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Background & Aims: Exogenous intravenous glutamine supplementation to head trauma patients leaves intracerebral glutamate concentration unaffected. The effect of an exogenous supply upon glutamine and glutamate exchange across the brain has still not been characterised., Methods: A prospective randomised cross-over study, where i.v. glutamine dipeptide was compared with placebo. Arterio-venous concentration differences of free amino acids across the brain and amino acid flux across the leg were measured. In addition, the endogenous glutamine production was calculated. Fifteen mechanically ventilated head trauma patients with GCS < or =8 were included and studied during two consecutive 24-h periods on days 2-5 following head trauma., Results: Glutamine was continuously released from both the brain and the leg. The arterio-venous (a-v) concentration differences over the brain were calculated to be -49+/-26 and -27+/-14 micromol/L during the treatment and control periods respectively, showing a continuous release of glutamine (p<0.0001). On the other hand, the a-v difference of glutamate was not different from zero (p>0.2). The whole-body glutamine rate of appearance (R(a)) was calculated to be 218+/-75micromol/kg body weight/h., Conclusion: Intravenous glutamine supplementation to head trauma patients was associated with an unaffected amino acid exchange pattern across head and leg, without any measurable uptake of glutamate across the brain. Endogenous glutamine production was in the normal range despite the low plasma glutamine concentration. This pilot study opens the possibility to perform prospective clinical trials in head trauma patients to evaluate the clinical efficacy of exogenous glutamine supplementation.

A novel method is described to follow known and unknown compounds in biological processes using microdialysis sampling and mass spectrometric detection. By implementation of internal standard, desalting/enrichment for the sample work-up, and multivariate data analysis, this methodology is a basis for future applications in early diagnosis of diseases and organ damage, as a complement to the routinely used clinical methods for biological samples. The present study includes screening without specific target analytes, of samples collected by microdialysis from liver of anaesthetized rats before and after local damage to this organ. Sample series were classified by principal component analysis, and the stimulation was identified in the chemical patterns produced by the presented analytical tool.

Background: Microdialysis-based glucose sensors have recently been introduced for monitoring glucose levels in diabetic patients. The flow rate by which the fluid sample is pumped through the microdialysis catheter varies in different studies., Aim: To study the effects of various flow rates on glucose and its metabolites sampled by microdialysis during an oral glucose tolerance test., Material, Methods: Glucose, lactate, pyruvate and glycerol were measured with microdialysis in interstitial fluid of subcutaneous adipose tissue in twelve healthy young subjects before and during an oral glucose tolerance test using four different flow rates (0.3, 1, 2 and 5 microL/min) and a 30 mm dialysis membrane., Results: At the basal fasting state the dialysate glucose obtained by 0.3 microL/min was equal to capillary glucose concentration. A decrease in dialysate glucose levels during the basal state was observed for higher flow rates but not for 0.3 microL/min, which indicates a depleting effect. The relative increase after OGTT was similar for capillary glucose and flow rate 0.3 microL/min but not for higher flow rates., Conclusion: The low microdialysis flow rate (0.3 microL/min) facilitates the capture of true interstitial glucose concentrations during glucose fluctuations. Thus this low flow rate is preferred in studies of local tissue metabolism.

Objective: investigate the metabolic response of the spinal cord and the effect of allopurinol following cross clamping of the descending thoracic aorta in a porcine model., Design: experimental animal study., Materials: twelve domestic swine. Six pigs were pre-treated with allopurinol, while six pigs served as controls., Methods: measurement of extracellular concentrations of glucose, pyruvate, lactate, glycerol and glutamate using microdialysis in the lumbar spinal cord. Measurement of lumbar spinal blood flow using laser Doppler technique., Results: for all animals there was a significant decrease in concentrations of glucose and pyruvate together with a significant increase in the lactate-pyruvate ratio during aortic cross clamping. There was also a significant increase in glycerol concentrations 60 min after cross clamping, and a significant decrease in glutamate concentrations after 50 min. No differences in concentrations of glucose, pyruvate, lactate and glutamate or the glutamate-pyruvate ratio were observed between animals used as controls and those treated with allopurinol. The laser Doppler flux decreased to 40% of pre cross-clamp level, returning to normal values at declamping., Conclusion: the changes in energy-related metabolites reflect a considerable ischaemia in the spinal cord tissue but there was no convincing effect of allopurinol on the lumbar spinal cord metabolism during thoracic aortic cross clamping in this model., (Copyright 2001 Harcourt Publishers Limited.)

The possibility of using microdialysis catheters with a large pore size dialysis membrane (100 kDa) to investigate the action of macromolecules perfused into the interstitial space of peripheral tissues was explored. This was made possible by increasing the colloid osmotic pressure of the perfusate with 40 g/l of dextran-70 to prevent perfusate loss across the dialysis membranes. Microdialysis catheters were inserted into the quadriceps femoris muscle of 13 human subjects. With different perfusion flow rates (1. 33, 0.66, 0.33 and 0.16 microl/min) the recorded concentrations of glucose, lactate, and urea were in agreement with values previously obtained using a conventional membrane with a smaller pore size (20 kDa) [Rosdahl H, Hamrin K, Ungerstedt U, Henriksson. J Am J Physiol 1998;274:E936-45.]. When insulin was added to the perfusate, the concentration of glucose was significantly reduced, indicating that insulin diffuses across the dialysis membrane and has cellular effects that can be simultaneously recorded. The present findings are the first documentation on the use of microdialysis to study the local metabolic action of large peptide molecules in human tissues and may open new avenues for in-vivo metabolic research.

Objective: to investigate the feasibility of using microdialysate glucose, lactate and pyruvate concentrations for grading the severity of blood flow reduction in patients with critical limb ischaemia., Patients and Methods: microdialysis catheters were inserted (two subcutaneously and one intramuscularly) in the symptomatic limb of ten patients. To further reduce limb perfusion, the lower leg was elevated during part of the experiment., Results: elevation reduced ankle and toe blood pressure and transcutaneous oxygen tension. Microdialysate glucose concentration decreased at all three catheter sites, while lactate increased in the intramuscular catheter. Two patients interrupted the elevated position prematurely due to severe pain in the foot. They had among the highest lactate levels in the horizontal position and the most marked increases following provocation. Neither initial metabolite concentrations nor concentration changes during elevation were shown to correlate to conventional methods used to assess limb perfusion., Conclusions: in patients with critical limb ischaemia microdialysis can be used without complications. A significant decrease in glucose concentration may reflect lowered blood flow in the elevated position. Metabolic response, i.e. increase in lactate concentration during profoundly reduced limb perfusion was heterogeneous, indicating an overestimation of the presence of ischaemia in some patients using current diagnostic methods., (Copyright 2000 Harcourt Publishers Ltd.)

The present study tested the hypotheses that spontaneous gamma-aminobutyric acid (GABA) efflux in anterior rat striatum is 1) independent of intra- and extracellular calcium; and 2) is physiologically relevant. Extracellular dopamine (DA) and GABA were sampled from striatum of awake, freely moving rats using in vivo microdialysis. Although dialysate concentrations of DA were 2 to 3 times greater than GABA and were decreased by at least 70% by removal of calcium, GABA was unaffected even in the presence of EGTA or the intracellular calcium chelator APTRA-AM. Functional significance of this non-exocytotic pool of GABA was tested by injecting 3-mercaptopropionic acid (3-MPA), an inhibitor of GABA synthesis, into the striatum via a guide cannula sidled alongside a microdialysis probe and measuring subsequent effects on behavior and perfusate concentrations of GABA. Results show that 3-MPA increases gnawing behavior suggesting that basal, non-exocytotic GABA overflow normally functions to suppress gnawing.

Subcutaneous microdialysis was used to monitor the immediate metabolic changes with respect to glucose, glycerol and lactate in the extracellular space of adipose tissue among five small-for-gestational-age (SGA) and two appropriate-for-gestational-age (AGA) newborns. There was a good correlation between glucose levels in blood and dialysate (r = 0.97, n = 14). The infants showed rapid rises and falls in dialysate glucose levels that are not seen among older children and adults. The levels of lactate were higher than those reported in blood. Lactate may serve as an alternative source of energy for the neonate. Microdialysis is of potential value in increasing our understanding of metabolic events. It provides a safe on-line means of making continuous measurements in these fragile patients in order to detect periods of hypoglycaemia, at least in infants with a birth weight > 1000 g.

An exposure chamber for microdialysis on awake, freely moving rats during exposure to volatile agents is described. Inhalation exposure to 1000 and 2000 ppm toluene for 2 h was accompanied by an increase in extracellular dopamine levels within the striatum, but did not affect the homovanillic acid level. Neither the dopamine nor the homovanillic acid level was affected by toluene 500 ppm or isoamylacetate. It is suggested that the action of inhaled toluene on the dopamine neuron differs from that of the anaesthetic halothane, possibly by interfering with dopamine reuptake. Microdialysis seems to be a useful tool for studying the effects of volatile agents on brain neurotransmission.

The glucose concentration in the extracellular space of subcutaneous adipose tissue closely mirrors the blood glucose concentration. With a microdialysis technique, we undertook continuous ambulatory monitoring of adipose tissue glucose in 17 insulin-dependent diabetic patients with labile glycaemic control. The aims of the study were to investigate performance of the microdialysis device and to evaluate whether consecutive 24 h glucose profiles could be used to adjust insulin therapy. A microdialysis probe was implanted subcutaneously, perfused by a portable microinfusion pump, and dialysate fractions were collected every 1 or 2 h for 75 h. The mean (SE) tissue dialysate glucose concentration was 93 (3)% of the concentration in venous plasma, and variations in adipose tissue glucose closely paralleled changes in plasma glucose. Mean 24 h tissue glucose concentration correlated significantly with glycosylated haemoglobin (HbA1c; r = 0.62, p < 0.01). Most patients had a reproducible daily pattern of glucose swings, and in more than half the patients consecutive ambulatory glucose profiles were almost superimposed. When patients' insulin therapy was adjusted on the basis of ambulatory glucose monitoring, HbA1c decreased by almost 2% (p < 0.01), and this decrease lasted for at least 9 months. Microdialysis of adipose tissue can be used for continuous long-term monitoring of glucose concentrations in diabetic patients during ordinary daily life. Daily glucose profiles are often reproducible and the recordings may thus be used for individual tailoring of insulin therapy to improve glycaemic control.

Aluminum (Al) mobilization by i.v. desferrioxamine (DFO) into blood and into brain and liver extracellular space was followed in the Al-loaded rat using microdialysis probes. Dialyzable extracellular liver Al peaked at an estimated 1360 microgram/l whereas dialyzable blood and extracellular brain Al peaked at 860 and 155-175 microgram/l, respectively. Estimated Al concentrations were derived from dialysate Al from microdialysis probes which was corrected for probe efficiency. Liver Al after DFO was generally greater than blood Al. Both declined over time. The rapid increase in extracellular liver Al above blood Al suggests the ability of DFO to rapidly distribute out of the vascular compartment to mobilize Al from hepatocytes. Presumably an Al-DFO complex was formed and released into blood, from which it was eliminated. Extracellular brain Al and DFO was less than blood Al and did not decline during 5 h. After an i.v. Al-DFO injection to non-Al-loaded rats, blood Al was greater than liver much greater than brain Al, suggesting Al-DFO diffusion down a concentration gradient and demonstrating the ability of Al-DFO to permeate vascular membranes and the blood-brain barrier. The lack of decline of brain extracellular Al after DFO was presumably due to its inability to diffuse into the blood against the concentration gradient. The DFO-induced mobilization of Al in the brain may explain the reports of sudden onset or worsening of encephalopathy associated with DFO treatment of Al-loaded humans.

Microdialysis was applied to estimate the pharmacologically active concentration of caffeine and theophylline in blood, adipose tissue, muscle, liver and brain of rats. The concentration of the drugs in the extracellular space was estimated by perfusion with varying concentrations of the drug through the microdialysis probe (difference method). Caffeine (20 mg/kg) was found to be evenly distributed with a free concentration of approximately 120 microM. Theophylline concentration in the brain was 91 microM and in other tissues approximately 120 microM. The rate of penetration into brain extracellular space was higher for caffeine than for theophylline. It is suggested that the lower levels of theophylline attained in the brain may to some extent explain the differences in clinical action profile between caffeine and theophylline.

Microdialysis was applied to estimate the extracellular concentration of caffeine in subcutaneous abdominal adipose tissue of five healthy volunteers after oral administration of approximately 5 mg/kg (300 or 400 mg) of caffeine. The peak extracellular levels were in the range of 20 - 80 microM. The time-course in blood and in extracellular fluid was similar but the plateau concentrations were not closely correlated. The estimated mean concentration of five individuals was similar in blood and extracellular fluid. The intraindividual variation between probes was found to be small compared to the interindividual variation (8% versus 43%). It is concluded that microdialysis yield useful data on drug distribution in man and that distribution to adipose tissue may not strictly follow the concentrations in blood. A comparison with available information of the in vitro properties of caffeine shows that the levels attained in the extracellular fluid were too small to significantly affect phosphodiesterase but sufficiently high to block adenosine receptors.

The present study was aimed at testing the hypothesis that yawning and suppression of exploration induced by low doses of dopamine agonists in the rat are caused by a reduction of synaptic dopamine levels. The decrease in extracellular levels of dopamine in the corpus striatum induced by alpha-methyl-p-tyrosine (alpha MPT, 50-200 mg/kg IP), reserpine (2-5 mg/kg SC) and apomorphine (APO, 0.05 mg/kg SC) was measured in microdialysis experiments. Reserpine and alpha MPT reduced the dopamine levels to the same extent as APO. Exploratory behaviour was suppressed by APO, but not by alpha MPT (50 and 100 mg/kg) when tested in a separate experiment. Reserpine (2 mg/kg) suppressed exploration after 4 hr, but not after 3 hr. Changes in extracellular levels of dopamine were tested simultaneously with changes in yawning in another group of rats implanted with guide cannulae for microdialysis probes. There was a discrepancy in the time-course for the induction of yawning as compared to the changes in extracellular dopamine levels after APO (0.05 mg/kg) as well as after pergolide (0.02 mg/kg SC). Yawning appeared before and lasted shorter than the decrease in dopamine. The time-courses for APO-induced suppression of exploration and yawning were similar. The dose-response curve for APO-induced yawning was not changed by alpha MPT (200 mg/kg), while the suppression of exploration induced by APO, but not by pergolide, was enhanced by pretreatment with alpha MPT. The results show that yawning and suppression of exploration induced by dopamine agonists are not related to changes in extracellular levels of dopamine. It is proposed that these behaviours may be mediated by postsynaptic receptors.

Microdialysis in the human brain has been performed for the first time during thalamotomy intended to relieve tremor in patients with Parkinson's disease. The aim was to test the reliability of the microdialysis technique for biochemical characterization of a target area in the human brain during a routine operation. Microdialysis probes were introduced through the same trajectory as the lesioning electrode thus causing no additional damage to the brain. Dopamine, DOPAC, HVA, 5-HIAA, hypoxanthine, inosine, guanosine, adenosine, GABA, taurine, aspartate and glutamate were measured in the perfusate from the target region - the Vim nucleus. The results show initial high levels that reach baseline levels after 10-20 minutes. Surprisingly, consistent and reproducible levels were found, the only exception being one patient on 1-DOPA therapy who had elevated DA and metabolite levels.

In response to the increasing demand for refined techniques to record drug induced changes in motor activity we have designed and evaluated against observations an automatic test box that quantifies eight defined components of behaviour in rats. Activity, corresponding to the recordings from the commonly used photocell activity boxes. Total, and forward locomotion, expressing the actual distance the rat walks. Corner count, and corner time, reflecting the position of the animal in the box. Hole count, and hole time, expressing the reaction of the rat to an environmental stimulus i.e. holes in the bottom of the test box. Gnawing, which is a direct counting of the number of gnaws made by the animal. The recording parameters relate to our interest in behaviour influenced by monoamine neurotransmission and the result shows that the selected parameters are recorded with high reliability.

The present study characterized the voltage and calcium dependence of gamma-aminobutyric acid and dopamine overflow after the acute implantation of a microdialysis probe. Probes were implanted in dorsolateral striatum and globus pallidus. Experiments were performed under light halothane anesthesia. Basal, extracellular levels of GABA were not affected by tetrodotoxin (TTX) and were increased to 140 percent of basal values by calcium free Ringer. Basal, extracellular levels of dopamine were reduced to 14 percent of basal values by the addition of TTX and to 30 percent of basal values by the removal of calcium from the Ringer solution. The results suggest that in this in vivo preparation basal extracellular dopamine is largely of vesicular origin while GABA is not.

The effect of an ibotenic acid lesion in rostral striatum on tissue levels of neurokinin A and substance P in striatum and substantia nigra was studied in rat brain. A total of 32 micrograms (10 mg/ml) ibotenic acid was injected in four positions to lesion striatal cell bodies rostral to bregma. The neurokinin A level was reduced to a third of the control value in striatum and to less than half of the control level in substantia nigra. Neurokinin A, in addition to substance P, is shown to be possibly present in striato-nigral neurons, which provides further evidence for the existence of a striato-nigral tachykinin pathway.

A simple and rapid screening method, where the ability of neuroleptic drugs to antagonise the abnormal pattern of exploration induced by a low dose of d-amphetamine in a 10 min test, was evaluated. The d-amphetamine 2 mg/kg pretreatment induced both an increased locomotion, thought to reflect an increased dopamine transmission in the nucleus accumbens, and weak stereotypies, thought to reflect an increased dopamine transmission in the neostriatum. Haloperidol, chlorpromazine and thioridazine blocked all ongoing behaviours while clozapine and sulpiride, regarded as causing less extrapyramidal side effects in the clinic, only antagonised the d-amphetamine induced locomotion. The findings support the notion that the common site of action for anti-psychotic drugs is blockade of dopamine receptors outside the neostriatum while the blockade of dopamine receptors within the striatum probably are related to the propensity of these drugs to induce the extrapyramidal side effects. It seems possible with this method to screen neuroleptic drugs for their relative potency in blocking limbic and striatal dopamine receptors simultaneously in one short experiment. The method might be used when new anti-psychotic drugs with low incidences of extrapyramidal side effects are sought for.

The release of the tachykinins neurokinin A (NKA) and neurokinin B (NKB) from superfused slices of rat brain was studied. For radioimmunoassay of superfusates and tissue extracts an antiserum which reacts with both NKA and NKB but not with substance P was used. The released immunoreactive material, as well as the immunoreactive material in extracts of the tissue slices, was characterized by cation exchange chromatography and reversed phase high performance liquid chromatography (reversed phase-HPLC). Potassium (50 mM) evoked a calcium-dependent release of tachykinin-like immunoreactivity from slices of frontal cortex and striatum. Reversed phase-HPLC accumbens, striatum and the ventral part of the mesencephalon revealed two major immunoreactive components which co-eluted with synthetic NKA and NKB. Furthermore, the superfusates also contained three minor unidentified immunoreactive components.

Apomorphine in low doses inhibits spontaneous exploratory behaviour in rats. This effect is commonly referred to as an expression of selective stimulation of dopaminergic autoreceptors. The aim of the present study was to investigate the influence of neuroleptic drugs with different pharmacological profiles on this apomorphine induced inhibition of exploration using techniques for detailed recording of behaviour and multivariate statistical analysis of the results. By comparison with dose response analyses of apomorphine it was possible to determine whether a neuroleptic specifically antagonised the apomorphine effect or if the pattern of behaviour was qualitatively changed in some way. Apomorphine (0.05 mg/kg) was tested against cis-flupenthixol (0.01-0.5 mg/kg), haloperidol (0.01-0.1 mg/kg), metoclopramide (0.2-5 mg-kg), sulpiride (0.5-50 mg/kg) and SCH 23390 (0.005-0.05 mg/kg). Metoclopramide and haloperidol had weak antagonising effects against apomorphine while cis-flupenthixol and SCH 23390 was completely inefficient in this respect. The multivariate analysis indicated that the effects of haloperidol was restricted to only some aspects of the behavioural effects of apomorphine. Only sulpiride did selectively and dose-dependently antagonise the apomorphine induced behavioural suppression. The data provide evidence for a functional subdivision of dopamine receptors at the behavioural level.

Microdialysis is a new technique to monitor levels of chemical compounds in the extracellular space over time. It involves the implantation of a microdialysis probe into the brain tissue. The probe is similar to a push-pull probe but the perfusate is contained inside a semi-permeable membrane located at the tip of the probe. Substances in the extracellular fluid will diffuse into the perfusate while substances included in the perfusate will diffuse into the tissue. This principle opens up a wealth of possibilities to monitor chemical events within the brain and to study the working mechanism of various drugs. The perfusate may be analysed by a number of different techniques. In this paper we give a short summary of various HPLC techniques that have proven particularly useful.

A miniaturized liquid chromatographic system with electrochemical detection (LC-ED) was developed and applied to the analysis of dopamine and its metabolites in dialysate samples collected from the rat brain in vivo. An existing LC-ED system was down-scaled using a 1 mm I.D. small-bore column operated at a reduced flow-rate and with an injection volume of 1 microliter. With the small-bore system the limit of detection for dopamine of ca. 0.06 pg was almost 50 times less than with the conventional system, which represents a two-fold improvement in concentration sensitivity. Miniaturization was accomplished with negligible loss in resolution by using a conventional commercial amperometric detector with minor modifications. The results indicate that a number of useful advantages could be realized by the combination of this small-bore LC-ED system and the in vivo brain dialysis method.

The effects of 10 days treatment with haloperidol or sulpiride on tissue levels of neurokinin A-like and substance P-like immunoreactivity (NKA-LI and SP-LI) in various regions of rat brain were studied using reversed phase HPLC and radioimmunoassay. The most marked effect was a decrease in NKA-LI levels in n.accumbens after treatment with both sulpiride and haloperidol. Striatal NKA-LI and SP-LI levels were not clearly affected. NKA-LI levels but not SP-LI levels were decreased in substantia nigra by haloperidol. A low dose of sulpiride increased both NKA-LI and SP-LI levels in ventral tegmental area/n.interpeduncularis. In conclusion, region-specific changes of NKA-LI and SP-LI were seen after subchronic treatment with neuroleptics. It seems likely that NKA and SP are involved in the neuronal adaptation to the repeated treatment with neuroleptics.

This paper describes a method for monitoring drug-induced changes in brain dopamine (DA) release and metabolism in the awake rat simultaneous to measurements of behavioural activation. Intracerebral dialysis was combined with a novel activity box based on a circular running track and a simple method for perfusate collection. "Locomotor" and "general" activity were automatically monitored using a system of photobeams lining the track, and stereotyped behaviours were scored by direct observation. Striatal perfusates were analysed for endogenous DA and its main metabolites, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), using HPLC with electrochemical detection. This methodology was tested by comparing the behavioural effects of systemically administered amphetamine (2 mg/kg SC) and its central DA releasing action. Amphetamine caused a marked increase in striatal DA release (16-fold) which was followed over the time course by increased locomotion and stereotyped sniffing and head and forepaw movements. The limitations and wider applications of the methodology are discussed.

Methylxanthines, such as caffeine and theophylline, potentiate the rotation behaviour induced by dopamine receptor agonists in rats with unilateral lesions of the nigro-striatal pathway. In the present study we have examined the possibility that interaction with central adenosine mechanisms could influence rotation behaviour. Under in vitro conditions adenosine and N6-phenylisopropyl-adenosine (PIA) stimulate cyclic AMP accumulation. This effect was enhanced by the phosphodiesterase inhibitor rolipram, but blocked by alkylxanthines such as caffeine, theophylline and, particularly, 8-phenyl-theophylline. Rotation behaviour induced by apomorphine (0.05 mg/kg), was inhibited by PIA and rolipram and by a low dose of the adenosine deaminase inhibitor EHNA (2 mg/kg). By contrast, theophylline and 8-phenyl-theophylline caused a potentiation. The former drug stimulated rotation behaviour per se, while the latter did not. 8-Phenyl-theophylline entered the brain poorly and its concentration in brain it was less than 1/10 of theophylline. It is concluded that theophylline does not potentiate rotation behaviour secondarily to inhibition of phosphodiesterase. Antagonism of endogenous adenosine may partly explain the effect of methylxanthines. Possibly, some as yet unknown mechanism may also contribute to the effects of xanthine-derivatives on rotation behaviour.

This short article reviews HPLC-EC methodology that we are currently applying to measure DA, 5-HT and their acid metabolites in rat brain dialysates collected in vivo. HPLC-EC systems based on standard-bore HPLC columns are described which are sufficiently sensitive to allow detection of the monoamine transmitters and their metabolites in regional brain dialysates collected at 10-20 min intervals. A large reduction in sample requirement was achieved by "down-scaling" the conventional HPLC-EC assay to incorporate small-bore HPLC columns. The small-bore systems allowed monoamines to be detected in samples collected over 1 to a few minutes.

Exploratory behaviour was recorded in an automatic holeboard apparatus measuring activity, locomotion, rearing, hole exploration and corner restricted behaviour. Multivariate statistical methods were used to analyse the results. Low doses of the dopamine agonist apomorphine (0.01-0.1 mg/kg SC) dose-dependently reduced most variables measured, although the pattern of behaviour resembled that of normal animals. High doses of apomorphine (0.2-1.0 mg/kg SC) induced a qualitative change in the pattern of behaviour with a stereotyped locomotion and a reduced mean time of exploration of holes. The described methods for detection and statistical analysis of behaviour differentiates between the behavioural patterns induced by high and low doses of apomorphine and may be useful in finding and analysing patterns of drug induced behaviour related to various mechanisms of action.

In vitro and in vivo techniques were used to examine the production and subsequent fate of the endogenous excitotoxin quinolinic acid (QUIN) following administration of its bioprecursor L-kynurenine (KYN). Incubation of liver slices in the presence of 10-1000 microM KYN resulted in a dose- and time-dependent release of QUIN into the incubation medium. Less than 15% of total QUIN produced was recovered from the tissue. In vivo experiments, performed with a microdialysis probe inserted in the jugular vein of anesthetized rats, showed that injection of KYN (20-600 mg/kg, i.v.) causes rapid and dose-dependent increases in the serum level of QUIN. Peak QUIN concentrations in serum dialysates were reached 75 minutes following KYN administration. Longer lasting increases were detected following the administration of pyrazinamide (20 mg/kg, i.p.), an indirectly acting stimulator of QUIN biosynthesis in the periphery. The data demonstrate the feasibility of assessing the mechanisms of QUIN production and disposition in experimental paradigms which can be expected to allow insights into the function and possible dysfunction of QUIN in the brain.

When injected into the substantia nigra reticulata of naive rats, dynorphin-(1-17) caused a dose dependent contralateral rotation which was inhibited by naloxone in a dose dependent manner. Met-enkephalin injected in the same coordinates as the ones used with dynorphin also induced rotation but of lower intensity, while Leu-enkephalin did not elicit any significant response at the doses tested. D-amphetamine, injected subcutaneously 60 min after dynorphin, significantly enhanced the contralateral rotation induced by the intranigral administration of dynorphin. Experiments with dynorphin injections combined with D-amphetamine suggest that the dynorphin effects are mediated by neuronal loops involving dynorphin dopamine interactions.

In vivo microdialysis was utilized to evaluate the role of extracellular Na+ in regulating dopamine (DA) neurotransmission in the caudate-putamen of halothane-anaesthetized rats. Reduction of the extracellular Na+ concentration by introduction of a perfusion media containing 50mM Na+ (with choline replacement) produced an excessive release of DA that could be effectively blocked by nomifensine and Lu 19-005, potent inhibitors of an amine transport carrier. These results substantiate reports of a carrier-mediated efflux of DA from presynaptic terminals. Pretreatment with amphetamine, considered both a DA uptake inhibitor and releaser, did not, however, influence the efflux of DA induced by the low extracellular Na+ environment. Although the release of DA from an apparent non-granular cytosolic pool was greatly enhanced by the low extracellular Na+ environment, 3,4-dihydrophenylacetic acid (DOPAC) levels, which supposedly reflect metabolism of non-vesicular DA, were minimally effected. In contrast, homovanillic acid (HVA) was sensitive to extracellular Na+ and not directly related to extracellular levels of either DA or DOPAC, suggesting the possibility of a Na+-sensitive (carrier-mediated?) process involved in the formation of HVA. Overall, the results of this paper cannot be completely reconciled with the traditional concept of intracellular organization of DA pools and suggests the possibility of various non-granular pools being differentially sensitive to efflux and metabolism.