Transmitter-dependent involvement of the respiratory interneurons in the locomotor rhythm in the pulmonate mollusc Lymnaea

Pedal cell RPeD1 of the pond snail L. stagnalis becomes involved in a central rhythm identified as an activity of the central pattern generator (CPG) for locomotion. The RPeD1 rhythm developed as driven by a synaptic input in isolated CNS preparations treated with 0.05 mM serotonin (5HT) or 0.1 mM 5-hydroxytryptophan (5HTP). The 5HT-induced co-ordinated rhythmic activity was retained by each of two pedal ganglia after complete isolation thus suggesting that the respective CPG lies entirely within the pedal portion of the CNS and is paired. The findings suggest that the RPeD1 switching from one network to another represents a neurotransmitter-dependent phenomenon.     

Neuroanatomical,immunocytochemical, and physiological studies of the pharyngeal retractor muscle and its putative regulatory neurons playing a role in withdrawal and feeding in the snail, <Emphasis Type="Italic">Helix pomatia</Emphasis>

We describe the neurons regulating two separate functions of the pharyngeal retractor muscle (PRM), namely sustained contraction during body withdrawal and rhythmic phasic contractions during feeding, in the snail, Helix pomatia. The distribution of central neurons innervating the PRM is organized into two main units; one in the buccal-cerebral ganglion complex, the other in the subesophageal ganglion complex. Serotonin- (5-HT-), FMRFamide- (FMRFa-), and tyrosine-hydroxylase-immunostained neurons are present among the PRM neurons that densely innervate the PRM. 5HT both decreases and increases the amplitude of the electrically evoked contraction between concentrations of 0.1 M and 1 M. Dopamine (DA) only decreases the amplitude of contraction at a 1-M threshold concentration. In contrast, FMRFa increases the amplitude of the contraction and slightly elevates the tone of the PRM but requires a higher threshold (10 M). Assay by high-performance liquid chromatography of 5HT and DA in the PRM has shown that the 5HT level decreases during locomotion but increases during feeding, whereas the DA level increases during locomotion but slightly decreases during feeding. Thus, different segments of the PRM are innervated by neurons from different loci within the central nervous system. The segments of the PRM distal to the pharynx are innervated from loci of the subesophageal ganglion complex suggesting that they mediate withdrawal. The proximal segment of the PRM is innervated from cerebral and buccal loci indicating that these neurons mediate the feeding rhythm produced by buccal and cerebral feeding central pattern generators to induce rhythmic phasic contractions in the PRM during feeding.This work was supported by Hungarian Scientific Research Fund (OTKA) grants (T034106, T037389, T037505), the Wellcome Trust CRIG Programme, and the Wellcome Trust Travel Grant.     

Involvement of pedal peptide in locomotion in Aplysia: modulation of foot muscle contractions

Pedal peptide (Pep) is a 15-amino-acid neuropeptide that is localized within the Aplysia central nervous system (CNS) predominantly to a broad band of neurons in each pedal ganglion. Pep-neurons were identified by intracellular staining and immunocytology or by radioimmunoassay (RIA) of extracts from identified neurons. RIA reveals that 97% of all Pep-like immunoreactivity (IR-Pep) in pedal nerves is found in the three nerves that innervate the foot. Nearly every Pep-neuron sends an axon out at least one of these three nerves. Application of Pep to foot muscle causes an increase in the amplitude and relaxation rate of contractions driven by nerve stimulation or intracellular stimulation of pedal motor neurons. The increase in relaxation rate was the predominant effect. Intracellular recording in "split-foot" preparations reveals that Pep-neurons increase their overall firing rates and fire in bursts with each step during locomotion. Recovery of IR-Pep from foot perfusate following pedal nerve stimulation increases in a frequency-dependent fashion. Thus it appears that one function of Pep-neurons is to modulate foot muscle contractility during locomotion in Aplysia.     

Branchial musculature of a venerid clam: pharmacology,distribution, and innervation

This study was meant to analyze the neural control of the branchial muscles of the clam Mercenaria mercenaria. Gills isolated from the animal contract in response to 5-hydroxytryptamine (5HT), dopamine (DA), and acetylcholine (ACh); but the ACh contraction occurred only if the gills had been pretreated with the cholinesterase inhibitor eserine. The 5HT antagonists cyproheptadine and mianserin blocked the contractile effects of all of the agonists. However, gills exposed to the 5HT antagonists and eserine relaxed in response to ACh. The DA antagonist SCH-83566 inhibited the effects of DA, but had no effect on contractions induced by 5HT and ACh. The ACh antagonist hexamethonium inhibited both the excitatory and inhibitory effects of ACh, but had no effect on contractions induced by 5HT and DA. 5HT and DA in gill tissue were visualized by using immunohistochemistry. Within each gill filament are dorsoventral neurons running adjacent to the epithelium and containing immunoreactive 5HT and DA. A complex network of 5HT-positive fibers is associated with the septa, blood vessels, and muscles, whereas DA-positive fibers are restricted to the septa. We propose that 5HT is the excitatory transmitter to the gill muscles, and that DA and ACh exert their excitatory effects by stimulating 5HT motor nerves. ACh may also be an inhibitory transmitter of the muscles.     

Disruption of 5-hydroxytryptaminergic neuronal function blocks the action of morphine on tuberoinfundibular dopaminergic neurons

Subcutaneous injections of morphine to male rats reduced dopamine(DA) turnover (α-methyltyrosine-induced decline of DA concentrations) in the median eminence, and increased DA turnover in the striatum. Selective destruction of central 5-hydroxytryptamine(5HT)-neurons with intracerebroventricular injections of 5,7-dihydroxytryptamine, or the administration of metergoline, a putative 5HT antagonist, blocked the inhibitory effects of morphine on DA turnover in the median eminence. In the same experiments disruption of 5HT neurotransmission processes caused a similar but less dramatic antagonism of the stimulatory actions of morphine on DA turnover in the striatum. Thus, 5HT neurons play a role in mediating the effects of morphine on tuberoinfundibular and possibly on nigrostriatal DA neurons.     

Effect of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) on monoamine neurotransmitters in mouse brain & heart

The effect of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was studied on dopamine (DA), norepinephrine (NE), serotonin (5HT) and γ-aminobutyric acid (GABA) neurons in mouse brain and on NE neurons of mouse heart. MPTP (45 mg/kg) was administered s.c. to mice twice daily for 2 consecutive days. This dosage regimen produced a decrease in the forebrain concentrations of DA and NE at 7 and 20 days after injection. In contrast, the forebrain concentrations of 5HT and GABA were not significantly decreased at either time. MPTP administration also produced a marked decrease in the uptake of ³H-DA into striatal slices and ³H-NE into cerebral cortical slices. In contrast, the uptake of ³H-NE into hypothalamic slices and the uptake of ³H-5HT into slices from several brain regions were not altered. MPTP initially reduced the concentration of NE in the heart, but unlike the persistent decreases in the forebrain concentrations of NE and DA, the NE concentration in the heart returned to control levels at approximately 20 days after MPTP administration. These results, showing that MPTP can produce a long lasting and selective decrease in the forebrain concentrations of NE and DA and in the uptake of radioactive DA and NE into brain slices, suggest that MPTP can cause the destruction of catecholamine neurons in mouse brain. In contrast, MPTP administration does not appear to produce long term changes in either 5HT or GABA neurons.     

Effects of Chronic Restraint Stress on Feeding Behavior and on Monoamine Levels in Different Brain Structures in Rats

Monoaminergic systems are important modulators of the responses to stress. Stress may influence feeding behavior, and the involvement of monoamines in the control of food intake is well recognized. We investigated the effects induced by chronic-restraint stress, 1 h a day, for 40 days, on eating behavior and on monoamines in distinct brain structures. Increased consumption of sweet pellets, and not of peanuts, was observed. Dopamine (DA), serotonin (5–HT), and their metabolites were measured by HPLC-EC. After chronic restraint, the results observed were decreased 5–HT in hippocampus, with increased 5–HIAA/5–HT; decreased 5–HIAA levels in cortex; reduction in DA in hippocampus, and increased levels in amygdala and hypothalamus; HVA increased in cortex, as well as HVA/DA ratio, while DOPAC/DA decreased. HVA decreased in hypothalamus, as well as HVA/DA, and DOPAC/DA and HVA/DA decreased in the amygdala. These results suggest that restraint stress differentially affects the activity of central dopaminergic and serotonergic neurons, and this may be related to the effects observed in eating behavior.     

Locomotion induced by non-contingent intracranial stimulation: comparison to psychomotor stimulant

Non-contingent experimenter-applied stimulation (nEAS) to the ventral mesencephalon, unlike contingent intracranial self-stimulation (ICSS), elicits high rates of general locomotion. This locomotion may be due to the nature of the presentation of stimulation, in that nEAS is non-contingent, while ICSS depends on a specific and focused response (e.g., bar pressing). Psychomotor stimulants also elicit high amounts of general locomotion, with the locomotion attributed to increased dopamine release. Interestingly, dopamine release decreases or is absent with repeated ICSS, but not nEAS. This suggests that the locomotion elicited by nEAS may be the result of DA release similar to that observed with psychomotor stimulants. To determine the relationship between locomotion induced by nEAS and psychomotor stimulants, locomotion elicited by nEAS was directly compared to that produced by cocaine, a psychomotor stimulant and indirect DA agonist. Six groups of rats were examined: (1) DA+ group: rats were implanted with a stimulating electrode in the ventral mesencephalon and activation of DA neurons was verified during surgery by monitoring DA release in the striatum; (2) DA- group: rats were also implanted with stimulating electrodes, but the location in the ventral mesencephalon did not elicit DA release; (3) 10-mg/kg cocaine group: rats were exposed to a low dose (10 mg/kg) of cocaine; (4) 40-mg/kg cocaine group: rats were exposed to a high dose (40 mg/kg) of cocaine; (5) saline group: rats were injected with saline; and (6) naive group: rats received no treatment. The topography of behavior was assessed in all rats during four periods: a pre-treatment baseline, treatment, early post-treatment, and a late post-treatment end point. The results suggest that locomotion elicited by nEAS was stereotypic, dependent upon DA release and similar, but not identical, to psychomotor stimulant-induced locomotion.     

Distribution of tyrosine-hydroxylase-immunoreactive and dopamine-immunoreactive neurons in the central nervous system of the snail Helix pomatia

The distribution and characterization of dopamine-containing neurons are described in the different ganglia of the central nervous system of Helix on the basis of the distribution of tyrosine hydroxylase immunoreactive (TH-ir) and dopamine immunoreactive (DA-ir) neurons. Both TH-ir and DA-ir cell bodies of small diameter (10–25 m) can be observed in the buccal, cerebral and pedal ganglia, dominantly on their ventral surface, and concentrated in small groups close to the origin of the peripheral nerves. The viscero-parietal-pleural ganglion complex is free of immunoreactive cell bodies but contains a dense fiber system. The largest number of TH-ir and DA-ir neurons can be detected in the pedal, and cerebral ganglia. The average number of TH-ir and DA-ir neurons significantly differs but all the identifiable groups of TH-ir neurons also show DA-immunoreactivity. Therefore, we consider the TH-ir neurons in those groups as being DA-containing neurons. The amounts of DA in the different ganglia assayed by high performance liquid chromatography correspond to the distribution and number of TH-ir and DA-ir neurons in the different ganglia. The axon processes of the labeled small-diameter neurons send thin proximal branches toward the cell body layer but only rarely surround cell bodics, whereas distally they give off numerous branches in the neuropil and then leave the ganglion through the peripheral nerves. In the cerebral ganglia, the analysis of the TH-ir pathways indicates that the largest groups of labeled neurons send their processes through the peripheral nerves in a topographic order. These results furnish morphological evidence that DA-containing neurons of Helix pomatia have both central and peripheral roles in neuronal regulation.     

Dopamine and 5-hydroxytryptamine actions on the cardiac ganglion of the lobster Homarus americanus

The cardioexcitor monoamines dopamine (DA) and 5-hydroxytryptamine (5HT) accelerate bursting by isolated cardiac ganglia of the lobster Homarusamericanus most effectively when they act on a region of the ganglionic trunk anterior to the small cells which have been considered the pacemakers of the system. 5HT may exert its acceleratory action by depolarizing cell processes. Neither the somata nor the spike-initiating zones of the small cells have to be directly exposed to 5HT or DA in order for acceleration to occur. When 5HT is applied selectively to the small cells bursts are prolonged, probably as a result of increases in the duration of the endogenous burst-organizing potentials (driver potentials) generated by these neurons. This action on the small cells can lead to prolonged and intensified bursts of the full ganglion during the onset of 5HT action when the whole ganglion is exposed to the monoamine. Neither DA nor 5HT has a direct effect on the characteristics of large cell (motorneuron) driver potentials. Accepted: 3 September 1997     

Distribution of serotonin-containing neurons in the central nervous system of the snail Helix pomatia

Summary The distribution of serotonin (5HT)-containing neurons in the central nervous system of the snail Helix pomatia has been determined in whole-mount preparations by use of immunocytochemical and in vivo 5,6-dihydroxy-tryptamine labelling. 5HT-immunoreactive neuronal somata occur in all but the buccal and pleural ganglia. Immunoreactive fibres are present throughout the central nervous system. The 5HT-immunoreactive neuronal somata characteristically appear in groups, located mainly in the cerebral, pedal, visceral and right parietal ganglia. The majority of 5HT-immunoreactive neurons is located in the pedal ganglia. Additionally a dense network of 5HT-immunoreactive varicose fibres is found in the neural sheath of the central nervous system including all the nerves and ganglia. The number and distribution of 5HT-immunoreactive neurons correlates with that demonstrated by 5,6-dihydroxytryptamine labelling method.     

Reconstruction of dopaminergic neural network and locomotion function in planarian regenerates

Planarian, an invertebrate flatworm, has a high capacity for regeneration when compared with other worms and animals. We show here for the first time that the reconstructed dopamine (DA) neural network regulates locomotion and behavior in planarian regenerates. The gene encoding tyrosine hydroxylase in the planarian Dugesia japonica (DjTH) was identified. DjTH protein was coexpressed with aromatic amino acid decarboxylase-like A (DjAADCA) in the planarian central nervous system (CNS). In addition, DjTH-knockdown planarians lost the ability to synthesize DA, but showed no change in 5-hydroxytryptamine synthesis. When the planarian body was amputated, DjTH-positive neurons were regenerated in the brain newly rebuilt from the tail piece at Day 3, and the DjTH-positive axonal and dendritic neural network in the CNS (dopaminergic tiara) was reconstructed at Days 5-7. At that time, autonomic locomotion and methamphetamine-induced hyperkinesia were also suppressed in DjTH-knockdown planarians. Planarian locomotion and behavior seem to be regulated in both cilia- and muscle-dependent manners. In DjTH-knockdown planarians, muscle-mediated locomotion and behavior were significantly attenuated. These results suggest that DA neurons play a key role in the muscle-mediated movement in planarians.     

Aspects of the neuroendocrine control of ovulation and broodiness in the domestic hen

The neuroendocrine control of ovulation and broodiness in the domestic hen involves complex interactions between hypothalamic neuropeptides, neurotransmitters, and ovarian steroids which regulate the secretion of luteinizing hormone (LH) and prolactin. Nuclear progesterone receptor is localized in many neurons throughout the hypothalamus but is absent from LHRH neurons. Hence, the positive feedback action of progesterone on LH release is not mediated by a genomic mechanism within the LHRH neuron. Precursors of 5-hydroxytryptamine (5HT) and dopamine (DA) inhibit the preovulatory release of LH, while the turnover rates of these neurotransmitters in the anterior hypothalamus decrease when preovulatory levels of LH are at their highest. Further, a population of receptors for 5HT which occurs in the anterior hypothalamus in laying birds is absent in nonlaying, incubating hens. Taken together, these observations suggest that the preovulatory surge of LH is mediated by a transitory decrease in the inhibitory action of 5HT and possibly DA, on the secretion of LHRH. Neurons containing 5HT may play a role in the regulation of prolactin release and, more specifically, in the control of broodiness. Drugs which enhance the function of 5HT neurons stimulate prolactin release while increased prolactin secretion in incubating hens is associated with an increase in the turnover of 5HT in the anterior hypothalamus. No receptors for 5HT were demonstrable in the anterior pituitary gland, showing that the prolactin-releasing activity of 5HT must be mediated by a prolactin-releasing factor (PRF). A candidate for a physiological PRF is vasoactive intestinal polypeptide (VIP).(ABSTRACT TRUNCATED AT 250 WORDS)     

The serotonin receptor SER-1 (5HT2ce) contributes to the regulation of locomotion in Caenorhabditis elegans

Serotonin (5-hydroxytryptamine: 5HT) is an important neuroactive substance in the model roundworm, Caenorhabditis elegans. Aside from having effects in feeding and egg-laying, 5HT inhibits motility and also modulates several locomotory behaviors, notably food-induced slowing and foraging. Recent evidence showed that a serotonergic 5HT2-like receptor named SER-1 (also known as 5HT2ce) was responsible for the effect of 5HT on egg-laying. Here we confirm this observation and show that SER-1 also plays an important role in locomotion. A mutant lacking SER-1 was found to be highly resistant to exogenous 5HT in the absence of food and this resistant phenotype was rescued by reintroducing the SER-1 gene in a mutant background. Pharmacological studies showed that the same antagonists that blocked the activity of recombinant SER-1 in vitro also inhibited the effect of 5HT on motility, suggesting the same receptor was responsible for both effects. When tested for locomotory behaviors, the SER-1 mutant was found to be moderately defective in food-induced slowing. In addition, the mutant changed direction more frequently than the wildtype when searching for food, suggesting that SER-1 may play a role in navigational control during foraging. Both these effects required the presence of MOD-1, a 5HT gated chloride channel, and the results indicate that SER-1 and MOD-1 modulate these behaviors through a common pathway. On the basis of expression analysis of a ser-1::GFP translational fusion, SER-1 is prominently located in central, integrating neurons of the head ganglia (RIA and RIC) but not the body wall musculature. The evidence suggests that SER-1 controls locomotion through indirect modulation of neuromuscular circuits and has effects both on speed and direction of movement.     

Altered serotonin and dopamine metabolism in the CNS of serotonin 5-HT(1A) or 5-HT(1B) receptor knockout mice

Measurements of serotonin (5-HT), dopamine (DA), and noradrenaline, and of 5-HT and DA metabolites, were obtained by HPLC from 16 brain regions and the spinal cord of 5-HT(1A) or 5-HT(1B) knockout and wild-type mice of the 129/Sv strain. In 5-HT(1A) knockouts, 5-HT concentrations were unchanged throughout, but levels of 5-HT metabolites were higher than those of the wild type in dorsal/medial raphe nuclei, olfactory bulb, substantia nigra, and locus coeruleus. This was taken as an indication of increased 5-HT turnover, reflecting an augmented basal activity of midbrain raphe neurons and consequent increase in their somatodendritic and axon terminal release of 5-HT. It provided a likely explanation for the increased anxious-like behavior observed in 5-HT(1A) knockout mice. Concomitant increases in DA content and/or DA turnover were interpreted as the result of a disinhibition of DA, whereas increases in noradrenaline concentration in some territories of projection of the locus coeruleus could reflect a diminished activity of its neurons. In 5-HT(1B) knockouts, 5-HT concentrations were lower than those of the wild type in nucleus accumbens, locus coeruleus, spinal cord, and probably also several other territories of 5-HT innervation. A decrease in DA, associated with increased DA turnover, was measured in nucleus accumbens. These changes in 5-HT and DA metabolism were consistent with the increased aggressiveness and the supersensitivity to cocaine reported in 5-HT(1B) knockout mice. Thus, markedly different alterations in CNS monoamine metabolism may contribute to the opposite behavioral phenotypes of these two knockouts.     

Effect of Aging on 24-Hour Changes in Serotonin and Dopamine Turnover, and Somatostatin and Amino Acid Content, of the Rat Hypothalamus and Pituitary Gland

The effect of aging on neurotransmitter and peptide content in the hypothalamichypophysial unit has commonly been analyzed at single time points in the 24-h cycle. Since significant changes in circadian rhythmicity occur during aging, this study aimed to examine 24-h rhythmicity in hypothalamic and pituitary serotonin (5HT) and dopamine (DA) turnover and content, and somatostatin and amino acid content in 2 months-old and 18-20 months-old rats, killed at 6 different time intervals throughout the light-dark cycle. Aged rats showed suppressed or disrupted 24-h rhythms of 5HT and DA turnover and of somatostatin, glutamate, aspartate and taurine content (anterior hypothalamus), of 5HT and DA turnover and of somatostatin, glutamate, taurine and glycine content (medial hypothalamus) and of DA turnover and amino acid content (posterior hypothalamus). Twenty-four h variations in DA, somatostatin, aspartate, GABA and glycine content of the anterior hypophysis and in all parameters tested in the neurointermediate lobe became suppressed or disrupted in aged rats. Mean values generally decreased with age, except for DA content in the anterior pituitary lobe and aspartate content in the neurointermediate lobe. Conclusions: Examination of neurotransmitter and neuropeptide content at different times of the day is needed to analyze the effects of aging in the hypothalamic-hypophysial unit.     

D-2 dopamine antagonist-like effects of SCH 23390 on A9 and A10 dopamine neurons

The effects of SCH 23390 on d-amphetamine-induced suppression of A9 and A10 DA neuronal firing were determined. SCH 23390 potently reversed d-amphetamine on both A9 and A10 DA neurons. Compared to haloperidol, SCH 23390 was 5 times more potent on A9 DA neurons and 20 times more potent on A10 DA neurons. However, the magnitude of the reversal effect was greater with haloperidol than SCH 23390. In addition, haloperidol produced a further increase in firing of both A9 and A10 DA neurons after SCH 23390 maximally increased firing. It was concluded that SCH 23390 has D-2 DA antagonist-like properties, possibly mediated via an interaction at D-1 DA receptors, which may be functionally linked with D-2 DA receptors. The marked potency of SCH 23390 in reversing d-amphetamine could be due to its combined antagonist effects at 5HT2 and D-1 DA receptor sites.     

Conjunctive processing of locomotor signals by the ventral tegmental area neuronal population

The ventral tegmental area (VTA) plays an essential role in reward and motivation. How the dopamine (DA) and non-DA neurons in the VTA engage in motivation-based locomotor behaviors is not well understood. We recorded activity of putative DA and non-DA neurons simultaneously in the VTA of awake mice engaged in motivated voluntary movements such as wheel running. Our results revealed that VTA non-DA neurons exhibited significant rhythmic activity that was correlated with the animal's running rhythms. Activity of putative DA neurons also correlated with the movement behavior, but to a lesser degree. More importantly, putative DA neurons exhibited significant burst activation at both onset and offset of voluntary movements. These findings suggest that VTA DA and non-DA neurons conjunctively process locomotor-related motivational signals that are associated with movement initiation, maintenance and termination.     

Effects of haloperidol and phentolamine on the crustacean cardiac ganglion

Haloperidol (a dopamine D₂ blocker in vertebrates) and phentolamine (an α-adrenergic blocker) alter the pattern of bursting by the isolated cardiac ganglion of the lobster when perfused at concentrations of 10^?6–10^?5 mol/l. Both drugs decrease the frequency of bursting and increase burst duration. They are most effective in slowing the ganglion when applied selectively to the anterior ganglionic trunk, the same region of the ganglion where dopamine (DA) and 5-hydroxytryptamine (5HT) are most effective in speeding up bursting. When exogenous monoamine transmitters are applied in the presence of 3×10^?6 mol/l haloperidol, the effect of 5HT, but not of DA, is significantly reduced. At the same concentration, phentolamine does not suppress the actions of DA, 5HT or noradrenaline (NA). Both haloperidol and phentolamine significantly alter the properties of endogenous burst-organizing potentials (driver potentials) generated by motorneurons in the ganglion. It is possible that the effects of these drugs on bursting reflect alteration of endogenous electrical properties of the constituent neurons, rather than receptor antagonism.     

Regulation of dopaminergic neuron firing by heterogeneous dopamine autoreceptors in the substantia nigra pars compacta

Dopamine (DA) receptors generate many cellular signals and play various roles in locomotion, motivation, hormone production, and drug abuse. According to the location and expression types of the receptors in the brain, DA signals act in either stimulatory or inhibitory manners. Although DA autoreceptors in the substantia nigra pars compacta are known to regulate firing activity, the exact expression patterns and roles of DA autoreceptor types on the firing activity are highly debated. Therefore, we performed individual correlation studies between firing activity and receptor expression patterns using acutely isolated rat substantia nigra pars compacta DA neurons. When we performed single-cell RT-PCR experiments, D(1), D(2)S, D(2)L, D(3), and D(5) receptor mRNA were heterogeneously expressed in the order of D(2)L > D(2)S > D(3) > D(5) > D(1). Stimulation of D(2) receptors with quinpirole suppressed spontaneous firing similarly among all neurons expressing mRNA solely for D(2)S, D(2)L, or D(3) receptors. However, quinpirole most strongly suppressed spontaneous firing in the neurons expressing mRNA for both D(2) and D(3) receptors. These data suggest that D(2) S, D(2)L, and D(3) receptors are able to equally suppress firing activity, but that D(2) and D(3) receptors synergistically suppress firing. This diversity in DA autoreceptors could explain the various actions of DA in the brain.