Kehong Zhang

Animal models of attention-deficit hyperactivity disorder.

Attention-deficit hyperactivity disorder (ADHD) involves clinically heterogeneous dysfunctions of sustained attention, with behavioral overactivity and impulsivity, of juvenile onset. Experimental models, in addition to mimicking syndromal features, should resemble the clinical condition in pathophysiology, and predict potential new treatments. One of the most extensively evaluated animal models of ADHD is the spontaneously hypertensive rat. Other models include additional genetic variants (dopamine transporter gene knock-out mouse, coloboma mouse, Naples hyperexcitable rat, acallosal mouse, hyposexual rat, and population-extreme rodents), neonatal lesioning of dopamine neurons with 6-hydroxydopamine, and exposure to other neurotoxins or hippocampal irradiation. None is fully comparable to clinical ADHD. The pathophysiology involved varies, including both deficient and excessive dopaminergic functioning, and probable involvement of other monoamine neurotransmitters. Improved models as well as further testing of their ability to predict treatment responses are required.

Long-term effects of olanzapine, risperidone, and quetiapine on serotonin 1A, 2A and 2C receptors in rat forebrain regions

Abstract.Rationale: Serotonin (5-HT) and its receptors have been implicated in various neuropsychiatric disorders. Altered serotonergic neurotransmission and interactions between 5-HT and dopamine (DA) systems may contribute to the pathophysiology of idiopathic psychotic or manic disorders. Interactions with 5-HT receptors may also contribute to special properties of modern antipsychotic drugs not yet evaluated for long-term effects on 5-HT receptors. Objective and methods: We surveyed effects of newer atypical antipsychotics on 5-HT receptor types 1A, 2A, and 2C in rat forebrain regions by quantitative receptor autoradiography with selective radioligands following 28 days of continuous infusion of drugs or control vehicle. Results: Infusion of olanzapine, risperidone, and quetiapine increased 1A, but decreased 2A receptor labeling in frontal cerebral cortex. Olanzapine decreased binding at 2C receptors in hippocampal CA1 and CA3 regions and perhaps entorhinal cortex; olanzapine, but neither risperidone nor quetiapine, also decreased 2C labeling in caudate-putamen. Conclusions: The findings suggest that altered 5-HT1A and 5-HT2A receptor levels in frontal cortex, and 5-HT2C receptors in other forebrain regions, may contribute to psychopharmacological properties of these novel atypical antipsychotic agents, perhaps including their antipsychotic or antimanic actions, and low risk of adverse extrapyramidal effects.

Role of Dopamine D4 Receptors in Motor Hyperactivity Induced by Neonatal 6-Hydroxydopamine Lesions in Rats

The role of dopamine D4 receptors in behavioral hyperactivity was investigated by assessing D4 receptor expression in brain regions and behavioral effects of D4 receptor-selective ligands in juvenile rats with neonatal 6-hydroxydopamine lesions, a laboratory model for attention deficit-hyperactivity disorder (ADHD). Autoradiographic analysis indicated that motor hyperactivity in lesioned rats was closely correlated with increases in D4 but not D2 receptor levels in caudate-putamen. D4-selective antagonist CP-293,019 dose-dependently reversed lesion-induced hyperactivity, and D4-agonist CP-226,269 increased it. These results indicate a physiological role of dopamine D4 receptors in motor behavior, and may suggest much-needed innovative treatments for ADHD.

Regulation of working memory by dopamine D4 receptor in rats.

Working memory is regulated by neurotransmitters in prefrontal cortex (PFC), including dopamine and norepinephrine. Previous studies of dopamine function in working memory have focused on the D1 and D2 receptors, with most evidence suggesting a dominant role for the D1 receptor. Since the dopamine D4 receptor is highly expressed in PFC, we hypothesize that it may also contribute to working memory. To test this hypothesis, we examined behavioral effects of L-745,870, a highly selective, centrally active, D4 antagonist, using a delayed alternation task in rats. Task performance was dose-dependently affected by the D4 antagonist, depending on individual baseline functional status of working memory. In rats with good baseline performance, the D4 antagonist had no effects at low doses, whereas high doses disrupted working memory. In rats with poor baseline working memory, the D4 antagonist significantly improved working memory at low doses, and higher doses were not distinguishable from vehicle controls. Effects of the D4 antagonist among poor performers were most robust when task demand for working memory was high, with lesser effects at lower demand level, suggesting that such effects were selective for working memory. The present findings indicate a significant role of the D4 receptor in working memory, and suggest innovative, D4-based, treatment of cognitive deficits associated with neuropsychiatric disorders.

Dopamine D4 Receptors: Beyond Schizophrenia

Dopamine D4 receptors mediate a wide range of neuronal signal transduction cascades. Malfunctions of these mechanisms may contribute to the pathophysiology of neuropsychiatric disorders, and their modification underlies the actions of many psychotropic drugs. Postmortem neuropathological and genetic studies provide inconclusive associations between D4 receptors and schizophrenia. Clinical trials of partially selective lead D4 antagonists have proved them to be ineffective against psychotic symptoms in patients diagnosed with schizophrenia. However, associations are emerging between D4 receptors and other neuropsychiatric disorders, including attention-deficit hyperactivity disorder as well as specific personality traits such as novelty seeking. Preclinical studies indicate that D4 receptors play a pivotal role in the cellular mechanisms of hyperactivity, impulsivity, and working memory. Accordingly, D4 receptors have broader implications for human illnesses than has been suggested by early focus on psychotic illness as a clinical target, and selective D4 agents may yield clinically useful drugs for several neuropsychiatric disorders that require improved treatments.

Plasticity of Dopamine D4 Receptors in Rat Forebrain: Temporal Association with Motor Hyperactivity Following Neonatal 6-Hydroxydopamine Lesioning

Genetic studies suggest that dopamine D4 receptor polymorphism is associated with attention deficit hyperactivity disorder (ADHD). We recently reported that motor hyperactivity in juvenile male rats with neonatal 6-hydroxydopamine lesions of the central dopamine system can be reversed by dopamine D4 receptor-selective antagonists. In this study, effects of such lesions on D4 as well as other dopamine receptors (D1 and D2) were autoradiographically quantified at selected developmental stages. Neonatal lesions resulted in motor hyperactivity at postnatal day (PD) 25, but not at PD 37 or 60. Correspondingly, D4 receptor levels in lesioned rats were substantially increased in caudate-putamen and decreased in nucleus accumbens at PD 25, but not at PD 37 or 60. Neonatal lesions also led to relatively minor changes in D1 and D2 receptor binding in various forebrain regions. However, the time-course of lesion-induced motor hyperactivity correlated only with changes in D4, but not D1 and D2 receptors. These results further support the hypothesis that D4 receptors may play a pivotal role in lesion-induced hyperactivity, and possibly in clinical ADHD.

Enhanced expression of dopamine D(1) and glutamate NMDA receptors in dopamine D(4) receptor knockout mice.

Expression of dopamine ([DA] D1 and D2) and glutamate ([Glu]), (N-methyl-d-aspartic acid [NMDA], α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid [AMPA], and kanaic acid [KA]) receptor types were analyzed autoradiographically in forebrain regions of D4 receptor knockout mice and their wild-type controls. Selective radioligand binding to D4 receptors was virtually absent in D4 receptor knockout mouse brain in contrast to significant specific D4 binding in forebrain tissue of wild-type controls. Labeling of D1 receptors was significantly increased in nucleus accumbens (NAc; 39%) and caudate putamen (CPu; 42%) of D4-knockout mice vs wild-type controls. In addition, NMDA receptor labeling was significantly increased in NAc (31%), CPu (40%), and hippocampal CA1 (21%) and CA3 (25%) regions of D4 knockouts vs wild-type controls. No changes in D2, AMPA or KA receptors were found. The findings suggest that D1, D4, and NMDA receptors might interact functionally and that developmental absence of D4 receptors might trigger compensatory mechanisms that enhance expression of D1 receptors in NAc and CPu, and NMDA receptors in NAc, CPu, and hippocampus. The findings also encourage cautious interpretation of results in knockout mice with targeted absence of specific genes, as complex adaptive changes not directly related to the missing gene might contribute to physiological and behavioral responses.

Dopamine depletion abolishes apomorphine- and amphetamine-induced increases in extracellular serotonin levels in the striatum of conscious rats: A microdialysis study

We investigated how serotonergic neurotransmission was affected by 6-hydroxydopamine (6-OHDA) lesioning of the adult rat brain dopamine (DA) system. In this animal model for Parkinsons disease (PD), the effect of destroying ascending DA pathways on extracellular levels of serotonin (5-HT) and 5-HT innervation in rat striatum were examined. Profound unilateral lesions of the nigrostriatal DA pathways were made by infusing 6-OHDA unilaterally into either the right medial forebrain bundle or the right substantia nigra. At 5 weeks after lesioning extracellular levels of DA and 5-HT were determined with microdialysis and high-pressure liquid chromatography under basal conditions and after systemic injections of apomorphine or amphetamine. DA nerve-terminal destruction and 5-HT innervation were determined with quantitative autoradiography. 6-OHDA lesioning reduced extracellular levels of DA below detection limits and led to statistically significant increases in extracellular 5-HT. Apomorphine, and amphetamine, respectively increased extracellular 5-HT to 8.2- and 2.2-fold above baseline levels in intact animals; these effects were absent in 6-OHDA-lesioned animals. Basal levels of [(3)H]paroxetine binding to 5-HT transporters in caudate-putamen increased by 41% in 6-OHDA-lesioned animals. These results suggest that 6-OHDA lesioning led to hyperinnervation of 5-HT nerve terminals and increases in basal extracellular 5-HT levels, but also to an unexplained loss of apomorphine and amphetamine-induced release of 5-HT. Addressing whether this impairment has significance in the onset of PD might lead to development of new strategies to manage parkinsonian symptoms.

GABAB receptors: altered coupling to G-proteins in rats sensitized to amphetamine

Modified dopamine and glutamate neurotransmission in discrete brain regions is implicated in stimulant-induced behavioral sensitization. Release of both neurotransmitters is influenced by GABA(B) metabotropic receptors for the principal inhibitory neurotransmitter GABA. Accordingly, GABA(B) receptors were examined in rats sensitized to amphetamine by measuring receptor density and coupling to G-proteins indicated as [(3)H]baclofen binding and baclofen-mediated [(35)S]GTP gamma S binding. Repeated treatment with (+)-amphetamine (5mg/kg per day, i.p., for five days) sensitized the rats to amphetamine challenge (1mg/kg) at 14 days, but not one day, later. GABA(B) receptor density was not altered at either time. Baclofen-mediated [(35)S]GTP gamma S binding, however, was selectively augmented in the prefrontal cortex and attenuated in the nucleus accumbens at 14 days, but not one day, after amphetamine treatment. Changes in GABA(B) receptor coupling to G-proteins in rats sensitized to amphetamine, but not in similarly treated but unsensitized rats, lead us to suggest that altered GABA(B) receptor functioning may contribute to the expression of amphetamine-induced behavioral sensitization.

Long-term effects of newer antipsychotic drugs on neuronal nitric oxide synthase in rat brain.

Neuronal nitric oxide synthase (nNOS) catalyzes the synthesis of neuronal nitric oxide from L-arginine. Behavioral and neurochemical studies implicate neuronal nitric oxide in the pathophysiology of schizophrenia and in the actions of standard antipsychotic drugs. However, involvement of nNOS in the actions of newer antipsychotic drugs requires further investigation. Accordingly, density levels of nNOS, a marker for neuronal nitric oxide production, were examined in rat forebrain regions by computed autoradiography after repeated treatment (28 days) with three newer antipsychotic agents, olanzapine, risperidone, and quetiapine. No significant differences in nNOS levels were detected in representative cortical, limbic, and extrapyramidal brain regions of drug-treated vs vehicle-treated animals. The findings suggest that nNOS plays a minimal role in mediating the long-term actions of newer antipsychotic drugs.