Christopher Pycock

Effect of 6-Hydroxydopamine Lesions of the Medial Prefrontal Cortex on Neurotransmitter Systems in Subcortical Sites in the Rat

Abstract: The effect of lesions of the catecholamine nerve terminals in the medial prefrontal cortex of the rat on neurotransmitter mechanisms within the basal ganglia has been investigated. Bilateral 6‐hydroxydopamine lesions were stereotaxically placed in the dopamine‐rich (DA) area of the frontal cortex. Animals were pretreated with desmethylimipramine to block the uptake of neurotoxin into noradrenergic (NA) terminals and to make it more selective for DA terminals. The lesion produced a selective reduction of both NA and DA from the medial prefrontal cortex, a result related to falls in tyrosine hydroxylase activity at this site. Lesioned animals showed enhanced DA turnover and utilisation in striatal and limbic regions. There was no change in subcortical tyrosine hydroxylase activity. In addition there were significant falls in other putative neurotransmitters within basal ganglia sites, including 5‐hydroxytryptamine and GABA. Decreased activity of the neurotransmitter‐synthesizing enzymes glutamate decarboxylase and choline acetyltransferase was also recorded in certain regions of the basal ganglia. The results suggest that frontal cortical catecholamine systems may serve to regulate various neurotransmitter mechanisms in the basal ganglia.

Behavioural and biochemical effects of dopamine and noradrenaline depletion within the medial prefrontal cortex of the rat

The effects of 6-hydroxydopamine (6-OHDA) lesions of catecholamine terminals within the medial prefrontal cortex on spontaneous motor activity, dopamine (DA)-dependent stereotyped behaviour and subcortical dopamine turnover were investigated in the rat. Two types of lesions were examined, bilateral injection of 6-OHDA into the medial prefrontal cortex of untreated rats (6-OHDA alone), and bilateral injection of 6-OHDA into the medial prefrontal cortex of animals pretreated with the noradrenaline (NA) uptake blocking agent desmethylimipramine (6-OHDA/-DMI). Ten days after surgery the 6-OHDA lesions produced no significant change in spontaneous motor activity and had no overall effects on stereotyped behaviour induced by apomorphine or (+)-amphetamine. This lesion caused gross depletion of NA within the medial prefrontal cortex and curiously, elevated DA concentrations within this site. No changes in DA concentration were recorded within subcortical sites, although concentrations of DA metabolites within striatum and nucleus accumbens were reduced. In contrast, the 6-OHDA/DMI lesion of the medial prefrontal cortex significantly enhanced spontaneous motor activity and amphetamine-induced stereotyped behaviour. Apomorphine-induced stereotypy, on the other hand, was significantly reduced. Biochemically the lesion caused a large depletion of DA with relatively little loss of NA within the medial prefrontal cortex. In addition, from this and another study (ref. 33), increases in DA and its metabolite concentrations were measured in striatum and nucleus accumbens, together with an apparent increase in DA turnover within these subcortical sites. It is thus apparent that in the absence of a substantial portion of the DA innervation of the medial prefrontal cortex, with a largely intact NA innervation, there is an increase in motor activity and amphetamine-induced stereotypy which may be related to functional changes in DA activity within subcortical telencephalic structures. Such a finding might suggest that DA within the frontal cortex has a behaviourally inhibitory role in the rat, although further work is required to substantiate this.

Specific stimulating effect of glycine on 3H-dopamine efflux from substantia nigra slices of the rat.

The effect of glycine was studied on the efflux of tritium-labelled dopamine, 5-HT and GABA from small slices of rat substantia nigra in vitro. A depolarising stimulus (50 mM KCl) stimulated the efflux of 3H-5HT, 3H-dopamine and 3H-GABA in a calcium-dependent fashion. Glycine (50 and 100 micromoles) stimulated the spontaneous efflux of 3H-dopamine but not that of 3H-5HT or 3H-GABA. Taurine, GABA and beta-alanine, all at 100 micromoles, had no effect on release of radioactivity after prelabelling nigral slices with 3H-dopamine. In keeping with a transmitter role for glycine at this site, 3H-glycine was taken up by small slices of rat substantia nigra by both high affinity (Km = 2.4 micromoles) and low affinity systems (Km = 5.96 mM). However 50 mM KCl was without effect on the efflux of radioactivity from nigral slices prelabelled with 3H-glycine.

Behavioural and biochemical changes following chronic administration of L-DOPA to rats

Rats were given powdered diet containing L-DOPA (together with the peripheral decarboxylase inhibitor carbidopa) for a period of 6 months. The estimated daily intake was in the range 20-30 mg/kg. Initially, at 1 week and 1 month, L-DOPA-fed rats exhibited enhanced spontaneous locomotor activity, but this fell to within the control range by 3 and 6 months, although (+)-amphetamine-induced hyperactivity was greater at 6 months in L-DOPA-treated animals than in control rats. Six months after receiving L-DOPA in their diet rats showed enhanced stereotypy scores to a series of dopamine agonists administered acutely including (+)-amphetamine, nomifensine, L-DOPA, apomorphine and piribedil compared with the control animals. In another behaviour test L-DOPA administration reduced the cataleptic potency of both fluphenazine and haloperidol was increased. Biochemically 6 months treatment of rats with L-DOPA was associated with significantly increased plasma concentrations of L-DOPA, enhanced striatal levels of L-DOPA, dopamine and dopamine metabolites, enhanced specific binding (as indicated by increased Bmax values) of [3H] spiroperidol, [3H] ADTN and [3H] 5-HT to striatal membranes, and increased basal and dopamine-stimulated striatal adenylate cyclase activity. The results are discussed in the light of changes of sensitivity of cerebral dopamine receptors, an increase in receptor numbers, and the tolerance to L-DOPA which often develop in the treatment of Parkinsons disease.

Potassium-Stimulated Release of Radiolabelled Taurine and Glycine from the Isolated Rat Retina

Abstract: The release of preloaded [3H]glycine and [3H]taurine in response to a depolarising stimulus (12.5‐50 mM KCl) has been studied in the superfused rat retina. High external potassium concentration immediately increased the spontaneous efflux of [3H]glycine, the effect of 50 mM K+ apparently being abolished by omitting calcium from the superfusing medium. In contrast, although high potassium concentrations increased the spontaneous emux of [3H]taurine from the superfused rat retina, this release was not evident until the depolarising stimulus was removed from the superfusing medium. The magnitude of this “late” release of [3H]taurine was dependent on external K+ concentrations, and appeared immediately after cessation of the stimulus irrespective of whether it was applied for 4, 8, or 12 min. Potassium (50 mM)‐induced release of taurine appeared partially calcium‐dependent, being significantly reduced (p < 0.01) but not abolished by replacing calcium with 1 mM EDTA in the superfusate. High‐affinity uptake systems for both [3H]glycine and [3H]taurine were demonstrated in the rat retina in vitro (Km values, 1.67 μM and 2.97 μM; Vmax values, 19.3 and 23.1 nmol/g wet weight tissue/h, respectively). The results are discussed with respect to the possible neuro‐transmitter roles of both amino acids in the rat retina.

Differential actions of diazepam on the release of [3H]5-hydroxytryptamine from cortical and midbrain raphe slices in the rat

The possible interaction of the benzodiazepine diazepam with synaptic mechanisms of the central neurotransmitter 5-hydroxytryptamine (5-HT) has been studied in the rat using the superfusion in vitro release technique. The effect of diazepam (1-100 microM) on the spontaneous and potassium-induced release of radioactivity from superfused slices preloaded with [3H]5-HT prepared from cerebral cortex (fronto-parietal) and midbrain raphe region was monitored and compared. The results demonstrate a differential action of diazepam on the release of cortical and raphe 5-HT. In cerebral cortex diazepam significantly reduces both spontaneous and potassium-evoked release of [3H]5-HT. Conversely in raphe slices diazepam significantly enhances both spontaneous and potassium-evoked release of [3H]5-HT. In both tissues the effect of diazepam on K+-stimulated neurotransmitter release was abolished in the presence of picrotoxin. The experiments suggest that the effects of benzodiazepines on 5-HT mechanisms may be manifested at least in part through gamma-aminobutyric acid receptors.

Effects of ω-amino acids on tritiated dopamine release from rat striatum: Evidence for a possible glycinergic mechanism

Abstract The effects of GABA and glycine on the release of tritiated dopamine from prelabelled slices of rat striatum have been compared. Both GABA (>50 μM ) and glycine (>200 μM ) released tritiated dopamine, but had no effect on the release of radiolabelled 5-hydroxytryptamine and GABA. The GABA antagonist picrotoxin (50 μM) markedly reduced the ability of GABA to release [ 3 H]dopamine, but had no effect on the glycine response. Conversely strychnine (0.5 μM), a specific glycine receptor antagonist at low concentrations, abolished both the GABA and the glycine response on [ 3 H]dopamine release. Two other ω-amino acids, β-alanine and taurine, both at 500 μM, had no effect on [ 3 H]dopamine release from rat striatal slices. In additional experiments, release of radioactivity was demonstrated from neonatal rat spinal cord and striatal slices after prelabelling with [ 3 H]glycine. This release was calcium-dependent. The possibility that glycine may function as a neurotransmitter substance within the striatum is considered, and the hypothesis that GABA may partially exert some of its pharmacological effects through the glycine receptor is discussed.

Regional changes in the concentrations of cerebral monoamines and their metabolites after ethanolamine-O-sulphate induced elevation of brain γ-aminobutyric acid concentrations

Abstract The effect of ethanolamine- O -sulphate induced elevation of cerebral GABA concentrations on monoamine and their metabolites levels has been studied in various regions of the rat brain. Increased GABA concentrations were associated with a decrease in turnover of dopamine in limbic regions: striatal dopamine was not significantly affected. An increased turnover of 5-hydroxytryptamine was also observed in other brain areas. Increased cerebral GABA concentrations had no effect on regional noradrenaline turnover. The possible sites of interaction between the neurotransmitters are discussed.

Evidence that thalamic efferent neurones are non-cholinergic: a study in the rat with special reference to the thalamostriatal pathway

Controversy surrounds the question as to whether some fibres of the thalamostriatal projection, are cholinergic. The present experiments show that lesions of the parafascicular-intralaminar thalamus produced no reductions in choline acetyltransferase (ChAT) activities in any area of microdissected rat caudate-putamen complex or dorsolateral frontal cortex. We conclude that thalamostriatal projections are entirely non-cholinergic. Furthermore, lesions of the mediodorsal or periventricular thalamus resulted in no change in ChAT activities in their terminal projection areas (medio-/orbitofrontal cortices and nucleus accumbens, respectively). The probability that all thalamic outputs are non-cholinergic is discussed.

Lesions of the superior colliculus in the rat differentiate between nigrostriatal and mesolimbic dopamine systems

Bilateral electrolytic lesions of the superior colliculus in rats increased spontaneous locomotor activity, enhanced amphetamine-induced hyperactivity and attenuated apomorphine-induced biting. These lesions were associated with an increased rate of turnover of dopamine in the nucleus accumbens, but not in the striatum. Similarly concentrations of the dopamine metabolites homovanillic acid and 3,4-dihydroxyphenylacetic acid were elevated in accumbens tissue but not in striatum in rats with bilateral collicular lesions. The results indicate that lesions of the superior colliculus cause differentiation between hyperactivity and stereotypy, and that this may be related to blockade of a nigrostriatal outflow, and relief of inhibition on mesolimbic systems.