A test of the catecholamines hypothesis for an acute exercise-cognition interaction.

The purpose of the study was to examine the usage of norepinephrine (NE) and dopamine (DA) in the brain when exercising while simultaneously undertaking cognitive tests. Plasma concentrations of the NE metabolite 3-methoxy 4-hydroxyphenylglycol (MHPG) and the DA metabolite homovanillic acid (HVA) showed a linear increase from rest to exercising at 40% and 80% maximum power output (W.max) while simultaneously undertaking cognitive tasks (random number generation (RNG) and response time). Delta plasma concentrations of MHPG and HVA at each exercise intensity while undertaking cognitive tasks and while exercising without cognitive tasks did not differ. Taking blood samples at 0, 1, 3, and 5 min following cessation of exercise did not affect results. Regression correlations showed that delta MHPG and HVA plasma concentrations at the 1 and 3 min sampling times were strong predictors of delta RNG, response time and movement time. Reaction time at 80% W.max significantly increased, while movement time at 80% W.max significantly decreased. It was concluded that these results provide no support for a direct effect of increased catecholamines concentrations on cognitive performance during exercise. The regression data suggest that there is some relationship between exercise, catecholamines concentrations and cognition.

Effect of creatine supplementation and sleep deprivation, with mild exercise, on cognitive and psychomotor performance, mood state, and plasma concentrations of catecholamines and cortisol.

RATIONALE: Sleep deprivation has a negative effect on cognitive and psychomotor performance and mood state, partially due to decreases in creatine levels in the brain. Therefore, creatine supplementation should lessen the negative effects of sleep deprivation. OBJECTIVES: The objective of this study was to examine the effect of creatine supplementation and sleep deprivation, with mild exercise, on cognitive and psychomotor performance, mood state, and plasma concentrations of catecholamines and cortisol. METHOD: Subjects were divided into a creatine group (n=10) and a placebo group (n=9). They took 5 g of creatine monohydrate or a placebo, dependent on their group, four times a time a day for 7 days, immediately prior to the experiment. The study was double blind. Subjects undertook tests of random movement generation (RMG), verbal and spatial recall, choice reaction time, static balance and mood state pre-test (0 h), after 6, 12 and 24 h of sleep deprivation, with intermittent exercise. They were tested for plasma concentrations of catecholamines and cortisol at 0 and 24 h. RESULTS: At 24 h, the creatine group demonstrated significantly less change in performance from 0 h (delta) in RMG, choice reaction time, balance and mood state. There were no significant differences between groups in plasma concentrations of catecholamines and cortisol. Norepinephrine and dopamine concentrations were significantly higher at 24 h than 0 h, but cortisol were lower. CONCLUSIONS: Following 24-h sleep deprivation, creatine supplementation had a positive effect on mood state and tasks that place a heavy stress on the prefrontal cortex.

Pulmonary antioxidant concentrations and oxidative damage in ventilated premature babies.

OBJECTIVE: To determine the relation between lipid peroxidation and the antioxidants ascorbate, urate, and glutathione in epithelial lining fluid in ventilated premature babies, and to relate the biochemical findings to clinical outcome. DESIGN: A cohort study conducted between January 1999 and June 2001. SETTING: A NHS neonatal intensive care unit. PATIENTS: An opportunity sample of 43 ventilated babies of less than 32 weeks gestation. MAIN OUTCOME MEASURES: The duration of supplementary oxygen according to the definition of bronchopulmonary dysplasia (BPD; oxygen dependency at 36 weeks gestational age). METHODS: Epithelial lining fluid was sampled by bronchoalveolar lavage. Ascorbate, urate, glutathione, and malondialdehyde (a marker of lipid peroxidation) were measured. RESULTS: Babies who developed BPD had significantly lower initial glutathione concentrations (mean (SEM) 1.89 (0.62) v 10.76 (2.79) microM; p = 0.043) and higher malondialdehyde concentrations (mean (SEM) 1.3 (0.31) v 0.345 (0.09) microM; p < 0.05) in the epithelial lining fluid than those who were not oxygen dependent. These variables were poor predictors of the development of BPD. Gestational age, endotracheal infection, and septicaemia had good predictive power. The level of oxidative damage was associated with the presence of endotracheal infection/septicaemia rather than inspired oxygen concentration. CONCLUSIONS: Endotracheal infection, septicaemia, and gestational age, rather than antioxidant concentrations, are the most powerful predictors of the development of BPD.

The characteristics of arginine transport by rat cerebellar and cortical synaptosomes.

The uptake of L-[3H]arginine into synaptosomes prepared from rat cerebellum and cortex occurred by a high-affinity carrier-mediated process. The uptake of arginine appeared to be potentiated by removal of extracellular Na+, inhibited by high levels of extracellular K+, but not by depolarization with veratridine or 4-amino pyridine. The effect of Na+ removal or K+ elevation did not seem to be due to changes in intracellular Ca2+ or pH. In both brain regions, uptake was significantly inhibited by L-arginine, L-lysine, L-ornithine, and L-homoarginine, but not by D-arginine nor L-citrulline. Uptake was also inhibited by NG-monomethyl-L-arginine acetate, but not by NG-nitro-L-arginine methyl ester nor NG-nitro-L-arginine except in the cortex at a concentration of 1 mM. The results indicate that the carrier system operating in synaptosomes showed many of the characteristics of the ubiquitous y+ system seen in many other tissues, although its apparent sensitivity to variations in extracellular Na+ was unusual.

Changes in brain glutathione levels during postnatal development in the rat.

The concentration of glutathione in the rat cerebral cortex, cerebellum and liver was examined during postnatal development. The glutathione level in both brain regions was low at the earliest age studied (postnatal day (PND) 3) and peaked at PND7. In the liver, the glutathione level was maximal at PND30. The glutathione peak at PND7 occurs during a period of intense synaptogenesis and may be related to a neuroprotective role during brain development.

On the significance of perfusion rate in the study of glutamate release from superfused synaptosomes.

The effect of perfusion rate on the apparent release of [(3)H]glutamate from prelabelled and superfused rat cortical synaptosomes was examined. The proportion of tissue [(3)H]glutamate released in response to a 4 ml depolarizing pulse of 15 mM K+ increased almost linearly with perfusion rates from 1 ml min(-1) to 10 ml min(-1). Release did not increase markedly between 10 ml min(-1) and 20 ml min(-1). The basal efflux of [(3)H]glutamate also increased with perfusion rate. The increase in both basal efflux and (K+)-induced release is interpreted as being due to a greater amount of released transmitter avoiding recapture by uptake processes as perfusion rate increases. This is supported by the observation that increasing the potential number of uptake sites in the tissue decreases both the basal and (K+)-evoked release of the transmitter. The significance of this with respect to optimal perfusion rates for studies on the regulation of glutamate release is discussed.

Acquisition of conditioned inhibition in rats is impaired by ablation of serotoninergic pathways.

The effects of chronically ablating the serotoninergic inputs to various regions of the rat brain on the ability to solve a feature-negative discrimination was measured. After intracerebroventricular administration of the specific neurotoxin 5,7-dihydroxytryptamine, the rats exhibited an impaired capacity to solve such a discrimination, irrespective of whether auditory or visual stimuli were used. Further behavioural analysis revealed that this effect was not due to a reduced capacity to form excitatory associations, since both groups responded equally to reinforced stimuli. By contrast, the lesion more likely resulted in a failure to endow the non-reinforced stimuli with inhibitory properties. This suggestion was supported by the observation that, in a retardation test, the conditioned inhibitor aroused less inhibition in the lesioned group than in vehicle-injected controls. Furthermore, the conditioned inhibitor failed to pass a summation test in lesioned animals, again indicating that their hampered ability to master the discrimination was the result of an impairment in the formation of inhibitory associations. It is concluded that destruction of central 5-hydroxytryptamine-containing pathways impairs the functioning of brain areas underlying inhibitory associative learning.

The uptake of gamma-aminobutyric acid and glutamate by synaptosomes from the visual cortex of albino and pigmented rabbits.

The synaptosomal uptake of glutamate and gamma aminobutyric acid [GABA] in the visual cortex of albino and pigmented rabbits was compared. GABA uptake was similar in both pigmented and albino rabbits, but glutamate uptake was greater in the pigmented rabbit. The kinetics of glutamate uptake in albino and pigmented rabbits suggested that the number of functioning glutamate synapses may be lower in the albino. The significance of this with respect to the differences in visual processing in the two types of rabbit is discussed.

Postnatal development of the calcium-dependency of glutamate release from rat cortical synaptosomes: comparison with 5-hydroxytryptamine release.

The ontogeny of the Ca(2+)-dependency of the depolarisation-induced release of preloaded [3H]glutamate and [3H]5-hydroxytryptamine [5-HT] from rat cortical synaptosomes was examined. 5-HT release was found to be exclusively Ca(2+)-dependent at all ages studied. In contrast, glutamate release only showed a significant Ca(2+)-dependent component from postnatal day 10 [PND 10] onwards. This correlated with the ontogeny of the glutamate accumulating activity of synaptic vesicles, a finding consistent with vesicles being the site of Ca(2+)-dependent release. The effectiveness of K(+)-depolarisation in inducing the Ca(2+)-dependent release of both transmitters increased during the early neonatal period, reaching near adult levels at PND20 for 5-HT and PND30 for glutamate.

On the role of nitric oxide as a cellular messenger in brain.

The characteristics of the high-affinity uptake of [3H]-L-arginine into cerebellar and cortical synaptosomes were investigated. Uptake into cerebellar synaptosomes was often greater than seen in cortical synaptosomes under similar experimental conditions, and this was reflected by a higher Vmax in synaptosomes from this brain region. Uptake into synaptosomes prepared from both brain regions was markedly enhanced by removing extracellular Na+, adn inhibited by high concentrations of extracellular K+. Depolarisation with 4-aminopyridine or veratridine has no effect on uptake. Uptake was also unaffected by hyperpolarisation. The profile of inhibition of arginine uptake by related amino acids was similar to that seen for the y+ carrier, but the other characteristic alluded to above suggest that the carrier is distinct from the classical y+ system. The possible relationship between the carrier and the metabolism of arginine through the nitric oxide [NO] pathway, and the role of NO in the central nervous system is discussed.

Effects of short-term hypoxia on [3H]glutamate release from preloaded hippocampal and cortical synaptosomes.

The effect of short-term hypoxia on the release of [3H]glutamate from preloaded hippocampal and cortical synaptosomes was studied in a rapid superfusion system. The technique minimised the loss of released glutamate by reuptake. The results indicated that the effects of short term hypoxia were qualitatively similar to those reported in previous studies using more long-term hypoxia, but were significantly smaller. The non-Ca(2+)-dependent efflux of glutamate from cortical synaptosomes was increased by hypoxia as was the Ca(2+)-dependent release from hippocampal tissue. Possible mechanisms for these findings were discussed. The small amplitude of these changes in comparison to the effects seen in slowly perfused tissue in vitro and in vivo indicated that the contribution made by changes in neuronal efflux to the overall increase in extracellular glutamate seen in hypoxia is relatively minor.

Changes in synaptosomal glutamate release during postnatal development in the rat hippocampus and cortex.

The effectiveness of K+ depolarisation in inducing the release of [3H]L-glutamate from preloaded hippocampal and cortical synaptosomes was examined in rats aged from postnatal day 4 (PND 4) to adult. In the lower age groups studied (PND 4-PND 15), the response to depolarisation was always smaller than that seen in the adult. From PND 15, the sensitivity of the release process increased steadily to a maximum level in the adult. The relatively small amounts of glutamate released in response to K(+)-depolarisation in the younger age groups may be a factor which contributes to the relative insensitivity of neonatal brain to ischaemic damage. Discrete variations in the sensitivity to K+ depolarisation observed in animals aged from PND 4 to PND 15 may be involved in plastic changes in neural activity which are known to occur during this important development period.

Synaptosomal tryptophan uptake and efflux following lesion of central 5-hydroxytryptaminergic neurones.

1. This study attempted to determine whether the activation of the tryptophan carrier in rat forebrain synaptosomes caused by depolarization or by extracellular sodium depletion occurred exclusively in 5-hydroxytryptaminergic nerve endings. 2. Ascending 5-hydroxytryptaminergic neurones were lesioned either electrolytically or by intraventricular administration of 5,7-dihydroxytryptamine. The extent of the lesion was assessed by comparing the uptake of [3H]-5-hydroxytryptamine (5-HT) in lesioned animals and in sham-operated controls. [3H]-5-HT uptake was reduced by 85.9 +/- 1.63% (mean +/- s.e. mean) in animals receiving electrolytic lesions, and by 87.4 +/- 4.51% in those receiving 5,7-dihydroxytryptamine. 3. The uptake of [3H]-tryptophan by synaptosomes from lesioned animals incubated in standard Na(+)-rich media was slightly lower (278.8 +/- 27.3 pmol mg-1 protein min-1) than that observed in sham-operated controls (360.6 +/- 30.3 pmol mg-1 protein min-1). However, uptake in the absence of extracellular Na+ was increased to a similar extent in both the sham-operated (539 +/- 54.5 pmol mg-1 protein min-1) and lesioned animals (507.2 +/- 42.4 pmol mg-1 protein min-1). 4. The efflux of [3H]-tryptophan in response to extracellular Na+ depletion was similar in sham-operated and lesioned animals. Release expressed as a percentage of tissue [3H]-tryptophan released in response to the pulse of Na(+)-free medium was 6.691 +/- 0.585 (n = 4) in sham-operated controls and 8.195 +/- 0.906 in lesioned animals. 5. The efflux of [3H]-tryptophan in response to K+ depolarization was also unchanged in lesioned animals when compared with sham-operated controls. Release, expressed as described above was, in sham-operated controls 3.76 +/- 0.41 (n = 4) and 4.09 +/- 0.30 in lesioned animals. 6. The results of this study show that the tryptophan carrier which is activated by depolarization or by extracellular Na+ depletion is not located exclusively on 5-hydroxytryptaminergic nerve endings. Moreover the contribution made by 5-hydroxytryptaminergic neurones appears to be only minor.

On the mechanism by which extracellular sodium depletion causes 5-hydroxytryptamine release from rat brain synaptosomes.

The release of 3H-labelled 5-hydroxytryptamine (5-HT) from preloaded and superfused rat forebrain synaptosomes in response to extracellular Na+ depletion was studied. In the absence of monoamine oxidase inhibitors, the release of [3H]-5-HT caused by Na+ depletion was not affected by immobilizers of the plasma membrane 5-HT carrier. The release of [3H]-5-HT in response to Na+ depletion was also either independent of, or inversely related to the concentration of extracellular Ca2+ depending on the degree to which extracellular Na+ was reduced. The efflux of 45Ca2+ from prelabelled synaptosomes was decreased by Na+ reduction but the amplitude of the changes in 45Ca2+ efflux did not totally correlate with the changes in [3H]-5-HT efflux under the same experimental conditions. These results suggest that the release of [3H]-5-HT caused by Na+ depletion in drug-free synaptosomes is not mediated by 5-HT efflux through the plasma membrane carrier, nor to changes in cytosolic Ca2+ consequent to changes in Ca2+ fluxes across the plasma membrane. The results have been tentatively explained as an elevation of spontaneous 5-HT efflux caused by an increase in membrane fluidity mediated by the ionic manipulations used to produce the Na+-depleted media.

The mechanism by which monoamine oxidase inhibitors give rise to a non-calcium-dependent component in the depolarization-induced release of 5-HT from rat brain synaptosomes.

1. The effects of the monoamine oxidase inhibitors pargyline and nialamide on the Ca2+-dependency of [3H]-5-hydroxytryptamine release from superfused rat brain synaptosomes has been studied in order to evaluate the discrepancies that have occasionally been observed in studying transmitter release by in vivo and in vitro techniques. 2. The application of K+ pulses of low concentration (12.5-20 mM) caused an essentially Ca2+-dependent release of [3H]-5-HT. However, at K+ concentrations above 30 mM, a small non-Ca2+-dependent component appeared. 3. At high concentrations of K+ (30-55 mM), nialamide (18 microM) or pargyline (7 microM) increased the amount of [3H]-5-HT released which could be accounted for by an increase in the non-Ca2+-dependent component of release. 4. The elevation of the non-Ca2+-dependent component of release caused by the monoamine oxidase inhibitors was totally abolished by the inhibitors of the plasma membrane 5-HT carrier, chlomipramine (500 nM), citalopram (50 nM) and fluoxetine (1 microM). 5. The results suggest that the non-Ca2+-dependent component of release seen with high depolarizing concentrations of K+, particularly in the presence of monoamine oxidase inhibitors, is caused by the efflux of [3H]-5-HT through the plasma membrane carrier which seems to be activated during depolarization. 6. The significance of these findings to the physiological in vivo situation, and to the use of in vitro preparations in the study of transmitter release is discussed.

Studies on the mechanism by which tryptophan efflux from isolated synaptosomes is stimulated by depolarization.

1. The efflux and influx of tryptophan across the synaptosomal plasma membrane has been studied under a variety of experimental conditions, in order to examine the mechanism by which depolarization enhances the efflux of tryptophan from superfused synaptosomes. 2. Efflux of [3H]-tryptophan from preloaded superfused synaptosomes was found to be enhanced by K+ depolarization in a Ca2+ and dose-dependent manner. In contrast, [3H]-phenylalanine efflux was only poorly stimulated by depolarization and only by very high concentrations of K+. 3. Tryptophan efflux was also enhanced by decreasing the extracellular Na+ concentration, but this effect was not dependent on extracellular Ca2+. 4. Influx of [3H]-tryptophan into synaptosomes was stimulated by extracellular Na+ removal, but the uptake of [3H]-phenylalanine was unaffected by this procedure. 5. Both the induced influx and efflux of tryptophan observed under these experimental conditions was inhibited by immobilizing the plasma membrane carrier with parachlorophenylalanine. This implied that both the enhanced influx and efflux arose as a consequence of the activation of the membrane tryptophan carrier, the direction of the observed effect being dependent upon the manner in which the experiments were conducted. 6. The relationship between depolarization, the activation of the membrane tryptophan carrier and the significance of this to the in vivo situation is discussed.