Stephen N. Mitchell

Acute administration of nicotine increases the in vivo extracellular levels of dopamine, 3,4-dihydroxyphenylacetic acid and ascorbic acid preferentially in the nucleus accumbens of the rat: Comparison with caudate-putamen

Using in vivo dialysis and voltammetry, the effect of acute administration of (-)-nicotine (0.8 mg/kg free base, s.c.) on extracellular levels of dopamine, 3,4-dihydroxyphenylacetic acid, 5-hydroxy-indoleacetic acid and ascorbic acid in the nucleus accumbens and caudate-putamen of chloral hydrate-anaesthetised rats has been examined. Nicotine stimulated release of dopamine only in the nucleus accumbens, measured using dialysis. After a short time delay levels of 3,4-dihydroxyphenylacetic acid in both the nucleus accumbens and caudate-putamen also increased. In both regions, 5-hydroxyindoleacetic acid was unaffected by nicotine. Using voltammetry the effect of nicotine on extracellular levels of 3,4-dihydroxyphenylacetic acid and ascorbic acid was examined. An increase in 3,4-dihydroxyphenylacetic acid was observed in both regions after nicotine. This increase was blocked by pretreatment with the central nicotinic receptor antagonist mecamylamine (5 mg/kg). Nicotine increased the level of ascorbic acid in the nucleus accumbens and caudate-putamen; while in animals pretreated with mecamylamine, nicotine decreased levels of ascorbate. These results show that acute administration of nicotine stimulated release of dopamine in the nucleus accumbens and increased the levels of DOPAC and ascorbic acid in the nucleus accumbens and caudate-putamen. This effect is probably mediated by nicotinic receptors as it was antagonised by mecamylamine.

Effect of acute administration of nicotine on in vivo release of noradrenaline in the hippocampus of freely moving rats: A dose-response and antagonist study

The effect of systemic administration of (-)-nicotine on release of noradrenaline in the hippocampus was studied by in vivo microdialysis in freely moving rats, using dialysate containing nomifensine (5 microM). (-)-Nicotine, at both 0.4 and 0.8 mg/kg but not 0.2 mg/kg, rapidly and significantly increased extracellular levels of noradrenaline. Extracellular levels of dopamine were also increased, but this was only significant after the larger dose. Both 0.4 and 0.8 mg/kg also produced a significant increase in extracellular levels of the metabolites of dopamine, 3,4-dihydroxyphenylacetic acid and homovanillic acid. Extracellular levels of the metabolite of 5-hydroxytryptamine, 5-hydroxyindoleacetic acid, increased after 0.8 mg/kg but this effect was only apparent much later. Injection of a second 0.8 mg/kg challenge of (-)-nicotine, 150 min after the first, produced similar increases in extracellular levels of noradrenaline, dopamine, 3-4-dihydroxyphenylacetic acid and homovanillic acid. Over the experimental period, there was no further increase in extracellular levels of 5-hydroxyindoleacetic acid. Increases in extracellular levels of noradrenaline, dopamine, 3,4-dihydroxyphenylacetic acid and homovanillic acid, in response to 0.8 mg/kg (-)-nicotine, were prevented by the systemic administration of mecamylamine, but not hexamethonium (both at 5 mg/kg). Mecamylamine also inhibited the delayed increase in extracellular levels of 5-hydroxyindoleacetic acid, produced by the first injection of (-)-nicotine. These results suggest that (-)-nicotine, dose-dependently stimulated the release and metabolism of amine transmitters by an action at central nicotinic receptors. However, the precise site of action, i.e. at nerve terminals, cell bodies or both, requires further elucidation.

Effects of Chronic and Subchronic Nicotine on Tyrosine Hydroxylase Activity in Noradrenergic and Dopaminergic Neurones in the Rat Brain

: Chronic nicotine (0.8 mg/kg by daily subcutaneous injection) over a 7 to 28‐day period was found to increase the activity of tyrosine hydroxylase in predominantly noradrenergically innervated regions but not in dopaminergic projection areas. Increases in tyrosine hydroxylase activity were observed in dopaminergic cell body regions only after nicotine treatment for 3 to 5 days. The increase in tyrosine hydroxylase activity in noradrenergic neurones was evident first in the cell bodies in the locus coeruleus from 3 to 7 days, reaching 223% of control activities, and was followed by increases of up to 205% in the terminals up to 3 weeks later. It was then established that nicotine for 7 days was sufficient to increase the activity of the enzyme to the same extent in the terminals at 21 days even without further nicotine administration. This is consistent with axonal transport preceded by induction of the enzyme in noradrenergic cell bodies, whereas “delayed activation” might account for the transient effect seen in dopaminergic cell body regions. The response in the locus coeruleus to nicotine for 7 days was completely blocked by daily preinjection with mecamylamine but not with hexamethonium, which is consistent with the effect of nicotine on tyrosine hydroxylase being mediated by central nicotinic receptors.

Regionally specific effects of acute and chronic nicotine on rates of catecholamine and 5-hydroxytryptamine synthesis in rat brain

Acute (-)-nicotine administration (0.4 and 0.8 mg/kg s.c.) produced a regionally specific increase in the rate of catecholamine synthesis in the rat nucleus accumbens, hypothalamus and hippocampus but not elsewhere, including the caudate-putamen. In all regions rates of 5-hydroxytryptamine synthesis were unaffected. (-)-Cotinine (0.4 and 0.8 mg/kg), the major metabolite of (-)-nicotine was without effect. (-)-Nicotine-induced increase in catecholamine synthesis occurred by a direct stimulation of central nicotinic receptors, as mecamylamine (5 mg/kg) but not hexamethonium (5 mg/kg) was an effective antagonist. Following repeated daily injections of (-)-nicotine (0.8 mg/kg) for up to 28 days, the induced catecholamine response following a subsequent challenge was unaffected in the nucleus accumbens and hypothalamus, but was increased in the hippocampus. This effect persisted for up to 14 days following withdrawal. Rates of 5-hydroxytryptamine synthesis remained unaltered after chronic pretreatment.

5,7-Dihydroxytryptamine lesions in the fornix—fimbria attenuate latent inhibition

When animals are preexposed to a stimulus without consequence they are subsequently slower to associate this stimulus with an important event, such as footshock. This retarding effect of stimulus preexposure is called latent inhibition and can be demonstrated in a variety of classical and instrumental paradigms and in a wide range of species, including man. Latent inhibition is disrupted in acute schizophrenics and by amphetamine treatment in both rat and man. The present study investigated the role of hippocampal 5HT terminals in latent inhibition using a conditioned suppression procedure with male Sprague-Dawley rats. Microinjections of 5,7-dihydroxytryptamine in the fornix-fimbria significantly reduced hippocampal indoleamine levels and attenuated latent inhibition of conditioned suppression. This finding supports the hypothesis that the destruction of mesolimbic 5-hydroxytryptamine terminals reduces latent inhibition. This result is discussed in terms of the possible involvement of reduced serotonergic function in schizophrenic attentional disorder. In addition to the predicted lesion effect, biochemical analyses indicated that experimental treatments in the latent inhibition procedure altered neurotransmitter turnover: utilization ratios for 5-hydroxytryptamine and/or dopamine were increased in preexposed relative to nonpreexposed animals in four of the six brain regions sampled.

Investigation of the SSRI augmentation properties of 5-HT2 receptor antagonists using in vivo microdialysis

Recent evidence that 5-HT(2) receptors exert a negative influence on central 5-hydroxytryptamine (5-HT) neurones suggests that 5-HT(2) receptor antagonists may augment the effects of serotonin selective reuptake inhibitors (SSRIs). The present study investigated whether pre-treatment with 5-HT(2) receptor antagonists enhances the effect of SSRI administration on hippocampal extracellular 5-HT of freely moving rats. Administration of the SSRI citalopram at a low (2mg kg(-1)) and higher (4 mg kg(-1)) dose, increased dialysate 5-HT by 5- and 8-fold, respectively. Pre-treatment with the 5-HT(2) receptor antagonist ketanserin (4 mg kg(-1)) augmented the effect of 4 mg kg(-1) but not 2mg kg(-1) citalopram. The effect of 4 mg kg(-1) citalopram was also augmented by pre-treatment with either the 5-HT(2C) receptor antagonist SB 242084 (0.5mg kg(-1)) or the 5-HT(2A) receptor antagonist MDL 100907 (0.5mg kg(-1)). As with citalopram, fluoxetine elevated dialysate 5-HT at both a low (5mg kg(-1)) and higher (20mg kg(-1)) dose. However, neither dose of fluoxetine was augmented by ketanserin (4 mg kg(-1)). These results confirm recent findings that 5-HT(2) receptor antagonists augment the effect of citalopram on extracellular 5-HT, and indicate the involvement of 5-HT(2C) and possibly 5-HT(2A) receptors. The lack of augmentation of fluoxetine might reflect the intrinsic 5-HT(2) receptor antagonist properties of this drug.

Selective action of (−)-2-oxa-4-aminobicyclo[3.1.0]hexane-4,6-dicarboxylate (LY379268), a group II metabotropic glutamate receptor agonist, on basal and phencyclidine-induced dopamine release in the nucleus accumbens shell

The effect of the group II metabotropic receptor agonist (-)-2-oxa-4-aminobicyclo[3.1.0]hexane-4,6-dicarboxylate (LY379268), on basal and phencyclidine-induced dopamine efflux were measured in the shell and core subdivisions of the nucleus accumbens--regions which are associated with limbic and motor functions, respectively. Extracellular levels of dopamine were measured using microdialysis in conscious animals, and LY379268 was delivered locally by inclusion in the artificial cerebrospinal fluid (aCSF) flowing through the microdialysis probe. Local administration of LY379268 in the concentration range 10 nM-10 microM reduced basal levels of dopamine in the nucleus accumbens shell, whilst having no effect in the nucleus accumbens core. In the nucleus accumbens shell, basal levels were reduced to approximately 60% compared to the pre-injection control, with a maximal reduction occurring at concentrations of LY379268 > or =100 nM. The response to LY379268 (100 nM) was reversible, with levels returning to baseline following its removal from the aCSF. In a separate experiment, local perfusion of the nucleus accumbens shell with LY379268 (at both 1 and 10 microM) reduced the magnitude of the response to a subsequent systemic administration of phencyclidine (5 mg/kg i.p.). The reduction in the peak dopamine response was only evident with doses of LY379268 that also reduced basal dopamine efflux--LY379268 being ineffective against PCP at 10 nM. However, in animals pre-treated with LY379268 at 1 or 10 microM, PCP still evoked a dopamine response, and in these animals the relative extent of the response was not significantly different between the respective treatment groups. In contrast, in the nucleus accumbens core the magnitude of the dopamine response to PCP was unaffected by local application of LY379268 (at 1 or 10 microM). Our data suggest that within the nucleus accumbens, there exists a distinct regional difference in the control of dopamine release by group II mGluRs, with the nucleus accumbens shell being preferentially affected. Moreover, the selective action of LY379268 on dopamine levels in the nucleus accumbens shell may have implications for the potential antipsychotic activity of group II mGluR agonists.

Activation of the retrohippocampal region in the rat causes dopamine release in the nucleus accumbens: disruption by fornix section

There is a well-described projection from the retrohippocampus (subiculum and entorhinal cortex) to the nucleus accumbens that is involved in the control of psychomotor behaviour, and is implicated in the aetiology of schizophrenia. Cortical abnormalities are widely reported in the brains of schizophrenic patients, but it is unclear whether they are the cause or consequence of those changes in subcortical systems that are treated with neuroleptic drugs. We have, therefore, conducted a series of microdialysis experiments in anaesthetized rats to determine whether infusion of the excitotoxin, N-methyl-D-aspartate, into the retrohippocampus increases mesolimbic dopamine release. We found a clear and reproducible increase in extracellular dopamine in the nucleus accumbens following N-methyl-D-aspartate (2.5 microg), that was abolished when we sectioned the fimbria-fornix. Furthermore, inhibition of gamma-aminobutyric acid receptors following retrohippocampus administration of bicuculline (4 microg), also increased dopamine in the nucleus accumbens. The dopamine response to bicuculline was accompanied by an increase in dopamine metabolism, was long lasting, and also reduced by fornix section.The response to both N-methyl-D-aspartate and bicuculline depends on the integrity of the projection from the retrohippocampus to the nucleus accumbens. The results provide an underlying mechanism whereby a primary insult in the temporal cortex, caused by excessive N-methyl-D-aspartate receptor stimulation, can produce a hyperdopaminergic state. In addition, an increase in subiculo-accumbens activity evoked by bicuculline may also explain why patients with limbic epilepsy can develop a psychosis.

Are the cognitive-enhancing effects of nicotine in the rat with lesions to the forebrain cholinergic projection system mediated by an interaction with the noradrenergic system?

Experiments were conducted to test the hypothesis that the enhancing effect of nicotine on water maze performance in rats with lesions of the forebrain cholinergic projection systems (FCPS) is mediated by an interaction with the noradrenergic system, in particular the ascending dorsal noradrenergic bundle (DNAB) and its projection areas. Three groups of rats received lesions of either: i) the nucleus basalis (NBM) and medial septal area/diagonal band (MSA/DB) by infusion of alpha-amino-3-hydroxy-4-izoxazole propionic acid (AMPA) (FCPS group), ii) DNAB, by infusion of 6-hydroxydopamine (6-OHDA) (NOR group), or iii) both FCPS plus DNAB (COMB group). Control animals received vehicle. Choline acetyltransferase activity was reduced in the cortex and hippocampus of the FCPS and COMB groups and in the hippocampus of the NOR group. NA level was reduced in the cortex and hippocampus of the FCPS and COMB groups, but not the FCPS group. In a reference memory task, the performance of both the NOR and COMB groups, but not the NOR group, was significantly worse than that of controls; there was no effect of nicotine administration (0.1 mg/kg) on escape latency or other measures in this task. In a working memory task, FCPS and COMB rats took longer to find the submerged platform on the second and following trials, and there was a significant enhancement of performance by nicotine in both groups, but not in controls. These results indicate that the enhancing effects of nicotine in rats with FCPS lesions are not mediated by an interaction with the DNAB.

Temporally distinct cognitive effects following acute administration of ketamine and phencyclidine in the rat

Non-competitive N-methyl-D-aspartate receptor (NMDAR) antagonists such as phencyclidine (PCP) and ketamine are commonly and interchangeably used to model aspects of schizophrenia in animals. We compared here the effects of acute administration of these compounds over a range of pre-treatment times in tests of instrumental responding (VI 30s response schedule), simple reaction time (SRT) and cognitive flexibility (reversal learning and attentional set shifting digging task) in rats. At standard pre-treatment times (15-30 min), both ketamine and PCP produced overall response suppression in VI 30 and increased reaction times in SRT suggesting that any concomitant cognitive performance deficits are likely to be confounded by motor and/or motivational changes. However, the use of extended pre-treatment times produced deficits in cognitive flexibility measured up to 4h after drug administration in the absence of motor/motivational impairment. Generally, PCP increased impulsive responding in the SRT indicating a possible loss of inhibitory response control that may have contributed to deficits observed in reversal learning and attentional set-shifting. In contrast to PCP, ketamine did not have the same effect on impulsive responding, and possibly as a consequence produced more subtle cognitive deficits in attentional set-shifting. In summary, acute treatment with NMDAR antagonists can produce cognitive deficits in rodents that are relevant to schizophrenia, provided that motor and/or motivational effects are allowed to dissipate. The use of longer pre-treatment times than commonly employed might be advantageous. Also, ketamine, which is more frequently used in clinical settings, did not produce as extensive cognitive deficits as PCP.