and recuperates neurological deficits in MCAO/R rats

Purpose of researchSince, transient focal cerebral ischaemia exhibits detrimental effect not only during the course of ischaemia but also after the onset of reperfusion, the current study is focussed on identifying the appropriate therapeutic time point at which NG-nitro-l-arginine methyl ester (l-NAME) exerts better neuroprotection. Principal resultsPre-ischaemic administration of l-NAME ameliorated neurological deficits much better than the during ischaemic and post-ischaemic groups. Pre-ischaemic l-NAME has also mitigated glutamate excitotoxicity, increased glutamine synthetase activity, ATP and NAD levels, decreased nitrate/nitrite content, down regulated TNF-α and upregulated IL-10 expressions and reduced the cerebral infarction significantly than the during ischaemic and post-ischaemic groups. Major conclusionCurrent study revealed that l-NAME improved neurological deficit at the pre-ischaemic state in transient focal cerebral ischaemia and has also significantly ameliorated glutamate excitotoxicity. Though l-NAME showed neuroprotective effects when administered at during and post-ischaemia (during reperfusion), it exerts considerable neuroprotection when administered pre-ischaemically.

Ceftriaxone attenuates ethanol drinking and restores extracellular glutamate concentration through normalization of GLT-1 in nucleus accumbens of male alcohol-preferring rats

Alteration of glutamatergic-neurotransmission is a hallmark of alcohol dependence. We have previously reported that chronic ethanol-drinking downregulated glutamate transporter 1 (GLT-1) in nucleus accumbens (NAc) in male P rats in a manner that was reversed by ceftriaxone treatment. However, the effect of ceftriaxone on extracellular glutamate concentrations in NAc after chronic ethanol-drinking has not yet been studied. In the present study, male P rats were treated with ceftriaxone (100 mg/kg/day, i.p.) for five consecutive days following five-weeks of free choice ethanol (15% and 30%) drinking. In vivo microdialysis was performed to measure the extracellular glutamate concentrations in NAc and the effect of blockade of GLT-1 with dihydrokainic acid (DHK) on extracellular glutamate in NAc of ceftriaxone-treated rats was determined. Ceftriaxone treatment attenuated ethanol intake as well as ethanol preference. Extracellular glutamate was significantly higher in NAc after five-weeks of ethanol drinking in saline-treated compared to water control rats. Ceftriaxone treatment blocked the increase extracellular glutamate produced by ethanol intake. Blockade of GLT-1 by DHK reversed the effects of ceftriaxone on glutamate and implicated the role of GLT-1 in the normalization of extracellular glutamate by ceftriaxone. In addition, GLT-1 protein was decreased in ethanol exposed animals and ceftriaxone treatment reversed this deficit. Ceftriaxone treatment also increased glutamine synthetase activity in NAc but not in PFC as compared to ethanol drinking saline-treated rats. Our present study demonstrates that ceftriaxone treatment prevents ethanol drinking in part through normalization of extracellular glutamate concentrations in NAc of male P rats via GLT-1.

Cross-Effects of Nitrogen and Sulphur Starvation in <i>Chlorella sorokiniana</i> 211/8K

Nitrogen (N) and sulphur (S), being essential macronutrients, have important roles in microalgae metabolism. Effects of N- or S-shortage were investigated in the green microalgae Chlorella sorokiniana subjected to 24 h of starvation, by measuring the glutamine synthetase (GS) and O-ace- tylserine(thiol)lyase (OASTL) activities, proteins and amino acids levels. To test possible metabolic impact related to carbon (C) metabolism in response to N- or S-deprivation, starch and total C, N and S contents were also determined. The growth of C. sorokiniana cells was affected by N or S availability. The algae cultured for 24 h in a medium deprived of nitrogen or sulphur showed a decrease in the growth rate and changes in the average volume cell. Nitrogen starvation affected proteins level in the algae cells more than S-deprivation did. The decline in the protein levels observed under S-deficient conditions was coupled with the accumulation of the amide glutamine and with OASTL activity increase; additionally, N-deficiency promoted a decrease in cysteine (Cys) levels (50%) and an increase in GS activity. Nevertheless, S-deprivation had negligible effects on GS activity, while N-deprivation significantly affected OASTL activity. Total C was also estimated in cells N- or S-deprived; nitrogen deprivation strongly affected total C content more than S-deprivation, which in addition reduced the content of C and N, but leaves intact their ratios. Our results support the hypothesis that in Chlorella sorokiniana cells a reciprocal influence of N, S and C assimilation occurs.

Dexamethasone enhances glutamine synthetase activity and reduces N-methyl-D-aspartate neurotoxicity in mixed cultures of neurons and astrocytes

Astrocytes are claimed to protect neurons against excitotoxicity by clearing glutamate from the extracellular space and rapidly converting it into glutamine. Glutamine, is then released into the extracellular medium, taken up by neurons and transformed back into glutamate which is then stored into synaptic vesicles. Glutamine synthetase (GS), the key enzyme that governs this glutamate/glutamine cycle, is known to be upregulated by glucocorticoids. In the present work we have thus studied in parallel the effects of dexamethasone on glutamine synthetase activity and NMDA-induced neuronal death in cultures derived from the brain cortex of murine embryos. We showed that dexamethasone was able to markedly enhance GS activity in cultures of astrocytes but not in near pure neuronal cultures. The pharmacological characteristics of the dexamethasone action strongly suggest that it corresponds to a typical receptor-mediated effect. We also observed that long lasting incubation (72 h) of mixed astrocyte-neuron cultures in the presence of 100 nM dexamethasone significantly reduced the toxicity of NMDA treatment. Furthermore we demonstrated that methionine sulfoximine, a selective inhibitor of GS, abolished the dexamethasone-induced increase in GS activity and also markedly potentiated NMDA toxicity. Altogether these results suggest that dexamethasone may promote neuroprotection through a stimulation of astrocyte glutamine synthetase.

Ornithine phenylacetate targets alterations in the expression and activity of glutamine synthase and glutaminase to reduce ammonia levels in bile duct ligated rats

In liver failure, ammonia homeostasis is dependent upon the function of the ammonia metabolising enzymes, glutamine synthetase (GS) and glutaminase (GA) but data about their protein expression and activity are lacking. The aims of this study were to determine the protein expression and activity of GS and GA in individual organs in a rat model of chronic liver disease and to test whether the treatment with the ammonia-lowering agent ornithine phenylacetate (OP) modulates their activities. 49 SD rats were studied 35 days after sham-operation or bile duct ligation (BDL). The BDL group received: L-ornithine (0.6 mg/kg/day), Phenylacetate (0.6 mg/kg/day), OP (0.6 mg/kg/day) or placebo (saline) for 5 days prior to sacrifice. Arterial ammonia, amino acids and liver biochemistry were measured. Expressions of GS and GA were determined by Western-blotting and activities by end-point methods in liver, muscle, gut, kidney, lung, and frontal cortex. In BDL rats, hepatic GS enzyme activity was reduced by more than 80% compared to sham rats. Further, in BDL rats GA activity was reduced in liver but increased in the gut, muscle and frontal cortex compared to sham rats. OP treatment resulted in a reduction in hyperammonemia in BDL rats, associated with increased GS activity in the muscle and reduced gut GA activity. In a rat model of chronic liver failure, hyperammonemia is associated with inadequate compensation by liver and muscle GS activity and increased gut GA activity. OP reduces plasma ammonia by increasing GS in the muscle and reducing GA activity in the gut providing additional insights into its mechanism of its action. GS and GA may serve as important future therapeutic targets for hyperammonemia in liver failure.

Treadmill Exercise Induces Hippocampal Astroglial Alterations in Rats

Physical exercise effects on brain health and cognitive performance have been described. Synaptic remodeling in hippocampus induced by physical exercise has been described in animal models, but the underlying mechanisms remain poorly understood. Changes in astrocytes, the glial cells involved in synaptic remodeling, need more characterization. We investigated the effect of moderate treadmill exercise (20 min/day) for 4 weeks on some parameters of astrocytic activity in rat hippocampal slices, namely, glial fibrillary acidic protein (GFAP), glutamate uptake and glutamine synthetase (GS) activities, glutathione content, and S100B protein content and secretion, as well as brain-derived neurotrophic factor (BDNF) levels and glucose uptake activity in this tissue. Results show that moderate treadmill exercise was able to induce a decrease in GFAP content (evaluated by ELISA and immunohistochemistry) and an increase in GS activity. These changes could be mediated by corticosterone, whose levels were elevated in serum. BDNF, another putative mediator, was not altered in hippocampal tissue. Moreover, treadmill exercise caused a decrease in NO content. Our data indicate specific changes in astrocyte markers induced by physical exercise, the importance of studying astrocytes for understanding brain plasticity, as well as reinforce the relevance of physical exercise as a neuroprotective strategy.

Nitrogen metabolism of the intestine during digestion in a teleost fish, the plainfin midshipman (Porichthys notatus)

Digestion affects nitrogen metabolism in fish, as both exogenous and endogenous proteins and amino acids are catabolized, liberating ammonia in the process. Here we present a model of local detoxification of ammonia by the intestinal tissue of the plainfin midshipman (Porichthys notatus) during digestion, resulting in an increase in urea excretion of gastrointestinal origin. Corroborating evidence indicated whole-animal ammonia and urea excretion increased following feeding, and ammonia levels within the lumen of the midshipman intestine increased to high levels (1.8±0.4 μmol N g(-1)). We propose that this ammonia entered the enterocytes and was detoxified to urea via the ornithine-urea cycle (O-UC) enzymes, as evidenced by a 1.5- to 2.9-fold post-prandial increase in glutamine synthetase activity (0.14±0.05 and 0.28±0.02 μmol min(-1) g(-1) versus 0.41±0.03 μmol min(-1) g(-1)) and an 8.7-fold increase in carbamoyl phosphate synthetase III activity (0.3±1.2 versus 2.6±0.4 nmol min(-1) g(-1)). Furthermore, digestion increased urea production by isolated gastrointestinal tissue 1.7-fold, supporting our hypothesis that intestinal tissue synthesizes urea in response to feeding. We further propose that the intestinal urea may have been excreted into the intestinal lumen via an apical urea transporter as visualized using immunohistochemistry. A portion of the urea was then excreted to the environment along with the feces, resulting in the observed increase in urea excretion, while another portion may have been used by intestinal ureolytic bacteria. Overall, we propose that P. notatus produces urea within the enterocytes via a functional O-UC, which is then excreted into the intestinal lumen. Our model of intestinal nitrogen metabolism does not appear to be universal as we were unab le to activate the O-UC in the intestine of fed rainbow trout. However, literature values suggest that multiple fish species could follow this model.

Regulation of ammonium assimilation in Haloferax mediterranei: Interaction between glutamine synthetase and two GlnK proteins

GlnK proteins belong to the PII superfamily of signal transduction proteins and are involved in the regulation of nitrogen metabolism. These proteins are normally encoded in an operon together with the structural gene for the ammonium transporter AmtB. Haloferax mediterranei possesses two genes encoding for GlnK, specifically, glnK1 and glnK2. The present study marks the first investigation of PII proteins in haloarchaea, and provides evidence for the direct interaction between glutamine synthetase and both GlnK1 and GlnK2. Complex formation between glutamine synthetase and the two GlnK proteins is demonstrated with pure recombinant protein samples using in vitro activity assays, gel filtration chromatography and western blotting. This protein–protein interaction increases glutamine synthetase activity in the presence of 2-oxoglutarate. Separate experiments that were carried out with GlnK1 and GlnK2 produced equivalent results.

Salinity affects indirectly nitrate acquisition associated with glutamine accumulation in cowpea roots

The aim of this study was to test the hypothesis that salinity can affect indirectly the nitrate acquisition by a negative modulation triggered by glutamine accumulation. Cowpea plants were exposed to a mild NaCl concentration (50 mM) in order to restrict growth and N-demand. After 21 d, pretreated plants and control plants were supplied with 0, 5 and 10 mM of Ca(NO3)2 for 3 d in absence of NaCl. Salt pretreated plants showed a great limitation in acquisition of NO3 -, indicated by decline in the nitrate uptake rate, NO3 - accumulation, nitrate reductase activity and protein content. The restriction of NO3 - utilization was positively associated with increased glutamine synthetase activity and glutamine accumulation, especially in roots.

Salinity affects indirectly nitrate acquisition associated with glutamine accumulation in cowpea roots

The aim of this study was to test the hypothesis that salinity can affect indirectly the nitrate acquisition by a negative modulation triggered by glutamine accumulation. Cowpea plants were exposed to a mild NaCl concentration (50 mM) in order to restrict growth and N-demand. After 21 d, pretreated plants and control plants were supplied with 0, 5 and 10 mM of Ca(NO3)2 for 3 d in absence of NaCl. Salt pretreated plants showed a great limitation in acquisition of NO3 -, indicated by decline in the nitrate uptake rate, NO3 - accumulation, nitrate reductase activity and protein content. The restriction of NO3 - utilization was positively associated with increased glutamine synthetase activity and glutamine accumulation, especially in roots.

The neuroprotective effect of two statins: simvastatin and pravastatin on a streptozotocin-induced model of Alzheimer’s disease in rats

Astrocytes play a fundamental role in glutamate metabolism by regulating the extracellular levels of glutamate and intracellular levels of glutamine. They also participate in antioxidant defenses, due to the synthesis of glutathione, coupled to glutamate metabolism. Although the cause of Alzheimer's disease (AD) remains elusive, some changes in neurochemical parameters, such as glutamate uptake, glutamine synthetase activity and glutathione have been investigated in this disease. A possible neuroprotective effect of two statins, simvastatin and pravastatin (administered p.o.), was evaluated using a model of dementia, based on the intracerebroventricular (ICV) administration of streptozotocin (STZ), and astrocyte parameters were determined. We confirmed a cognitive deficit in rats submitted to ICV-STZ, and a prevention of this deficit by statin administration. Moreover, both statins were able to prevent the decrease in glutathione content and glutamine synthetase activity in this model of AD. Interestingly, simvastatin increased per se glutamate uptake activity, while both statins increased glutamine synthetase activity per se. These results support the idea that these drugs could be effective for the prevention of alterations observed in the STZ dementia model and may contribute to reduce the cognitive impairment and brain damage observed in AD patients.

Glutamine synthetase gene expression and glutamate transporters in C6-glioma cells

Glutamine synthetase (GS) is the major glutamate-forming enzyme of vertebrae and is accepted to be a marker of astroglial cells. Maturation of astroglial cells is characterized by an increase in GS activity, and the regulation of this enzyme is the topic of many publications. The amino acid glutamate is the major excitatory neurotransmitter in the brain and mediates normal excitatory synaptic transmission by interaction with postsynaptic receptors. Glutamate also acts as a potent neurotoxin when present at high concentration. Glutamate neurotoxicity plays an important role in the pathophysiology of many neurological disorders, such as Alzheimer's disease, Huntington's disease and amyotrophic lateral sclerosis. In the normal condition, L-glutamate is predominantly taken up, metabolized and recycled by astrocytes through the glutamate transporters (GLAST/GLT1) and glutamine synthetase (GS) catalytic activity. Because of the fundamental role of these glutamate transporters and the glutamine synthetase enzyme in controlling cerebral glutamate level, regulation of GS and studying of the glutamate transporters in glial cells is important. Astrocytes are supportive cells and act as the site of detoxification of glutamate in the brain. However, their isolation from the brain is a tedious, costly and time consuming procedure. On the other hand, the C6-glioma cells are readily available on the market. They are well characterized and have been a useful model for CNS glia in many laboratories. For this study, we used the C6-glioma cell line as a model system. We examined the presence or absence of glial specific glutamate transporters (GLTI and GLAST) in C6-glioma cells, which was done by immunocytochemistry. We also examined glutamine synthetase gene expression in these cells by treatment of the C6-glioma cells with estrogen (17ß estradiol). The findings from this study provide useful information about C6-glioma cells which makes the study of the CNS tremendously inexpensive.

Pharmacological evaluation of glutamate transporter 1 (GLT-1) mediated neuroprotection following cerebral ischemia/reperfusion injury

Recently glutamate transporters have emerged as a potential therapeutic target in a wide range of acute and chronic neurological disorders, owing to their novel mode of action. The modulation of GLT-1, a major glutamate transporter has been shown to exert neuroprotection in various models of ischemic injury and motoneuron degeneration. Therefore, an attempt was made to explore its neuroprotective potential in cerebral ischemia/reperfusion injury using ceftriaxone, a GLT-1 modulator. Pre-treatment with ceftriaxone (100 mg/kg. i.v) for five days resulted in a significant reduction ( P < 0.01) in neurological deficit as well as cerebral infarct volume after 1 h of ischemia followed by 24 h of reperfusion injury. It also caused a significant ( P < 0.05) upregulation of GLT-1 mRNA, protein and glutamine synthetase (GS) activity. Furthermore, inhibition of ceftriaxone-mediated increased glutamine synthetase activity by dihydrokainate (DHK), a GLT-1 specific inhibitor, confirms the specific effect of ceftriaxone on GLT-1 activity. In addition, ceftriaxone also induced a significant ( P < 0.01) increase in [ 3H]-glutamate uptake, mediated by GLT-1 in glial enriched preparation, as evidenced by use of DHK and DL-threo-beta-benzyloxyaspartate (DL-TBOA). Thus, the present study provides overwhelming evidence that modulation of GLT-1 protein expression and activity confers neuroprotection in cerebral ischemia/reperfusion injury.

Salt tolerant mutant of Anabaena doliolum exhibiting efficient ammonium uptake and assimilation.

Effect of salinity (NaCl, 100 mM) on growth, nitrate reductase (NR) and glutamine synthetase (GS) activities, and uptake of NH4 (+) was studied in the wild type (WT) and the NaCl-tolerant mutant type (MT) of cyanobacterium Anabaena doliolum. Results obtained in the presence of salt showed significant reduction in the growth rate of both WT and MT cells of A. doliolum by about 77.8 and 40 %, respectively over without NaCl. Similarly rate of NR activity in both WT and MT strains was reduced by 45.5 and 44.5 %, respectively. On the contrary, rate of GS activity of both the WT and MT strains in the presence 100 mM of NaCl increased by 34 and 159 %, respectively. The results of this study indicate that tolerance to NaCl in A. doliolum is more dependent on NH4 (+) assimilation rather than on nitrate assimilation in relation to N-metabolism. The increased GS activity in MT cells of the cyanobacterium is possibly because of high rate of energy dependent NH4 (+) uptake.

Determining potential glutamine synthetase and glutamate dehydrogenase activity in soil

Ammonium (NH 4 +), an important nitrogen (N) source for microorganisms, is assimilated via two major pathways. One route is catalyzed by glutamate dehydrogenase (GDH), while the other mechanism involves two enzymes, glutamine synthetase (GS) and glutamate synthase (GOGAT). The GS/GOGAT enzyme system requires more energy to operate, but has a much higher affinity for NH 4 + than GDH. We describe procedures to determine potential GS and GDH activity in soil samples. GS and GDH are intracellular enzymes. We used chloroform fumigation to make cell membranes permeable for substrates and products of the enzymes. Fumigation for 4 h increased GS activity almost ten-fold compared to the unfumigated control. Under optimized assay conditions, GS activity increased linearly for at least 80 min, indicating that the substrates were not limiting. In contrast to what was found for GS activity, direct addition of substrates to the soil to assay GDH activity did not result in a linear increase in GDH activity over time. A linear response for 3 h, however, resulted when the soil samples were first extracted with buffer solution and the reagents were added after centrifugation. The differences between the assays explain why fumigation for 3 d prior to the assay increased GDH activity by only 60%. In a microcosm study with glucose and NH 4 + addition, the activity of the two enzymes depended on the carbon (C) to N ratio of the amendment. With increasing C to N ratios from 5 to 120, GS activity doubled, while C to N ratios higher than 120 did not further increase GS activity. In contrast, GDH activity decreased by 13% with increasing C to N ratios from 5 to 200. The GDH to GS activity ratio in soil may therefore yield valuable information about the availability of N relative to C at a specific time.

A combination of metabolome and transcriptome analyses reveals new targets of the Corynebacterium glutamicum nitrogen regulator AmtR

The effects of a deletion of the amtR gene, encoding the master regulator of nitrogen control in Corynebacterium glutamicum, were investigated by metabolome and transcriptome analyses. Compared to the wild type, different metabolite patterns were observed in respect to glycolysis, pentose phosphate pathway, citric acid cycle, and most amino acid pools. Not all of these alterations could be attributed to changes at the level of mRNA and must be caused by posttranscriptional regulatory processes. However, subsequently carried out transcriptome analyses, which were confirmed by gel retardation experiments, revealed two new targets of AmtR, the dapD gene, encoding succinylase involved in m-diaminopimelate synthesis, and the mez gene, coding for malic enzyme. The regulation of dapD connects the AmtR-dependent nitrogen control with l-lysine biosynthesis, the regulation of mez with carbon metabolism. An increased l-glutamine pool in the amtR mutant compared to the wild type was correlated with deregulated expression of the AmtR-regulated glnA gene and an increased glutamine synthetase activity. The glutamate pool was decreased in the mutant and also glutamate excretion was impaired.

Effect of Nitrate on Activities and Transcript Levels of Nitrate Reductase and Glutamine Synthetase in Rice

Real-time polymerase chain reaction analysis was used to compare the effect of NO − 3 on the activities of nitrate reductase (NR) and glutamine synthetase (GS), and the transcript levels of two NR genes, OsNia1 and OsNia2, two cytosolic GS1 genes, OsGln1;1 and OsGln1;2, and one plastid GS2 gene OsGln2, in two rice ( Oryza sativa L.) cultivars Nanguang (NG) and Yunjing (YJ). Both cultivars achieved greater biomass and higher total N concentration when grown in a mixed N supply than in sole NH + 4 nutrition. Supply of NO − 3 increased NR activity in both leaves and roots. Expression of both NR genes was also substantially enhanced and transcript levels of OsNia2 were significantly higher than those of OsNia1. NO − 3 also caused an increase in GS activity, but had a complex effect on the expression of the three GS genes. In roots, the OsGln1;1 transcript increased, but OsGln1;2 decreased. In leaves, NO − 3 had no effect on the GS1 expression, but the transcript for OsGln2 increased both in the leaves and roots of rice with a mixed supply of N. These results suggested that the increase in GS activity might be a result of the complicated regulation of the various GS genes. In addition, the NO − 3-induced increase of biomass, NR activity, GS activity, and the transcript levels of NR and GS genes were proportionally higher in NG than in YJ, indicating a stronger response of NG to NO − 3 nutrition than YJ.

Chronic exposure to glufosinate-ammonium induces spatial memory impairments, hippocampal MRI modifications and glutamine synthetase activation in mice

Glufosinate-ammonium (GLA), the active compound of a worldwide-used herbicide, acts by inhibiting the plant glutamine synthetase (GS) leading to a lethal accumulation of ammonia. GS plays a pivotal role in the mammalian brain where it allows neurotransmitter glutamate recycling within astroglia. Clinical studies report that an acute GLA ingestion induces convulsions and memory impairment in humans. Toxicological studies performed at doses used for herbicidal activity showed that GLA is probably harmless at short or medium range periods. However, effects of low doses of GLA on chronically exposed subjects are not known. In our study, C57BL/6J mice were treated during 10 weeks three times a week with 2.5, 5 and 10 mg/kg of GLA. Effects of this chronic treatment were assessed at behavioral, structural and metabolic levels by using tests of spatial memory, locomotor activity and anxiety, hippocampal magnetic resonance imaging (MRI) texture analysis, and hippocampal GS activity assay, respectively. Chronic GLA treatments have effects neither on anxiety nor on locomotor activity of mice but at 5 and 10 mg/kg induce (1) mild memory impairments, (2) a modification of hippocampal texture and (3) a significant increase in hippocampal GS activity. It is suggested that these modifications may be causally linked one to another. Since glutamate is the main neurotransmitter in hippocampus where it plays a crucial role in spatial memory, hippocampal MRI texture and spatial memory alterations might be the consequences of hippocampal glutamate homeostasis modification revealed by increased GS activity in hippocampus. The present study provides the first data that show cerebral alterations after chronic exposure to GLA.