Metabolite Changes in the Cerebral Cortex of Treated and Untreated Infant Hydrocephalic Rats Studied Using In Vitro 31P-NMR Spectroscopy

Abstract: The effect of hydrocephalus on cerebral energy metabolites and on intermediates of membrane phospholipid metabolism has been studied in H-Tx rats with inherited infantile hydrocephalus. Hydrocephalic rats and rats with shunts placed at 4–5 days or at 10 days after birth were subjected to magnetic resonance imaging in vivo before 21 days of age to determine the dimensions of the ventricles and cortex. At 21 days, the brains from the three groups of rats, together with age-matched control littermates, were frozen in situ, and chloroform/methanol extracts of cerebral cortex were prepared for high-resolution ³¹P-NMR spectroscopy. Hydrocephalus resulted in modest decreases in most metabolites quantified. Levels of phosphocreatine, ATP, and diphosphodiesters plus NAD were significantly reduced by 23–32%, and inorganic phosphate content was reduced but not significantly. Levels of the membrane phospholipid intermediates phosphorylethanolamine, glycerophosphorylethanolamine, and glycerophosphorylcholine were also significantly reduced by 30–33%, indicating changes in membrane metabolism. These general decreases are consistent with a loss of cell contents, possibly due to changes in dendrite structure in hydrocephalus. Rats shunt-treated at 4–5 days were similar to control rats for all energy metabolites, but those treated later at 10 days had reduced phosphocreatine and ATP levels. Shunt-treated rats also had reductions in levels of membrane phospholipids, some of which occurred in sham-operated rats. It is concluded that hydrocephalus leads to reductions in levels of energy metabolites and in levels of membrane phospholipids and that the changes in energy metabolites can be reversed by early, but not by later, shunt treatment.     

Tricarboxylic Acid Cycle Activity Measured by <Superscript>13</Superscript>C Magnetic Resonance Spectroscopy in Rats Subjected to the Kaolin Model of Obstructed Hydrocephalus

Evaluating early changes in cerebral metabolism in hydrocephalus can help in the decision making and the timing of surgical intervention. This study was aimed at examining the tricarboxylic acid (TCA) cycle rate and ¹³C label incorporation into neurotransmitter amino acids and other compounds 2 weeks after rats were subjected to kaolin-induced progressive hydrocephalus. In vivo and ex vivo magnetic resonance spectroscopy (MRS), combined with the infusion of [1,6-¹³C]glucose, was used to monitor the time courses of ¹³C label incorporation into the different carbon positions of glutamate in the forebrains of rats with hydrocephalus as well as in those of controls. Metabolic rates were determined by fitting the measured data into a one-compartment metabolic model. The TCA cycle rate was 1.3 ± 0.2 μmoles/gram/minute in the controls and 0.8 ± 0.4 μmoles/gram/minute in the acute hydrocephalus group, the exchange rate between α-ketoglutarate and glutamate was 4.1 ± 2.5 μmoles/gram/minute in the controls and 2.7 ± 2.6 μmoles/gram/minute in the hydrocephalus group calculated from in vivo MRS. There were no statistically significant differences between these rates. Hydrocephalus caused a decrease in the amounts of glutamate, alanine and taurine. In addition, the concentration of the neuronal marker N-acetyl aspartate was decreased. ¹³C Labelling of most amino acids derived from [1,6-¹³C]glucose was unchanged 2 weeks after hydrocephalus induction. The only indication of astrocyte impairment was the decreased ¹³C enrichment in glutamine C-2. This study shows that hydrocephalus causes subtle but significant alterations in neuronal metabolism already early in the course of the disease. These sub-lethal changes, however, if maintained and if ongoing might explain the delayed and programmed neuronal damage as seen in chronic hydrocephalus.     

Changes in Regional Brain Levels of Amino Acid Putative Neurotransmitters After Prolonged Treatment with t he Anticonvulsant Drugs Diphenylhydantoin, Phenobarbitone, Sodium Valproate, Ethosuximide, and Sulthiame in the Rat

The effect of prolonged treatment (10 days) with the anticonvulsant drugs diphenylhydantoin (DPH), phenobarbitone, sodium valproate, ethosuximide and sulthiame, both singly and in combination, on regional rat brain amino acid neurotransmitter concentrations (GABA, glutamate, aspartate and taurine) were assessed. DPH had a major effect in the cerebellum and hypothalamus in that it significantly reduced cerebellar GABA, taurine and aspartate and hypothalamic GABA and aspartate. Sodium valproate significantly elevated GABA and taurine in most regions. Aspartate and glutamate were less affected. Phenobarbitone significantly elevated GABA concentrations in all brain regions, while taurine concentration was only elevated in the cerebral cortex. Ethosuximide induced changes were small compared to the other anticonvulsants while sulthiame produced complex changes. Anticonvulsant drugs administered in combination resulted in complex changes, suggesting that their mode of action is different.     

Change in glucose metabolism with time in hydrocephalic rats

Energy metabolism was studied by measurements of the activity and isozyme pattern of lactate dehydrogenase (LDH) in rats with kaolin-induced hydrocephalus at 1, 2, 4, and 6 weeks after the induction, as well as in normal, control rats. The total LDH activity in hydrocephalic rats was 1.5 times that in controls throughout the examination period. The main LDH isozyme in hydrocephalic rats was LDH 5(M4) at 2 weeks, but the isozyme pattern was the same as in controls at 4 and 6 weeks. These results suggest that the ratio of anaerobic to aerobic glycolysis increases in the acute stage of hydrocephalus and gradually lowers to become similar to that in controls with time.     

Changes in brain temperature and free amino acids in normal and protein deprived suckling rats exposed to room temperature

Previous reports on early-induced protein-calorie malnutrition (PCM) in rats have indicated alterations in the concentration of free amino acids and of protein synthesis in the brain. Recently it was shown that early-induced protein deprivation (PD) retards the development of thermoregulation. This resulted in a failure to maintain a normal rectal temperature after short exposure to room temperature (+22°C) still at the age of 20–25 days corresponding to changes seen in normal rats at an age of 10–15 days. In the present study, 20-day old PD and normal rats where examined with regard to the effect of exposure to room temperature on brain temperature and on brain free amino acids. The results show a similar reduction in brain and rectal temperature of the PD rats occuring within 30 minutes after exposure to room temperature. The reduction was in the range of 5°C. PD rats kept in room temperature for 5 hours and then allowed to recover at 32.5°C showed a slow increase in brain and rectal temperature but normal temperatures were not reached even after 1 hour. The concentration of free amino acids in the brain was examined in rats kept for 1 hour at room temperature or at 32.5°C. In the PD rats kept at 32.5°C, free aspartate and glutamate were reduced whereas taurine, GABA and glycine were increased as compared to their corresponding control rats. As a result of the reduced brain temperature in PD rats exposed to room temperature there was a reduction in free asparagine. The lability of the pool of asparagine may be related to the low levels of aspartate and glutamate in PD rats. On the basis of the present findings it is recommended that temperature-sensitive parameters are examined in PCM rats at a normal body temperature.Special Issue dedicated to Prof. Holger Hydén     

Maternal Undernutrition During Lactation: Effect on Amino Acids in Brain Regions of Offspring

Sprague-Dawley dams were fed either a protein-calorie deficient or control diet from day 5 to day 21 after parturition. The concentrations of seven amino acids (aspartate, glutamate, gamma-aminobutyric acid, glycine, glutamine, serine, and taurine) were determined in brain regions from 17-day-old undernourished offspring and from 35-day-old rehabilitated rats. The brain regions examined were the cortex, cerebellum, corpus striatum, hippocampus, hypothalamus, brainstem, and midbrain. At 17 days of age, taurine was the amino acid with the highest concentration, whereas at 35 days glutamate had the highest concentration. This change was due to the fact that the concentration of taurine decreased significantly in all brain regions between 17 and 35 days, whereas the concentration of glutamate remained high or increased somewhat in all brain regions except the hypothalamus and brainstem. When the age-matched offspring of control and undernourished rats were compared, several interesting and significant differences were found. The concentrations of glutamate and aspartate were significantly lower (decreased 16-34%) in the cerebellum, brainstem, cortex, and midbrain in 17-day-old undernourished rats. The aspartate level was also significantly decreased in the corpus striatum and hypothalamus in 17-day-old offspring. However, the deficiencies of aspartate and glutamate were transient and reversible. In contrast, the concentration of taurine was increased in the hypothalamus (31%) and hippocampus (12-33%) at both 17 and 35 days of age and in the midbrain (17%) at 17 days. Other transient abnormalities in amino acid levels were found in undernourished offspring. The results of these experiments suggest that undernutrition during lactation causes delayed CNS development, which is manifested in altered concentrations of the neurotransmitters aspartate, glutamate, and taurine.     

Ultrastructural features of the human frontal cortex neurons of maturing and hydrocephalic cerebrum

Human frontal cortex neurons of hydrocephalic infants were studied with electron microscope. A number of maturing neurones exhibited shrinkage and increased electron density of cytoplasmic matrix in the midst of gross hydrocephalic interstitial oedema. Remarkable preservation of growth cones and arrays of microtubules in a few neuronal perikarya and axons were discernible. Rows of normal looking neurones with parallel arrays of cisternae of rough endoplasmic reticulum were also detected. Thus, preservation of significant number of neurones was apparent even in cases of extreme hydrocephalus.     

Astrocyte metabolism is disturbed in the early development of experimental hydrocephalus

The proper diagnosis of the arrested or the progressive form of hydrocephalus has a critical impact on treatment, but remains difficult. The assessment of early changes in cerebral metabolism might help in the development of adequate non-invasive diagnostic tools. This study examined the alterations in label incorporation in neurotransmitter amino acids and other compounds in kaolin-induced progressive hydrocephalus in rats by means of magnetic resonance spectroscopy (MRS) combined with the administration of [1-13C]glucose and [1,2-13C]acetate. Some 2, 4 and 6 weeks after kaolin injection into the cisterna magna, cerebrum, brainstem and cerebellum were dissected. Interestingly, labelling of most amino acids derived from [1-13C]glucose showed no alterations, whereas labelling from [1,2-13C]acetate was affected. Two weeks after induction of hydrocephalus the taurine concentration was decreased, whereas the concentration of [1,2-13C]lactate was increased in the cerebrum and that of [1,2-13C]GABA in the brainstem. Furthermore, labelling from [1,2-13C]acetate was significantly decreased in [4,5-13C]glutamate, [1,2-13C]glutamate and [1,2-13C]GABA in cerebrum from 4 weeks after hydrocephalus induction. The concentration of N-acetylaspartate, a neuronal marker, was unchanged. However, labelling of the acetyl group from [1-13C]glucose was decreased in cerebellum and brainstem at 6 weeks after the induction of hydrocephalus. As glucose is metabolized predominately by neurones, whereas acetate is exclusively taken up by astrocytes, these results indicate that mostly astrocytic, and only later neuronal, metabolism is disturbed in the kaolin model of hydrocephalus. If verified in patients using in vivo MRS, impaired astrocyte metabolism might serve as an early indication for operative treatment.     

Effects of 3-acetylpyridine on the levels of several amino acids in different CNS regions of the rat

The effects of one intraperitoneal injection of 60–65 mg/kg of 3-acetylpyridine (3-AP) on the levels of aspartate, glutamate, GABA, taurine, glycine, and alanine in the cerebellum, medulla, telencephalon, and diencephalon-mesencephalon of the rat were studied at various times (4–28 days) after injection. In the first 4–7 days, the levels of glutamate, GABA, glycine, and alanine in the cerebellum were 10–30% higher in the 3-AP-treated rats than in the control animals. By day 14, the levels of these four amino acids were normal (in the case of glutamate and glycine) or below normal (for GABA and alanine). By day 21, the values for GABA and alanine returned to normal. In the first 7 days, the level of aspartate in the cerebellum was the same in both the 3-AP- and saline-injected groups. From days 14 to 28, the level of aspartate in the cerebellum was 10–20% lower in the 3-AP-injected group than in the saline-treated animals. The level of taurine in the cerebellum was 15–30% lower in the 3-AP group than in the control group from days 7 to 28. The pattern of changes observed in the medulla in the first 7 days was similar to that found in the cerebellum for this period. However, unlike the data for the cerebellum, the level of aspartate in the medulla was unchanged by the 3-AP injection from day 14 to day 28, and the level of glutamate in the medulla remained higher (10–15%) from days 14 to 28 in the 3-AP-injected animals with respect to control values. The levels of taurine in the medulla were lower (10–15%) from day 7 to day 28 in the 3-AP injected group with respect to control values. The injection of 3-AP did not alter the levels of aspartate, glutamate, GABA, taurine, glycine, or alanine in the telencephalon on days 7, 14, 21, or 28 and in the diencephalon-mesencephalon on day 21 with respect to control levels.     

Does urinary taurine reflect changes in protein metabolism? A study with cycloheximide in rats

Abstract

We have previously reported on the changes in urinary taurine levels in rats following treatment with some hepatotoxic agents and compounds reported to affect protein synthesis. This study follows the time course of the elevation of urinary taurine after treatment of rats with cycloheximide which was maximal 8–12 h alter dosing and was dose related. [³H]-leucine incorporation into proteins was used as an indicator of protein synthesis. There was a significant reduction in [³H]-leucine incorporation into acid precipitable proteins 8 h but not 24 h after dosing. The reduction in incorporation was negatively correlated with the raised levels of both serum and urinary taurine 8 h after dosing. Liver glutathione was raised both 8 and 24 h after dosing rats and liver taurine was significantly reduced at 8 h. It is suggested that measuring urinary taurine in collections made continuously might provide a simple, non-invasive biomarker for monitoring the effects of xenobiotics or other external stimuli on the status of protein synthesis.     

Possible role of glutamate, aspartate, glutamine, GABA or taurine on cadmium toxicity on the hypothalamic pituitary axis activity in adult male rats

This work was designed to evaluate the possible changes in glutamate, aspartate, glutamine, GABA and taurine within various hypothalamic areas the striatum and prefrontal cortex after oral cadmium exposure in adult male rats, and if these changes are related to pituitary hormone secretion. The contents of glutamine, glutamate, aspartate, GABA and taurine in the median eminence, anterior, mediobasal and posterior hypothalamus, and in prefrontal cortex in adult male rats exposed to 272.7 mol l^–1 of cadmium chloride (CdCl₂) in the drinking water for one month. Cadmium diminished the content of glutamine, glutamate and aspartate in anterior hypothalamus as compared to the values found in the untreated group. Besides, there is a decrease in the content of glutamate, aspartate and taurine in the prefrontal cortex. The amino acids studied did not change in median eminence, mediobasal and posterior hypothalamus or the striatum by cadmium treatment. Plasma prolactin and LH levels decreased in rats exposed to the metal. These results suggest that (1) cadmium differentially affects amino acid content within the brain region studied and (2) the inhibitory effect of cadmium on prolactin and LH secretion may be partially explained by a decrease in the content of both glutamate and aspartate in anterior hypothalamus, but not through changes in GABA and taurine.     

Neuropeptide changes and neuroactive amino acids in CSF from humans and sheep with neuronal ceroid lipofuscinoses (NCLs, Batten disease)

Anomalies in neuropeptides and neuroactive amino acids have been postulated to play a role in neurodegeneration in a variety of diseases including the inherited neuronal ceroid lipofuscinoses (NCLs, Batten disease). These are often indicated by concentration changes in cerebrospinal fluid (CSF). Here we compare CSF neuropeptide concentrations in patients with the classical juvenile CLN3 form of NCL and the classical late infantile CLN2 form with neuropeptide and neuroactive amino acid concentrations in CSF from sheep with the late infantile variant CLN6 form.A marked disease related increase in CSF concentrations of neuron specific enolase and tau protein was noted in the juvenile CLN3 patients but this was not observed in an advanced CLN2 patient nor CLN6 affected sheep. No changes were noted in S-100b, GFAP or MBP in patients or of S-100b, GFAP or IGF-1 in affected sheep. There were no disease related changes in CSF concentrations of the neuroactive amino acids, aspartate, glutamate, serine, glutamine, glycine, taurine and GABA in these sheep.The changes observed in the CLN3 patients may be progressive markers of neurodegeneration, or of underlying metabolic changes perhaps associated with CLN3 specific changes in neuroactive amino acids, as have been postulated. The lack of changes in the CLN2 and CLN6 subjects indicate that these changes are not shared by the CLN2 or CLN6 forms and changes in CSF concentrations of these compounds are unreliable as biomarkers of neurodegeneration in the NCLs in general.     

Neuropeptide changes and neuroactive amino acids in CSF from humans and sheep with neuronal ceroid lipofuscinoses (NCLs,Batten disease)

Anomalies in neuropeptides and neuroactive amino acids have been postulated to play a role in neurodegeneration in a variety of diseases including the inherited neuronal ceroid lipofuscinoses (NCLs, Batten disease). These are often indicated by concentration changes in cerebrospinal fluid (CSF). Here we compare CSF neuropeptide concentrations in patients with the classical juvenile CLN3 form of NCL and the classical late infantile CLN2 form with neuropeptide and neuroactive amino acid concentrations in CSF from sheep with the late infantile variant CLN6 form.A marked disease related increase in CSF concentrations of neuron specific enolase and tau protein was noted in the juvenile CLN3 patients but this was not observed in an advanced CLN2 patient nor CLN6 affected sheep. No changes were noted in S-100b, GFAP or MBP in patients or of S-100b, GFAP or IGF-1 in affected sheep. There were no disease related changes in CSF concentrations of the neuroactive amino acids, aspartate, glutamate, serine, glutamine, glycine, taurine and GABA in these sheep.The changes observed in the CLN3 patients may be progressive markers of neurodegeneration, or of underlying metabolic changes perhaps associated with CLN3 specific changes in neuroactive amino acids, as have been postulated. The lack of changes in the CLN2 and CLN6 subjects indicate that these changes are not shared by the CLN2 or CLN6 forms and changes in CSF concentrations of these compounds are unreliable as biomarkers of neurodegeneration in the NCLs in general.     

Cadmium effects on 24 h changes in glutamate,aspartate, glutamine,GABA and taurine content of rat striatum

This work evaluates the possible changes in 24 h variations of striatal aspartate, glutamate, glutamine, gamma-aminobutyric acid (GABA) and taurine content after oral cadmium treatment. Male rats were submitted to cadmium exposure at two doses (25 and 50 mg/L of cadmium chloride (CdCl₂)) in the drinking water for 30 days. Control rats received cadmium-free water. After the treatment, rats were killed at six different time intervals throughout a 24 h cycle. Differential effects of cadmium on 24 h amino acid fluctuations were observed. Metal exposure modified the daily pattern of the amino acids concentration found in control animals, except for GABA and taurine with the lowest dose used. Exposure to 25 mg/L of CdCl₂ decreased mean content of aspartate, as well as GABA concentration. These results suggest that cadmium exposure affects 24 h changes of the studied amino acids concentration in the striatum, and those changes may be related to alterations in striatal function.     

Neurochemical Changes in the Rat Occipital Cortex and Hippocampus after Repetitive and Profound Hypoglycemia During the Neonatal Period: An Ex Vivo 1H Magnetic Resonance Spectroscopy Study

The brain of a human neonate is more vulnerable to hypoglycemia than that of pediatric and adult patients. Repetitive and profound hypoglycemia during the neonatal period (RPHN) causes brain damage and leads to severe neurologic sequelae. Ex vivo high-resolution ¹H nuclear magnetic resonance (NMR) spectroscopy was carried out in the present study to detect metabolite alterations in newborn and adolescent rats and investigate the effects of RPHN on their occipital cortex and hippocampus. Results showed that RPHN induces significant changes in a number of cerebral metabolites, and such changes are region-specific. Among the 16 metabolites detected by ex vivo ¹H NMR, RPHN significantly increased the levels of creatine, glutamate, glutamine, γ-aminobutyric acid, and aspartate, as well as other metabolites, including succine, taurine, and myo-inositol, in the occipital cortex of neonatal rats compared with the control. By contrast, changes in these neurochemicals were not significant in the hippocampus of neonatal rats. When the rats had developed into adolescence, the changes above were maintained and the levels of other metabolites, including lactate, N-acetyl aspartate, alanine, choline, glycine, acetate, and ascorbate, increased in the occipital cortex. By contrast, most of these metabolites were reduced in the hippocampus. These metabolic changes suggest that complementary mechanisms exist between these two brain areas. RPHN appears to affect occipital cortex and hippocampal activities, neurotransmitter transition, energy metabolism, and other metabolic equilibria in newborn rats; these effects are further aggravated when the newborn rats develop into adolescence. Changes in the metabolism of neurotransmitter system may be an adaptive measure of the central nervous system in response to RPHN.     

Abnormalities in Amino Acid Neurotransmitters in Discrete Brain Regions of Genetically Dystonic Hamsters

The concentrations of 11 amino acids, including the neurotransmitters gamma-aminobutyric acid, glutamate, aspartate, glycine, and taurine, were determined by HPLC in 12 brain regions of genetically dystonic (dtSZ) hamsters and age-matched nondystonic controls. Since dystonia in mutant dtSZ hamsters is transient and disappears after about 70 days of age, amino acids were determined at the age of maximum severity of dystonia (30-40 days) and after disappearance of the disease, to examine which neurochemical changes were related to dystonia. In dtSZ hamsters with the maximum severity of dystonia, significant changes in concentrations of the neurotransmitters gamma-aminobutyric acid, glutamate, aspartate, and taurine were found in several regions involved in motor functions, e.g., cerebellum, thalamus, and corpus striatum. Most of these changes were not permanent but disappeared in parallel with dystonia, implicating a causal relationship between altered aminoacidergic neurotransmission and dystonia in mutant dtSZ hamsters.     

The surface fine structure of the ependymal lining of the lateral ventricle in rats with hereditary hydrocephalus

Summary The ependyma of the lateral ventricle of rats with hereditary hydrocephalus was studied using scanning electron microscopy. Normal rats from the same litters were used as control animals. The surface morphology of the lateral ventricle of normal rats corresponded to results reported by other authors. The most prominent changes in the surface morphology of the ependyma of the hydrocephalic rats were seen in the cilia. They were shortened, fewer in number and clumped or matted. The surface of the ependymal cells was flattened and contained small, irregular projections. The number of large supraependymal cells, regarded as neurons, appeared to have diminished in the hydrocephalic rats. The number of supraependymal macrophages was greatly increased in these rats, suggesting the existence of an ependymitis. Send offprint requests to: College of Veterinary Medicine, Department of Anatomy and Embryology, Hämeentie 57, SF-00550 Helsinki 55, Finland Acknowledgement: This study was supported by a grant from the Sigrid Jusélius Foundation to S.T.     

Electroencephalography in the diagnosis of hydrocephalus in golden hamsters (Mesocricetus auratus)

The golden hamster (Mesocricetus auratus) is a popular laboratory animal and is used in a multitude of behavioural studies. However, it has been shown that it suffers from different forms of hereditary hydrocephalus, which may result in behavioural changes. This prospective study was designed to look into the usefulness of electroencephalography (EEG) measurements in the diagnosis of hydrocephalus in hamsters. The EEGs of the hydrocephalic hamsters were evaluated double-blind and showed a high-voltage slow wave activity, with a fast activity superimposed onto it. This pattern has already been well described in other hydrocephalic species and differed significantly from the EEGs that were obtained from the normal hamsters. It was concluded from our study that a background activity with an amplitude over 50 muV in combination with a frequency of < or =5 Hz was highly indicative of hydrocephalus in young hamsters. We believe that the EEG could be a very useful diagnostic tool in the screening for hydrocephalus in hamsters.     

Effects of selective doses of x-irradiation on the levels of several amino acids in the cerebellum of the rat

The cerebella of rats were exposed to selective doses of low levels of x-irradiation beginning on day 4, 8, or 12 following birth. The doses of x-irradiation given on days 12, 13, and 15 (12–15X group) resulted in a 24% reduction in the wet weight of the cerebella; the doses given on days 8, 9, 11, 13, and 15 (8–15X group) resulted in a 57% weight reduction; the doses given on days 4, 5, 7, 9, 11, 13, and 15 (4–15X group) resulted in a 67% weight reduction. The schedule of x-irradiation begun on day 12, which prevented the acquisition of the late-forming granule cells, reduced the levels (nmole/mg dry tissue weight) of alanine (22%) and glutamate (10%), and increased the levels of glycine (15%), GABA (13%), and taurine (71%), with respect to control values. The schedule begun on day 8, which prevented the acquisition of stellate and granule cells, reduced the levels of alanine (15%), glutamate (12%), and taurine (21%), and increased the levels of glycine (102%) and GABA (56%). The schedule begun on day 4, which prevented the acquisition of basket, stellate, and granule cells, reduced the level of glutamate (15%) and increased the levels of glycine (186%) and GABA (78%). The levels of alanine and taurine in the cerebella of the 4–15X group were the same as control values. The level of aspartate in the cerebella of the 3 groups of x-irradiated animals was not significantly different from control values. The consistent reduction in the level of glutamate as a function of the number of doses of x-irradiation is suggestive that glutamate may have a higher level in the granule cells than in other cells in the cerebellum, and that the higher level may be a reflection of a possible excitatory transmitter role for glutamate. In addition, the data are interpreted in terms of taurine being associated with the stellate cells and possibly serving as a transmitter for these inhibitory interneurons.     

Tetanus Toxin-Induced Effects on Extracellular Amino Acid Levels in Rat Hippocampus: An In Vivo Microdialysis Study

Abstract: Tetanus toxin is a potent neurotoxin that is widely considered to produce its effect through impairment of inhibitory neurotransmission. We report the effect of a single unilateral intrahippocampal injection of tetanus toxin on extracellular levels of neuroactive amino acids in freely moving rats, at times ranging between 1 and 7 days posttreatment. Tetanus toxin treatment did not alter extracellular levels of aspartate, glutamate, and taurine at any time during the study. However, although extracellular GABA levels were unaffected by toxin injection 1, 2, and 3 days after treatment, they were reduced (45 ± 8% of contralateral vehicle-injected level) at day 7. Challenge with a high K⁺ concentration, 7 days after treatment, produced elevations in extracellular levels of taurine and GABA in both vehicle- and toxin-injected hippocampi, with evoked levels of GABA being lower in the toxin-treated side (39 ± 16% of contralateral vehicle-injected level). Aspartate and glutamate levels were not increased by high-K⁺ infusion. These findings are discussed in relation to the possible role that an imbalance in excitatory/inhibitory tone may play in the production of tetanus toxin-induced neurodegeneration.