Partial protection by desmethylimipramine of the mesocortical dopamine neurones from the neurotoxic effect of 6-hydroxydopamine injected in ventral mesencephalic tegmentum. The role of noradrenergic innervation

As shown in the rat by estimation of dopamine (DA) and noradrenaline (NA) levels, bilateral 6-hydroxydopamine (6-OHDA, 4 micrograms/microliter) lesions made in the ventral mesencephalic tegmentum (VMT) destroy both ascending DA and NA neurones. Pretreatment of rats with desmethylimipramine (DMI, 30 mg/kg, i.p.), 30 min before microinjection of 6-OHDA into the VMT partially prevented the destruction of the DA neurones innervating the prefrontal and cingulate cortices but not those innervating subcortical structures (nucleus accumbens, olfactory tubercles, septum). Results obtained from the prefrontal cortex of rats with extensive lesions of the ascending NA neurones performed 15 days prior to the 6-OHDA lesions of the VMT in the presence of DMI, imply that NA innervation of the VMT seems to be required for DMI to protect the cortical DA neurons from the neurotoxic effect of 6-OHDA.     

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.     

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.     

Body weight changes after neurochemical manipulations of lateral amygdala: Noradrenergic and dopaminergic mechanisms

The feeding behavior of rats was studied after neurochemical lesions of lateral amygdalar terminal fields of mesolimbic dopaminergic (DA) and coerular noradrenergic (NA) systems. 6-Hydroxydopamine (6-OHDA) with or without desmethylimipramine (DMI) premedication was bilaterally injected into the lateral amygdala or periamygdaloid-piriform area. Lateral amygdalar injections of 6-OHDA resulted in hyperdipsia, hyperphagia and body weight increase with a relative decrease in amygdalar NA concentration, while 6-OHDA plus DMI treatments were followed by weight decrease and a mild decrease in DA level. When the cannulas were placed into the periamygdaloid-piriform cortex nothing but body weight increase developed. The importance of the actual balance of the two amygdalar catecholaminergic (CA) systems in feeding is discussed.     

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.     

Selective lesioning of forebrain noradrenaline neurons at birth abolishes the improved maze learning performance induced by rearing in complex environment

The effect of selective destruction of forebrain noradrenaline (NA) neurons induced by 6-hydroxydopamine (6-OHDA) at Day 1 after birth on Hebb-Williams maze performance was investigated in adult rats housed after weaning in a complex environment (EC) or an isolated (IC) environment for 35 days. Saline treated control rats raised in the EC made fewer errors than those raised in the IC. This effect of EC was completely abolished in 6-OHDA treated rats; for these animals no improved performance due to the housing condition was obtained. Protection of the NA neurons against 6-OHDA neurotoxicity by pretreatment with desipramine (DMI) resulted in an effect of EC identical to that seen in saline-treated controls. Postweaning housing in the IC led to an increased locomotion as compared to housing in EC, but this effect was not affected by neonatal 6-OHDA and/or DMI treatment. Neurochemical analysis confirmed cortical NA and metabolite depletion as well as a good protection by the DMI pretreatment. The present results indicate that central NA neurons are involved critically in mediating mainly the cognitive components of behavioral alterations induced by EC.     

Desmethylimipramine promotes recovery of self-stimulation from the prefrontal cortex following footshock.

Intracranial self-stimulation (ICSS) was assessed from the prefrontal cortex in CD-1 mice immediately (0 h), 24 h and 168 h following exposure to uncontrollable footshock. Marked reductions in ICSS rates were observed in all mice immediately following the stressor. Although the ICSS alterations were transient in some animals, ICSS rates were reduced in the majority of animals 24-h and 168-h poststressor. Mice of either the shock or no shock treatment groups were administered either saline or desmethylimipramine (DMI, 5 mg/kg x 2) for 20 consecutive days. Chronic DMI ameliorated the stressor-induced ICSS deficits from the prefrontal cortex. Potential explanations for the stressor-provoked variations in ICSS and the effects of DMI are discussed.     

Dopamine depletion in the medial prefrontal cortex induces sensitized-like behavioral and neurochemical responses to cocaine

It has been postulated that behavioral sensitization to cocaine is associated with an attenuation of cocaine-induced dopamine (DA) transmission in the medial prefrontal cortex (mPFC). Hence, experiments were designed to examine the effects of chemically-induced cortical DA depletion on the acute behavioral and neurochemical responses to cocaine. One week following two bilateral 6-hydroxydopamine (6-OHDA) injections into the mPFC, animals received injections of cocaine (7.5, 15 or 30 mg/kg, i.p.) or saline (1 ml/kg, i.p.) in a randomized fashion with a minimum 3 day intertrial interval. Cocaine produced a dose-dependent increase in motor activity which was significantly enhanced in animals depleted (mean of 76%) of dopamine in the mPFC. Likewise, 6-OHDA lesions of the mPFC produced a significant enhancement of cocaine-induced DA transmission in the nucleus accumbens (NAC) as estimated by in vivo microdialysis. These data indicate a permissive involvement of cortical DA in mediating behavioral and neurochemical responses to cocaine, as well as confirm the ability of the mPFC to influence subcortical structures in response to an acute injection of cocaine. Collectively, the present findings suggest that alterations in cortical DA transmission may be a neural substrate mediating the development of sensitization to cocaine, and thus, may contribute to the addictive properties of cocaine.     

Effects of unilateral microinjections of sulpiride into the medial prefrontal cortex on circling behavior of rats

1. Bilateral 6-OHDA lesions of rats' medial prefrontal cortex increased locomotor activity after 7–10 days suggesting that cortical DA may normally inhibit motor behaviour. However, hyperactivity may have resulted from enhanced subcortical DA function.

2. Acute manipulation of frontal cortical DA neurotransmission in the present experiment avoided lesion-induced subcortical changes.

3. Sulpiride (0, 6, 12, 24 ug in 1 ul) was Injected unilaterallv into the medial prefrontal cortex of rats pretreated with (+)-amphetamine (1.5 mg/kg i.p.).

4. Circling behavior was scored during four 5-min intervals of a 60-min test session which began with injections and placement in a flat, circular arena.

5. SUL resulted in ipslversive circling whereas its vehicle did not. These results were consistent with those seen with other DA drugs and suggest an excitatory influence of frontal cortical DA on locomotor activity.     

Nondopaminergic prefrontocortical efferent fibers modulate D1 receptor denervation supersensitivity in specific regions of the rat striatum

A unilateral injection of 6-OHDA (6 microgram/1.5 microliter) was made into the fields of Forel in order to estimate the effects of the destruction of ascending dopaminergic (DA) pathways on the denervation supersensitivity of DA D1 receptors in the rat striatum. DA-sensitive adenylate cyclase activity was markedly enhanced in the anteromedian part of the striatum 3 weeks after the lesion (+68%) and remained elevated for several weeks thereafter (+36%). A different response occurred in the laterodorsal striatum, where the increase in DA-sensitive adenylate cyclase activity was less pronounced after 3 weeks (+40%) and no longer present after 7 weeks. Estimations of catecholamine levels indicated that the lesion made destroyed not only nigrostriatal DA neurons but other ascending catecholaminergic fibers projecting into the cerebral cortex as well. In addition, retrograde transport experiments made with wheat germ agglutinin coupled to horseradish peroxidase indicated that the anteromedian part of the striatum, but not the laterodorsal one, receives both an ipsi- and contralateral cortical projection originating in the prefrontocortical DA field. When the destruction by 6-OHDA of this contralateral DA innervation was combined to the unilateral lesion of the fields of Forel, the increase in DA-sensitive adenylate cyclase activity in each striatal area 3 or 7 weeks postlesion was prevented. This effect was due to DA denervation of the prefrontal cortex since striatal D1 denervation supersensitivity was still observed when contralateral ascending noradrenergic fibers were selectively destroyed by a 6-OHDA lesion made laterally to the pedunculus cerebellaris superior. These results suggest that, by controlling the activity of corticostriatal neurons, the mesocorticoprefrontal DA neurons exert a permissive role on the development of D1-receptor denervation supersensitivity in specific areas of the striatum.     

The origin and distribution of dopamine-containing afferents to the rat frontal cortex

The present study investigated distribution of the dopaminergic afferents to the rat frontal cortex by microdissection of the prefrontal and cingulate cortex areas and layers. Radiochemical assays for DA, NA and 5-HT revealed that, of these three amines, only DA was unevenly distributed in the frontal cortex, being concentrated especially in the medial prefrontal cortex and in the cingulate cortex. Parallel determinations of [³HDA uptake and tyrosine hydroxylase (TOH) showed that DA uptake and TOH activity were also concentrated in the DA-rich areas. Unilateral injections of 6-OHDA into the ascending noradrenergic bundle resulted in a dramatic fall (> 90%) in cortical noradrenaline. However, in the DA-rich areas, (i.e. areas containing 40–100 ng/g DA), the DA content was increased, whereas the DA content of areas with only low amounts of DA (10–25 ng/g DA) decreased. Lesions of the mesencephalic DA cells resulted in loss of the DA content of the DA-rich areas, whereas control lesions in the medial dorsal thalamic nucleus had no effect on any of the neurotransmitters or enzymes studied. Lesions confined to the ventral tegmental area (A10) cell group depleted the medial prefrontal cortex of DA, but only partially depleted the cingulate cortex. Lesions of the substantia nigra (A9 cell group) indicated that the more superficial layers (I–IV) of the cingulate cortex were partially depleted of DA, but the lower layers (IV–VI) were not affected. These results indicate that the dopaminergic afferents to the frontal cortex originate from the dopaminergic cell bodies in the A9 and A10 nuclei. The A9 cell group innervates the superficial layers of the cingulate cortex. The A10 cell group innervates the deeper layers of the medial prefrontal cortex and also innervates the lower layers of the cingulate cortex.     

Mesolimbicocortical dopamine terminal fields are necessary for normal locomotor and investigatory exploration in rats

Rats explore a novel open field or novel object less after denervation of mesolimbicocortical dopaminergic terminal fields produced by bilateral 6-hydroxy-dopamine (6-OHDA) microinjections into the anterolateral hypothalamus after pretreatment with desmethylimipramine (DMI). These behavioral deficits were correlated with complete or nearly complete loss of fluorescent dopaminergic (DA) terminals in the nucleus accumbens, olfactory tubercle, dorsal bed nucleus of the stria terminalis, lateral septal nucleus and the deep layers of the frontal and piriform cortices. There were also fewer A10, medial A9, and A8 DA fluorescent cells after the 6-OHDA-DMI injections; this suggests retrograde degeneration of the cells of origin of the mesolimbicocortical DA system. When the DMI pretreatment was omitted, identical bilateral 6-OHDA microinjections also produced severe loss of norepinephrine (NE) fibers in the neocortex, hippocampus, lateral hypothalamus and ventral bed nucleus of the stria terminalis. The addition of this noradrenergic damage did not change the exploratory deficits observed after mesolimbicocortical DA denervation alone.Systemic administration of the DA agonist apomorphine, but not the adrenergic agonist clonidine, to the 6-OHDA-DMI rats repaired the deficits in exploration of a novel open field or novel object. The increased locomotion in a novel open field and investigation of a novel object produced by apomorphine in 6-OHDA-DMI rats were blocked by the DA antagonist, pimozide. This is evidence that apomorphine restored exploratory responses by stimulating dopaminergic receptors. The exploratory responses produced by apomorphine were also blocked by testing rats in a familiar open field or with a familiar novel object. This is evidence that apomorphine facilitates exploratory responding to novel stimuli by 6-OHDA-DMI rats, but that the same dose of apomorphine does not increase activity when 6-OHDA-DMI rats are confronted by familiar stimuli.We conclude: (1) that mesolimbicocortical dopaminergic terminals are necessary for normal exploratory behavior in rats; and (2) that DA released by these terminals may facilitate optimal sensorimotor integration in these terminal fields during spontaneous exploratory behavior.     

Magnitude and duration of hyperactivity following neonatal 6-hydroxydopamine is related to the extent of brain dopamine depletion

This experiment examined the relationship between the extent of brain dopamine (DA) neuron destruction in the neonatal rat and locomotor hyperactivity during subsequent development. Brain DA neurons were destroyed selectively in neonatal rats by intraventricular injections of 6-hydroxydopamine (6-OHDA) following desmethylimipramine (DMI) pretreatment of both days 3 and 6 of life. Groups of rats received total doses of 50, 70, 100 or 200 microgram of 6HDA or the vehicle solution. Each group of rats given 6-OHDA displayed 3- to 5-fold increases in locomotor activity relative to vehicle control rats on days 16 and 18 of life. Rats given 50 or 70 microgram of 6-OHDA displayed hyperactivity that diminished during days 18-32 of life, approaching the level of activity seen in vehicle-treated rats. It contrast, rats given 100 or 200 microgram of 6-OHDA displayed consistently high levels of locomotion during days 18-32 of life. When tested as adults (days 55-66 of life) only those rats given 200 micrograms of 6-OHDA as neonates continued to display locomotor hyperactivity. The extent of 6-OHDA-induced depletion of DA was proportional to the magnitude of locomotor hyperactivity seen during neonatal life. Brain DA was depleted to the greatest extent in rats which displayed permanent hyperactivity. Regardless of the extent of depletion of brain DA, adult rats given intraventricular injections of 125, 200 or 275 micrograms of 6-OHDA at 48 days of age (following pargyline and DMI pretreatment) displayed no significant change in locomotor activity. These results indicate that the magnitude and duration of locomotor hyperactivity seen following neonatal 6-OHDA injections are correlated with the extent of loss of central DA neurons and suggest that brain DA projections exert important influences on the ontogeny of normal locomotion.     

6-OHDA lesions of the ventral tegmental area block morphine-induced but not amphetamine-induced facilitation of self-stimulation

The biphasic effect of morphine on intracranial self-stimulation (ICSS) (suppression followed by facilitation) was examined in rats following injections of 6-OHDA or vehicle into the ventral tegmental area (VTA). During 7 days of chronic administration of morphine, sham-lesioned animals gradually developed tolerance to the rate-reducing effects of the drug and a concurrent sensitization to its rate-enhancing effects (measured 1 and 3 h post-injection, respectively). VTA lesioned rats showed neither tolerance to the rate suppression nor any facilitation of ICSS throughout testing. Amphetamine's facilitation of ICSS remained intact in all subjects. The lesion was restricted to the VTA cell bodies but produced a significant depletion of dopamine (DA) in both the nucleus accumbens and in the striatum. Morphine facilitation of ICSS is suggested to be dependent on an indirect opiate receptor-VTA DA cell interaction. It is also proposed that amphetamine facilitation of ICSS is a 'mass action' effect involving the full DA terminal area of the forebrain rather than a subregion of that area.     

Distribution of tritium label in the neonate rat brain following intracisternal or subcutaneous administration of [3H]6-OHDA. An autoradiographic study

The present report describes the distribution of tritium label after injection of newborn rats with [3H]6-hydroxydopamine ([3H]6-OHDA). The animals were injected either intracisternally (i.c.) or subcutaneously (s.c.), with or without pretreatment with nomifensine, which blocks the high-affinity uptake of both noradrenaline (NA) and dopamine (DA), and sacrificed at intervals from 40 min to 24 h post-injection (p.i.). In i.c. injected animals, tritium label is demonstrable as early as 40 min p.i. in neurons of all known NA and DA cell groups. In NA neurons, it is taken up into cell body, dendrites, preterminal and terminal axons. The intensity of neuronal labeling is highest within the first 4 h p.i. and decreases in most neurons with longer postinjection intervals. A significant proportion of both NA and DA neurons degenerate beginning 6 h p.i., the majority show morphological signs of the axon reaction 24 h p.i. Uptake of [3H]6-OHDA into serotonergic and non-catecholaminergic neurons is not demonstrable. [3H]6-OHDA is accumulated by the following extraneuronal cells of the CNS: ependymal cells, epithelial cells of the choroid plexus, subependymal macrophages, smooth muscle cells in the wall of large intraparenchymal blood vessels, meningeal cells and glial cells. The time course of accumulation and disappearance of the label varies among these extraneuronal elements. The meningeal cells show the highest labeling intensity and degenerate within 24 h p.i. After pretreatment of the animals with nomifensine, the uptake of [3H]6-OHDA into NA and DA neurons is totally blocked; by contrast uptake of the labeled drug into extraneuronal cells is not prevented. These findings show that [3H]6-OHDA is not only accumulated by neurons possessing the high-affinity uptake for NA or DA, but by numerous other, extraneuronal cells which also participate in the metabolism of catecholamines.     

Decreased norepinephrine in the ventral lamina terminalis region is associated with angiotensin II drinking response deficits following local 6-hydroxydopamine injections

Previous studies have demonstrated that punctate injections of 6-hydroxydopamine (6-OHDA) into structures along the ventral lamina terminalis reduce drinking responses elicited by angiotensin II (ANG II). The purpose of the present study was to extend these findings by quantifying the catecholamine depletions that are associated with the drinking response deficits. Male Sprague-Dawley rats were given injections of 6-OHDA into the median preoptic nucleus (MnPO) and the organum vasculosum of the lamina terminalis (OVLT) after pretreatment with pargyline. Controls were injected with vehicle and a third group received 6-OHDA after pretreatment with desmethylimipramine (DMI). All subjects were tested for drinking responses to centrally injected ANG II and carbachol. Changes in catecholamine content of the MnPO, OVLT, supraoptic nucleus, paraventricular nucleus, caudate and cortex were determined using HPLC with electrochemical detection after behavioral testing. Only rats injected with 6-OHDA without DMI pretreatment showed a significant reduction in ANG II-induced drinking responses. This behavioral deficit was associated with a significant norepinephrine decrease in the ventral lamina terminalis region. All 3 groups showed comparable drinking responses to carbachol. These data support the hypothesis that noradrenergic innervation of the ventral lamina terminalis region is necessary for ANG II-induced drinking responses.     

Dopamine and skilled limb use in the rat: more severe bilateral impairments follow substantia nigra than sensorimotor cortex 6-hydroxydopamine injection.

The experiments examined the suggestion that the dopaminergic (DA) projection to the motor cortex are involved in the motor impairments that follow complete hemitelencephalic DA depletions. The neurotoxin, 6-hydroxydopamine (6-OHDA), was injected unilaterally into the sensorimotor cortex (MCtx), the ventral tegmental area (VTA), or into the substantia nigra pars compacta (SN) of rats trained to reach for food with either forelimb. The SN injections produced large (greater than 95%) unilateral striatal dopamine (DA) depletions and severe bilateral impairments in limb use. VTA and MCtx injections did not produce impairments in limb use or severe depletions of cortical DA. An effective test of the contribution of cortical DA to skilled limb use must await a more effective technique for producing selective cortical DA depletion. Nevertheless, the results suggest that the severe impairments of skilled forelimb use that follow hemitelencephalic DA depletions may stem primarily from depletion of the nigrostriatal DA projection.     

Prefrontal cortex and neostriatum self-stimulation in the rat: Differential effects produced by apomorphine

In a dose-response experiment, the effects of intraperitoneal injections of the dopamine receptor agonist, apomorphine (0.075, 0.15, 0.3, 0.6 and 1.2 mg/kg) were studied on self-stimulation elicited from electrodes implanted in the medial and sulcal prefrontal cortex and caudate-putamen in the rat. From the medial and sulcal prefrontal cortex electrodes, apomorphine produced a dose-related decrease of self-stimulation rate which was consistent across animals. From the caudate-putamen electrodes, on the contrary, apomorphine produced a facilitatory effect in the majority of the animals at one or more doses, however, at other doses a decreased self-stimulation rate was observed. The clear and consistent effects of apomorphine on self-stimulation of the prefrontal cortex, together with other experimental evidence in the same line, suggest that dopamine is mediating self-stimulation of this cortical area.     

Distribution of tritium label in the neonate rat brain following intracisternal or subcutaneous administration of [H]6-OHDA. An autoradiographic study

The present report describes the distribution of tritium label after injection of newborn rats with [³H]6-hydroxydopamine ([³H]6-OHDA). The animals were injected either intracisternally (i.c.) or subcutaneously (s.c.), with or without pretreatment with nomifensine, which blocks the high-affinity uptake of both noradrenaline (NA) and dopamine (DA), and sacrificed at intervals from 40 min to 24 h post-injection (p.i.). In i.c. injected animals, tritium label is demonstrable as early as 40 min p.i. in neurons of all known NA and DA cell groups. In NA neurons, it is taken up into cell body, dendrites, preterminal and terminal axons. The intensity of neuronal labeling is highest within the first 4 h p.i. and decreases in most neurons with longer postinjection intervals. A significant proportion of both NA and DA neurons degenerate beginning 6 h p.i., the majority show morphological signs of the axon reaction 24 h p.i. Uptake of [³H]6-OHDA into serotonergic and non-catecholaminergic neurons is not demonstrable.[³H]6-OHDA is accumulated by the following extraneuronal cells of the CNS: ependymal cells, epithelial cells of the choroid plexus, subependymal macrophages, smooth muscle cells in the wall of large intraparenchymal blood vessels, meningeal cells and glial cells. The time course of accumulation and disappearance of the label varies among these extraneuronal elements. The meningeal cells show the highest labeling intensity and degenerate within 24 h p.i.After pretreatment of the animals with nomifensine, the uptake of [³H]6-OHDA into NA and DA neurons is totally blocked; by contrast uptake of the labeled drug into extraneuronal cells is not prevented.These findings show that [³H]6-OHDA is not only accumulated by neurons possessing the high-affinity uptake for NA or DA, but by numerous other, extraneuronal cells which also participate in the metabolism of catecholamines.