Insulin affects dopamine overflow in the nucleus accumbens and the striatum

To better characterize the central nervous system response to peripheral insulin administration, male Sprague-Dawley rats were fitted with microdialysis probes in the nucleus accumbens (NAC; n = 23) and striatum (STR; n = 22). Awake intact rats were injected with either 0, 200, 400, or 600 mU regular insulin i.p. Dopamine overflow was measured for at least 2 h postinjection. In the NAC, four postinjection samples were collected once every 30 min. In the STR, eight postinjection samples were collected, once every 20 min. Dopamine baselines in the NAC and STR were 9.22 pg +/- 2.02 and 10.33 pg +/- 2.22 per sample, respectively. In the nucleus accumbens, dopamine release was significantly greater in the group treated with 600 mU insulin (203 +/- 38% of baseline at 30 min). In the STR, increased dopamine release was observed in the groups treated with 200 and 400 mU insulin, whereas a suppression of a dopamine release was observed in the group treated with 600 mU. These data demonstrate that the metabolic state induced by peripheral insulin injection causes dopamine metabolism to change in both the NAC and STR, and at least in part support the hypothesis that insulin may have reinforcing properties in its effect on NAC dopamine release.     

Age-related changes in potassium-evoked overflow of dopamine in the striatum of the rhesus monkey

Rapid (5Hz) chronoamperometric recordings using Nafion-coated carbon fiber electrodes (30–90 microns o.d.) combined with pressure-ejection of potassium from micropipettes were used to investigate potassium-evoked overflow of dopamine (DA) in the striatum of young (5 to 10 years old) and middle-aged (19 to 23 years old) anesthetized rhesus monkeys. The potassium-evoked DA-like signals from the 19- to 23-year-old animals were significantly lower in amplitude than those recorded in the young animals. In addition, the temporal dynamics of DA signals in the caudate nucleus of middle-aged animals were faster, while the time courses of the signals recorded in the putamen of middle-aged monkeys were significantly longer as compared to the signals recorded from young animals. Moreover, home cage activity levels of the middle-aged animals were significantly lower. Taken together, these data support age-related changes in the output of DA from DA fibers in the striatum of middle-aged monkeys.     

The glutamate-mediated release of dopamine in the rat striatum: further characterization of the dual excitatory-inhibitory function

A push-pull cannula supplied with an artificial cerebrospinal fluid containing the tritiated precursor of dopamine, [3H]tyrosine, was implanted in the caudate nucleus of rats anesthetized with halothane. The extracellular dopamine and dihydroxyphenylacetic acid were measured in successive 20 min fractions (both in their tritiated and unlabeled form) and the ratio between the two forms calculated. Glutamate was added to the superfusing cerebrospinal fluid to investigate its role in the process of dopamine release. The release of dopamine and the efflux of dihydroxyphenylacetic acid were activated by a low concentration (10(-8) M) of glutamate. In contrast, a higher concentration (10(-4) M) of the amino acid reduced the release of dopamine. These results first confirmed the presence of a dual mechanism of control, by glutamate, of the dopamine release in the striatum depending on the extracellular concentration. Secondly, these treatments affected the dihydroxyphenylacetic acid amount and predominantly the tritiated form of dopamine, suggesting that the glutamate induces an important increase of the amine synthesis, in spite of a moderate effect on the release. The reversal of the inhibition by applications of tetrodotoxin (5 x 10(-7) M) and bicuculline (10(-4) M) confirmed that it was mediated by an indirect mechanism involving a GABAergic neurotransmission. In addition, the increase of the spontaneous dopamine release during bicuculline application suggested the existence of a tonic mechanism of inhibition of dopamine release in the striatum. This was confirmed by the fact that local xylocaine-induced anesthesia of the sensory motor cortex increased the spontaneous release of dopamine in the striatum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Time course of postmortem modifications in synaptosomal [3H]dopamine uptake and [3H]GBR 12783 binding to the dopamine uptake complex in the rat striatum

The present study focuses on the changes of two biochemical markers of the striatal dopaminergic innervation evaluated after different postmortem storage periods of rat heads either at room temperature (21 degrees C) or at 4 degrees C: (i) the uptake of [3H]dopamine (DA) into striatal synaptosomes and (ii) the specific binding of [3H]GBR 12783, a selective ligand for the neuronal dopamine uptake sites, to a striatal membrane fraction. The uptake of [3H]DA was completely abolished after 24 and 72 h storage of the tissue at 21 degrees C and 4 degrees C, respectively, whereas in the same conditions, the binding of [3H]GBR 12783 was slightly decreased. The Km and Kd of these two processes were virtually unchanged after the different storage periods considered, whereas the Vmax and Bmax were markedly decreased.     

Real-time voltammetric detection of cocaine-induced dopamine changes in the striatum of freely moving mice

In the present voltammetric study, we have characterized cocaine-induced changes in evoked dopamine release and uptake in the striatum of freely moving mice in real time. Cocaine induced marked dopamine uptake inhibition measured as apparent K_m changes, producing a maximal effect 20 min following a single injection (15 mg/kg, i.p.). Changes in uptake were paralleled by increases in evoked dopamine release per stimulus pulse, revealing a high correlation between these two parameters following cocaine administration. This initial characterization of cocaine effects on striatal dopamine transmission in the commonly used C57BL/6 mouse strain provides a basis for future voltammetric studies using genetic mouse models.     

Glutamate regulates the spontaneous and evoked release of dopamine in the rat striatum

Resting and evoked extracellular dopamine levels in the striatum of the anesthetized rat were measured by fast-scan cyclic voltammetry in conjunction with carbon fiber microelectrodes. Identification of the substance detected in vivo was achieved by inspection of background-subtracted voltammograms. Intrastriatal microinfusion of kynurenate, a broad-spectrum antagonist of ionotropic glutamate receptors, caused a decrease in the resting extracellular level of dopamine. The kynurenate-induced decrease was unaffected by systemic pretreatment with pargyline, an inhibitor of monoamine oxidase, but was significantly attenuated by systemic pretreatment with alpha-methyl-p-tyrosine, an inhibitor of tyrosine hydroxylase. Although glutamate by itself did not affect resting extracellular dopamine levels, glutamate did attenuate the kynurenate-induced decrease. Kynurenate decreased dopamine release in response to electrical stimulation of the medial forebrain bundle, an effect that was also attenuated by glutamate. These results suggest that both spontaneous and evoked dopamine release in the rat striatum are under the local tonic excitatory influence of glutamate. Interactions between central dopamine and glutamate systems that have been implicated in the etiologies of Parkinson's disease, schizophrenia, stress, and substance abuse. The precise nature of those interactions, however, remains a matter of some controversy.     

Foetal ventral mesencephalic cell suspension grafts to the 6-hydroxydopamine-lesioned rat reduce the rate of dopamine uptake in the contralateral striatum

The present study employed fast cyclic voltammetry, at carbon-fibre microelectrodes, to monitor and compare the rate of dopamine uptake in the rat striatum contralateral to (a) the 6-hydroxydopamine (6-OHDA)-lesioned/grafted striatum and (b) the 6-OHDA-lesioned/sham grafted striatum. Cell suspensions of foetal rat ventral mesencephalic tissue were grafted into the dopamine-depleted striatum of unilaterally 6-OHDA-lesioned rats. Six weeks after grafting, animals with functional, mature grafts were monitored for dopamine elimination in the contralateral striatum following electrical stimulation of the median forebrain bundle, before and after treatment with the dopamine uptake inhibitor GBR 12909. Compared to animals with sham grafts, amphetamine-amplified rotational behaviour was significantly reduced in animals with grafts of foetal ventral mesencephalic tissue. Fast cyclic voltammeric measurements followed by evaluation with the aid of a kinetic model revealed that in grafted animals, the rate of dopamine uptake via the high affinity uptake mechanism, following treatment with GBR 12909, was significantly reduced when compared to sham grafted animals.     

Haloperidol prevents the methamphetamine-induced apomorphine subsensitivity of dopamine metabolism in rat striatum

Our previous experiment showed that the sensitivity of striatal dopaminergic metabolism to apomorphine was lowered after pretreating rats with methamphetamine. This study further demonstrated that haloperidol administered with methamphetamine prevented the effect of the methamphetamine pretreatment. Since gamma-butyrolactone was administered in combination with apomorphine so as to remove complications which might arise from the modification of impulse flow through the striatonigral feedback loop, it is possible that the observed effect of apomorphine upon striatal dopaminergic metabolism and its antagonism with haloperidol may be mediated by autoreceptors of dopaminergic axon terminals in rat striatum.     

CPP antagonizes hypoxia-induced changes in dopamine metabolism in the striatum of newborn rat

Treatment with 3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP), a potent and competitive N-methyl-D-aspartate antagonist, is able to reduce the hypoxia-induced increase in striatal dopamine level by 26% even after the hypoxic insult has occurred. The hypoxia-induced decrease of the striatal 3,4-dihydroxyphenylacetic acid level can also be reversed by CPP. This study demonstrates that CPP can antagonize the hypoxia-induced changes in the dopamine metabolism in the striatum of the newborn rat.     

Prenatal cocaine exposure alters potassium-evoked dopamine release dynamics in rat striatum

The emerging profile for the effects of prenatal cocaine exposure presents two prominent features in the exposed offspring: cognitive/attention deficits and an age-associated trend toward motor/tone abnormalities up to 2 years of age. One candidate mechanism underlying these clinical features is long-lasting alterations to dopamine (DA) neuron function. However, the impact of prenatal cocaine exposure on DA release in dopaminergic terminal fields in vivo in mature offspring is poorly understood. Long-Evans female rats were implanted with an i.v. access port, bred, and given saline or cocaine-HCl (3 mg/kg/ml) for gestational days (GD) 8-14 (1x/day), GD 15-21 (2x/day), or GD 8-21 (1x/day-GD 8-14, 2x/day-GD 15-21). Using in vivo high-speed chronoamperometric recordings, potassium-stimulated DA release was measured in striatum of anesthetized male offspring 90-150 days after birth. There was a trend toward increased potassium-evoked DA signal amplitudes in offspring exposed to cocaine at any time period examined. In offspring exposed to cocaine during GD 8-21 and GD 15-21, but not at GD 8-14, there were significant decreases in the clearance capacity of the potassium-evoked DA signal compared with control offspring. The time required to clear 80% of the evoked DA signal (T(80)) in striatum for DA was significantly prolonged (approximately 150% of control) and this effect was further increased in the mean-evoked DA concentration range for these two groups. We also measured total dopamine transporter (DAT) and tyrosine hydroxylase protein levels in these offspring by blot immunolabeling and found a small, but significant, decrease in DAT protein in striatum from offspring exposed at GD 8-21 and GD 15-21. Collectively, these data demonstrate that prenatal cocaine exposure during dopamine neuron neurogenesis has long-lasting effects on DA neuron function lasting into early adulthood which may be related in part to steady state DAT protein levels. These molecular events may be associated with established cognitive deficits and perhaps the trends seen in altered motor behavior.     

MK-801, a non-competitive antagonist of NMDA receptor, prevents methamphetamine-induced decrease of striatal dopamine uptake sites in the rat striatum.

We investigated the effects of MK-801, a non-competitive antagonist of NMDA receptor, on methamphetamine-induced decrease in dopamine (DA) uptake sites in the rat striatum. Repeated administrations of an escalating dose of methamphetamine (2.5, 5, 7.5, 10 mg/kg s.c. x2, every other day for a week) produced decreased DA uptake sites assayed by binding with [3H]GBR 12935 in the striatum. Co-administration of MK-801 and methamphetamine significantly prevented the methamphetamine-induced decrease in striatal [3H]GBR 12935 binding. Administration of MK-801 alone did not affect [3H]GBR 12935 binding. These results suggest that some neurochemical effects of methamphetamine may be mediated via mechanism involving excitatory amino acids.     

Concentration-dependent actions of stimulated dopamine release on neuronal activity in rat striatum

Voltammetric analysis was combined with single unit recording to measure the effects of endogenous dopamine, released by electrical stimulation of the median forebrain bundle, on neuronal activity in the rat striatum in vivo. Fast differential ramp voltammetry, a more sensitive form of fast cyclic voltammetry, was used to measure extracellular dopamine levels during a 50-ms scan epoch every 500 ms. Using the same carbon fibre microelectrode, neuronal activity was recorded in between the electrochemical epochs. A steady-state electrochemical signal equivalent to about 100 nM dopamine was seen in the unstimulated striatum. The responses of 122 striatal units to stimulated dopamine release were recorded in 37 acute experiments. Ninety-one units which displayed a large spike amplitude (greater than or equal to 50 microV) were recorded during stimulated release of dopamine initially to levels of between 100 and 500 nM. The majority (49) showed a profound excitation, 23 showed inhibition, and nine units gave complex responses. Only 10 units were unresponsive. All the responses of these large units outlasted the transient increase in dopamine levels, often for more than 1 min. In contrast, all the 31 units which displayed a small spike amplitude (less than 50 microV) were powerfully activated by dopamine release within this range. Administration of alpha-methyl-para-tyrosine (250 mg/kg i.p.) abolished both dopamine release and the response of the five large units and four small units examined, indicating that the neuronal response was directly attributable to dopamine. Dopamine release was increased by increasing the stimulus duration over the range 0.25-10 s. With increasing levels of dopamine release the excitatory response of large units rose to a maximum and then decreased until it was eventually transformed entirely into an inhibition at dopamine levels above 1 microM. In contrast, the excitatory response of small units always increased in magnitude with increasing dopamine release to levels greater than 1 microM. The large units that showed inhibition at low levels of dopamine were also inhibited at high levels. Tail-pinch stimuli excited 21/23 large units and all seven small units tested, although this stimulus did not evoke a detectable rise in dopamine levels. We suggest that the fundamental action of dopamine in the striatum is excitation, whether involving D1 or D2 receptors. The small units described here could be inhibitory interneurons which convert the excitatory response of large units into inhibition. Dopamine may regulate striatal function by enhancing particular input-output pathways while also activating lateral inhibitory mechanisms serving to "gate-out" alternative outputs.     

Neuronal bungarotoxin blocks the nicotinic stimulation of endogenous dopamine release from rat striatum

Nicotinic receptors in the brain are receiving increased attention due in part to the recent cloning of receptor subunits and to postmortem studies revealing alterations in receptor density associated with Alzheimer's disease. The peptide neurotoxin neuronal bungarotoxin (NBT) has been shown to block nicotinic cholinergic responses in autonomic ganglia and in retinal ganglion cells. These findings suggest that NBT may be a useful probe for studying nicotinic receptors in the brain. Therefore, we have investigated the effects of NBT on the nicotine-mediated enhancement of endogenous dopamine release from rat striatal slices. It was found that the transient increase in dopamine release caused by 100 microM nicotine was completely blocked by 100 nM NBT, indicating that NBT is a functional nicotinic antagonist in this system.     

Functions of dopamine in the dorsal and ventral striatum

Manipulations of dopamine levels in the dorsal and ventral striatum are shown to affect the activation of behaviour in distinct, yet parallel ways, which depend upon the nature of the neocortical and limbic input to these structures. Whereas dopamine in the dorsal striatum contributes to the sensorimotor co-ordination of consummatory behaviour and the development of a ‘response set’ in motor preparatory processes for skilled responses, dopamine in the ventral striatum influences the impact of reward-related stimuli on appetitive aspects of behaviour. The circumstances under which the striatal dopamine projections are normally active to effect these functions are defined by studies which attempt to correlate firing in single units or neurochemical indices of dopamine activity with environmental conditions, internal states and behaviour.     

Ultrastructural evidence for co-localization of dopamine D2 and micro-opioid receptors in the rat dorsolateral striatum

Previous studies have shown significant changes in dopamine and opioid receptors in the basal ganglia following administration of cocaine. Cocaine administration results in a significant increase in the number of opioid receptors in dopamine-enriched brain regions. The aim of this study was to determine if dopamine D2 receptors (D2r) and micro-opioid receptors (microOr) are localized to the same neurons in the dorsolateral striatum. Immunoperoxidase and immunogold-silver labeling combined with electron microscopy was used to examine the ultrastructural localization of both receptors in the dorsolateral striatum. Approximately half of the microOr-labeled somatodendritic processes showed immunolabeling for the D2r. Similarly, about half of the D2r-labeled dendrites and cell bodies showed immunolabeling for the microOr. In conclusion, our results indicate that individual neurons in the rat dorsolateral striatum may be directly modulated by both dopaminergic and opioid ligands. These data also suggest that the molecular mechanism responsible for the up-regulation of microOrs in the caudate and putamen following cocaine exposure may depend, in part, on the co-existence of D2rs and micro-Ors in these cells.     

Irradiation inactivation studies of the dopamine D1 receptor and dopamine-stimulated adenylate cyclase in rat striatum

In frozen rat striatal tissue, exposed to 10 MeV electrons from a linear accelerator, the sizes of the dopamine (DA) D1 receptor and the DA-sensitive adenylate cyclase complex were determined using target size analysis. The number of D1 receptors (labelled by [3H]SCH 23390) declined monoexponentially with increasing radiation intensity, yielding a molecular weight (mol.wt.) of 80 kDa. Also the activity of the catalytic unit (C) of the adenylate cyclase (as measured by forskolin stimulation), decreased monoexponentially, however with a mol.wt. of 145 kDa. Both basal, DA- and fluoride (F-)-stimulated activity declined in a concave-downward fashion with a limiting mol.wt. of 134, 138 and 228 kDa, respectively. It was estimated that the basal and DA-stimulated activity originated from an enzyme complex with a mol.wt. of 325 kDa, a value close to the combined size of RGs and C. These data suggest that F- stimulation of the adenylate cyclase, which occurs by a Gs activation, does not cause dissociation of Gs into the alpha s and beta gamma subunits. Further, the DA-regulated adenylate cyclase apparently exists as a complex consisting of RGs and C; the mechanism of hormonal activation is a dissociation of C from this complex.     

Preservation of dopamine release in the denervated striatum

Dopamine metabolism and release were determined in the striata of rats sustaining varying damage to the nigrostriatal dopamine (DA) projection. DA metabolism, inferred from concentrations of dihydroxy-phenylacetic acid (DOPAC) or homovanillic acid (HVA), decreased with DA denervation of more than 20%. Dopamine release, inferred from the concentration of 3-methoxytyramine (3-MT), did not decrease unless the denervation was at least 80%. The amount of 3-MT per surviving neuron exceeded that for DOPAC over most of the denervation range. Thus, striatal DA release is preserved at normal levels with the survival of only 20% of the striatal DA innervation. Decreases in DA release, rather than decreases in DA metabolism or the density of dopamine innervation, coincide with the appearance of behavioral impairments.     

Effect of chronic calcium antagonist treatment on dopamine recognition sites in rat striatum

The effects of nimodipine and flunarizine administration (18 days 15 mg/kg/day p.o.) on striatal dopamine recognition sites in rats were investigated in vivo. In vitro flunarizine but not nimodipine displaces [3H]spiroperidol binding. After in vivo treatment both drugs induce a significant increase in the number of sulpiride displaceable spiroperidol binding sites (flunarizine, +114%, nimodipine +61%) concomitant with an increase in the dissociation constant. Binding parameters return toward control values after 1 week of suspension of the treatment. The results suggest that the repeated in vivo treatment with nimodipine and flunarizine may significantly interact with dopaminergic transmission leading to adaptive changes of the dopamine recognition sites.