Differential time-course profiles of dopamine release and uptake changes induced by three dopamine uptake inhibitors

Using real-time voltammetry, we compared the effects of cocaine (1.0, 3.0, or 10 microM), WIN 35428 (0.1, 0.5, or 2.0 microM), and nomifensine (0.2, 1.0, or 5.0 microM) on electrically evoked dopamine release and uptake in the rat accumbens slice. The time course for onset and offset of the drug effects were determined by perfusing single drug concentration for 30 min, followed by a 60-min washout. Cocaine elicited a rapid, concentration-independent increase in dopamine release and a more gradual, concentration-dependent inhibition of uptake. During washout, uptake inhibition rapidly abated to near baseline values. During the same period, the potentiation of dopamine release exhibited a slower offset for all concentrations and, for 10 microM cocaine, was even greater than that observed during drug perfusion ("rebound" increase). The release rebound was not observed during continuous 90-min perfusion, verifying that cocaine washout per se was a sufficient condition. Selective D1 or D2 antagonists (0.5 microM SCH 39166 or 2 microM sulpiride, respectively) were without effect on cocaine-induced release alterations. WIN 35428 and nomifensine induced similar changes in dopamine kinetics during perfusion. However, in contrast to cocaine, no consistent release rebound was observed during their washout. For 2 microM WIN 38425, washout and continuous perfusion groups exhibited similar changes in dopamine release and uptake. The time-course mismatch between uptake inhibition and DA release potentiation as well as release rebound during washout suggests that altered dopamine release might play a role in behavioral effects of cocaine.     

A novel biosensor with high signal‐to‐noise ratio for real‐time measurement of dopamine levels in vivo

Fast‐scan cyclic voltammetry (FSCV) is an established method for measuring dopamine (DA) levels in the brain in real time. However, it is difficult to discriminate DA from other monoamines such as serotonin (5‐hydroxytryptamine, 5‐HT) and norepinephrine (NE). We report a novel DA‐specific biosensor consisting of a carbon‐fiber electrode coated with an ion‐exchange membrane, a layer containing monoamine oxidase B, and a cellulose membrane. We performed FSCV using the probe to monitor the amount of DA in vitro and in vivo. First, we measured currents in vitro in phosphate‐buffered saline as we added one micromole each of DA, 5‐HT, and NE. The results confirmed that the biosensor selectively detected DA. Next, we implanted the probe in the striatum of male rats to investigate whether it could selectively detect changes in the DA content in vivo. The probe detected both the tonic change induced by methamphetamine administration and the phasic change induced by electrical stimulation of the medial forebrain bundle. In contrast, the electrode in the 6‐hydroxydopamine–lesioned striatum did not respond to systemic selective serotonin or serotonin/norepinephrine reuptake inhibitors, confirming its selectivity. Furthermore, the probe in the striatum could still detect changes in the DA level 1 week after electrode implantation. The results suggest that the novel biosensor can measure real‐time changes in DA levels in vivo with a relatively high signal‐to‐noise ratio.     

Probing presynaptic regulation of extracellular dopamine with iontophoresis

Iontophoresis allows for localized drug ejections directly into brain regions of interest driven by the application of current. Our lab has previously adapted a method to quantitatively monitor iontophoretic ejections. Here those principles have been applied in vivo to modulate electrically evoked release of dopamine in anesthetized rats. A neutral, electroactive marker molecule that is ejected purely by electroosmotic flow (EOF) was used to monitor indirectly the ejection of electroinactive dopaminergic drugs (raclopride, quinpirole, and nomifensine). Electrode placements were marked with an iontophoretically ejected dye, pontamine sky blue. We show that EOF marker molecules, acetaminophen (AP) and 2-(4-nitrophenoxy) ethanol, have no effect on electrically evoked dopamine release in the striatum or the sensitivity of electrode. Additionally, we establish that a short, 30 second ejection of raclopride, quinpirole, or nomifensine with iontophoresis is sufficient to affect autoreceptor regulation and the re-uptake of dopamine. These effects vary in lifetime, indicating that this technique can be used to study receptor kinetics.     

Effect of moderate ethanol dose on dopamine uptake in rat nucleus accumbens in vivo

The present study was designed to evaluate the effects of a moderate dose of ethanol (1 g/kg) on dopamine (DA) dynamics in rat nucleus accumbens (NAc) using fast-scan cyclic voltammetry. Voltammetric recordings were made every 100 ms at a carbon fiber microelectrode, positioned in the NAc core. Acute ethanol did not significantly alter DA uptake parameters (K(m) and V(max)), but amplitudes of the DA signals were decreased after the drug in both freely moving and anesthetized rats. Therefore, the present in vivo voltammetry results suggest that DA uptake changes are not involved in ethanol-induced increases in extracellular DA concentrations.     

Terminal effects of ethanol on dopamine dynamics in rat nucleus accumbens: an in vitro voltammetric study

To assess the direct effects of acute ethanol on dopamine (DA) terminals, evoked DA release and uptake were measured in rat nucleus accumbens slices using fast-scan cyclic voltammetry. Low and moderate concentrations of ethanol (20, 45 and 100 mM) did not alter evoked DA release, while high concentrations (150 and 200 mM) significantly decreased DA release (18 and 36%, respectively) in a calcium-dependent manner. No significant difference was found between the rate of DA disappearance measured before and after the drug. These data indicate that uptake of DA through the dopamine transporter is unaffected by ethanol, even at high concentrations. Therefore, low to moderate concentrations of ethanol have no effect on DA dynamics at the level of the nerve terminal in the nucleus accumbens. This is consistent with the hypothesis that cell body regions of DA neurons are the primary target for the stimulating and reinforcing effects of ethanol. High concentrations of ethanol can locally depress DA release, and this may correlate with the sedative actions of the drug.     

Functional and anatomical evidence for different dopamine dynamics in the core and shell of the nucleus accumbens in slices of rat brain

The characteristics of dopamine (DA) uptake and release were compared in the core and shell of the nucleus accumbens (NAc). DA release was elicited from rat brain slices by local electrical stimulation, and its extracellular concentration was monitored with fast-scan cyclic voltammetry using Nafion-coated, carbon-fiber microelectrodes. The voltammetric results show that the values of DA release and uptake in the shell NAc are approximately one-third of those measured in the core region, and DA uptake in the shell was less sensitive than the core to inhibition by either cocaine or nomifensine. The density of [³H]mazindol binding sites in the NAc was examined by autoradiography and the shell was found to have an average of half the number of DA uptake sites measured in the core region. This combination of anatomical and functional results shows that DA neurotransmission in the shell NAc is distinct from that in the core region. These data are consistent with the view that multiple functional forms of the DA transporter, exhibiting disparate kinetics and pharmacology, exist in different brain regions that exhibit disparate kinetics and pharmacology. Different forms of the transporter, combined with different release kinetics and auto- and heteroreceptor activity, give a vast range of possibilities for regional variation in DA neurotransmission. © 1996 Wiley-Liss, Inc.     

L-arginine increases dopamine transporter activity in rat striatum via a nitric oxide synthase-dependent mechanism

Literature reports suggest that nitric oxide (NO) participates in the regulation of dopaminergic neurotransmission, possibly through interaction with cysteine residues of the dopamine transporter (DAT). Rotating disk electrode voltammetry was used to measure dopamine (DA) transport in rat striatum to determine if 1) the nitric oxide synthase (NOS) substrate, L-arginine (L-Arg), could affect DAT activity; 2) L-Arg-dependent effects on DAT activity could be blocked by NOS and guanylate cyclase inhibitors, a NO scavenger, DA, and cocaine; 3) a NO donor could affect DAT activity; and 4) L-Arg could protect the DAT from a sulfhydryl agent. L-Arg increased DAT activity by increasing V(max). NOS inhibitors (S-ethylisothiourea and S-isopropylisothiourea), a NO scavenger (2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide), DA, and cocaine blocked the L-Arg effect. The guanylate cyclase inhibitor, 1H-(1,2,4)-oxadiazolo[4,3a]quinoxalin-1-one, did not. The NO donor, S-nitroso-N-acetylpenicillamine, decreased DAT activity and L-Arg protected the DAT from the effects of the sulfhydryl agent N-ethylmaleimide. These results suggest that L-Arg, via NO, may play a role in regulating DAT activity in rat striatum by increasing the V(max) of DA transport. Furthermore, it is suggested that the effects of L-Arg on DAT activity may be due to modification of the DAT itself, possibly via the NO-mediated modification of DAT cysteine residues. Finally, NO produced from L-Arg may affect the DAT differently than NO from NO donors. These results further the notion that dopaminergic neurotransmission may be regulated by changes in DAT activity caused by L-Arg and NOS.     

In vivo measurement of somatodendritic release of dopamine in the ventral tegmental area

The ventral tegmental area (VTA), the locus of mesolimbic dopamine cell bodies, contains dopamine. Experiments in brain slices have demonstrated that VTA dopamine can be released by local electrical stimulation. Measurements with both push‐pull cannula and microdialysis in intact animals have also obtained evidence for releasable dopamine. Here we demonstrate that dopamine release in the VTA can be evoked by remote stimulations of the medial forebrain bundle (MFB) in the anesthetized rat. In initial experiments, the MFB was electrically stimulated while a carbon‐fiber electrode was lowered to the VTA, with recording by fast‐scan cyclic voltammetry. While release was not observed with the carbon fiber 4–6 mm below dura, a voltammetric response was observed at 6–8 mm below dura, but the voltammogram was poorly defined. At lower depths, in the VTA, dopamine release was evoked. Immunohistochemistry experiments with antibodies for tyrosine hydroxylase (TH) confirmed that dopamine processes were primarily found below 8 mm. Similarly, tissue content determined by liquid chromatography revealed serotonin but not dopamine dorsal to 8 mm with both dopamine and serotonin at lower depths. Evaluation of the VTA signal by pharmacological means showed that it increased with inhibitors of dopamine uptake, but release was not altered by D2 agents. Dopamine release in the VTA was frequency dependent and could be exhausted by stimulations longer than 5 s. Thus, VTA dopamine release can be evoked in vivo by remote stimulations and it resembles release in terminal regions, possessing a similar uptake mechanism and a finite releasable storage pool. Synapse 63:951–960, 2009. © 2009 Wiley‐Liss, Inc.     

Increase of dopamine turnover in bilateral striata after unilateral injection of haloperidol into substantia nigra of unrestrained rats

In order to investigate self-regulation of dopamine (DA) neurons, the effects of intranigrally administered haloperidol (Hal), a DA receptor antagonist, on nigrostriatal DA systems were examined using differential pulse voltammetry with carbon fiber electrode. The measurements were achieved in the bilateral caudate-putamen (CP) of behaving rats, in the region of which DA or 3,4-dihydroxyphenylacetic acid made an oxidative current peak (P2) spontaneously. Unilateral injection of Hal (5 micrograms in 1 microliter) into the substantia nigra of rat increased P2 in a time-dependent manner. This phenomenon was observed in both CP, but a more significant increase was in the ipsilateral side (156 +/- 2% of spontaneous height 2.75 h after injection) than in the contralateral side (129 +/- 7%). These effects enlarged in a dose-dependent manner. The same results were found in tissue homogenates determined by high-performance liquid chromatography with electrochemical detection. In the latter case, however, no significant difference was observed between the left and right sides. The present results suggest that Hal, attaching nigral autoreceptors on the cell bodies and dendrites, blocks inhibitory influence of endogenous DA and then activates the nigrostriatal DA neurons, while the contribution of non-dopaminergic neurons is also possible.     

Methylphenidate analogs with behavioral differences interact differently with arginine residues on the dopamine transporter in rat striatum

The methylphenidate analogs N-methyl-4-methyl-methylphenidate and N-benzylmethylphenidate are believed to interact differently with the dopamine transporter (DAT) in vitro and in vivo. Herein, we report that methylphenidate and N-methyl-4-methyl-methylphenidate, but not N-benzylmethylphenidate, protect the rat striatal DAT from the arginine-selective chemical modifying agent, phenylglyoxal. This suggests that methylphenidate and N-methyl-4-methyl-methylphenidate, but not N-benzylmethylphenidate, interact with the guanidine groups of arginine residues in the DAT of rat striatum. This differential interaction may, at least in part, explain the in vitro and in vivo differences between N-methyl-4-methyl-methylphenidate and N-benzylmethylphenidate.     

Anatomical and pharmacological characterization of catecholamine transients in the medial prefrontal cortex evoked by ventral tegmental area stimulation

Voltammetric measurements of catecholamines in the medial prefrontal cortex (mPFC) are infrequent because of lack of chemical selectivity between dopamine and norepinephrine and their overlapping anatomical inputs. Here, we examined the contribution of norepinephrine to the catecholamine release in the mPFC evoked by electrical stimulation of the ventral tegmental area (VTA). Initially, electrical stimulation was delivered in the midbrain at incremental depths of ?5 to ?9.4 mm from bregma while catecholamine release was monitored in the mPFC. Although catecholamine release was observed at dorsal stimulation sites that may correspond to the dorsal noradrenergic bundle (DNB, containing noradrenergic axonal projections to the mPFC), maximal release was evoked by stimulation of the VTA (the source of dopaminergic input to the mPFC). Next, VTA‐evoked catecholamine release was monitored in the mPFC before and after knife incision of the DNB, and no significant changes in the evoked catecholamine signals were found. These data indicated that DNB fibers did not contribute to the VTA‐evoked catecholamine release observed in the mPFC. Finally, while the D2‐receptor antagonist raclopride significantly altered VTA‐evoked catecholamine release, the α₂‐adrenergic receptor antagonist idazoxan did not. Specifically, raclopride reduced catecholamine release in the mPFC, opposite to that observed in the striatum, indicating differential autoreceptor regulation of mesocortical and mesostriatal neurons. Together, these findings suggest that the catecholamine release in the mPFC arising from VTA stimulation was predominately dopaminergic rather than noradrenergic. Synapse 68:131–143, 2014. © 2013 Wiley Periodicals, Inc.     

Voltammetry of extracellular dopamine in rat striatum during ICSS-like electrical stimulation of the medial forebrain bundle

Fast-scan voltammetry cyclic in the striatum of anesthetized rats has been used to monitor extracellular dopamine during forced electrical stimulation of the media forebrain bundle using parameters that mimic intracranial self-stimulation. The temporal resolution provided by microelectrodes positioned very near sites of dopamine release allows resolution of the response to individual 500-ms stimulation trains separated by 500-ms intervals. Uptake inhibition by Nomifensine alters the resolution obtained at short times after initiation of stimulation.     

Regional variations in the physiology of the rat caudate-putamen. 2. Effects of amphetamine and amphetamine induced dopamine release on basal and cortical stimulation evoked multiple unit activity

Summary. Dopaminergic terminals within the caudate-putamen are located in an ideal position to modulate the corticostriatal system. Since this is the major afferent system of the striatum, dopamine has very powerful effects on striatal electrophysiological activity. The striatum is a regionally specialized multifunctional nucleus. It is therefore important to determine if dopamine has the same modulatory effects within different areas of the nucleus. The effects of 2.5 mg/Kg D-amphetamine (IP) on cortical stimulation evoked and basal multiple unit activity (MUA) was measured in 7 dorsal and 7 ventral striatal areas of the urethane anaesthetized rat. In general, amphetamine caused an increase in the basal activity and a decrease in the cortical stimulation evoked activity. However, there were both qualitative and quantitative regionally dependent differences in these responses. The effect on basal MUA was more pronounced in the dorsal and caudal areas whereas the effect on cortical stimulation evoked MUA was more pronounced in the ventral areas. The electrophysiological effects of amphetamine within the striatum were correlated with its regionally dependent effects on extracellular dopamine. This produced a measure of the effects of striatal dopamine on regional electrophysiological activity. This information was also used to determine the mathematical relationship between dopamine concentration change and the change in MUA. These data indicate that the excitatory effects of amphetamine-induced dopamine release on the non-stimulated MUA progressively increase along the rostro-caudal axis of the nucleus. In addition, the effects were more pronounced in the ventromedial as compared to the ventrolateral areas. These effects correlated best with the rate of change in dopamine concentration. In the dorsal striatum amphetamine-induced increases in dopamine had a regionally homogeneous inhibitory effect on the stimulated MUA. In the ventral striatum however, it had a progressively stronger effect along the rostro-caudal axis. These effects correlated best with the absolute change in dopamine concentration. Received April 17, 2002; accepted December 2, 2002 Published online March 5, 2003 Authors' address: Dr. G. Glynn, School of Pharmacy and Allied Health Professions, Creighton University, 2500 California Plaza, Omaha, NE 68178, U.S.A., e-mail: GGlynn@creighton.edu     

The ontogeny of amphetamine-induced dopamine release in the caudate-putamen of the rat

Amphetamine-induced dopamine (DA) release in the caudate-putamen of adult rats was compared with that in the 35-36-day-old and 21-22-day-old rat pup, using in vivo voltammetry. In the adult and 35-36-day groups, 1.0 mg/kg amphetamine (AMP) produced a significant increase in DA release, while 0.1 mg/kg produced no significant change in DA release. In the 21-22-day group, 1.0 mg/kg AMP produced a slight increase, followed immediately by a significant decrease in DA release. Similarly, at a dose of 0.1 mg/kg, AMP produced a significant decrease in DA release. This decrease was greater than that seen after the 1.0 mg/kg dose of AMP. Tyramine produced no significant change in DA release, however, it served as a control for peripheral cardiovascular effects. These data suggest that AMP-induced DA release in the caudate-putamen is mature by postnatal day 35. The AMP-induced decrease in DA release found in the 21-22-day group is not due to either the cardiovascular effects of AMP or to a depletion of DA content. This decrease in neostriatal DA release may be due to a decrease in the neuronal firing of nigrostriatal DA neurons that is caused by an AMP-induced increase in dendritic DA release in the substantia nigra, exerting an inhibitory effect through DA autoreceptors.     

Regional variations in the physiology of the rat caudate-putamen. 1. In vivo subregional effects of amphetamine on extracellular dopamine concentration

Summary. The caudate putamen is a neurochemically and functionally heterogeneous nucleus. Understanding the correlation between these regional variations in neurochemistry and function could greatly aid in the treatment of neurological disorders associated with this area of the brain. Since dopaminergic dysfunction has been implicated in some of these disorders, regional variations in the neurochemistry of this transmitter system are of particular interest. The dopaminergic response to 2.5 mg/Kg D-amphetamine was examined by in vivo voltammetry in 7 dorsal and 7 ventral regions of the caudate-putamen in the urethane anaesthetized rat. Extracellular dopamine concentration increased in all the areas examined. However, the effect was regionally heterogeneous-areas separated by as little as 1 mm showing significantly different responses in terms of both the absolute change and the rate of change in dopamine concentration. A significant general trend was also evident. Amphetamine produced an increasing effect in extracellular dopamine concentrations in the dorsal and lateral areas of the nucleus. It was concluded that the regionally heterogeneous effects of amphetamine on extracellular dopamine could be attributed to regional variations in the density of dopamine transport sites within the caudate-putamen. Since this transport site is the site of entry of a number of neurotoxins this finding may contribute to our understanding of the functional loss associated with disorders such as Parkinson's disease. Received April 17, 2002; accepted December 2, 2002 Published online March 5, 2003 Acknowledgements We thank Dr. B. K. Yamamoto for assistance reviewing an early draft of the paper. All experiments were performed strictly adhering to the policy on the use of animals in neuroscience research as drafted by the Society for Neuroscience. Authors' address: Dr. G. Glynn, School of Pharmacy and Allied Health Professions, Creighton University, 2500 California Plaza, Omaha, NE 68178, U.S.A., e-mail: GGlynn@creighton.edu     

Regional specificity in the real‐time development of phasic dopamine transmission patterns during acquisition of a cue–cocaine association in rats

Drug seeking is significantly regulated by drug‐associated cues and associative learning between environmental cues and cocaine reward is mediated by dopamine transmission within the nucleus accumbens (NAc). However, dopamine transmission during early acquisition of a cue–cocaine association has never been assessed because of the technical difficulties associated with resolving cue‐evoked and cocaine‐evoked dopamine release within the same conditioning trial. Here, we used fast‐scan cyclic voltammetry to measure sub‐second fluctuations in dopamine concentration within the NAc core and shell during the initial acquisition of a cue–cocaine Pavlovian association. Within the NAc core, cue‐evoked dopamine release developed during conditioning. However, within the NAc shell, the predictive cue appeared to cause an unconditioned decrease in dopamine concentration. The pharmacological effects of cocaine also differed between sub‐regions, as cocaine increased phasic dopamine release events within the NAc shell but not the core. Thus, real‐time measurements not only revealed the initial development of a conditioned neurochemical response but also demonstrated differential phasic dopamine transmission patterns across NAc sub‐regions during the acquisition of a cue–cocaine association.     

Striatal dopamine uptake and swim performance of the aged rat

The striatum and olfactory tubercle of 30-month-old F344 rats contain significantly (21-24%) less dopamine compared with young adult (8-month) animals. However, rats of the two age groups show identical Km and Vmax values for the kinetics of [3H]dopamine uptake into striatal homogenates; uptake into the olfactory tubercle also appeared unaffected in old age. The preservation of dopamine uptake despite reductions in content of the transmitter suggests that the forebrain dopaminergic nerve terminals are intact, but that reduced dopamine synthesis and/or enhanced degradation may occur in the existing terminals. Administration to senescent animals of the dopamine uptake blockers nomifensine or bupropion (but not the norepinephrine uptake blocker desmethylimipramine) improved their swim performance to levels comparable with young adult animals. The findings suggest that amine reuptake may limit the synaptic effectiveness of dopamine released in the aged striatum.     

Selective subregional dopamine depletions in the rat caudate-putamen following nigrostriatal lesions.

Previous work has demonstrated a complex neurochemical and neuroanatomical heterogeneity of the striatum in normal brains. The present research investigated whether the heterogeneous distribution of dopamine would be altered following unilateral injections of the neurotoxin 6-hydroxydopamine into the substantia nigra of the rat. Four weeks following injection, the nucleus accumbens and subregions of the caudate-putamen and substantia nigra were dissected and analyzed by HPLC with electrochemical detection for dopamine, 5-hydroxytryptamine, and their respective metabolites. Levels of dopamine and its metabolites in the anterodorsolateral caudate-putamen were depleted more than medial, posterior, and ventral, striatal areas in partially lesioned animals (less than 90% dopamine depletion). This resulted in an alteration of striatal heterogeneity such that a mediolateral gradient of dopamine tissue content was now superimposed on the normal rostrocaudal gradient observed in controls. Paralleling these findings, dopamine was more depleted in the lateral, as opposed to the medial, substantia nigra. These results indicate that the nigrostriatal dopamine system degenerates in a heterogeneous fashion following 6-hydroxydopamine administration. It is speculated that the differential loss of dopamine neurons observed in the nigra of Parkinson's patients may be due to a differential sensitivity to toxins within the nigra.     

The dopamine patchwork of the rat nucleus accumbens core

The dopamine (DA) terminal field in the rat dorsal striatum is organized as a patchwork of domains that show distinct DA kinetics. The rate and short‐term plasticity of evoked DA release, the rate of DA clearance and the actions of several dopaminergic drugs are all domain‐dependent. The patchwork arises in part from local variations in the basal extracellular concentration of DA, which establishes an autoinhibitory tone in slow but not fast domains. The present study addressed the hypothesis that a domain patchwork might also exist in the nucleus accumbens core (NAcc), a DA terminal field that is deeply involved in reward processing and the mechanisms underlying substance abuse. DA recordings in the NAcc by fast‐scan voltammetry during electrical stimulation of the medial forebrain bundle confirmed that the NAcc contains a patchwork of fast and slow domains showing significantly different rates of evoked DA release and DA clearance. Moreover, the NAcc domains are substantially different from those in the dorsal striatum. There were no signs in the NAcc of short‐term plasticity of DA release during multiple consecutive stimuli, and no signs of a domain‐dependent autoinhibitory tone. Thus, the NAcc domains are distinct from each other and from the domains of the dorsal striatum.     

Adenosine A1 receptor-mediated inhibition of dopamine release from rat striatal slices is modulated by D1 dopamine receptors

Dopamine release is regulated by presynaptic dopamine receptors and interactions between adenosine and dopamine receptors have been well documented. In the present study, dopamine release from isolated striatal slices from Wistar rats was measured using fast cyclic voltammetry. Single-pulse stimulation (0.1 ms, 10 V) was applied every 5 min over a 2-h period. Superfusion with the adenosine (A)(1) receptor agonist N(6)-cyclopentyladenosine (CPA), but not the A(2) receptor agonist 3-[4-[2-[[6-amino-9-[(2R,3R,4S,5S)-5-(ethylcarbamoyl)-3,4-dihydroxy-oxolan-2-yl]purin-2-yl]amino]ethyl] phenyl]propanoic acid (CGS 21680), inhibited dopamine release in a concentration-dependent manner (IC(50) 3.80 x 10(-7) m; n = 10). The dose-response curve to CPA was shifted to the right (IC(50) 6.57 x 10(-6) m; n = 6, P < 0.05 vs. control) by the A(1) receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX). Neither the D(1) agonist 6-chloro-APB nor the D(1) antagonist R-(+)-8-chloro-2,3,4,5-tetrahydro-3-methyl-5-phenyl-1H-3- benzazepine-7-ol (SCH 23390) altered dopamine release on their own. However, SCH 23390 (3 microm) significantly attenuated the response to CPA (IC(50) 1.44 x 10(-5) m; n = 6, P < 0.01 vs. control). Furthermore, the inhibitory effect of CPA was significantly increased in the presence of 6-chloro-APB (1 microm). In radioligand binding experiments, CPA interacted with high- and low-affinity states of [(3)H]DPCPX-lableled A(1) receptors. The high-affinity agonist binding to A(1) receptors was inhibited by the stable guanosine triphosphate analogue Gpp(NH)p. In contrast, neither the proportion nor the affinity of high-affinity A(1) receptors was altered by dopamine or SCH 23390. These results provide evidence that the inhibition of dopamine release by adenosine A(1) receptors is dependent, at least in part, on the simultaneous activation of D(1) dopamine receptors. While the mechanism underlying this interaction remains to be determined, it does not appear to involve an intramembrane interaction between A(1) and D(1) receptors.