Peggy Schneider

A Novel Elevated Plus-Maze Procedure to Avoid the One-Trial Tolerance Problem

The elevated plus-maze (EPM) test is one of the most commonly used behavioral assays to evaluate anxiety-related behavior in rodents. It is an economic test (5 min duration) without prior conditioning of the animals. The critical measure for anxiety is the time spent in the open arms of the maze. A confounding problem of the EPM is the so called one-trial tolerance (OTT), characterized by a marked decrease of open arm exploration in spite of treatment with anxiolytic acting benzodiazepines upon re-exposure to the EPM. This consistent finding is often raised as an evidence for the inappropriateness to re-test rodents in the EPM. However, a reliable re-test paradigm would broaden the usability and effectiveness of this test. Therefore, we tested how an extension of the inter-trial interval to 28 days (instead of the usual 24 h), and an additional change of the testing room would affect the open arm time and other behaviors on the EPM. In two experiments, drug-naive Wistar rats were exposed to the EPM on trial 1, and treated intraperitoneally with either vehicle or midazolam (0.25 mg/kg) 30 min before trial 2. Then, trial 2 (28 days after trial 1) was carried out in either the same testing room (Experiment 1) or in another unfamiliar room (Experiment 2). Twenty-eight days after trial 1 the open arm time of the rats in the vehicle treated control rats of both experimental groups was comparable to that of the first trial, independent of the testing room. Most importantly, we found that the treatment with the benzodiazepine midazolam had a significantly anxiolytic-like (i.e., increase of open arm time) effect in trial 2 only when conducted in the previously unfamiliar testing room (Experiment 2). We suggest that in order to reliably re-test the EPM and to prevent confounding effects due to the OTT, an inter-trial interval of 28 days and a change in testing rooms reinstates anxiolytic-like actions of benzodiazepines.

The Elevated Plus-Maze Test: Differential Psychopharmacology of Anxiety-Related Behavior

The role of individual factors in behavioral neuroscience is an important, but still neglected, area of research. For example, the Elevated Plus-Maze Test has been one of the most used paradigms to gauge unconditioned aversively motivated behavior in rodents. However, despite a great number of experiments with this test there have been only few efforts to assess systematic individual variations in the elevated plus-maze and related neurobiological functions. The present review aims to give, first, a general overview and introduction about the test, and second, an animal model of anxiety based on natural variance of plus-maze behavior within a given unselected population of rats. Finally, critical aspects of such approaches in animal research are discussed, and suggestions are given as to where to go from here.

Adolescent peer-rejection persistently alters pain perception and CB1 receptor expression in female rats.

Peer-interactions are particularly important during adolescence and teenagers display enhanced sensitivity toward rejection by peers. Social rejection has been shown to induce alterations in pain perception in humans. However, the neurobiological consequences of adolescent social rejection have yet to be extensively characterized, and no appropriate animal model is available. Here, we propose inadequate playful interactions in adolescent rats as a novel animal model for social peer-rejection and examine potential long-term consequences into adulthood. Acute social pairing of female adolescent Wistar rats with an age-matched rat from the less playful Fischer344 strain was found to alter social play and decrease pain reactivity, indicating Fischer rats as inadequate social partners for Wistar animals. Therefore, in a second experiment, adolescent female Wistar rats were either reared with another Wistar rat (adequate social rearing; control) or with a Fischer rat (inadequate social rearing; play-deprived). Beginning on day 50, all Wistar rats were group housed with same-strain partners and tested for behavioral, neurobiological and endocrine differences in adulthood. Playful peer-interactions were decreased during adolescence in play-deprived animals, without affecting social contact behavior. Consequently, adult play-deprived rats showed decreased pain sensitivity and increased startle reactivity compared to controls, but did not differ in activity, anxiety-related behavior or social interaction. Both groups also differed in their endocrine stress-response, and expression levels of the cannabinoid CB1 receptor were increased in the thalamus, whereas FAAH levels were decreased in the amygdala. The present animal model therefore represents a novel approach to assess the long-term consequences of peer-rejection during adolescence.

Central metabolite changes and activation of microglia after peripheral interleukin-2 challenge.

Interleukin (IL)-2 regulates the immune response through the proliferation of activated T-cells and also exerts effects on the central nervous system (CNS). Alongside having marked neurobehavioral effects, IL-2 has been suggested to impact on various psychiatric disorders. The immune-CNS communication of IL-2 remains unclear, although, it is suggested that microglia are the source and target of IL-2. Here, we analyzed changes in brain metabolites following a peripheral IL-2 challenge and examined the contribution of microglia in mediating these effects. Rats were assessed by magnetic resonance spectroscopy (MRS) in a 9.4 T scanner for baseline metabolite levels in the prefrontal cortex (PFC) and the hippocampus. After 7 days animals were scanned again following a single injection of IL-2 (2.5 μg/kg) and then tested on the elevated plus-maze for the correlation of IL-2-induced brain metabolites and measures of anxiety. In another experiment CD25(+) microglia cells were determined. A separate group of rats was injected either with IL-2 or vehicle, and afterward the PFC and hippocampus were dissected and fluorescence activated cell sorting (FACS) analysis was performed. The MRS scans in the intra-individual study design showed a significant increase in myo-inositol in the analyzed regions. A significant correlation of anxiety-like measures and myo-inositol, a marker for microglia activity, was found in the hippocampus. The FACS analysis showed a significant increase in CD25(+) microglia in the hippocampus compared to controls. The results support the role of microglia as a mediator in the immune-CNS communication and the effects of peripheral IL-2.

Adverse Social Experiences in Adolescent Rats Result in Enduring Effects on Social Competence, Pain Sensitivity and Endocannabinoid Signaling

Social affiliation is essential for many species and gains significant importance during adolescence. Disturbances in social affiliation, in particular social rejection experiences during adolescence, affect an individual’s well-being and are involved in the emergence of psychiatric disorders. The underlying mechanisms are still unknown, partly because of a lack of valid animal models. By using a novel animal model for social peer-rejection, which compromises adolescent rats in their ability to appropriately engage in playful activities, here we report on persistent impairments in social behavior and dysregulations in the endocannabinoid (eCB) system. From postnatal day (pd) 21 to pd 50 adolescent female Wistar rats were either reared with same-strain partners (control) or within a group of Fischer 344 rats (inadequate social rearing, ISR), previously shown to serve as inadequate play partners for the Wistar strain. Adult ISR animals showed pronounced deficits in social interaction, social memory, processing of socially transmitted information, and decreased pain sensitivity. Molecular analysis revealed increased CB1 receptor (CB1R) protein levels and CP55, 940 stimulated [35S]GTPγS binding activity specifically in the amygdala and thalamus in previously peer-rejected rats. Along with these changes, increased levels of the eCB anandamide (AEA) and a corresponding decrease of its degrading enzyme fatty acid amide hydrolase (FAAH) were seen in the amygdala. Our data indicate lasting consequences in social behavior and pain sensitivity following peer-rejection in adolescent female rats. These behavioral impairments are accompanied by persistent alterations in CB1R signaling. Finally, we provide a novel translational approach to characterize neurobiological processes underlying social peer-rejection in adolescence.

Adolescent social rejection alters pain processing in a CB1 receptor dependent manner

Experiences of social rejection represent a major source of distress and in particular peer rejection during adolescence has been implicated in various psychiatric disorders. Moreover, experimentally induced acute social rejection alters pain perception in humans, implicating overlapping neurocircuits for social and physical pains. We recently demonstrated that rearing of adolescent Wistar rats with inadequate, less playful play partners (Fischer 344) persistently decreases pain sensitivity, although the detailed mechanisms mediating the aversiveness during the social encounter remained unsettled. With the present study we examined the behavioral performance during acute interaction of female adolescent Wistar rats with either age-matched same-strain partners or rats from the Fischer 344 strain. We here identify the low responsiveness upon playful attacks, which appears to be characteristic for social play in the Fischer 344 strain, as one of the main aversive components for adolescent Wistar animals during cross-strain encounters, which subsequently diminishes thermal pain reactivity. A detailed behavioral analysis further revealed increased ultrasonic vocalization at 50kHz and an increased frequency of playful attacks for adolescent Wistar animals paired with a Fischer 344 rat compared to same-strain control pairs. Finally, an acute injection of a subthreshold dose of the cannabinoid type 1 receptor inverse agonist/antagonist SR141716 before the social encounter abolished enhanced play-soliciting behavior in Wistar/Fischer 344 pairs as well as the behavioral consequences of the rejection experience in adolescent Wistar rats, further emphasizing an important modulatory role of the endocannabinoid system in mediating the effects of social behavior and social pain.

Towards trans-diagnostic mechanisms in psychiatry: Neurobehavioral profile of rats with a loss of function point mutation in the dopamine transporter gene

ABSTRACT The research domain criteria (RDoC) matrix has been developed to reorient psychiatric research towards measurable behavioral dimensions and underlying mechanisms. Here, we used a new genetic rat model with a loss-of-function point mutation in the dopamine transporter (DAT) gene (Slc6a3_N157K) to systematically study the RDoC matrix. First, we examined the impact of the Slc6a3_N157K mutation on monoaminergic signaling. We then performed behavioral tests representing each of the five RDoC domains: negative and positive valence systems, cognitive, social and arousal/regulatory systems. The use of RDoC may be particularly helpful for drug development. We studied the effects of a novel pharmacological approach metabotropic glutamate receptor mGluR2/3 antagonism, in DAT mutants in a comparative way with standard medications. Loss of DAT functionality in mutant rats not only elevated subcortical extracellular dopamine concentration but also altered the balance of monoaminergic transmission. DAT mutant rats showed deficits in all five RDoC domains. Thus, mutant rats failed to show conditioned fear responses, were anhedonic, were unable to learn stimulus-reward associations, showed impaired cognition and social behavior, and were hyperactive. Hyperactivity in mutant rats was reduced by amphetamine and atomoxetine, which are well-established medications to reduce hyperactivity in humans. The mGluR2/3 antagonist LY341495 also normalized hyperactivity in DAT mutant rats without affecting extracellular dopamine levels. We systematically characterized an altered dopamine system within the context of the RDoC matrix and studied mGluR2/3 antagonism as a new pharmacological strategy to treat mental disorders with underlying subcortical dopaminergic hyperactivity. Summary: The first systematic RDoc study of a disease mechanism proposes dopamine transporter DAT mutant rats as a model for drug development, targeting a hyperdopaminergic state.

37. Interleukin-2 affects social interaction behavior in relation to anxiety-like trait behavior in the elevated plus-maze in adult Wistar rats

epinephrine (E) and norepinephrine (NE) for 18 h. Tumor-necrosisfactor (TNF) alpha and interleukin (IL)-6 were then measured in culture supernatants. Overall, incubation with E and NE induced dose-dependent suppression of TNF (F = 77.66; p < 0.001), and IL-6 production (F = 28.79; p < 0.001), with significantly stronger suppression of TNF (F = 26.36; p < 0.001). Acute stress, however, induced relative resistance of TNF (F = 11.38; p < 0.001), but not IL-6 (F = 1.56; p = 0.22) to the inhibitory signals of E and NE. The present findings show that changes in inflammatory target tissues sensitivity in response to acute psycho-social stress are not restricted to glucocorticoids, but that the catecholamine sensitivity of inflammatory cells is stress-responsive as well. Most importantly, catecholamine sensitivity of immune cells appears to display a higher cytokine specificity compared to GC sensitivity.

Systemic interleukin-2 affects individual anxiety-related social behaviour, gene-transcription and metabolites in the brain

294 Prolonged stress disrupts IL-12 and CpG immunostimulatory effects B. Levi, Y. Goldfarb, L. Sorski, E. Rosenne, R. Melamed, S. Ben-Eliyahu Neuroimmunology Research Unit, Dept. of Psychology, Tel Aviv University, Tel-Aviv, Israel IL-12 and CpG are immunostimulators vastly investigated in cancer patients, and our animal studies indicated their efficacy in reducing postoperative immunosuppression. However, it is important to indicate that immunostimulation in animal studies is commonly initiated under no-stress conditions, unlike in the clinical settings where anxiety and stress overlap immunotherapy. In the current study we tested whether exposure to prolonged stress could impair the beneficial effects of immunostimulation. Two hours following initiation of various stress paradigms (for 24 h), rats were treated with IL-12 or PBS, and compared to non-stressed animals. IL-12 elevated NK cell numbers and activity under control conditions significantly more than in animals subjected to pharmacological stress. Additionally, the improvement of in vivo lung tumor clearance by IL-12 was attenuated in animals subjected to either pharmacological or behavioral stress. In mice, CpG or PBS was administered under prolonged stress or control conditions. CpG elevated IL-12 mRNA and plasma levels, as well as numbers of various dendritic cell subsets expressing maturation and activation markers (e.g., CD80 or CD69). Exposure to prolonged behavioral stress markedly attenuated or abolished the increase in these indices. Interestingly, CORT levels under stress conditions were higher in CpG-treated animals than in PBS-treated animals. Overall, our findings indicate that continuous stress can reduce the efficacy of immunostimulation and attenuate its beneficial effects on immunity and cancer resistance. doi:10.1016/j.bbi.2010.07.077 Abstract # 295 A novel neuronal mechanism for transition from acute to chronic pain: Epinephrine and prostaglandin-E2 as models for stress and inflammation-associated hyperalgesia N. Eijkelkamp , H. Wang , J.N. Wood , R. Dantzer , K.W. Kelley , C.J. Heijnen , A. Kavelaars a,b a University Medical Center Utrecht, Laboratory for Neuroimmunology and Developmental Origins of Disease (NIDOD), Lundlaan 6, Utrecht 3584 EA, Netherlands b Integrative Immunology and Behavior Program, University of Illinois at Urbana-Champaign, Urbana, IL, USA c Molecular Nociception group, University College London, London WC1E