Effect of chlorpromazine on central nervous system concentrations of manganese,iron, and copper

Chronic administration of chlorpromazine produced alterations in trace metal concentrations in caudate nucleus, frontal cortex, and cerebellar hemisphere of guinea pigs. Manganese concentration following chronic chlorpromazine rose significantly in the caudate nucleus and cerebellar hemisphere whereas iron concentration rose most significantly in the caudate nucleus. Copper content was decreased in all regions examined. The possible significance of increased manganese and iron in the caudate nucleus is discussed in relationship to the clinical problems of chlorpromazine-induced dyskinesias.     

Histochemical distribution of non-haem iron in the human brain.

The detailed anatomical distribution of iron in the post-mortem human brain has been studied using Perl's and Turnbull's methods with the diaminobenzidine intensification procedure for the demonstration of non-haem Fe3+ and Fe2+, respectively. Attention to methodological procedures has revealed that even brief immersion of tissue in routinely used fixatives causes a reduction of staining intensity in areas of high iron content and, often, loss of staining in areas of low iron content. Optimal staining is obtained using frozen section briefly fixed for 5 min in 4% formalin and Perl's stain (Fe3+) with diaminobenzidine intensification. Highest levels of stainable iron were found in the extrapyramidal system with the globus pallidus, substantia nigra zona reticulata, red nucleus and myelinated fibres of the putamen showing highest staining reactivity. Moderate staining intensity with Perl's technique was found in the majority of forebrain, midbrain and cerebellar structures with the striatum, thalamus, cortex and deep white matter, substantia nigra zona compacta, and cerebellar cortex showing consistent staining patterns with intensification of Perl's stain. The brain-stem and spinal cord generally only showed staining with the intensification procedure and even this was of low intensity. Microscopically the non-heam iron appears to be found predominantly in glial cells as fine cytoplasmic granules which in heavily stained areas coalesce to fill the entire cell. Iron-positive granules appear to be free in the neuropil and also around blood vessels in the globus pallidus, striatum and substantia nigra. The neuropil shows a fibrous impregnation when stained for iron which is, in part, derived from glial processes, myelinated fibres and fibre bundles. Neurones, in general, show only very low reactivity for iron, and this is difficult to discern due, often, to the higher reactivity of the surrounding neuropil. In the globus pallidus and substantia nigra zona reticulata, neurones with highly stainable iron content are found with granular cytoplasmic iron reactivity similar to that seen in the local glial cells. Our results are comparable with those of early workers, but with the use of intensification extend the distribution of non-haem iron to areas previously reported as negative. No apparent correlation of iron staining with known neurotransmitter systems is seen and the predilection for the extrapyramidal system is not easily explained, though the non-haem iron in the brain appears to be as a storage form in the iron storage protein ferritin. The localization of iron in the brain provides a foundation for the study of iron in certain neurodegenerative diseases such as Parkinson's disease, where iron has been implicated in the pathogenesis.     

Effect of Allopurinol on the Hepatic and Cerebellar Iron,Selenium, Zinc and Copper Status Following Acute Ethanol Administration to Rats

An acute ethanol load (50mmol/kg, i.p.) induces an increase in the total non-heme iron and in the low-molecular-weight non-heme iron complexes (LMW-Fe) content both in liver and cerebellum. This increase in LMW-Fe is associated with a decrease in some essential trace elements (selenium, zinc, copper) playing a role in the anti-oxidant system. These changes could contribute to the enhancement in lipid peroxidation which occurs at the hepatic and cerebellar level following the ethanol administration.

The administration of allopurinol prior to the ethanol load prevents the changes in non-heme iron and trace elements. This prevention may contribute to the protective effects of allopurinol on the ethanol-induced oxidative stress.     

Effects of dopaminergic drugs on the concentrations of PGE2 and PGF2alpha in various brain regions

It has long been shown by Biggio and Guidotti that multisynaptic nigro-cerebellar pathway of dopaminergic origin can control cerebellar cyclic guanosinmonophosphate (cGMP) content, a good index of the activity of Purkinje cells. In this line, it has been reported that haloperidol and sulpiride, significantly decrease cerebellar cGMP content while opposite changes are observed with apomorphine. In an attempt to establish whether other cerebellar cGMP-related parameters may be influenced by dopamine drugs. Authors have investigated the effects of haloperidol, sulpiride and apomorphine on cerebellar PGE2 and PGF2alpha. Results obtained indicate that haloperidol and sulpiride significantly reduce cerebellar PGE2 and PGF2alpha content while opposite changes are induced by apomorphine. Similar results have been observed in substantia nigra but not in other brain regions, such as corpus striatum and medial basal hypothalamus. The possibility that the observed changes in cerebellar PG-content may result from the modulation of striatal dopamine receptors is discussed.     

Contents of Cystathionine and Taurine in Various Cerebellar Regions of dl-Propargylglycine-Treated Rats

The contents of cystathionine and taurine, as well as cystathionine beta-synthase activity, in various cerebellar regions and pineal body of normal and DL-propargylglycine-treated rats were measured. The contents of cystathionine and taurine were found to be distributed unevenly in cerebellar regions of brain of both normal and DL-propargylglycine-treated rats. The content of cystathionine in each cerebellar region and pineal body increased gradually when the dose of DL-propargylglycine was increased from 10 mg to 30 mg per 200 g body weight. On the other hand, taurine content in each cerebellar region and pineal body decreased with the administration of 30 mg of DL-propargylglycine per 200 g body weight. The contents of cystathionine and taurine were greater in the pineal body than in various cerebellar regions. The activity of cystathionine beta-synthase was also distributed unevenly in various cerebellar regions of normal rat brain, and was unaltered following treatment of rats with DL-propargylglycine.     

Genetic ablation of PLA2G6 in mice leads to cerebellar atrophy characterized by Purkinje cell loss and glial cell activation

Infantile neuroaxonal dystrophy (INAD) is a progressive, autosomal recessive neurodegenerative disease characterized by axonal dystrophy, abnormal iron deposition and cerebellar atrophy. This disease was recently mapped to PLA2G6, which encodes group VI Ca(2+)-independent phospholipase A(2) (iPLA(2) or iPLA(2)β). Here we show that genetic ablation of PLA2G6 in mice (iPLA(2)β(-/-)) leads to the development of cerebellar atrophy by the age of 13 months. Atrophied cerebella exhibited significant loss of Purkinje cells, as well as reactive astrogliosis, the activation of microglial cells, and the pronounced up-regulation of the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). Moreover, glial cell activation and the elevation in TNF-α and IL-1β expression occurred before apparent cerebellar atrophy. Our findings indicate that the absence of PLA2G6 causes neuroinflammation and Purkinje cell loss and ultimately leads to cerebellar atrophy. Our study suggests that iPLA(2)β(-/-) mice are a valuable model for cerebellar atrophy in INAD and that early anti-inflammatory therapy may help slow the progression of cerebellar atrophy in this deadly neurodegenerative disease.     

Vestibulo-cerebellar participation in protection of duodenal mucosa: possible role of neurotransmitters

Vestibulo cerebellar lesion in rats produced a decrease in the intracellular presecreted mucus together with a decrease in the norepinephrine (NE) and serotonin (5HT) content of the duodenal tissue. Whereas vestibulo cerebellar stimulation by rotation produced an increase in the intracellular presecreted mucus and an increase in the NE content of the duodenal tissue but very little increase in 5HT content of the duodenum. The results suggest that the vestibulo cerebellum by modulating the tissue content of the neurotransmitter NE and 5HT has a direct influence in the protective mechanism through the intracellular mucus content.     

First direct demonstration of extensive GABA synthesis in mouse cerebellar neuronal cultures

Culturing mouse cerebellar neurones (predominantly glutamatergic) in the presence of [1-(13)C]glucose for 7 days resulted in a surprisingly extensive labelling of the inhibitory neurotransmitter GABA, the average content and labelling of which were 20 +/- 4 nmol/mg protein and 20 +/- 4%, respectively. Cultures of neocortical neurones (predominantly GABAergic) had under similar conditions a GABA content and labelling of 32 +/- 2 nmol/mg protein and 21 +/- 2%. The cerebellar cultures contained only 6% glutamate decarboxylase (GAD)-positive neurones when immunolabelled using a GAD67 antibody, while a dense network of neurones in the neocortical cultures stained positively for GAD67. Exposure of the cerebellar cultures to 50 microm kainic acid (KA) which is known to eliminate vesicular release of GABA, only marginally affected GABA labelling and cellular content. Likewise this treatment had no effect on the number of GAD67-positive neurones but a massive punctate immunostaining observed in control cultures was essentially eliminated. Increasing the KA concentration to 0.5 mm in the culture medium for 7 days led to a reduction of GABA labelling and content compared to cerebellar cultures not exposed to KA. Although it is likely that this large capacity for GABA synthesis resides in the relatively few GAD-positive neurones, it seems unlikely that they could account for the large average GABA content in the cultures. Therefore it must be concluded that the newly synthesized GABA is redistributed among the majority of the cells in these cultures, i.e. the glutamatergic neurones.     

Iron content and bioavailability in rice

Iron deficiency is probably the most widespread micronutrient deficiency in humans. Since rice is the main staple food for more than half of the global population, improving the iron content and bioavailability in rice is a perspective and an effective way to alleviate or even solve this problem. The present paper evaluates the iron content in different cereal foods (black rice, rice, red rice, sticky rice and millet) and different rice seeds as well as in the milling products, and the iron bioavailability of different forms. The data show that the iron content in black rice is higher than in the other rice types, and in rice chaff and husk the content is still fairly high. However, the iron content in rice and fine rice, which are the people's main staple food, is fairly low. As to the bioavailability of iron, it is fairly low in vegetable foods, almost at the level of 10%. Several methods have been applied to improve iron content and bioavailability in rice seed. Apart from breeding and genetic engineering, biochemical and physical approaches have frequently been used as prospective methods to regulate iron content and bioavailability in rice grains.     

Iron content and bioavailability in rice

Iron deficiency is probably the most widespread micronutrient deficiency in humans. Since rice is the main staple food for more than half of the global population, improving the iron content and bioavailability in rice is a perspective and an effective way to alleviate or even solve this problem. The present paper evaluates the iron content in different cereal foods (black rice, rice, red rice, sticky rice and millet) and different rice seeds as well as in the milling products, and the iron bioavailability of different forms. The data show that the iron content in black rice is higher than in the other rice types, and in rice chaff and husk the content is still fairly high. However, the iron content in rice and fine rice, which are the people's main staple food, is fairly low. As to the bioavailability of iron, it is fairly low in vegetable foods, almost at the level of 10%. Several methods have been applied to improve iron content and bioavailability in rice seed. Apart from breeding and genetic engineering, biochemical and physical approaches have frequently been used as prospective methods to regulate iron content and bioavailability in rice grains.     

Comparison of Proteins Involved with Cyclic AMP Metabolism Between Synaptic Membrane and Postsynaptic Density Preparations Isolated from Canine Cerebral Cortex and Cerebellum

Synaptic membrane and postsynaptic density (PSD) fractions isolated from canine cerebral cortex and cerebellum were assayed for the following proteins: adenylate cyclase and phosphodiesterase (PDE) activities against cyclic AMP and cyclic GMP, the regulatory subunit of the cyclic AMP-dependent protein kinase, and the substrate proteins for this kinase. The results were expressed on the basis of both the protein content of the fractions and the number of synapses in the synaptic membrane fractions. The number of synapses on a constant protein content basis was about three times higher in the cerebral cortex synaptic membrane fraction than in the comparable cerebellar fraction. Adenylate cyclase activity was from 3.4 to 5.6 times higher in the cerebral cortex membrane fraction than in the cerebellar membrane fraction based on protein content but only slightly higher based on synapse counts. PSD fractions had no adenylate cyclase activity. The cyclic AMP-PDE activity was from 17 to 27 times higher in the cerebral cortex membrane fraction than in the cerebellar membrane fraction based on protein content, and about five times higher based on synapse counts. By doing PDE histochemistry at the electron microscopy level it was found that all the cerebral cortex PSDs in the isolated fraction contained PDE activity, none being found associated with the broken-up material in the fraction. The amount of the regulatory subunit of the cyclic AMP-dependent protein kinase was about equal in the two fractions based on protein, but about one-third lower in cerebral cortex fraction than in cerebellar fractions. In the cerebral cortex membrane fraction the primary substrate for the cyclic AMP-dependent protein kinase is synapsin I, with much lower amounts in the cerebellar membrane fraction. The PSD fraction from the two sources also showed these differences in synapsin I content. In the cerebellar membrane fraction, the primary substrate for the enzyme is a approximately 245,000 Mr protein not found in the cerebral cortex membrane fraction. The findings that the turnover of cyclic AMP is much higher in cerebral cortex synapses than in cerebellar synapses, and that differences are found between the cerebral cortex and cerebellum with regard to the substrate proteins for the cyclic AMP-dependent protein kinase indicate a divergence in the effect of cyclic AMP between cerebral cortex and cerebellar synapses.     

Iron in Oxidative and Reduction Processes in Marine Sands

The iron content and the ratio of bivalent to total iron in the labile acid-soluble fraction of iron were studied in sublittoral sands of Vostok Bay, Sea of Japan. Iron oxidation and reduction rates in sand sediment were measured in the laboratory. Trivalent iron almost always prevailed in the acid-extractable fraction in the natural environment. The most oxidized state of iron occurred in spring and was associated with a low water temperature and a high seawater oxygen content; the most reduced iron was found in fall during periods of low hydrodynamics and low oxygen concentration. In sand, iron is mainly reduced by bacteria; this process is slow and can be inhibited by adding chloroform. Oxidation of iron is mainly a chemical process and cannot be stopped by chloroform. In sand, the content of redox equivalents such as trivalent iron is much greater than in dissolved oxygen of pore water. It is assumed that labile iron in sands acts as a redox buffer, is oxidized by dissolved pore water oxygen at the periods of advective mixing, and is slowly reduced by benthic bacteria during anaerobic conditions.     

Recovery of Motor Deficit,Cerebellar Serotonin and Lipid Peroxidation Levels in the Cortex of Injured Rats

The sensorimotor cortex and the cerebellum are interconnected by the corticopontocerebellar (CPC) pathway and by neuronal groups such as the serotonergic system. Our aims were to determine the levels of cerebellar serotonin (5-HT) and lipid peroxidation (LP) after cortical iron injection and to analyze the motor function produced by the injury. Rats were divided into the following three groups: control, injured and recovering. Motor function was evaluated using the beam-walking test as an assessment of overall locomotor function and the footprint test as an assessment of gait. We also determined the levels of 5-HT and LP two and twenty days post-lesion. We found an increase in cerebellar 5-HT and a concomitant increase in LP in the pons and cerebellum of injured rats, which correlated with their motor deficits. Recovering rats showed normal 5-HT and LP levels. The increase of 5-HT in injured rats could be a result of serotonergic axonal injury after cortical iron injection. The LP and motor deficits could be due to impairments in neuronal connectivity affecting the corticospinal and CPC tracts and dysmetric stride could be indicative of an ataxic gait that involves the cerebellum.     

A probable site of action of diazepam in rat cerebellar GABA system

Diazepam and some neurotropic drugs were examined for their effects on the cyclic guanosine 3′, 5′-monophosphate (cyclic GMP) content in rat cerebellum. Diazepam caused a marked decrease of cerebellar cyclic GMP content in a dose-dependent manner. Elevation of cerebellar gamma-aminobutyric acid (GABA) level by aminooxyacetic acid also caused a decrease in the cyclic GMP content. On the other hand, dl-amphetamine, oxotremorine, picrotoxin, and GABA-reducing agents such as isoniazid or thiosemicarbazide increased the content of this nucleotide. The increase of cyclic GMP content elicited by isoniazid was blocked completely by the premedication of diazepam in doses causing partial reduction of dl-amphetamine or oxotremorine action. Changes of cerebellar GABA level, which were caused by aminooxyacetic acid or thiosemicarbazide, did not influence the effect of diazepam. Moreover, the inhibitory action of diazepam on the picrotoxin-induced increase of cyclic GMP was a competitive type. These results suggest that diazepam facilitates the GABAergic function by acting on a picrotoxin-sensitive site of the GABA receptor complex in vivo.     

1-Methyl-4-phenylpyridinium-induced apoptosis in cerebellar granule neurons is mediated by transferrin receptor iron-dependent depletion of tetrahydrobiopterin and neuronal nitric-oxide synthase-derived superoxide

In this study, we investigated the molecular mechanisms of toxicity of 1-methyl-4-phenylpyridinium (MPP(+)), an ultimate toxic metabolite of a mitochondrial neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, that causes Parkinson-like symptoms in experimental animals and humans. We used rat cerebellar granule neurons as a model cell system for investigating MPP(+) toxicity. Results show that MPP(+) treatment resulted in the generation of reactive oxygen species from inhibition of complex I of the mitochondrial respiratory chain, and inactivation of aconitase. This, in turn, stimulated transferrin receptor (TfR)-dependent iron signaling via activation of the iron-regulatory protein/iron-responsive element interaction. MPP(+) caused a time-dependent depletion of tetrahydrobiopterin (BH(4)) that was mediated by H(2)O(2) and transferrin iron. Depletion of BH(4) decreased the active, dimeric form of neuronal nitric-oxide synthase (nNOS). MPP(+)-mediated "uncoupling" of nNOS decreased *NO and increased superoxide formation. Pretreatment of cells with sepiapterin to promote BH(4) biosynthesis or cell-permeable iron chelator and TfR antibody to prevent iron-catalyzed BH(4) decomposition inhibited MPP(+) cytotoxicity. Preincubation of cerebellar granule neurons with nNOS inhibitor exacerbated MPP(+)-induced iron uptake, BH(4) depletion, proteasomal inactivation, and apoptosis. We conclude that MPP(+)-dependent aconitase inactivation, Tf-iron uptake, and oxidant generation result in the depletion of intracellular BH(4), leading to the uncoupling of nNOS activity. This further exacerbates reactive oxygen species-mediated oxidative damage and apoptosis. Implications of these results in unraveling the molecular mechanisms of neurodegenerative diseases (Parkinson's and Alzheimer's disease) are discussed.     

A computational study of synaptic mechanisms of partial memory transfer in cerebellar vestibulo-ocular-reflex learning

There is a debate regarding whether motor memory is stored in the cerebellar cortex, or the cerebellar nuclei, or both. Memory may be acquired in the cortex and then be transferred to the cerebellar nuclei. Based on a dynamical system modeling with a minimal set of variables, we theoretically investigated possible mechanisms of memory transfer and consolidation in the context of vestibulo-ocular reflex learning. We tested different plasticity rules for synapses in the cerebellar nuclei and took robustness of behavior against parameter variation as the criterion of plausibility of a model variant. In the most plausible scenarios, mossy-fiber nucleus-neuron synapses or Purkinje-cell nucleus-neuron synapses are plastic on a slow time scale and store permanent memory, whose content is passed from the cerebellar cortex storing transient memory. In these scenarios, synaptic strengths are potentiated when the mossy-fiber afferents to the nuclei are active during a pause in Purkinje-cell activities. Furthermore, assuming that mossy fibers create a limited variety of signals compared to parallel fibers, our model shows partial memory transfer from the cortex to the nuclei.     

Comparison of injectable iron complexes in their ability to iron load tissues and to induce oxidative stress

Iron and copper homeostasis have been studied in various tissues after iron-loading with the polynuclear ferric hydroxide carbohydrate complexes, iron dextran, iron polymaltose, iron sucrose and iron gluconate for four weeks. There were significant increases in the iron content of the different rat tissues compared to controls, with the exception of the brain, which showed no change in its iron content following iron loading. However, the level of iron loading in the different tissues varied according to the preparation administered and only iron dextran was able to significantly increase the iron content of both broncho-alveolar macrophages and heart. The hepatic copper content decreased with iron loading, although this did not reach significance. However the copper content did not alter in the iron loaded broncho-alveolar macrophages. Despite such increases in hepatic iron content, there was little evidence of changes in oxidative stress, the activities of cytosolic (apart from iron dextran) or mitochondrial hepatic superoxide dismutase, SOD, were similar to that of the control rats, confirming the fact that the low reduction potential of these compounds prevents the reduction of the ferric moiety. It was not necessary for macrophages to significantly increase their iron content to initiate changes in NO^. release. Iron gluconate and iron sucrose increased NO^. release, while iron polymaltose and iron dextran decreased NO^. release although only the latter iron preparation significantly increased their iron content. It may be that the speciation of iron within the macrophage is an important determinant in changes in NO^. release after ex vivo stimulation. We conclude that tissues loaded with iron by such polynuclear iron complexes have variable loading despite the comparable iron dose. However, there was little evidence for participation of the accumulated iron in free radical reactions although there was some evidence for alteration in immune function of broncho-alveolar macrophages.     

Neuroprotective effects of iron chelator Desferal on dopaminergic neurons in the substantia nigra of rats with iron-overload

The aim of the present study was to investigate whether the iron chelator Desferal prevents the degeneration of dopaminergic neurons in the substantia nigra (SN) induced by iron-overload in rats. Using fast cyclic voltammetry, tyrosine hydroxylase (TH) immunohistochemistry, Perls' iron staining, and high-performance liquid chromatography-electrochemical detection, we measured the degeneration of dopaminergic neurons and increased iron content in the SN of rats overloaded with iron dextran and assessed the effects of treatment with Desferal. The results showed that iron dextran overload increased the iron content in the SN, decreased dopamine release and content, and reduced the numbers of TH-immunoreactive neurons. Treatment with Desferal prevented the increased iron content in the SN. As a result, dopamine release and content remained at almost normal levels, while the numbers of TH-immunoreactive neurons remained at control values. This study suggests that the iron chelator Desferal is neuroprotective against iron-overload, so iron chelators that can cross the blood-brain barrier may have the potential to treat cases where abnormal iron accumulation in the brain is associated with the degenerative processes, as in Parkinson's disease.     

Free iron in bacteria

Free iron content has been estimated in autotrophic and heterotrophic bacteria. It constituted 40-50 micrograms/g dry weight as compared to 15 micrograms/g dry weight in animal cells. A method for estimation of free iron has been proposed. It is based on formation of paramagnetic dinitrosyl iron complexes by free iron and protein thiol groups or low molecular weight thiol ligands. The reasons for high iron content in bacteria have been discussed.     

Neurochemical Changes in Cerebellum of Goldfish Exposed to Various Temperatures

Acclimation of goldfish at 35°C increased the cerebellar content of aspartate, glutamate, and taurine and [³H]glutamate uptake. Acclimation at 4°C increased the levels of glutamine, serine, and alanine and glutamine synthetase (GS) activity. Adenosine content increased in cerebellum of fish acclimated to warm temperature. K⁺-evoked release of endogenous and exogenous glutamate from cerebellar slices increased in fish acclimated at 35°C compared to 4°C. The basal level of cyclic adenosine 3:5-monophosphate (cAMP) in perfused cerebellar slices in fish acclimated at 35°C was much higher than in fish acclimated at 5° and 22°C. It is concluded that variations of environmental temperature produces large neurochemical changes in goldfish cerebellum.