Sprouting of striatal serotonin nerve terminals following selective lesions of nigro-striatal dopamine neurons in neonatal rat

The effects of neonatal intracisternal 6-hydroxydopamine (6-OHDA; 50 micrograms) treatment on striatal serotonin (5-HT) nerve terminals in rat have been characterized using histo- and neurochemical methods. The 6-OHDA lesion caused a 60% reduction of striatal dopamine (DA) concentration when analyzed in the adult stage, while 5-HT levels were increased by about 40% and 3H-5-HT uptake in vitro was increased by about 60%. Using computerized image analysis, a marked increase in 5-HT-like immunoreactive terminal density was found in both rostral (+200%) and caudal (+50%) striatum. Pretreatment with the DA uptake blocker amfolenic acid completely counteracted the 6-OHDA-induced alterations in both DA and 5-HT neurons in the striatum, while pretreatment with the noradrenaline uptake blocker desipramine had no significant effects. Regional analysis of 5-HT levels in the CNS after neonatal 6-OHDA treatment or the combined desipramine + 6-OHDA treatment showed no significant effect in any of the brain areas analyzed, apart from the observed 5-HT increase in striatum. It was furthermore observed that the striatal 5-hydroxyindoleacetic acid (5-HIAA)/5-HT ratio was decreased, while the 3,4-dihydroxyphenylacetic acid (DOPAC)/DA ratio was increased following the 6-OHDA lesion, indicating compensatory mechanisms in turnover of transmitters. These alterations were completely reversed after pretreatment with amfolenic acid. The present results support the view that the 5-HT hyperinnervation following neonatal 6-OHDA treatment is a collateral sprouting response induced by lesioning of the striatal DA neurons.     

GBR12909 stimulates hypothalamo-pituitary-adrenal activity by inhibition of uptake at hypothalamic dopamine neurons

We have previously demonstrated that the inhibition of neurotransmitter uptake at dopamine (DA) terminals stimulates the hypothalamo-pituitary-adrenal axis. In the present study we investigated the role of central DA neuronal systems in the regulation of this axis. Administration of the DA uptake inhibitor GBR12909 (3–30 μg) into the third ventricle dose-dependently elevated serum adrenocorticotropin hormone (ACTH) levels in rats. GBR12909 (10 μg) elevated serum ACTH levels following administration into the paraventricular nucleus of the hypothalamus but not into the lateral ventricle. The administration of 6-OHDA into the third ventricle significantly decreased DA content in the hypothalamus and striatum and significantly attenuated the ACTH response to GBR12909 (10 μg, third ventricle or 10 mg/kg, i.p.). Conversely, 6-OHDA lesions of the medial forebrain bundle, which depleted 99% of DA in the caudate but did not decrease DA content in the hypothalamus, and did not attenuate the ACTH response to i.p. GBR12909. Measurement of GBR12909-induced inhibition of [³H]DA uptake into synaptosomal preparations indicates the presence of GBR12909-sensitive DA transporters in the region of the hypothalamus surrounding the thrid ventricle. The present findings suggest that an endogenous DA neuronal system terminating in the hypothalamus mediates the effects of stimulants on hypothalamo-pituitary-adrenal function and might play a role in the ongoing regulation of hypothalamo-pituitary-adrenal activity.     

Neonatal 6-hydroxydopamine-induced hypo/hyperactivity: blockade by dopamine reuptake inhibitors and effect of acute D-amphetamine

Five experiments were performed to assess the changes in motor activity resulting from neonatal administration of 6-hydroxydopamine (6-OHDA) on Days 1 or 2 postnatal, at doses of either 75 or 100 μg in a volume of 10 μl vehicle, following pretreatment with either GBR 12909 (40 mg/kg, s.c.) or amphonelic acid (4.0 mg/kg, s.c.) or saline. Motor activity was measured either over 60-min test periods on five consecutive days of testing or at 12-min intervals within a single 60-min test session. The initial extent of locomotor hyperactivity was dependent upon the neonatal dose of 6-OHDA: the 100 μg, but not 75 μg, dose induced marked hyperactivity from test day 1 onwards whereas the 75 μg dose did so from test day 3 onwards. The initial hypoactivity for rearing behaviour was observed for both doses of 6-OHDA: this hypoactivity was altered over successive test days so that by test day 5 an hyperactivity by the 75 μg, but not 100 μg, was observed. Pretreatment with either GBR 12909 or amphonelic acid abolished the effects of both doses of 6-OHDA. In the within 60-min test session procedure, 6-OHDA treated rats (both 75 and 100 μg) showed initial hyperactivity for locomotion that intensified, in relation to the other groups, over each 12-min interval and initial hypoactivity for rearing that developed into hyperactivity over each 12-min interval. Pretreatment with either GBR 12909 or amphonelic acid again abolished the effects of both doses of 6-OHDA (75 and 100 μg) rats, compared to the control groups in all four experiments. In Experiment V, a low dose of D-amphetamine abolished the hyperactivity of 6-OHDA (75μg) treated rats whereas a higher dose did so only transiently. Pretreatment with GBR 12909 abolished these effects. These findings underline the neuropharmacological utility of the neonatal 6-OHDA treatment for studying brain receptor system adaptive changes underlying the respective functional alterations and as a possible laboratory model for clinical disorders.     

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 increased 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 innvervation, there is an increase in motor activity and amphetamine-induced stereotypy which may be related to functional changes in DA activity within subcortical telecephalic 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.     

Contribution of Blockade of the Noradrenaline Carrier to the Increase of Extracellular Dopamine in the Rat Prefrontal Cortex by Amphetamine and Cocaine

This study was performed to investigate the relative role of noradrenaline (NA) and dopamine (DA) carrier blockade in the effects of psychostimulants on DA transmission in the rat prefrontal cortex (PFCX). To this end, changes of extracellular DA and NA in the PFCX and of extracellular DA in the nucleus accumbens (NAc) were measured following the administration of amphetamine and cocaine, which are known to bind to both DA and NA carriers, or GBR 12909, a selective DA carrier blocker. After non-intravenous injection, amphetamine (0.25 and 0.5 mg/kg, s.c.) and cocaine (5 and 10 mg/kg, i.p.) increased extracellular DA in the PFCX to a larger extent than in the NAc, while the reverse applied to GBR 12909 (2.5 and 5 mg/kg, i.p.). These differences were obtained in spite of the fact that the three drugs elicited at each dose level a similar peak increase of extracellular DA in the NAc. Amphetamine and cocaine also increased extracellular NA in the PFCX and this effect was quantitatively similar to that on extracellular DA in the same area. Intravenous doses of cocaine and GBR 12909, corresponding to those which maintain self-administration in the rat, while equieffective in raising extracellular DA in the NAG, had different effects on extracellular DA in the PFCX. In fact, in contrast to cocaine, GBR 12909 increased extracellular DA in the PFCX to a lesser extent than in the NAc or did not modify it at all. The peak increase of extracellular DA in the PFCX was highly correlated to that of NA in the same area but was poorly correlated to the increase of extracellular DA in the NAc. These results suggest that amphetamine and cocaine increase extracellular DA in the PFCX largely through the blockade of the NA carrier. Direct evidence for this hypothesis was provided by the observation that, when the NA carrier was blocked by reverse dialysis of the PFCX with desipramine (1 μM), cocaine and GBR 12909 lost their differences in the ability to increase extracellular DA in the PFCX.     

Suppression of noradrenergic innervation compensates for behavioral deficits induced by lesion of dopaminergic terminals in the lateral septum

Spontaneous alternation which is disrupted by lesion of septal dopaminergic (DA) afferents was chosen as a behavioral marker for the study of functional interactions between DA and noradrenergic (NA) innervation of the lateral septum. Three groups of rats were studied: a solvent group which received only vehicle injection, and two lesioned groups, one with DA lesion and the second with simultaneous DA + NA lesion of the septal innervation. DA lesion was produced by infusing 6-hydroxydopamine (6-OHDA) into the lateral septum after pretreatment with desmethylimipramine (DMI) injected intraperitoneally. The DA + NA lesion was produced by infusing 6-OHDA without DMI pretreatment. The lesion of DA innervation alone led to a disturbance of alternation behavior in a Y-maze, but performance was not affected by the combined DA + NA lesion. The group with septal DA lesion was then injected with 6-OHDA into the pedunculus cerebellaris superior (PCS) in order to destroy NA efferents from the locus coeruleus. The two other groups were sham-operated. After post-operative recovery, the rats were retested for spontaneous alternation. The rats with the PCS NA lesion subsequent to the DA septal lesion displayed normal alternation behavior. Their performance was not different from that of animals with both NA and DA lesions in the septum. Thus the NA lesion appears to prevent the alternation deficits induced by the DA septal lesion, and also abolishes the deficits induced by the prior DA lesions. These results may have therapeutic implications.     

Functional recovery after destruction of dopamine systems in the nucleus accumbens of rats. II. Facilitation by the ACTH-(4-9) analog ORG 2766

Functional recovery from motor hypoactivity of rats with 6-OHDA lesions in the nucleus accumbens is accelerated by intra-accumbal or subcutaneous treatment with the ACTH-(4-9) analog ORG 2766. The spontaneous recovery period of 3 weeks is shortened to 7 days by daily treatment with this peptide during the first 6 days after the lesion. The 6-OHDA lesion induced a decrease of about 30-40% in the levels of dopamine, HVA and DOPAC as well as in the uptake of [3H]dopamine in nucleus accumbens tissue in vitro. Treatment with ORG 2766 during the first 6 days following the lesion did not affect the lesion-induced changes in these biochemical parameters. Binding studies with [3H]haloperidol in nucleus accumbens tissue of placebo or ORG 2766-treated sham-lesioned rats revealed a linear Scatchard plot 7 days after the sham lesion. In tissue of placebo-treated 6-OHDA lesioned animals a similar linear Scatchard plot was found but in tissue of ORG 2766-treated 6-OHDA-lesioned rats the Scatchard plot was curvilinear in shape indicating two types of binding sites. In the 6-OHDA-lesioned rats treated with ORG 2766 the behavioral response upon apomorphine challenge was enhanced suggesting the existence of functional supersensitivity of the DA system. Similar changes in Scatchard plots and apomorphine-induced behavioral changes have been previously reported after spontaneous recovery. The present study indicates that ORG 2766 accelerates the process of functional recovery from impaired motor behavior of rats with 6-OHDA lesions in the nucleus accumbens, which may be due to development of denervation supersensitivity.     

Cross-sensitization between cocaine and GBR 12909, a dopamine uptake inhibitor.

In the present experiment, rats which had received repeated injections of 20 mg/kg GBR 12909, a selective dopamine uptake inhibitor, were subjected to challenge doses of GBR 12909 and cocaine which were near or below the threshold for locomotor activation in drug-naive animals. The behavior measured was general motor activity, monitored in photocell testing cages. Results verify earlier findings which showed that rats treated chronically with GBR 12909 display a potentiated locomotor response to low doses of the drug (behavioral sensitization). In addition, it was found that GBR 12909-sensitized rats show a potentiated locomotor response to 5 mg/kg cocaine (cross-sensitization). These results are discussed with regard to various mechanisms which may underlie the sensitization phenomena observed, including possible neurochemical changes at the presynaptic level.     

rAAV-mediated nigral human parkin over-expression partially ameliorates motor deficits via enhanced dopamine neurotransmission in a rat model of Parkinson's disease

We hypothesized that over-expressing the E3 ligase, parkin, whose functional loss leads to Parkinson's disease, in the nigrostriatal tract might be protective in the unilateral 6-hydroxydopamine (6-OHDA) rat lesion model. Recombinant adeno-associated virus (rAAV) encoding human parkin or green fluorescent protein (GFP) was injected into the rat substantia nigra 6 weeks prior to a four-site striatal 6-OHDA lesion. Vector-mediated parkin over-expression significantly ameliorated motor deficits as measured by amphetamine-induced rotational behavior and spontaneous behavior in the cylinder test but forelimb akinesia as assessed by the stepping test was unaffected. rAAV-mediated human parkin was expressed in the nigrostriatal tract, the substantia pars reticulata, and the subthalamic nucleus. However, in lesioned animals, there was no difference between nigral parkin and GFP-transduction on lesion-induced striatal tyrosine hydroxylase (TH) innervation or nigral TH positive surviving neurons. A second lesion experiment was performed to determine if striatal dopamine (DA) neurotransmission was enhanced as measured biochemically. In this second group of parkin and GFP treated rats, behavioral improvement was again observed. In addition, striatal TH and DA levels were slightly increased in the parkin-transduced group. In a third experiment, we evaluated parkin and GFP transduced rats 6 weeks after vector injection without DA depletion. When challenged with amphetamine, parkin treated rats tended to display asymmetries biased away from the treated hemisphere. Nigral parkin over-expression induced increases in both striatal TH and DA levels. Therefore, while parkin over-expression exerted no protective effect on the nigrostriatal DA system, parkin appeared to enhance the efficiency of nigrostriatal DA transmission in intact nigral DA neurons likely due to the observed increases in TH.     

Amygdalar noradrenergic and dopaminergic mechanisms in the regulation of hunger and thirst-motivated behavior

The feeding behavior of rats was studied after neurochemical damage of the amygdalar terminal fields of mesolimbic dopaminergic (DA) and coerular noradrenergic (NA) pathways. 6-Hydroxydopamine (6-OHDA) or 6-hydroxydopa (6-OHDOPA) were injected bilaterally into the central part of amygdala. 6-OHDA was also injected after desmethylimipramine (DMI) pretreatment in order to study the selective destruction of DA terminals. The body weight increased after 6-OHDA injection and a mild hyperphagia and hyperdipsia developed. The 6-OHDA plus DMI treatment resulted in body weight decrease, hypophagia and hypodipsia. These effects were dose-dependent. While a high dose of 6-OHDOPA (15 mug/mul) decreased the body weight, an increase of weight was observed after a low dose (4 mug/0.5 mul). After 6-OHDA, 6-OHDA plus DMI or the high dose of 6-OHDOPA the DA concentration dropped significantly in the amygdala while low-dose 6-OHDOPA resulted in DA increase. In every case there was a parallel change in striatal DA content. The amygdalar NA concentration decreased after both 6-OHDA and the high dose of 6-OHDOPA. There was no change in NA levels after 6-OHDA plus DMI treatment and the NA concentration increased after the injection of a low dose of 6-OHDOPA. When DA/NA ratio was calculated the results showed that body weight increases were accompanied by a relative deficit in NA while a relative deficit of DA was present if body weight decreased. Our results suggest that the amygdalar balance of these transmitters may play an important role in the regulation of body weight and the contradictions of results with electrolytic lesions in the amygdala can be resolved at transmitter level.     

GBR12909 attenuates amphetamine-induced striatal dopamine release as measured by [(11)C]raclopride continuous infusion PET scans.

Major neurochemical effects of methamphetamine include release of dopamine (DA), serotonin (5-HT), and norepinephrine (NE) via a carrier-mediated exchange mechanism. Preclinical research supports the hypothesis that elevations of mesolimbic DA mediate the addictive and reinforcing effects of methamphetamine and amphetamine. This hypothesis has not been adequately tested in humans. Previous in vivo rodent microdialysis demonstrated that the high affinity DA uptake inhibitor, GBR12909, attenuates cocaine- and amphetamine-induced increases in mesolimbic DA. The present study determined the ability of GBR12909 to attenuate amphetamine-induced increases in striatal DA as measured by [(11)C]raclopride continuous infusion positron emission tomography (PET) scans in two Papio anubis baboons. [(11)C]Raclopride was given in a continuous infusion paradigm resulting in a flat volume of distribution vs. time for up to 45 min postinjection. At that time, a 1.5 mg/kg amphetamine i.v. bolus was administered which caused a significant (30.3%) reduction in the volume of distribution (V(3)"). The percent reduction in the volume of distribution and, hence, a measure of the intrasynaptic DA release ranged between 22-41%. GBR12909 (1 mg/kg, slow i.v. infusion) was administered 90 min before the administration of the radiotracer. The comparison of the volume of distribution before and after administration of GBR12909 showed that GBR12909 inhibited amphetamine-induced DA release by 74%. These experiments suggest that GBR12909 is an important prototypical medication to test the hypothesis that stimulant-induced euphoria is mediated by DA and, if the DA hypothesis is correct, a potential treatment agent for cocaine and methamphetamine abuse. Furthermore, this quantitative approach demonstrates a way of testing various treatment medications, including other forms of GBR12909 such as a decanoate derivative.     

Real-time monitoring of endogenous noradrenaline release in rat brain slices using fast cyclic voltammetry: 1. Characterisation of evoked noradrenaline efflux and uptake from nerve terminals in the bed nucleus of stria terminalis, pars ventralis

Fast cyclic voltammetry (FCV) at carbon fibre microelectrodes was used to monitor endogenous noradrenaline (NA) efflux in superfused slices of bed nucleus of stria terminalis pars ventralis (BSTV) in 'real time'. NA efflux was evoked by local electrical stimulation at bipolar tungsten stimulating electrodes. Confirmation of the identity of the released species as NA was made on the basis of anatomical, electrochemical and pharmacological proofs. Firstly, the signal matched the NA innervation density; efflux of monoamine was greater in BSTV than in the pars dorsalis of the nucleus. Secondly, the voltammogram of the released species was indistinguishable from those of the catecholamines NA and dopamine (DA) but dissimilar to that of the indoleamine serotonin (5-hydroxytryptamine, 5-HT). Thirdly, amine efflux was influenced in a predictable fashion by the drugs tested. Tetrodotoxin (10(-6) M) or omission of Ca2+ from the superfusate reversibly reduced amine efflux by 90.2 and 88.0% respectively. Ro 4-1284 (10(-6) M) decreased amine efflux by 75.8%. Desipramine (5 x 10(-8) M), the selective NA uptake blocker, significantly increased amine efflux and uptake half-life (to 214.3 and 389.5% of control respectively). Fluvoxamine (5 x 10(-7) M) and GBR 12909 (3 x 10(-7) M), blockers of 5-HT and DA uptake respectively, had no effect on amine efflux, although fluvoxamine caused a modest (91.0%) increase in the uptake half-life. Pargyline (2 x 10(-6) M) affected neither efflux nor uptake. The combined anatomical, electrochemical and pharmacological data confirm that the monoamine detected in BSTV by local electrical stimulation was NA. Stimulated NA efflux was stable and reproducible over at least 2.5 h (longest period tested). This study demonstrates the ability of FCV to selectively monitor endogenous NA efflux and uptake in 'real time' and with high spatial resolution.     

Specificity of dopaminergic neuronal degeneration induced by intracerebral injection of 6-hydroxydopamine in the nigrostriatal dopamine system.

The neurotoxic specificity of injections of 6-hydroxydopamine (6-OHDA) into areas containing either dopamine (DA) cell bodies (substantia nigra) or DA axon terminals (striatum) was studied. This selective effect was compared to the unspecific effects of copper sulfate (CuSO4) injection and electrocoagulation. One to two days after unilateral nigral injection of 2 mug of either 6-OHDA or CuSO4 into the nigra the volume of the unspecific lesions around the tip of the cannula was very similar. Only the 6-OHDA-induced lesions were associated with elective degeneration of the nigral DA neurons. Ten days after the administration of the same compounds the gliosis in the substantia nigra was much more extensive in CuSO4-than in 6-OHDA-treated rats; however, the reduction of DA concentrations in the ipsilateral striatum was only noticeable after 6-OHDA (-62%). A somewhat similar decrease of striatal DA levels (-52%) was observed after large electrocoagulation of the substantia nigra. Ten days after 6-OHDA (8mug) or electrolytic lesion of the striatum the Km for DA, serotonin and choline uptakes were similar in the striata of both sides, suggesting that the uptake process in the non-damaged neurons of the lesioned side was functionally normal. Following electrolytic lesion of the striatum, serotonin and choline Vmax values were decreased to about the same extent as the striatal reduction in weight and DA levels. When directly administered into the striatum 6-OHDA also produced a decline in DA concentration and Vmax but in contrast did not affect serotonin and choline uptake (Vmax), suggesting that the drug specifically destroyed dopaminergic neurons. The present data confirm that selective DA denervation can be achieved when appropriate amounts of the drug are injected into brain tissue in order to limit the unspecific lesion.     

Role of ventral mesencephalic reticular formation and related noradrenergic and serotonergic bundles in turning behaviour as investigated by means of kainate, 6-hydroxydopamine and 5,7-Dihydroxytryptamine lesions

A unilateral kainate (KA) infusion (2 × 0.15 μg, 2 × 0.25 μg) in the ventral mesencephalic reticular formation (MRF) resulted in spontaneous contraversive turning lasting only a few days. Upon challenge with apomorphine (0.5 mg/kg s.c) or amphetamine (5 mg/kg i.p.) the contraversive turning could be reinstated. The incidence, as well as the intensity, of the drug-induced response decreased over the 45 days of observation. KA infused in the ventral MRF induced typical lesions after doses of2 × 0.15 μg but resulted in demyelination after2 × 0.25 μg. These lesions failed to reduce noradrenaline (NA), serotonin (5 HT) or dopamine (DA) in various forebrain areas. Unilateral lesion of ascending NA projections by 6-OHDA infusion (4 μg) within the NA bundles coursing in the mesencephalon or near the locus coeruleus, failed to induce motor asymmetries. Unilateral selective lesion of the ventral NA bundle by local 6-OHDA (2 μg) infusion also failed to induce motor asymmetries, either spontaneously or in response to dopaminergic drugs. Unilateral lesion of ascending 5-HT projections by the tegmental infusion of 5,7-dihydroxytryptamine (10 μg) also failed to induce motor asymmetries in response to dopaminergic drugs but resulted in contraversive circling in response to 5-hydroxytryptophan. These data indicate that intrinsic neurones of the ventral MRF play a role in turning behaviour and exclude, in contrast with previous studies, a role of NA or 5-HT projections in the contraversive turning responses to DA receptor agonists obtained after lesions of the ventral MRF.     

Chronic cocaine increases kappa-opioid receptor density: lack of effect by selective dopamine uptake inhibitors

Continuous infusion of cocaine or the selective dopamine uptake inhibitors GBR 12909 or RTI-117 increases locomotor stimulation, to which partial tolerance occurs. In addition, all three drugs produce significant decreases in tyrosine hydroxylase immunoreactivity in caudate putamen and nucleus accumbens core, suggesting a decreased dopaminergic tone. An interaction between cocaine and opioids has long been documented. Chronic cocaine significantly increases mu and kappa-opioid receptors and treatment with a kappa-opioid agonist markedly reduces the behavioral effects of cocaine. In addition, chronic cocaine, but not GBR 12909, increases prodynorphin gene expression in caudate putamen. To further understand the interaction between cocaine and the kappa-opioid system, the effects of a chronic continuous infusion for 14 days of cocaine or one of the selective dopamine uptake inhibitors GBR 12909 or RTI-117 via osmotic minipump were examined on kappa-opioid receptors using the selective kappa-opioid ligand [3H] U-69593. [3H] U-69593 binding density was significantly increased in caudate putamen, nucleus accumbens shell, claustrum, and endopiriform nucleus after cocaine, while neither GBR 12909 nor RTI-117 had any effect. The increased kappa-opioid receptor densities observed following cocaine are likely not related to dopamine uptake inhibition, since they were not produced by selective dopamine uptake inhibitors. These findings suggest that regulation of kappa-opioid receptors by cocaine may be via inhibition of serotonin or norepinephrine uptake, by a combination of effects on two or three monoamine transporters, or by a mechanism unrelated to transporter inhibition.     

Dopamine D1 receptor behavioral responsitivity following selective lesions of the striatal patch compartment during development.

The behavioral effects of selective destruction of the dopamine (DA) input to the patch compartment of rat striatum early in development was investigated. Rat pups were given bilateral intrastriatal (i.s.) injections of the neurotoxin 6-hydroxydopamine (6-OHDA) on day of birth (P0) or postnatal day 1 (P1), which resulted in selective behavioral alterations following DA agonist treatment in adulthood. Neonatally-lesioned rats exhibited self-biting behavior following treatment with the DA precursor L-dihydroxyphenylalanine (L-DOPA). In response to treatment with the selective D1 agonist SKF38393, there was an increased incidence of abnormal perioral movements. The cataleptogenic effects of the D1 antagonist SCH23390 and the D2 antagonist haloperidol were also studied. Neonatally-lesioned rats were significantly less cataleptic compared to control rats following D1 antagonist treatment, but not following D2 antagonist treatment. Autoradiographs of [3H]mazindol binding to DA uptake sites (a measure of DA terminal density) showed a 'patchy' loss of approx. 40-50% in striatal tissue sections derived from the i.s. lesioned rats. These data suggest that injections of 6-OHDA into the striatum during this early postnatal period cause a DA lesion that results in long-term effects on a D1 receptor system.     

GBR-12909-induced self-injurious behavior: role of dopamine

A regimen of repeated administration of GBR (10 or 20 mg/kg/day, i.p., for 4 days) to female Sprague-Dawley rats induced a dose-and time-related increase in the incidence of self-injurious behavior (SIB) that consisted of injury to body areas, paws and tail. The treatment regimen decreased striatal DA and DOPAC levels. Dopaminergic denervation with 6-hydroxydopamine (6-OHDA) or D1 DA antagonist, SCH-23390 or D2 DA antagonist, spiperone, blocked the GBR-induced SIB. Male rats were less sensitive than female rats to exhibit a comparable incidence of SIB. Taken together, the study reveals that repeated administration of GBR induces SIB that is dependent on the integrity of nigrostriatal dopaminergic system and the presence of D1 and/or D2 DA receptors.     

Intracranial self-stimulation from the sulcal prefrontal cortex in the rat: The effect of 6-hydroxydopamine or kainic acid lesions at the site of stimulation

An electrode cannula system was used to elicit intracranial self-stimulation (ICSS) from the sulcal prefrontal cortex in rats to test the behavioral effects of local infusions of 6-hydroxydopamine (6-OHDA) or kainic acid (KA) into the brain area surrounding the electrode tip. In experimental I sulcal ICSS animals received injections of 6-OHDA with or without desipramine (DMI) pretreatment to block 6-OHDA uptake into noradrenergic (NA) terminals. Those animals that received DMI pretreatment were subsequently shown to have sustained sulcal cortical dopaminergic (DA) denervation while sulcal molecular layer NA systems were spared as revealed with glyoxylic acid-induced catecholamine histofluorescence. Those animals not receiving DMI pretreatment sustained near-complete denervation of both NA and DA sulcal cortical systems. Neither treatment had a lasting effect on sulcal ICSS suggesting that sulcal ICSS is not dependent on the presynaptic release or DA of NA into that brain area.In experiment II KA injections that lesioned neurons in sulcal cortical layers V and VI resulted in the abolition of sulcal ICSS for the duration of a 21 day postlesion trial period. These results suggest that activation of a descending corticofugal system originating in the sulcal cortex is responsible for the mediation of sulcal prefrontal cortical ICSS. This system was mapped by the selective silver impregnation of degenerating neural elements resulting from effective lesions.     

Effects of 6-hydroxydopamine on central noradrenaline neurons during ontogeny.

The effects of the catecholamine neurotoxic compound, 6-hydroxydopamine (6-OHDA) have been investigated on central noradrenaline (NA) neurons after neonatal administration. In agreement with previous studies this treatment (1-3 X 100 mg/kg) led to a pronounced reduction of the in vitro uptake of [3H]NA and the endogenous NA in the cerebral cortex, while these parameters were markedly augmented in the pons and medulla oblongata, regions containing the NA perikarya. The 6-OHDA induced changes in the cerebral cortex and the pons-medulla could be completely prevented by the 'membrane pump' blocker desipramine, indicating that the effects are associated with a specific neurotoxic action of 6-OHDA on the NA neurons. Consistently, 6-OHDA acutely (within 2 h) produced a marked reduction of the [3H]NA uptake in both the cerebral cortex and pons-medulla. In the cerebral cortex the nadir (approximately 75% reduction) was reached within 6 h and remained so, while in the pons-medulla the [3H]NA uptake rapidly recovered, being maximally elevated after 14 days (50-80% increase) and remained so for at least 6 months. The [3H]NA uptake in the pons-medulla from 6-OHDA treated rats had the same kinetic and pharmacological properties as that of control. Thus the observed differences in [3H]NA uptake are most likely quantitatively related to actual changes in the number of NA nerve terminals. Treatment with lower 6-OHDA doses (10 or 50 mg/kg) resulted in less pronounced reduction of [3H]NA uptake initially, and there was a gradual recovery of tuptake with time in the cerebral cortex, which was more pronounced after the lower dose. These results are indications of regenerative growth, which may be possible when a critical part of the axon is spared from the neurotoxic effect of 6-OHDA. Administration of 6-OHDA on various days after birth disclosed that both the reduction of [3H]NA uptake in the cerebral cortex and the increase of [3H]NA uptake in the pons-medulla did not appear as permanent phenomena when 6-OHDA was given later than on the seventh postnatal day. This is most likely associated with the postnatal development of the blood-brain barrier. It may be concluded that the neonatal 6-OHDA treatment causes a marked NA denervation in the forebrain, e.g. the cerebral cortex, and an increased outgrowth of NA nerve terminals in the pons-medulla, which is preceded by a partial damage. This partial NA denervation is then followed by a regeneration (regenerative and/or collateral sprouting) and a stimulated outgrowth of NA nerve terminals.     

Comparative analysis of the effects of iontophoretically applied dopamine in different regions of the rat brain, with special reference to the cingulate cortex.

A systematic comparison of the effects of iontophoresed dopamine (DA) was carried out in the neostriatum (NS), nucleus accumbens (Acb) and anterior cingulate (ACg), prefrontal (PF) and parietal (Par) cortex of urethane-anesthetized rats, before and after treatment with the specific DA uptake blockers GBR 12909 and Bupropion. Similar experiments were also conducted after DA denervation with 6-hydroxydopamine and after DA depletion with alpha-methyl-p-tyrosine. The average rate of spontaneous neuronal firing was comparable in all regions, except in the NS after DA depletion. A majority of the units were inhibited by DA in every region and condition tested. As assessed with the IT50 index, the responsiveness to DA was not markedly different between regions, indicating that the postsynaptic sensitivity to this amine is independent of the density of DA receptors and of DA innervation. In contrast, the average duration of DA inhibitions (RT90) was considerably longer (5-fold) in the intact ACg than in the PF, Par, NS, or Acb. Moreover, treatment with both DA uptake blockers reduced the duration of DA inhibitions in ACg (4- to 9-fold); while lengthening it in PF, NS and Acb; and having no apparent effect in Par. DA depletion and DA denervation also reduced the duration of the DA inhibitions in ACg without effect in Par. Taken together, these results provide further evidence for the existence of a presynaptic, positive-feedback mechanism in ACg, triggered by DA, and favouring the further release of this transmitter upon its reuptake in DA nerve terminals.