Role of serotonin and dopamine receptor binding in antipsychotic efficacy.

In an effort to analyse the contribution of individual serotonin and dopamine receptor subtypes to antipsychotic medication response, we analysed the correlation between clinically effective antipsychotic drug dose and binding affinity to cloned serotonin and dopamine receptor subtypes. Clinically effective dosage and binding affinity to the D(2) dopamine receptor subtype were moderately correlated for typical antipsychotic medications (r=0.57, p=0.04), and were similarly modestly correlated for atypical antipsychotic drugs (r=0.66, p=0.07). Surprisingly for typical antipsychotic medications, a stronger inverse correlation was observed between drug dosage and 5-HT(2C) affinity (r=-0.65, p=0.03). The strongest correlation observed for typical antipsychotic medications was between medication dose and 5-HT(2C)/D(2) binding affinity ratio (r=-0.81, p=0.002). For atypical antipsychotic medications, highly significant correlations were observed between medication dose and receptor-binding affinity to D(3) dopamine receptor (r=0.78, p=0.02), and with the ratios of D(2)/5-HT(1A) (r=0.85, p=0.009), D(3)/5-HT(1A) (r=0.78, p=0.021), D(2) (5-HT(2A)/5-HT(1A)) (r=0.75, p=0.033) and D(3) (5-HT(2A)/5-HT(1A)) (r=0.75, p=0.03) receptor-binding affinities. The correlation between medication dose and D(2) (5-HT(2C)/5-HT(1A)) receptor-binding affinity ratio was of similar magnitude (r=0.70, p=0.055). No significant correlations were identified between atypical antipsychotic medication dose and 5-HT(1A), 5-HT(2A), 5-HT(2C), 5-HT(2C)/D(2) or 5-HT(2A)/D(2) receptor-binding affinities. These observations suggest an interaction between D(2) and 5-HT(2C) receptor-binding effects contributing to the therapeutic response achieved following treatment with typical antipsychotic medications. This suggests that for typical antipsychotic medications, constitutive serotonin 5-HT(2C) receptor signalling interacts with and facilitates the antipsychotic benefit achieved through dopamine D(2) receptor blockade. Additionally, this analysis demonstrates that, in contrast to typical antipsychotic medications, the therapeutic effectiveness of atypical antipsychotic medications results from opposing interactions among three distinct domains: (1) antipsychotic potency is enhanced by increased D(2) and D(3) dopamine receptor-binding affinity; (2) antipsychotic efficacy is also facilitated by increased binding affinity to serotonin 5-HT(2C) and 5-HT(2A) receptors; (3) in contrast, antipsychotic potency is reduced by elevations in 5-HT(1A) receptor-binding affinity.

The dopamine D3 receptor antagonist NGB 2904 increases spontaneous and amphetamine-stimulated locomotion.

The dopamine D3 receptor is believed to play an important role in regulation of rodent locomotor behavior, and has been proposed as a therapeutic target for substance abuse, psychotic disorders, and Parkinson's disease. One model of dopamine D3 receptor function, based on studies utilizing D3 receptor knockout mice and D3 receptor-preferring agonists, proposes that D3 receptor stimulation is inhibitory to psychostimulant-induced locomotion, in opposition to the effects of concurrent dopamine D1 and D2 receptor stimulation. Recent progress in medicinal chemistry has led to the development of highly-selective dopamine D3 receptor antagonists. In order to extend our understanding of D3 dopamine receptor's behavioral functions, we determined the effects of the highly-selective dopamine D3 receptor antagonist NGB 2904 on amphetamine-stimulated and spontaneous locomotion in wild-type and dopamine D3 receptor knockout mice. NGB 2904 (26.0 microg/kg s.c.) enhanced amphetamine-stimulated locomotion in wild-type mice, but had no measurable effect in dopamine D3 receptor knockout mice. Of a range of doses (0.026 microg-1.0 mg/kg) given acutely or once daily for seven days, the highest dose of NGB 2904 (1.0 mg/kg) stimulated spontaneous locomotion in wild-type mice, but was without measurable effect in dopamine D3 receptor knockout mice. These behavioral effects of NGB 2904 contrast with those described for other highly D3 receptor-selective antagonists, which have not previously demonstrated an effect on spontaneous locomotor activity. In combination, these data add to the behavioral profile of this novel D3 receptor ligand and provide further support for a role for dopamine D3 receptor inhibitory function in the modulation of rodent locomotion.

Dopamine and serotonin receptor binding and antipsychotic efficacy.

The relationship between clinically effective antipsychotic drug dosage and binding affinity to cloned dopamine (DA) and serotonin receptor subtypes was analyzed in an effort to elucidate the contribution of individual receptor subtypes to medication response. Clinically effective dose and binding affinity to D(2) DA receptor were modestly correlated for typical antipsychotic medications (r=0.54, p=0.046), but surprisingly were not correlated for atypical antipsychotics (r=0.41, p=0.31). For typical antipsychotics, a more robust inverse relationship was observed between medication dose and 5-HT(2C) affinity (r=-0.68, p=0.021). The strongest correlation for typical antipsychotics was observed between drug dosage and 5-HT(2C)/D(2) binding affinity ratio (r=-0.81, p=0.003). For atypical antipsychotics, no significant correlations were identified between medication dosage and 5-HT(2C), 5-HT(2A), 5-HT(2C)/D(2), or 5-HT(2A)/D(2) receptor-binding affinities. In contrast, atypical antipsychotic medication dosage was highly correlated with the ratios of D(2) (5-HT(2A)/5-HT(1A)) (r=0.80, p=0.031), and D(2) (5-HT(2C)/5-HT(1A)) (r=0.78, p=0.038) binding affinities. These observations demonstrate an interaction between D(2) and 5-HT(2C) receptor effects contributing to positive symptom response for typical antipsychotic medications, suggesting that signaling through 5-HT(2C) receptors interacts with and improves antipsychotic effects achieved via D(2) receptor blockade. This analysis also demonstrates that, in contrast to typical antipsychotics, therapeutic effects of atypical antipsychotic medications are determined by opposing interactions among three different domains: (1) increasing D(2) DA receptor-binding affinity enhances antipsychotic potency. (2) Increasing 5-HT(2C) and 5-HT(2A) receptor-binding affinities also facilitate antipsychotic efficacy. (3) Increasing 5-HT(1A) receptor-binding affinity, in contrast, reduces antipsychotic efficacy.

Relative expression of D3 dopamine receptor and alternative splice variant D3nf mRNA in high and low responders to novelty.

Studies in rodents suggest an important role for the D3 dopamine receptor in regulating locomotor responses to spatial novelty and psychostimulants. The D3 receptor alternatively spliced variant D3nf produces a non-dopamine binding protein that may alter D3 receptor localization by dimerizing with the full-length receptor. In the high responder/low responder (HR/LR) model, the locomotor response to an inescapable, novel spatial environment predicts individual differences in the locomotor and rewarding effects of psychostimulants. We hypothesized that individual differences in D3 receptor expression could contribute to individual differences in the locomotor response to novelty in the HR/LR model. To test this hypothesis, we screened rats for response to a novel spatial environment and analyzed brain tissue for mRNA levels of the D3 receptor and D3nf by real-time RT-PCR. The ratios of D3/D3nf mRNA in prefrontal cortex and substantia nigra/ventral tegmentum were significantly lower in HRs than in LRs. There were no differences in relative expression of D3/D3nf between HRs and LRs in nucleus accumbens. These data further support a role for the D3 dopamine receptor in behavioral responses to novelty and, given the established relationship between novelty and psychostimulant responses, suggest that the D3 receptor may be an important target for assessment of drug abuse vulnerability. Additionally, these findings are consistent with the hypothesis that alternative splicing may contribute to regulation of D3 dopamine receptor function.

Altered behavioral response to dopamine D3 receptor agonists 7-OH-DPAT and PD 128907 following repetitive amphetamine administration.

Behavioral sensitization, the progressive and enduring enhancement of certain behaviors following repetitive drug use, is mediated in part by dopaminergic pathways. Increased locomotor response to drug treatment, a sensitizable behavior, is modulated by an opposing balance of dopamine receptor subtypes, with D1/D2 dopamine receptor stimulation increasing and D3 dopamine receptor activation inhibiting amphetamine-induced locomotion. We hypothesize that tolerance of D3 receptor locomotor inhibition contributes to behavioral sensitization. In order to test the hypothesis that expression of behavioral sensitization results in part from release of D3 receptor-mediated inhibition, thereby resulting in decreased response to D3 receptor agonists, we examined the effect of repetitive amphetamine administration on the behavioral response to the D3 receptor preferring agonists 7-OH-DPAT and PD 128907. D3-selective effects have recently been described for both drugs at a low dose. At 1 week following completion of a repetitive treatment regimen, amphetamine-pretreated rats displayed a decreased response to D3-selective doses of both 7-OH-DPAT and PD 128907, when compared to animals receiving saline pretreatment. Moreover, in addition to the quantitative alteration in response, there was a change in the inter-relation between response to amphetamine and D3 agonist. A highly significant inverse relation between locomotor inhibitory response to PD 128907 and the locomotor-stimulant response to amphetamine was observed prior to amphetamine treatment. In contrast, 10 days following repetitive amphetamine treatment, the relation between response to PD 128907 and amphetamine was not detected. The observed behavioral alteration could not be accounted for by changes in D3 receptor binding in ventral striatum. These findings suggest a persistent release of D3 receptor-mediated inhibitory influence contributes to the expression of behavioral sensitization to amphetamine.

7-OH-DPAT and PD 128907 selectively activate the D3 dopamine receptor in a novel environment.

The D3 dopamine receptor is expressed primarily in limbic brain areas, and appears to play an inhibitory role in rodent locomotor behavior. Evidence suggests a potential role for the D3 receptor in the pathology of neuropsychiatric disease. Progress in elucidating D3 receptor function has been hampered, however, by a lack of well-characterized, selective ligands and by conflicting information regarding the behavioral phenotype of D3 receptor knockout mice. Here, we describe studies evaluating the behavioral effects of (+/-)-7-hydroxy-N,N-di-n-propyl-2-aminotetralin (7-OH-DPAT) and PD 128907, two D3 receptor agonists whose in vivo selectivity has been a topic of considerable controversy. We demonstrate that both compounds inhibit locomotion under novel environmental conditions in wild-type (WT) mice, but are without measurable behavioral effect under identical conditions in D3 receptor knockout mice. Additionally, we demonstrate that at low, D3 selective doses, these compounds are without behavioral effect in both WT and D3 receptor knockout mice that have acclimated to the testing environment. These findings suggest that D3 receptor stimulation inhibits novelty-stimulated locomotion, and establish conditions for the use of 7-OH-DPAT and PD 128907 as D3 receptor agonists in vivo. Potential implications of these observations are discussed.