Daria Peleg-Raibstein

Late Prenatal Immune Activation in Mice Leads to Behavioral and Neurochemical Abnormalities Relevant to the Negative Symptoms of Schizophrenia

Based on the human epidemiological association between prenatal infection and higher risk of schizophrenia, a number of animal models have been established to explore the long-term brain and behavioral consequences of prenatal immune challenge. Accumulating evidence suggests that the vulnerability to specific forms of schizophrenia-related abnormalities is critically influenced by the precise timing of the prenatal immunological insult. In the present study, we tested the hypothesis whether late prenatal immune challenge in mice may induce long-term behavioral and neurochemical dysfunctions primarily associated with the negative symptoms of schizophrenia. We found that prenatal exposure to the viral mimic polyriboinosinic-polyribocytidilic acid (Poly-I:C; 5 mg/kg, i.v.) on gestation day (GD) 17 led to significant deficits in social interaction, anhedonic behavior, and alterations in the locomotor and stereotyped behavioral responses to acute apomorphine (APO) treatment in both male and female offspring. In addition, male but not female offspring born to immune challenged mothers displayed behavioral/cognitive inflexibility as indexed by the presence of an abnormally enhanced latent inhibition (LI) effect. Prenatal immune activation in late gestation also led to numerous, partly sex-specific changes in basal neurotransmitter levels, including reduced dopamine (DA) and glutamate contents in the prefrontal cortex and hippocampus, as well as reduced γ-aminobutyric acid (GABA) and glycine contents in the hippocampus and prefrontal cortex, respectively. The constellation of behavioral and neurochemical abnormalities emerging after late prenatal Poly-I:C exposure in mice leads us to conclude that this immune-based experimental model provides a powerful neurodevelopmental animal model especially for (but not limited to) the negative symptoms of schizophrenia.

Maternal high-fat diet in mice programs emotional behavior in adulthood.

The maternal environment has a significant role in the normal development of the fetus and may have long-term impact on brain development including critical central pathways such as the gamma-aminobutyric acid (GABA), serotonergic and the neurotrophin systems. For example, maternal malnutrition plays an important role in programming many aspects of physiology and behavior including predisposition to mental-health related disorders such as anxiety. Here we investigated the effects of maternal high-fat diet or control diet for nine weeks (prior to gestation, gestation and lactation) on the adult offspring with respect to anxiety related behaviors as well as exploration and conditioned fear response. We found that offspring born to high-fat diet mothers showed increased anxiety-like behaviors, but intact conditioned fear response and exploratory behavior. In addition, brain-derived neurotrophic factor (BDNF) was significantly increased in the dorsal hippocampus, while GABA(A) alpha2 receptor subunit and 5-hydroxytryptamine 1A (5-HT1A) receptor showed increased levels in the ventral hippocampus. In summary, these findings suggest that maternal high-fat diet consumption during critical periods in the development of the fetus, might increase the risk of abnormal behaviors in adulthood related to anxiety.

Effects of dorsal and ventral hippocampal NMDA stimulation on nucleus accumbens core and shell dopamine release

This study has analysed the effects of infusing N-methyl-D-aspartate (NMDA) into either the ventral or dorsal hippocampus on dopamine (DA) transmission in the nucleus accumbens (NAC) core or shell for the first time. Dopamine was measured using in vivo microdialysis with high performance liquid chromatography with electrochemical detection (HPLC-EC). Unilateral NMDA infusion (0.5 microg) into the ventral hippocampus (VH) increased extracellular DA levels in NAC shell during the first 30 min following infusion compared to saline (SAL) infused animals. In contrast, NAC core DA levels were unaffected. NMDA infusion into the dorsal hippocampus (DH) led to a decrease in NAC core DA levels; this effect was not observed in the SAL-infused group. DA levels in NAC shell remained unaltered. At the end of the experiments, we examined the response to a systemic amphetamine (AMPH) injection of 1mg/kg on extracellular DA levels of the NAC core and shell. Interestingly, on2ly animals previously infused with NMDA into the VH exhibited a sensitized DA response in the NAC shell in response to the AMPH injection. We can conclude that VH activation has an acute stimulatory effect on DA release in the shell and that DH activation has a suppressive effect on extracellular DA levels in the core.

ACTIVATION OF DOPAMINERGIC NEUROTRANSMISSION IN THE MEDIAL PREFRONTAL CORTEX BY N-METHYL-D-ASPARTATE STIMULATION OF THE VENTRAL HIPPOCAMPUS IN RATS

Many behavioral functions-including sensorimotor, attentional, memory, and emotional processes-have been associated with hippocampal processes and with dopamine transmission in the medial prefrontal cortex (mPFC). This suggests a functional interaction between hippocampus and prefrontal dopamine. The anatomical substrate for such an interaction is the intimate interconnection between the ventral hippocampus and the dopamine innervation of the mPFC. The present study yielded direct neurochemical evidence for an interaction between ventral hippocampus and prefrontal dopamine transmission in rats by demonstrating that subconvulsive stimulation of the ventral hippocampus with N-methyl-d-aspartate (NMDA; 0.5 mug/side) activates dopamine transmission in the mPFC. Postmortem measurements revealed that bilateral NMDA stimulation of the ventral hippocampus, resulting in locomotor hyperactivity, increased the homovanillic acid/dopamine ratio, an index of dopamine transmission, in the mPFC; indices of dopamine transmission in any of five additionally examined forebrain regions (amygdala, nucleus accumbens shell/core, lateral prefrontal cortex, caudate putamen) were unaltered. In vivo microdialysis measurements in freely moving rats corroborated the suggested activation of prefrontal dopamine transmission by demonstrating that unilateral NMDA stimulation of the ventral hippocampus increased extracellular dopamine in the ipsilateral mPFC. The suggested influence of the ventral hippocampus on prefrontal dopamine may be an important mechanism for hippocampo-prefrontal interactions in normal behavioral processes. Moreover, it indicates that aberrant hippocampal activity, as found in neuropsychiatric diseases, such as schizophrenia and mood disorders, may contribute to disruption of certain cognitive and emotional functions which are extremely sensitive to imbalanced prefrontal dopamine transmission.

Prepulse inhibition predicts working memory performance whilst startle habituation predicts spatial reference memory retention in C57BL/6 mice.

Prepulse inhibition (PPI) of the acoustic startle reflex refers to the attenuation of the startle response to an intense pulse stimulus when it is shortly preceded by a weak non-startling prepulse stimulus. It is a well-established high-throughput translational measure of pre-attentive sensory gating, and its impairment is detected in several neuropsychiatric diseases including schizophrenia. It has been hypothesized that PPI might be associated with, or predictive of, cognitive deficiency in such diseases, and therefore provide an efficient assay for screening drugs with potential pro-cognitive efficacy. Free from any predetermined disease model, the present study evaluated in a homogeneous cohort of inbred C57BL/6 mice the presence of a statistical link between PPI expression and cognitive performance. Performance indices in a spatial reference memory test and a working memory test conducted in the Morris water maze, and contextual fear conditioning were correlated against pre-existing baseline PPI expression. A specific correlative link between working memory and PPI induced by weak (but not strong) prepulse was revealed. In addition, a correlation between habituation of the startle reflex and reference memory was identified for the first time: a stronger overt habituation effect was associated with superior spatial search accuracy. The PPI paradigm thus provides two independent predictors of dissociable cognitive traits in normal C57BL/6 mice; and they might serve as potential markers for high-throughput evaluation of potential cognitive enhancers, especially in the context of schizophrenia where deficits in startle habituation and PPI co-exist.

Withdrawal from repeated amphetamine administration leads to disruption of prepulse inhibition but not to disruption of latent inhibition

Summary.The present study represents a continuous effort to develop an animal model of schizophrenia based on the “endogenous dopamine sensitization” hypothesis. To achieve this goal, withdrawal from an escalating amphetamine (AMPH) regime administration [three injections per day over a period of 4 days and increasing doses from 1 to 10 mg/kg of AMPH or an equivalent volume of saline (SAL)] was employed. Animals exposed to this treatment were evaluated on their performance in attentional (Latent inhibition, LI) and sensorimotor gating (Prepulse inhibition, PPI) tasks in a drug free state and tested for locomotor sensitization following a low dose of AMPH challenge administration.LI using active avoidance, tested on withdrawal day 4, was unaffected. PPI of the acoustic startle response, measured on withdrawal days 6 and 70, was disrupted. On the 76th day of withdrawal, a low challenge dose of AMPH (1 mg/kg) led to a clear locomotor sensitization effect.

Differential effects on prepulse inhibition of withdrawal from two different repeated administration schedules of amphetamine.

In this study, rats were tested in behavioural paradigms relevant to schizophrenia during withdrawal from two different administration schedules of amphetamine (Amph). One of the escalating administration schedules, which has been employed in previous studies, consisted of three daily injections for 6 d with increasing dosages from 1 to 5 mg/kg Amph (Esc-5) and was compared to a hitherto never examined escalating administration schedule [three injections per day for 6 d escalating from 1 to 8 mg/kg Amph (Esc-8)]. Control animals received an equivalent volume of saline (Sal) injections according to the same schedule. Whereas rats treated with Esc-5, as reported before, failed to show an effect on prepulse inhibition (PPI), the Esc-8-treated rats exhibited a long-lasting disruption of PPI in a drug-free state on days 6, 13 and 55 of withdrawal. The Amph-pretreated animals demonstrated a similar magnitude of behavioural sensitization following an Amph challenge on withdrawal day 58 irrespective of the administration schedule. To evaluate if the withdrawal from the two Amph schedules led to a change in brain monoamine levels, a subgroup of animals was neurochemically examined in post-mortem for eight parameters in seven brain regions on withdrawal day 55. Withdrawal from the Esc-8 schedule induced reduced dopamine levels in the caudate putamen. Only this neurochemical finding and the PPI attenuation differentiated the Esc-8 animals from the Esc-5 and Sal animals. These data suggest that, based on the endogenous sensitization hypothesis of schizophrenia, the persistent disruption of PPI observed in animals withdrawn from Esc-8 can be used as a valid animal model of specific symptoms of schizophrenic patients.

Differential effects of post-weaning juvenile stress on the behaviour of C57BL/6 mice in adolescence and adulthood

RationaleThere is evidence that events early in post-weaning life influence brain development and subsequent adult behaviour and therefore play an important role in the causation of certain psychiatric disorders in later life. Exposing rodents to stressors during the juvenile period has been suggested as a model of induced predisposition for these disorders.ObjectiveThis is the first study to examine behavioural and pharmacological changes in adolescence and adulthood following juvenile stress in mice.Materials and methodsTwo cohorts of mice were simultaneously exposed to a stress protocol during postnatal days (PND) 25–30. Behavioural assessments reflecting emotional functions, cognitive functions, and psychostimulant sensitivity were then carried out at two time points: one cohort was tested during adolescence (PND 39–54; adolescent group), and the second cohort was tested during adulthood (PND 81–138; adult group).ResultsIn the adolescent mice, juvenile stress significantly attenuated conditioned freezing and led to decreased anxiety-like behaviour in the elevated plus maze, whereas no effect was observed on these tests in the adult mice. In contrast, adult mice exhibited poor avoidance learning following juvenile stress. When tested during adulthood, the mice stressed during the juvenile period showed a sensitised response to amphetamine compared to controls, whereas the response during adolescence was similar in stressed and control animals.ConclusionsOur results suggest that exposure to stressors during the juvenile period can exert long-term effects on the brain and behaviour and that these effects differ depending on whether the animals are tested during adolescence or adulthood.

Effects of withdrawal from an escalating dose of amphetamine on conditioned fear and dopamine response in the medial prefrontal cortex.

Neurochemical studies have shown that mesocortical dopamine projections are particularly responsive in aversive situations such as fear conditioning. The present study assessed behavioural and medial prefrontal cortex (mPFC) dopamine responses utilizing in vivo microdialysis during acquisition and expression of a conditioned fear response. In two independent experiments, rats were presented with either two or nine tone-shock pairings during formation of a conditioned fear response. In the second experiment, rats were pre-treated with repeated injections of either amphetamine or saline over a 6-day period and tested during withdrawal. Amphetamine pre-treatment as well as the conditioning procedure itself potentiated an increase in dopamine levels during formation, but not expression of a conditioned fear response. Locomotor activity induced by an amphetamine challenge (1mg/kg) was also enhanced in pre-treated amphetamine compared to saline pre-treated animals (experiment two). However, mPFC dopamine response to amphetamine challenge did not differ between treatment groups. We conclude that while the exact role of mPFC dopamine in behavioural sensitization is yet to be determined, mPFC dopamine release may underlie the increased fear response during acquisition but not expression of fear response.

Enhanced sensitivity to drugs of abuse and palatable foods following maternal overnutrition

Epidemiological studies have shown an association between maternal overnutrition and increased risk of the progeny for the development of obesity as well as psychiatric disorders. Animal studies have shown results regarding maternal high-fat diet (HFD) and a greater risk of the offspring to develop obesity. However, it still remains unknown whether maternal HFD can program the central reward system in such a way that it will imprint long-term changes that will predispose the offspring to addictive-like behaviors that may lead to obesity. We exposed female dams to either laboratory chow or HFD for a period of 9 weeks: 3 weeks before conception, during gestation and lactation. Offspring born to either control or HFD-exposed dams were examined in behavioral, neurochemical, neuroanatomical, metabolic and positron emission tomography (PET) scan tests. Our results demonstrate that HFD offspring compared with controls consume more alcohol, exhibit increased sensitivity to amphetamine and show greater conditioned place preference to cocaine. In addition, maternal HFD leads to increased preference to sucrose as well as to HFD while leaving the general feeding behavior intact. The hedonic behavioral alterations are accompanied by reduction of striatal dopamine and by increased dopamine 2 receptors in the same brain region as evaluated by post-mortem neurochemical, immunohistochemical as well as PET analyses. Taken together, our data suggest that maternal overnutrition predisposes the offspring to develop hedonic-like behaviors to both drugs of abuse as well as palatable foods and that these types of behaviors may share common neuronal underlying mechanisms that can lead to obesity.