Decrease In Extracellular Dopamine Research Articles

Dissecting the role of CB1 and CB2 receptors in cannabinoid reward versus aversion using transgenic CB1- and CB2-knockout mice

Cannabinoids produce both rewarding and aversive effects in humans and experimental animals. However, the mechanisms underlying these conflicting findings are unclear. Here we examined the potential involvement of CB1 and CB2 receptors in cannabinoid action using transgenic CB1-knockout (CB1-KO) and CB2-knockout (CB2-KO) mice. We found that Δ9-tetrahydrocannabinol (Δ9-THC) induced conditioned place preference at a low dose (1 mg/kg) in WT mice that was attenuated by deletion of the CB1 receptor. At 5 mg/kg, no subjective effects of Δ9-THC were detected in WT mice, but CB1-KO mice exhibited a trend towards place aversion and CB2-KO mice developed significant place preferences. This data suggests that activation of the CB1 receptor is rewarding, while CB2R activation is aversive. We then examined the nucleus accumbens (NAc) dopamine (DA) response to Δ9-THC using in vivo microdialysis. Unexpectedly, Δ9-THC produced a dose-dependent decrease in extracellular DA in WT mice, that was potentiated in CB1-KO mice. However, in CB2-KO mice Δ9-THC produced a dose-dependent increase in extracellular DA, suggesting that activation of the CB2R inhibits DA release in the NAc. In contrast, Δ9-THC, when administered systemically or locally into the NAc, failed to alter extracellular DA in rats. Lastly, we examined the locomotor response to Δ9-THC. Both CB1 and CB2 receptor mechanisms were shown to underlie Δ9-THC-induced hypolocomotion. These findings indicate that Δ9-THC's variable subjective effects reflect differential activation of cannabinoid receptors. Specifically, the opposing actions of CB1 and CB2 receptors regulate cannabis reward and aversion, with CB2-mediated effects predominant in mice.

Iron deficiency alters dopamine uptake and response to L‐DOPA injection in Sprague–Dawley rats

Iron deficiency (ID) disrupts brain dopamine (DA) and norepinephrine (NE) metabolism including functioning of monoamine transporters and receptors. We employed caudate microdialysis and no net flux (NNF) in post-weaning rats to determine if ID decreased the extraction fraction (E(d)). Five micromolar quinpirole, a dopamine D(2) receptor agonist, resulted in 80% decrease in extracellular DA and 45% higher E(d) in control animals. The D(2) agonist had no effect on E(d) in ID animals despite a reduction in basal DA. DAT mRNA levels were reduced by 58% with ID, while DAT protein in ventral midbrain and caudate and membrane associated DAT were also reduced by ID. Carbidopa/l-DOPA was administered to determine if elevated extracellular DA in ID was due to increased release. The DA response to l-DOPA in ID rats was 50% smaller and delayed, whereas the NE response was threefold higher. The caudate concentration of NE was also elevated in ID. Elevated dopamine-beta-hydroxylase activity in ID provides a tentative explanation for the increased NE response to l-DOPA. These experiments provide new evidence that ID results in altered synthesis and functioning of DAT and perhaps suggests some compensatory changes in NE metabolism.

Dexmedetomidine-Induced Decreases in Accumbal Dopamine in the Rat Are Partly Mediated via the Locus Coeruleus

We have demonstrated previously that the systemic administration of the selective alpha2-adrenoceptor agonist dexmedetomidine (Dex) decreases extracellular dopamine (DA) levels in the rat nucleus accumbens (NAcc). Because the locus ceruleus (LC) is a noradrenergic center linked to several of the pharmacological effects of Dex, we investigated the role of the LC in Dex-induced modulation of accumbal DA. Microdialysis probes were implanted in the NAcc and LC of Sprague-Dawley rats, and Dex 5 mM (Dex-High, n = 6), Dex 0.5 mM (Dex-Mid, n = 5), Dex 5 microM (Dex-Low, n = 6), or artificial cerebrospinal fluid (control, n = 5) was administered in the LC via retrograde microdialysis for 45 min. Extracellular DA levels were continuously measured in the NAcc dialysates using high-performance liquid chromatography coupled to electrochemical detection. Dex produced significant decreases in extracellular DA in the NAcc. Accumbal DA decreased maximally to 68.9% +/- 8.8%, 75.1% +/- 6.5%, and 77.04% +/- 12.8% of baseline in the Dex-High, Dex-Mid, and Dex-Low groups, respectively. No significant decrease in extracellular DA was observed in the control group. The coadministration of the highly selective alpha2-adrenoceptor antagonist (n = 6) RS 79948 20 mM prevented the Dex-induced decrease in accumbal DA. These data suggest that the LC plays a role in Dex-induced modulation of mesolimbic DA and support the hypothesis that noradrenergic systems can regulate remote dopaminergic sites in the central nervous system.

Dexmedetomidine decreases extracellular dopamine concentrations in the rat nucleus accumbens

The effects of dexmedetomidine, a highly selective α 2-adrenoceptor agonist, on extracellular dopamine (DA) concentrations in the nucleus accumbens of awake rats were collected via in vivo cerebral microdialysis and measured using HPLC with electrochemical detection. The administration of dexmedetomidine (DEX) at a low dose (2 μg/kg bolus i.v. over 2 min followed by a continuous infusion of 0.1 μg/kg per min) and a high dose (20 μg/kg bolus i.v. over 2 min followed by a continuous infusion of 1 μg/kg per min), significantly decreased extracellular dopamine concentrations in the nucleus accumbens. The observed decrease was dose-dependent, occurring sooner and to a greater magnitude in the rats receiving a high dose of DEX. This inhibitory modulation of accumbal dopamine was receptor-specific, as the decrease in extracellular DA produced by DEX was no longer evident following pre-treatment and co-infusion with the highly selective α 2-adrenoceptor antagonist, atipamezole (ATZ). Thus, these data suggest that adrenoceptor agonists and antagonists may modulate dopaminergic neurotransmission via mechanisms that are specific to the α 2-adrenoceptor.

Effects of nicotine and mecamylamine-induced withdrawal on extracellular dopamine and acetylcholine in the rat nucleus accumbens.

Prior research suggests that high levels of acetylcholine (ACh) in the nucleus accumbens (NAc) are associated with aversive states such as morphine withdrawal, but this has not been tested for nicotine withdrawal. The goal was to test the hypothesis that acute nicotine decreases extracellular ACh and increases extracellular dopamine (DA) in the NAc, while withdrawal from nicotine causes an opposite neurochemical imbalance with high extracellular ACh and low DA. Rats were prepared with a microdialysis probe in the NAc (primarily the shell region). They received one injection of nicotine (0.5 mg/kg, s.c.) or chronic nicotine (9 mg/kg per day via osmotic minipump). Naive animals receiving acute nicotine showed a mild, significant increase in both ACh (122% of baseline) and DA (124%). After chronic nicotine administration for 7 days, the nicotinic antagonist mecamylamine (1.0 mg/kg, s.c.) precipitated withdrawal with the appearance of somatic signs (teeth chattering and shakes/tremors) and a significant increase in extracellular ACh to 125% of baseline, while extracellular DA decreased to 65%. Control groups receiving saline in place of nicotine or mecamylamine did not show these effects. Earlier work suggests that the observed release of accumbens ACh and DA in response to acute nicotine administration may be a factor in nicotine-induced suppression of appetite. ACh release during withdrawal, coupled with the decrease in extracellular DA may play a role in the aversive aspects of nicotine withdrawal that contribute to dependency.

The regulation of NMDA-evoked dopamine release by nitric oxide in the frontal cortex and raphe nuclei of the freely moving rat

The role of nitric oxide (NO) in the N-methyl- d-aspartate (NMDA)-regulated release of dopamine (DA) in the frontal cortex and raphe nuclei of the freely moving rat was measured using in vivo microdialysis. The effects of infusing the NMDA antagonist 2-amino-5-phosphonopentanoic acid (AP5; 100 μM), neuronal nitric oxide synthase (nNOS) inhibitor 7-nitroindazole (7NI; 1 mM) or the nitric oxide donor S-nitroso- N-acetylpenicillamine (SNAP; 500 μM–5 mM) were studied. The infusion of NMDA caused a significant decrease in DA levels in both regions and these effects were reversed by AP5. AP5 alone was seen to increase DA, indicating that NMDA receptors tonically regulate DA release in these brain regions. 7NI also increased extracellular DA levels when administered alone and reversed the effects of NMDA in both regions. The NO donor SNAP (500 μM–1 mM) caused a dose-dependent decrease in extracellular DA in the RN. However in the frontal cortex the highest concentration of SNAP caused extracellular dopamine to increase. These results suggest that the regulation of NMDA-evoked DA release by NO occurs in both of the brain regions studied, although the manner in which the regulation occurs seems to differ between the two.

Halothane anesthesia decreases the extracellular level of dopamine in rat striatum: a microdialysis study in vivo.

In our previous microdialysis study, sevoflurane or isoflurane anesthesia significantly decreased the extracellular level of dopamine in rat striatum in vivo. On the other hand, other investigators demonstrated that halothane anesthesia either increased or did not affect the extracellular dopamine level. To explore the differences among these volatile anesthetics, the effects of halothane and nitrous oxide on the striatal dopamine level were reinvestigated. Halothane alone, nitrous oxide with or without halothane, or drugs known to affect the dopaminergic pathway were administered to rats. Microdialysates were collected every 20 min and directly applied to an on-line high-performance liquid chromatograph without any pretreatment. The effects of halothane on respiratory and cardiovascular variables were monitored. General anesthesia with halothane alone decreased the dialysate (extracellular) concentration of dopamine but increased that of dopamine metabolites. Nitrous oxide alone slightly increased dopamine metabolites in dialysates but did not affect the halothane-induced decrease in extracellular dopamine. Apomorphine and haloperidol reproduced reported results, confirming the adequacy of our methodology. Nomifensine- or methamphetamine-induced increase in extracellular dopamine was augmented by halothane. These results suggest that halothane potently enhances striatal dopamine release and activates the reuptake or metabolic process, which is consistent with our previous results for sevoflurane or isoflurane. Volatile anesthetics interfere with dopamine regulation, at least in the rat striatum.

Intrategmental infusion of cocaine decreases dopamine release and enhances norepinephrine release in the medial prefrontal cortex

We evaluated the effects of local cocaine infusion into the A10 (ventral tegmental area), the cell body of the mesocorticolimbic dopaminergic pathway, on the extracellular concentrations of dopamine and norepinephrine in the medial prefrontal cortex, one of its terminal fields. A 1-ml Hamilton syringe was used to infuse a cocaine solution, either 20 or 200 μM, into the ventral tegmental area of anesthetized rats for 120 min through a microdialysis probe. The pure artificial cerebrospinal fluid (0 μM cocaine) infusion served as a control and a lidocaine (100 μM) infusion was administered to prevent the local anesthetic effect of cocaine. After intrategmental cocaine infusion (either 20 or 200 μM), extracellular dopamine and norepinephrine in the ventral tegmental area both increased significantly to a steady state level (208±42 and 148±23% for low dose and 220±24 and 150±15% for high dose). Simultaneously, the 200-μM cocaine infusion caused a significant decrease in extracellular dopamine (77±5%) but an increase in norepinephrine (140±6%) in the medial prefrontal cortex. The local anesthetic, lidocaine, produced no effects on the dopamine or norepinephrine output (neither in the ventral tegmental area nor in the medial prefrontal cortex). This study not only supports recent findings of an increase in extracellular dopamine and norepinephrine in the ventral tegmental area on intrategmental cocaine infusion, but also demonstrates that cocaine infused locally in the ventral tegmental area can decrease dopamine and increase norepinephrine at a remote terminal area (medial prefrontal cortex). Finally, the introduction rate of cocaine into the ventral tegmental area by retrograde microdialysis was found to be 0.83 ng/min for the low dose and 8.14 ng/min for the high dose.

Glutamate receptor agonists decrease extracellular dopamine in the rat nucleus accumbens in vivo.

Intracerebral microdialysis was used to investigate the effects of local application of L-glutamate, N-methyl-D-aspartate, and the glutamate uptake inhibitor 1-trans-pyrrolidine-2,4-dicarboxylic acid (PDC) in the nucleus accumbens (NAc) on extracellular dopamine (DA) concentrations. The effects of locally applied PDC on extracellular glutamate concentrations were also examined. Glutamate produced a concentration-dependent decrease in extracellular DA that could be blocked by concurrent, local application of the broad spectrum ionotropic glutamate receptor antagonist kynurenic acid (KYN:1 mM). N-Methyl-D-aspartate had a concentration-dependent effect on DA release, with a low concentration (0.1 mM) producing a decrease and a higher concentration (1.0 mM) resulting in an increase. Both effects were blocked by KYN. PDC (1 mM) increased extracellular glutamate concentrations to 102% above baseline. The same concentration of PDC decreased extracellular DA concentrations, and coapplication of KYN attenuated this effect. These results indicate that glutamate receptor agonists can have both facilitatory and inhibitory effects on extracellular DA concentrations. However, the effects of PDC indicate that inhibition of DA release is the more physiologically relevant effect. Furthermore, the results of these and other experiments suggest that glutamate's inhibitory effects on DA release in the NAc are not due to direct actions of this excitatory amino acid on DA terminals. A multisynaptic model that accounts for glutamate's actions on DA release is proposed.

Chronic effects of single intrastriatal injections of 6-hydroxydopamine or 1-methyl-4-phenylpyridinium studied by microdialysis in freely moving rats

Extracellular dopamine (DA) and its main cerebral metabolites, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), were measured by bilateral striatal microdialysis in rats at different times (2, 7, 15 and 60 days) after unilateral administration into the right striatum of 1-methyl-4-phenylpyridinium ion (MPP +) or 6-hydroxydopamine (6-OHDA). In both cases the decrease in extracellular dopamine did not exceed 40% of control values. The response of DOPAC and HVA depended on the treatment: MPP + caused a marked acute decrease in the dopamine metabolites but allowed a progressive recovery that was very evident after 60 days; 6-OHDA caused a progressive decrease in the dopamine metabolites throughout the two months of the study. Tyrosine hydroxylase immunostaining revealed severe neuronal loss in substantia nigra two months after striatal administration of 6-OHDA, whereas no significant neuronal loss was found at the same time after MPP + administration. A bilateral challenge infusion of MPP + through the microdialysis probe was used to assess the dopaminergic capacity of both striata: at all the times studied there was a sharp depletion of DA on the non-lesioned side; both MPP +- and 6-OHDA-treated striata were unresponsive after a short time (2 days); after 2 months the response in MPP +-lesioned rats was similar on both sides, whereas 6-OHDA-lesioned striata were still unresponsive to MPP +. In rats, then, the effects of MPP + could be partly reversed whereas the effects of 6-OHDA were not. These results suggest that neurotoxins causing striatal dopamine loss may act through different mechanisms, which could be significant for the etiopathogenic development of Parkinson's disease.

α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptors modulate dopamine release in rat hippocampus and striatum

The effects of infusing the glutamate receptor agonist α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) on the release of dopamine (DA) has been studied in rat hippocampus and striatum in vivo. In hippocampus, AMPA (1–10 μM) produced a dose related increase in dialysate DA, but at 100 μM AMPA a sustained decrease in extracellular DA was observed. However, when samples were collected at 5-min intervals 100 μM AMPA infusion revealed a brief increase in hippocampal dialysate DA. Infusion of 100 μM AMPA and 100 μM diazoxide, which blocks AMPA receptor desensitization, led to a marked increase in extracellular DA, as did diazoxide alone, although to a lesser extent. The AMPA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3,-dione (CNQX; 200 μM) reversed the effect of AMPA and/or diazoxide infusion on dialysate DA and when infused alone, CNQX also decreased hippocampal dialysate DA. AMPA (50–500 μM) increased striatal DA release. The effect of AMPA on extracellular DA was reversed by CNQX (200 μM). Diazoxide infusion caused a decrease in striatal DA release, and this was not affected by CNQX. These data suggest that hippocampal, but not striatal AMPA receptor desensitization may play a role in regulating DA release.

Depletion of mesolimbic dopamine during behavioral despair: Partial reversal by chronic imipramine

Exposure of rate to the behavioral despair test (an animal model of depression) for 40 min resulted in a long-lasting depletion of mesolimbic dopamine output to about 40% of baseline values. The decrease in extracellular dopamine was partially prevented by chronic pretreatment with imipramine (20 mg/kg per day i.p. for 21 days). The results suggest that a fall in mesolimbic dopamine output may be associated with depressive states and indicate that changes in the functional status of the dopamine system contribute to the mechanism of action of imipramine.

Benzodiazepine-induced decreases in extracellular concentrations of dopamine in the nucleus accumbens after acute and repeated administration.

In vivo microdialysis was used to assess the effects of acute and repeated injections of the benzodiazepine midazolam on extracellular dopamine (DA) concentrations in the nucleus accumbens. Acute administration of midazolam (5 mg/kg, SC) elicited a 22% decrease in extracellular DA in the nucleus accumbens but failed to affect DA concentrations in the striatum. Similarly, six spaced intravenous infusions of midazolam, at a dose that has previously been found to support self-administration (0.05 mg per infusion), produced a 50% decrease in extracellular DA in the nucleus accumbens. In order to assess the effects of subchronic midazolam injections, two groups of rats were given injections of saline or midazolam (5 mg/kg, SC) for 14 days (two injections per day). A subsequent challenge injection of midazolam (5 mg/kg) decreased extracellular DA in the nucleus accumbens by 25% in both groups, indicating that neither tolerance nor sensitization occurred during the repeated drug administration. These experiments indicate (1) that midazolam differentially affects meso-accumbens and nigrostriatal DA neurons, and (2) that the midazolam-induced decrease in extracellular DA in the nucleus accumbens is not affected by repeated drug administration. The data further suggest that the rewarding effects of midazolam are not associated with increased release of DA in the nucleus accumbens.

Effects of nerve growth factor on cortical and striatal acetylcholine and dopamine release in rats with cortical devascularizing lesions

The effects of intraventricular nerve growth factor (NGF) or saline treatments on extracellular acetylcholine (ACh), dopamine (DA) and adenosine (Ade) levels in the cortex and striatum of rats with unilateral devascularizing cortical lesions were studied in vivo with microdialysis. The devascularizing cortical lesion produced a decrease in extracellular ACh levels in both cortex and striatum as compared to those in normal rats, while the NGF treatment produced a significant increase in ACh levels in both regions. NGF could even increase cortical ACh levels in normal rats. The cortical lesion produced a decrease in extracellular DA in the cortex, while the NGF treatment appeared to reverse this effect. No significant changes in DA were observed in the striatum. The present study gives evidence that a unilateral cortical devascularizing lesion leads to changes in extracellular ACh and DA levels in cortex and striatum and that these changes could be reversed with intraventricular NGF treatment.

Chronic nicotine treatment counteracts the decrease in extracellular neostriatal dopamine induced by a unilateral transection at the mesodiencephalic junction in rats: A microdialysis study

The effect of chronic treatment with (−)-nicotine on the decrease in extracellular dopamine (DA) levels in neostriatum induced by a unilateral transection at the meso-diencephalic junction in rats was studied. At the lesion time, Alzet minipumps filled with (−)-nicotine were implanted subcutaneously. Two weeks later, microdialysis probes were implanted bilaterally into the neostriatum. Perfusates were assayed for DA, acetylcholine (ACh) and metabolites in HPLC systems under basal and KCl stimulated conditions. The unilateral hemitransection produced an ipsilateral decrease in neostriatal extracellular DA, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), but not in ACh levels. Chronic nicotine treatment counteracted the lesion-induced decrease in DA, but had no effect on extracellular DA levels in the contralateral neostriatum or in normal rats. The results support the idea that chronic treatment may protect against degeneration of central DA neurons.

Normalization of extracellular dopamine in striatum following recovery from a partial unilateral 6-OHDA lesion of the substantia nigra: a microdialysis study in freely moving rats

It has been hypothesized that striatal dopamine (DA) terminals undergo compensatory changes in response to partial damage of the mesostriatal DA system, which results in higher concentrations of DA in the extracellular space than would be predicted by DA concentrations in post-mortem tissue. But, this hypothesis has never been tested directly in vivo, and therefore, the present study was designed to do so. Microdialysis was used in freely moving rats to estimate the concentrations of DA, dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) in striatal extracellular fluid; simultaneously from the hemisphere with a unilateral 6-hydroxydopamine (6-OHDA) lesion of the substantia nigra and from the intact hemisphere. It was found that following recovery from a 6-OHDA lesion, and during the resting state, the extracellular concentrations of DA were normal on the lesion side, even after that side was depleted of up to 99.0% of the DA measured in post-mortem tissue. Furthermore, the extracellular concentrations of DA were elevated in the intact hemisphere of animals with a >95% DA depletion. In rats with a <95% DA depletion amphetamine (1.5 mg/kg) caused a large increase in the extracellular concentration of DA in both the lesion and intact hemispheres (intact > lesion), but in rats with a > 95% tissue DA depletion amphetamine only enhanced extracellular DA on the intact side; on the lesion side amphetamine produced a progressive decrease in extracellular DA to nondetectable levels. Animals rotated towards the lesion side. Unlike DA, the extracellular concentrations of DOPAC and HVA were greatly reduced on the lesion side, and the extent of the depletion was highly correlated with lesion size. It is concluded that following partial unilateral damage to mesostriatal DA projections there are massive changes in the remaining DA terminals that are sufficient to normalize the extracellular (and presumably synaptic) concentrations of DA. The normalization of extracellular DA concentrations seen after extensive (but incomplete) damage to the mesostriatal system must play a major role in the sparing and recovery of behavioral function that is so characteristic of this system. After extensive damage the capacity of the remaining DA neurons to respond to increased demand is limited, however, and this may explain why behavioral deficits can be reinstated by stimuli that challenge the system.

Striatal neurochemistry of dynorphin-(1-13): in vivo electrochemical semidifferential analyses.

The striatal neurochemistry of dynorphin-(1-13) was studied by simultaneously measuring extracellular dopamine and serotonin voltammetrically and in vivo after the injection of dynorphin-(1-13) to male Sprague-Dawley rats. The subcutaneous administration of dynorphin-(1-13), at a dose (1.5 mg/kg), known to exert CNS mediated behavioral effects, caused a statistically significant decrease in extracellular dopamine and a statistically significant increase in extracellular serotonin from rat anterior striatum. These parallel and opposite effects of dynorphin-(1-13) on these biogenic amines occurred gradually during a three hour time course. Maximal effects on dopamine (55%) and on serotonin (62%) occurred at the end of the three hour period of study. Mean effects on dopamine and serotonin (35% and 42% respectively) were averaged from scan results over the three hour period of study; the results were significantly different from control values. Dose response studies showed that a lower dose of dynorphin-(1-13) (0.5 mg/kg sc) had little or no effect on the alteration of these biogenic amines from striatum. The highest dose of dynorphin-(1-13) studied, (3.0 mg/kg sc), predictably and significantly altered extracellular biogenic amines. The dose response, however, was not incremental. The results are consistent with the role of dynorphin-(1-13) as a neuromodulatory peptide. The results further support the concept that the neuromodulatory role of dynorphin-(1-13) may take place through neurotransmitter regulation. The data suggest that the function of dynorphin-(1-13) may be a presynaptic modulation of neurotransmission in striatum.