Differences in extracellular glutamate levels in striatum of rats with high and low exploratory activity.

BACKGROUND: Major inter-individual differences exist in vulnerability to anxiety and affective disorders, and the underlying neurobiology could help in understanding the predisposition to these disorders and treatment resistance. Recently the glutamatergic system has become a target in the development of novel antidepressants. METHODS: We compared extracellular glutamate levels in low (LE-) and high exploring (HE-) rats in hippocampus and striatum at baseline and after inhibition of re-uptake by perfusion with l-trans-pyrrolidine-2,4-dicarboxylate (PDC, 4mM). Glutamate levels in microdialysates were measured by HPLC after derivatization. RESULTS: In the striatum, baseline glutamate levels in young adult LE-rats and HE-rats were not significantly different, but the response to uptake inhibition was: perfusion with PDC increased extracellular glutamate levels in both LE- and HE-rats, but to a significantly lower extent in LE-rats. Although the characteristic levels of exploration of LE- and HE-rats had previously been shown to be stable up to 8 months of age, we identified a subgroup of HE-rats whose exploration levels had drastically dropped by age of 11 months (formerly HE-rats, HEF), and analyzed their data separately. There were no differences in the PDC-evoked striatal glutamate release between the three groups; however, the baseline glutamate levels were higher in the HEF-subgroup compared to the HE- and LE-animals. In the CA1 area of hippocampus, there were no differences in extracellular glutamate levels between the LE- and HE-rats either at baseline or after inhibition of uptake. CONCLUSION: These results suggest that inter-individual differences in exploratory behaviour may be related to striatal glutamatergic neurotransmission.

On the role of NOS1 ex1f-VNTR in ADHD-allelic, subgroup, and meta-analysis.

Attention deficit/ hyperactivity disorder (ADHD) is a heritable neurodevelopmental disorder featuring complex genetics with common and rare variants contributing to disease risk. In a high proportion of cases, ADHD does not remit during adolescence but persists into adulthood. Several studies suggest that NOS1, encoding nitric oxide synthase I, producing the gaseous neurotransmitter NO, is a candidate gene for (adult) ADHD. We here extended our analysis by increasing the original sample, adding two further samples from Norway and Spain, and conducted subgroup and co-morbidity analysis. Our previous finding held true in the extended sample, and also meta-analysis demonstrated an association of NOS1 ex1f-VNTR short alleles with adult ADHD (aADHD). Association was restricted to females, as was the case in the discovery sample. Subgroup analysis on the single allele level suggested that the 21-repeat allele caused the association. Regarding subgroups, we found that NOS1 was associated with the hyperactive/impulsive ADHD subtype, but not to pure inattention. In terms of comorbidity, major depression, anxiety disorders, cluster C personality disorders and migraine were associated with short repeats, in particular the 21-repeat allele. Also, short allele carriers had significantly lower IQ. Finally, we again demonstrated an influence of the repeat on gene expression in human post-mortem brain samples. These data validate the role of NOS-I in hyperactive/impulsive phenotypes and call for further studies into the neurobiological underpinnings of this association. © 2015 Wiley Periodicals, Inc.

Neuroinformatic analyses of common and distinct genetic components associated with major neuropsychiatric disorders.

Major neuropsychiatric disorders are highly heritable, with mounting evidence suggesting that these disorders share overlapping sets of molecular and cellular underpinnings. In the current article we systematically test the degree of genetic commonality across six major neuropsychiatric disorders-attention deficit hyperactivity disorder (ADHD), anxiety disorders (Anx), autistic spectrum disorders (ASD), bipolar disorder (BD), major depressive disorder (MDD), and schizophrenia (SCZ). We curated a well-vetted list of genes based on large-scale human genetic studies based on the NHGRI catalog of published genome-wide association studies (GWAS). A total of 180 genes were accepted into the analysis on the basis of low but liberal GWAS p-values (<10(-5)). 22% of genes overlapped two or more disorders. The most widely shared subset of genes-common to five of six disorders-included ANK3, AS3MT, CACNA1C, CACNB2, CNNM2, CSMD1, DPCR1, ITIH3, NT5C2, PPP1R11, SYNE1, TCF4, TENM4, TRIM26, and ZNRD1. Using a suite of neuroinformatic resources, we showed that many of the shared genes are implicated in the postsynaptic density (PSD), expressed in immune tissues and co-expressed in developing human brain. Using a translational cross-species approach, we detected two distinct genetic components that were both shared by each of the six disorders; the 1st component is involved in CNS development, neural projections and synaptic transmission, while the 2nd is implicated in various cytoplasmic organelles and cellular processes. Combined, these genetic components account for 20-30% of the genetic load. The remaining risk is conferred by distinct, disorder-specific variants. Our systematic comparative analysis of shared and unique genetic factors highlights key gene sets and molecular processes that may ultimately translate into improved diagnosis and treatment of these debilitating disorders.

Wfs1-deficient mice display altered function of serotonergic system and increased behavioral response to antidepressants.

It has been shown that mutations in the WFS1 gene make humans more susceptible to mood disorders. Besides that, mood disorders are associated with alterations in the activity of serotonergic and noradrenergic systems. Therefore, in this study, the effects of imipramine, an inhibitor of serotonin (5-HT) and noradrenaline (NA) reuptake, and paroxetine, a selective inhibitor of 5-HT reuptake, were studied in tests of behavioral despair. The tail suspension test (TST) and forced swimming test (FST) were performed in Wfs1-deficient mice. Simultaneously, gene expression and monoamine metabolism studies were conducted to evaluate changes in 5-HT- and NA-ergic systems of Wfs1-deficient mice. The basal immobility time of Wfs1-deficient mice in TST and FST did not differ from that of their wild-type littermates. However, a significant reduction of immobility time in response to lower doses of imipramine and paroxetine was observed in homozygous Wfs1-deficient mice, but not in their wild-type littermates. In gene expression studies, the levels of 5-HT transporter (SERT) were significantly reduced in the pons of homozygous animals. Monoamine metabolism was assayed separately in the dorsal and ventral striatum of naive mice and mice exposed for 30 min to brightly lit motility boxes. We found that this aversive challenge caused a significant increase in the levels of 5-HT and 5-hydroxyindoleacetic acid (5-HIAA), a metabolite of 5-HT, in the ventral and dorsal striatum of wild-type mice, but not in their homozygous littermates. Taken together, the blunted 5-HT metabolism and reduced levels of SERT are a likely reason for the elevated sensitivity of these mice to the action of imipramine and paroxetine. These changes in the pharmacological and neurochemical phenotype of Wfs1-deficient mice may help to explain the increased susceptibility of Wolfram syndrome patients to depressive states.

Evidence for impaired function of dopaminergic system in Wfs1-deficient mice.

Immunohistological studies suggest abundant expression of Wfs1 protein in neurons and nerve fibers that lie in the vicinity of dopaminergic (DA-ergic) fibers and neurons. Therefore, we sought to characterize the function of DA-ergic system in Wfs1-deficient mice. In wild-type mice, amphetamine, an indirect agonist of DA, caused significant hyperlocomotion and increase in tissue DA levels in the dorsal and ventral striatum. Both effects of amphetamine were significantly blunted in homozygous Wfs1-deficient mice. Motor stimulation caused by apomorphine, a direct DA receptor agonist, was somewhat stronger in Wfs1-deficient mice compared to their wild-type littermates. However, apomorphine caused a similar reduction in levels of DA metabolites (3,4-dihydroxyphenylacetic acid and homovanillic acid) in the dorsal and ventral striatum in all genotypes. Behavioral sensitization to repeated treatment with amphetamine (2.5 mg/kg) was observed in wild-type, but not in Wfs1-deficient mice. The expression of DA transporter gene (Dat) mRNA was significantly lower in the midbrain of male and female homozygous mice compared to wild-type littermates. Altogether, the blunted effects of amphetamine and the reduced gene expression of DA transporter are probably indicative of an impaired functioning of the DA-ergic system in Wfs1-deficient mice.

Lsamp⁻/⁻ mice display lower sensitivity to amphetamine and have elevated 5-HT turnover.

In mice, the limbic system-associated membrane protein (Lsamp) gene has been implicated in locomotion, anxiety, fear reaction, learning, social behaviour and adaptation. Human data links the LSAMP gene to several psychiatric disorders and completed suicide. Here, we investigated changes in major monoamine systems in mice lacking the Lsamp gene. First, the locomotor and rewarding effects of amphetamine were studied in Lsamp(-/-) mice and Lsamp(+/+) mice. Second, monoamine levels in major brain regions in response to saline and amphetamine injections were measured and, third, the expression levels of dopamine system-related genes in the brain were studied in these mice. Lsamp(-/-) mice displayed lower sensitivity to amphetamine in the motility box. Likewise, in the place preference test, the rewarding effect of amphetamine was absent in Lsamp(-/-) mice. In all brain regions studied, Lsamp(-/-) mice displayed lower serotonin (5-HT) baseline levels, but a greater 5-HT turnover rate, and amphetamine increased the level of 5-HT and lowered 5-HT turnover to a greater extent in Lsamp(-/-) mice. Finally, Lsamp(-/-) mice had lower level of dopamine transporter (DAT) mRNA in the mesencephalon. In conclusion, Lsamp-deficiency leads to increased endogenous 5-HT-ergic tone and enhanced 5-HT release in response to amphetamine. Elevated 5-HT function and reduced activity of DAT are the probable reasons for the blunted effects of amphetamine in these mice. Lsamp(-/-) mice are a promising model to study the neurobiological mechanisms of deviant social behaviour and adaptation impairment observed in many psychiatric disorders.

Brain responses to chronic social defeat stress: effects on regional oxidative metabolism as a function of a hedonic trait, and gene expression in susceptible and resilient rats.

Chronic social defeat stress, a depression model in rats, reduced struggling in the forced swimming test dependent on a hedonic trait-stressed rats with high sucrose intake struggled less. Social defeat reduced brain regional energy metabolism, and this effect was also more pronounced in rats with high sucrose intake. A number of changes in gene expression were identified after social defeat stress, most notably the down-regulation of Gsk3b and Map1b. The majority of differences were between stress-susceptible and resilient rats. Conclusively, correlates of inter-individual differences in stress resilience can be identified both at gene expression and oxidative metabolism levels.

Differential gene expression in a rat model of depression based on persistent differences in exploratory activity.

Affective disorders are often accompanied by changes in motivation and anxiety. We investigated the genome-wide gene expression patterns in an animal model of depression that separates Wistar rats belonging into clusters of persistently high anxiety/low motivation to explore and low anxiety/high motivation to explore (low explorers and high explorers, LE and HE, respectively), in three brain regions previously implicated in mood disorders (raphe, hippocampus and the frontal cortex). Several serotonin-, GABA-, and glutamatergic genes were differentially expressed in LE- and HE-rats. The analysis of Gene Ontology biological process terms associated with the differentially regulated genes identified a significant overrepresentation of genes involved in the neuron development, morphogenesis, and differentiation; the most enriched pathways from the Kyoto Encyclopedia of Genes and Genomes were the Wnt signalling, MAPK signalling, long-term potentiation, and long-term depression pathways. These findings corroborate some expression data from other models of depression, and suggest additional targets.

Rats with persistently high exploratory activity have both higher extracellular dopamine levels and higher proportion of D(2) (High) receptors in the striatum.

Increases in both striatal dopamine release and the proportion of the D(2) receptors in the high affinity state (D(2) (High)) accompany the behavioral sensitization to psychostimulants, but it is not known whether the physiological substrate of the interindividual differences locomotor and exploratory behavior is similar. Thus, we examined whether persistently high spontaneous exploratory activity is associated with extracellular dopamine as well as the proportion of D(2) (High) in the striatum. Extracellular dopamine levels were found to be significantly higher in rats with high exploratory activity (high explorers, HE) as compared with low explorers (LE) in baseline conditions as well as after administration of amphetamine (0.5 mg/kg, i.p.). Also, the HE animals had significantly higher proportion of striatal D(2) (High) receptors than the LE-rats (43.8 +/- 4.4% and 22.5 +/- 1.5%, respectively). Thus, the present findings support the notion that concomitant higher extracellular dopamine levels and the proportion of D(2) (High) receptors in the striatum, whether naturally occurring and persistent or pharmacologically induced, are causally related to high behavioral activity.

Amphetamine-induced locomotion, behavioral sensitization to amphetamine, and striatal D2 receptor function in rats with high or low spontaneous exploratory activity: differences in the role of locus coeruleus.

Individual differences in novelty-related behavior are associated with sensitivity to various neurochemical manipulations. In the present study the amphetamine-induced locomotor activity and behavioral sensitization to amphetamine (0.5 mg/kg) was investigated in rats with high or low spontaneous exploratory activity (HE- and LE-rats, respectively) after partial denervation of the locus coeruleus (LC) projections with a low dose of the selective neurotoxin DSP-4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine; 10 mg/kg). DSP-4 produced a partial depletion (about 30%) of noradrenaline in the frontal cortex of both HE- and LE-rats; additionally the levels of metabolites of dopamine and 5-HT were reduced in the frontal cortex and nucleus accumbens of the LE-rats. Amphetamine-stimulated locomotor activity was attenuated by the DSP-4 pretreatment only in the HE-rats and this effect persisted over repeated testing. Behavioral sensitization to repeated amphetamine was evident only in the LE-rats with intact LC projections. Repeated amphetamine treatment reduced D(2) receptor mediated stimulation of [(35)S]GTPgammaS-binding and dopamine-dependent change in GDP-binding affinity in the striatum, but only in HE-rats. The absence of amphetamine sensitization in HE-rats could thus be related to the downregulation by amphetamine of the G protein stimulation through D(2) receptors. Conclusively, acute and sensitized effects of amphetamine depend on the integrity of LC projections but are differently regulated in animals with high or low trait of exploratory activity. These findings have implications to the neurobiology of depression, drug addiction, and attention deficit hyperactivity disorder.

Rats with persistently low or high exploratory activity: behaviour in tests of anxiety and depression, and extracellular levels of dopamine.

Behaviour in novel environments is influenced by the conflicting motivators fear and curiosity. Because changes in both of these motivational processes are often simultaneously involved in human affective disorders, we have developed the exploration box test which allows separation of animals belonging to clusters with inherent high neophobia/low motivation to explore and low neophobia/high motivation to explore (LE and HE, respectively). In a novel home-cage, no behavioural differences were found between LE- and HE-rats, suggestive that it is not the general locomotor activity but specific features of the exploration box test that bring about the differences. In studies on both Wistar and Sprague-Dawley rats we found that the trait of exploratory activity remains stable over long periods of time and that LE and HE animals display differences in many other behavioural tests related to mood disorders. Namely, LE animals were found to display enhanced anxiety-like behaviour and to be generally less active in the elevated plus-maze, used more passive coping strategies in the forced swimming test, and acquired a more persistent association between neutral and stressful stimuli in fear conditioning test. LE animals consumed more sucrose solution in non-deprived conditions. We also found that both at baseline and in response to d-amphetamine (0.5mg/kg) administration, LE-rats had lower extracellular dopamine levels in striatum but not in nucleus accumbens. In conclusion, LE-rats appear more inhibited in their activity in typical animal tests of anxiety and are more susceptible to acute stressful stimuli.

Effect of CCK1 and CCK2 receptor blockade on amphetamine-stimulated exploratory behavior and sensitization to amphetamine.

Interactions between dopaminergic neurotransmission and cholecystokinin (CCK) in the CNS may be important in the pathogenesis of psychotic disorders and substance abuse. In this study, the effect of coadministration of the selective CCK receptor antagonists devazepide and L-365,260 (for selectively blocking CCK1 and CCK2 receptors, respectively), on the effect of amphetamine on the rat exploratory behavior, and on sensitization of locomotor response to amphetamine, were studied. Amphetamine (0.5 mg/kg) increased exploratory activity in the exploration box for 5 consecutive testing days, while devazepide (10 microg/kg) blocked and L-365,260 (10 microg/kg) enhanced amphetamine-induced stimulation of activity. Devazepide coadministration prevented the development of sensitization to amphetamine, while coadministration of L-365,260 with amphetamine potentiated the locomotor effect of a challenge dose of amphetamine. These results suggest that endogenous CCK, released during exploratory activity, shapes behavioral responses to amphetamine by acting on both receptor subtypes, and modulates the development of sensitization to amphetamine.