Imaging dopamine transmission in the frontal cortex: a simultaneous microdialysis and [11C]FLB 457 PET study.

In a recent human positron emission tomography (PET) study we demonstrated the ability to detect amphetamine-induced dopamine (DA) release in the prefrontal cortex as a reduction in the binding of the DA D(2/3) radioligand [(11)C]FLB 457. A key requirement for validating this paradigm for use in clinical studies is demonstrating that the changes in [(11)C]FLB 457 binding observed with PET following amphetamine are related to changes in dialysate DA concentration as measured with microdialysis. Microdialysis and PET experiments were performed to compare, in five rhesus monkeys, amphetamine-induced DA release and [(11)C]FLB 457 displacement in the frontal cortex after three doses of amphetamine (0.3 mg kg(-1), 0.5 mg kg(-1) and 1.0 mg kg(-1)). Amphetamine led to a significant dose-dependent increase in dialysate (0.3 mg kg(-1): 999±287%; 0.5 mg kg(-1): 1320±432%; 1.0 mg kg(-1): 2355±1026%) as measured with microdialysis and decrease in [(11)C]FLB 457 binding potential (BP(ND), 0.3 mg kg(-1): -6±6%; 0.5 mg kg(-1): -16±4%; 1.0 mg kg(-1): -24±2%) as measured with PET. The relationship between amphetamine-induced peak ΔDA and Δ[(11)C]FLB 457 BP(ND) in the frontal cortex was linear. The results of this study clearly demonstrate that the magnitude of dialysate DA release is correlated with the magnitude of the reduction in [(11)C]FLB 457 BP(ND) in the frontal cortex. The use of the [(11)C]FLB 457-amphetamine imaging paradigm in humans should allow for characterization of prefrontal cortical DA release in neuropsychiatric disorders such as schizophrenia and addiction.

Cognitive impact of genetic variation of the serotonin transporter in primates is associated with differences in brain morphology rather than serotonin neurotransmission.

A powerful convergence of genetics, neuroimaging and epidemiological research has identified the biological pathways mediating individual differences in complex behavioral processes and the related risk for disease. Orthologous genetic variation in non-human primates (NHPs) represents a unique opportunity to characterize the detailed molecular and cellular mechanisms that bias behaviorally and clinically relevant brain function. We report that a rhesus macaque orthologue of a common polymorphism of the serotonin transporter gene (rh5-HTTLPR) has strikingly similar effects on behavior and brain morphology to those in humans. Specifically, the rh5-HTTLPR (S)hort allele broadly affects cognitive choice behavior and brain morphology without observably affecting the 5-hydroxytryptamine (5-HT) transporter or 5-HT(1A) concentrations in vivo. Collectively, our findings indicate that 5-HTTLPR-associated behavioral effects reflect genotype-dependent biases in cortical development rather than static differences in serotonergic signaling mechanisms. Moreover, these data highlight the vast potential of NHP models in advancing our understanding of human genetic variation affecting behavior and neuropsychiatric disease liability.

Chronic cold exposure potentiates CRH-evoked increases in electrophysiologic activity of locus coeruleus neurons.

BACKGROUND: Chronic stress exposure can produce sensitization of norepinephrine release in the forebrain in response to subsequent stressors. Furthermore, the increase in norepinephrine release in response to the stress-related peptide corticotropin-releasing hormone (CRH) is potentiated by prior chronic stress exposure. To explore possible mechanisms underlying these alterations in norepinephrine release, we examined the effect of chronic stress on the electrophysiologic activity of locus coeruleus (LC) neurons in response to centrally applied CRH. METHODS: Single-unit recordings of LC neurons in halothane-anesthetized rats were used to compare the effect of intraventricular administration of CRH (0.3-3.0 microg) in control and previously cold-exposed (2 weeks at 5 degrees C) rats. RESULTS: The CRH-evoked increase in LC neuron activity was enhanced following chronic cold exposure, without alteration in basal activity of LC neurons. The enhanced CRH-evoked activation was apparent at higher doses of CRH but not at lower ones, resulting in an increased slope of the dose-response curve for CRH in previously cold-exposed rats. CONCLUSIONS: These data, in combination with previous data, suggest that the sensitivity of LC neurons to excitatory inputs is increased following chronic cold exposure. The altered functional capacity of LC neurons in rats after continuous cold exposure may represent an experimental model to examine the role of central noradrenergic neurons in anxiety and mood disorders.

Sensitization of norepinephrine release in medial prefrontal cortex: effect of different chronic stress protocols.

Previously, we demonstrated that continuous exposure of rats to cold (5 degrees C) for 2-3 weeks potentiates the increase in extracellular norepinephrine in the medial prefrontal cortex produced by acute tail shock. In the present study, we used in vivo microdialysis to examine whether this sensitization of evoked norepinephrine release also occurs in the medial prefrontal cortex following exposure to other chronic stress protocols. Rats exposed to 30 min of intermittent foot shock (0.6 mA) each day for 14 days, did not exhibit a greater increase in extracellular norepinephrine in response to acute tail shock. To determine whether this discrepancy between cold exposure and foot shock might be related to differences in the nature or the pattern of exposure to the chronic stressor, we also examined the effect of intermittent exposure to cold or continuous exposure to a foot shock protocol on tail shock-evoked norepinephrine release. Sensitized norepinephrine release did not develop following either intermittent exposure to cold (5 degrees C; 4 h/day for 14 days) or continuous exposure to a foot shock protocol (0.6 mA trains at random intervals 24 h/day for 14 days), suggesting that both the nature of the stressor as well as the pattern of exposure to the chronic stressor play a role in the development of sensitized norepinephrine release.

Clozapine preferentially increases dopamine release in the rhesus monkey prefrontal cortex compared with the caudate nucleus.

Despite substantial differences between species in the organization and elaboration of the cortical dopamine innervation, little is known about the pharmacological response of cortical or striatal sites to antipsychotic medications in nonhuman primates. To examine this issue, rhesus monkeys were chronically implanted with guide cannulae directed at the principal sulcus, medial prefrontal cortex, premotor cortex, and caudate nucleus. Alterations in dopamine release in these discrete brain regions were measured in response to administration of clozapine or haloperidol. Clozapine produced significant and long-lasting increases in dopamine release in the principal sulcus, and to a lesser extent, in the caudate nucleus. Haloperidol did not produce a consistent effect on dopamine release in the principal sulcus, although it increased dopamine release in the caudate. Clozapine's preferential augmentation of dopamine release in the dorsolateral prefrontal cortex supports the idea that clozapine exerts its therapeutic effects in part by increasing cortical dopamine neurotransmission.

Impact of corticotropin-releasing hormone on extracellular norepinephrine in prefrontal cortex after chronic cold stress.

We have previously demonstrated that exposing rats to cold (5 degrees C) for 3-4 weeks potentiates the increase in extracellular norepinephrine (NE) in the medial prefrontal cortex produced by acute tail shock. In the present study, we used microdialysis to determine the duration of cold exposure required to produce this sensitization and explored the mechanism of the phenomenon. Tail shock elicited a twofold greater increase in extracellular NE in the medial prefrontal cortex of rats exposed to cold for 2 weeks than in naive control rats or in rats exposed to cold for 1 week and tested either immediately or after a 2-week delay. Local infusion of 10 microM D-amphetamine or 30 mM K+ increased extracellular NE in the medial prefrontal cortex (approximately 350 and 190%, respectively) comparably in control rats and rats exposed to cold for 3 weeks. In contrast, intraventricular administration of 3.0 microg of corticotropin-releasing hormone increased extracellular NE in the medial prefrontal cortex by 65% in rats exposed to cold for 2 weeks, but only 35% in control rats. These results indicate that an enhanced responsiveness of noradrenergic neurons to acute tail shock (1) requires approximately 2 weeks of cold exposure to develop and (2) may be mediated by a change at the level of the noradrenergic cell bodies rather than the nerve terminals.

Characterization of excitatory amino acid modulation of dopamine release in the prefrontal cortex of conscious rats.

The effect of various classes of excitatory amino acid agonists on the release of dopamine in the medial prefrontal cortex (PFC) of awake rats was examined using intracerebral microdialysis. Local infusion of 20 muM alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA), through the microdialysis probe, produced a significant increase of more than twofold in extra-cellular levels of dopamine. Application of 100 microM AMPA increased these levels nearly 15 fold. The AMPA/kainate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (50microM) blocked the increase in dopamine release produced by 20 microM AMPA. Local infusion of kainate at concentrations of 5 and 20 microM increased dopamine release by nearly 150 and 500%, respectively. Local application of CMQX (50 microM) before 20 microM kainate significantly attenuated the stimulatory effect of kainate on dopamine levels. In contrast to AMPA and kainate, infusion of N-methyl-D-aspartate (NMDA) at 20 or 100 microM did not increase dopamine release. In fact, a trend toward a decrease in dopamine release was evident after 100 microM NMDA. The present study indicates that the in vivo release of dopamine in the PFC is facilitated by AMPA and kainate receptors. This modulation is more profound than that previously reported in basal ganglia. The lack of an excitatory effect of NMDA is in agreement with recent reports that the NMDA receptor may inhibit indirectly dopaminergic neurotransmission in the PFC.

Glutamatergic control of dopamine release during stress in the rat prefrontal cortex.

In vivo microdialysis was used to assess the hypothesis that the stress-induced increase in dopamine release in the prefrontal cortex is mediated by stress-activated glutamate neurotransmission in this region. Local perfusion of an alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA)/kainate receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione, blocked the stress-induced increase in dopamine levels, whereas an NMDA receptor antagonist, 2-amino-5-phosphonopentanoic acid, at the dose tested, was not able to alter this response significantly. These data indicate that the effect of stress on dopamine release in the prefrontal cortex is mediated locally by activation of AMPA/kainate receptors, which modulate the release of dopamine in this region.