Resident-intruder Test Research Articles

Regulation of stress-provoked aggressive behavior using endocannabinoids

Reactive impulsive aggression is characterized by outbursts of rage and violence when subjects encounter threatening stressful events. Although impulsive aggression and violence create a high-cost burden on health and society, relatively little is known about treatment. Early adolescent social isolation (SI) alters brain development and functions. It induces hyper-excitatory in the ventral hippocampus (vHip) to promote acute stress-provoked outbursts of aggression, referred to as impulsive aggression, in mouse models. Cannabinoid type 1 receptors (CB1Rs) act on presynaptic sites and suppress neurotransmitter release into synapses. Given that CB1R activation inhibits neurotransmitter releases and modulates excitatory network activity, we tested the hypothesis that CB1R activation reduces impulsive aggression in SI mice through decreasing excitatory activity in the vHip. Here, we report that CB1R agonists, WIN-552122 (WIN) or arachidonylcyclopropylamide (ACPA), ameliorated acute stress-provoked attack behavior in the resident-intruder test without affecting general locomotion activity. Increasing endocannabinoids (eCBs) by inhibiting degradation enzymes in the vHip reduced impulsive aggression, and the effect was blunted by administration of AM251, a CB1R antagonist. Acute stress in SI mice induced c-Fos expression, a marker of neuronal activation, on vHip neurons projecting to the ventromedial hypothalamus (VMH), a well-known brain area that controls attack behavior. eCB augmentation inhibited c-Fos expression in VMH-projecting vHip neurons surrounded by CB1Rs. These results suggest that enhancing eCB signaling in order to activate CB1Rs suppresses impulsive aggression via suppressing vHip→VMH neural activity and point to a role of CB1R activation in ameliorating impulsive aggression in adults who have had adverse experiences during early adolescence.

ACE2 Knock‐out Mice Show Decreased Blood Pressure Response and Increased Anxiety‐like Behavior to Acute Stress

Stress increases Ang-II production and elevates sympathetic outflow which results in anxiety-related behavior and hypertension. Here, we investigated a potential contribution of Angiotensin Converting Enzyme-2 (ACE2) in the regulation of stress and anxiety-related behavior, in conscious freely moving mice. Previous studies reported that overexpression of ACE2 attenuates HPA axis and decreases anxiety-related behavior (Wang et al., 2018); however, deletion of ACE2 and its implication for such responses are largely unknow. Male mice (30-33 g), 6 ACE2 knock-outs (KO) and 6 wild type (WT) controls were chronically implanted with blood pressure (BP) transducers for continuous recording of BP, heart rate (HR) and locomotor activity. Following a 1-week recovery, mice were subjected to resident-intruder test for 10 min, for the measurement of stress-induced cardiovascular changes. During the dark/active period, a naïve intruder-mouse was suddenly introduced in the experimental mice's cages and their cardiovascular responses were analyzed using the Ponemah software (DSI). In response to intruder-intrusion for 10 min, ACE2 KO mice showed significant (p<0.05) attenuation in BP increases (+15 ±7 mmHg) compared to their WT controls (+39 ±7 mmHg). However, no significant difference was observed in HR increase in ACE2 KO versus WT mice (+166 ±40 and +173±38 bpm, respectively, p=0.45). In contrast, ACE2 KO mice exhibited a significant increase in locomotor activity compared to controls (179 ±19 and 111 ±14 arbitrary units, respectively, p<0.05). These mice were later subjected to the elevated plus maze (EPM) and light-dark box (LDB) tests for 5 min to evaluate anxiety-related behavior. Interestingly ACE2 KO mice were observed to spend more time in closed arms (244 ±19 s) in EPM compared to WT controls (218 ±22 s, p<0.05). However, in the LDB test, ACE2 KO mice showed no difference (286 ±18 s) compared to WT (279 ±19 s, ns). All mice were then chronically implanted subcutaneously with osmotic minipumps filled with an AT1 receptor antagonist (losartan, 10 mg/kg). A week later, mice were again subjected to the intruder test. ACE2 KO mice showed substantial attenuation in BP increase to intruder-intrusion compared to WT mice (+6±3 and +19±6 mmHg respectively, p<0.05); with no significant difference in HR increase in both the groups. Interestingly, pre-treatment with losartan attenuated the intruder-induced locomotor activity in ACE2 KO mice (149±29, p=0.11). In addition, losartan treatment did not attenuate the time ACE2 KO mice spent in closed arms in EPM; but exhibited a decreased time spent in the dark compartment in the LDB test. Surprisingly, our results did not show exacerbated BP responses to acute stress in the absence of ACE2 but instead produced decreased BP responses suggesting other compensatory mechanisms involved. Losartan-mediated attenuation of the increased locomotor activity and anxiety-related behaviors in ACE2 KO mice strongly suggests an Ang-II mediated anxiety-related behaviors. Our experimental findings further extend the current notion that ACE2 potentially contributes to the regulation of stress and anxiety.

Low-normal doses of methiopropamine induce aggressive behaviour in mice.

Recreational use of illicit methiopropamine (MPA) is a public health concern because it produces neurochemical effects comparable with those induced by methamphetamine (METH). The present study investigated the effects of MPA on the expression of an aggressive behaviour. Eighty CD-1 male mice, after receiving intraperitoneal injection of saline, MPA (0.01-10 mg/kg), METH (0.01-10 mg/kg), or AMPH (0.01-10 mg/kg), once a week over a 5-week period, underwent the resident-intruder test and spontaneous locomotor activity measurement. Results showed that all psychostimulants induce aggressive behaviour even at low doses, with a dose-dependent increase and a time-dependent sensitisation. MPA potency was similar to METH and superior to AMPH. Therefore, MPA-induced aggressive behaviour may appear even at MPA dosages free of cardiovascular or other behavioural adverse effects and could become a non-intentional side effect that users experience after increasing and repeating MPA consumption.

Celastrus paniculatus oil ameliorates synaptic plasticity in a rat model of attention deficit hyperactivity disorder

Objective: To explore the effect and mechanism of action of Celastrus paniculatus oil on the treatment of perinatal rats with attention deficit hyperactivity disorder. Methods: In the perinatal stage, the rats were either isolated or administered with lead acetate to establish an animal model of attention deficit hyperactivity disorder. Atomoxetine served as the reference standard. Animals’ behaviours were assessed through Y-maze, novel object preference, fear conditioning and resident-intruder aggression tests. Oxidative stress parameters, bioamine concentration (dopamine, noradrenaline and 5-hydroxytryptamine), nerve growth factor, interleukin-6, nuclear factor-κB, and tumour necrosis factor (TNF)-α were estimated. Synaptophysin immunohistochemical assay was performed. Results: Celastrus paniculatus oil significantly improved behavioural parameters in Y maze, novel object preference, discrimination index, fear conditioning and resident intruder aggressive tests. The treatment groups showed a decrease in malondialdehyde level. Changes in the levels of dopamine, noradrenaline, and serotonin were restored by Celastrus paniculatus oil. Celastrus paniculatus oil increased nerve growth factor and decreased interleukin-6, nuclear factor-κB, and TNF-α. Synaptophysin immunoreactivity was also improved by Celastrus paniculatus oil with alleviated reactive gliosis, degeneration, and vascular proliferation. Conclusions: This research shows the therapeutic potential of Celastrus paniculatus oil for the treatment of attention deficit hyperactivity disorder.

Induction of aberrant agonistic behavior by a combination of serotonergic and dopaminergic manipulation in rats.

Serotonin (5-HT) and dopamine (DA) are involved in the regulation of social behaviors. However, the effects of their interactions on social behavior are not well understood. In this study, rats received a serotonergic neurotoxin injection into the raphe nuclei and/or systemic administration of L-3, 4-dihydroxyphenylalanine (L-DOPA), and their agonistic behaviors were investigated using the resident-intruder (RI) paradigm. Rats in the DA + /5-HT-group, which were administered both monoaminergic treatments, exhibited intense jump and flight responses to intruders. These behaviors were not observed in rats that received either 5-HT lesions or L-DOPA treatment only. To address the neural basis of these aberrant behaviors, we compared c-Fos immunoreactivity in the brain among the different groups. The DA + /5-HT-group had c-Fos activation in areas related to anti-predatory defensive behaviors, such as the ventromedial hypothalamic nucleus, premammillary nucleus, and periaqueductal gray. Moreover, this group had increased c-Fos expression in the ventroposterior part of the anterior olfactory nucleus (AOVP). To test the involvement of this area in the aberrant behaviors, cytotoxic lesions were performed in the AOVP prior to the monoaminergic treatments, and subsequent behaviors were examined using the RI test. The AOVP-lesioned DA + /5-HT-rats had attenuation of the aberrant behaviors. Together, these results suggest that the AOVP is involved in the generation of the aberrant defensive behaviors, and that 5-HT/DA balance is important in the regulation of social behaviors.

Territorial aggression by dwarf hamster females (Phodopus campbelli): A hormonal approach.

Females of many vertebrate species are territorial. However, few studies have been performed on territorial aggression and its hormonal bases in females. The present study determined whether dwarf hamster females (Phodopus campbelli) display territorial aggression and the role that estradiol (E2 ) and progesterone (P4 ) play in this behavior. Two experiments were carried out. First, 28 virgin females were mated with sexually inexperienced males. The females were submitted to oviduct ligation 2 weeks before mating to prevent pregnancy. After 15 days of mating, 16 out of 28 females were submitted to resident-intruder tests, whereas the remaining 12 females were nonconfronted. Blood samples were collected 30 min after the aggression tests to determine E2 and P4 by ELISA. In the second experiment, 40 females were mated with vasectomized males. Thirty days after mating, 30 dwarf hamsters were subjected to ovariectomy; 10 were nontreated, 10 were treated with E2 , and 10 with P4 . The remaining 10 were sham-operated. All females were submitted to resident-intruder tests, and blood samples were taken to quantify E2 and P4 . In the first experiment, 87.5% of dwarf hamster females displayed territorial aggression, and in the second, 100% of females were aggressive. Ovariectomy and ovariectomy plus E2 or P4 replacement did not affect aggressive territorial behavior. These results showed that this species' females are territorial, and this behavior is not dependent on ovarian steroid hormones.

Glutamate transporter-1 link astrocytes with heightened aggressive behavior induced by steroid abuse in male CF1 mice

The astrocytic glutamate transporter GLT-1 performs glutamate uptake thereby mediating NMDAr responses in neurons. Ceftriaxone (CEF) upregulates astrocytic GLT-1 expression/activity, which could counteract excessive glutamate levels and aggressive behavior induced by anabolic synthetic steroids such as nandrolone decanoate (ND). Here, adult male CF-1 mice were allocated to oil (VEH), ND, CEF, and ND/CEF groups. Mice were subcutaneously (s.c.) injected with ND (15 mg/kg) or VEH for 19 days, and received intraperitoneal (i.p.) injections of CEF (200 mg/kg) or saline for 5 days. The ND/CEF group received ND for 19 days plus coadministration of CEF in the last 5 days. On the 19th day, the aggressive phenotypes were evaluated through the resident-intruder test. After 24 h, cerebrospinal fluid was collected to measure glutamate levels, and the pre-frontal cortex was used to assess GLT-1, pGluN2BTyr1472, and pGluN2ATyr1246 by Western blot. Synaptosomes from the left brain hemisphere was used to evaluate mitochondrial function including complex II-succinate dehydrogenase (SDH), Ca2+ handling, membrane potential (ΔѰm), and H2O2 production. ND decreased the latency for the first attack and increased the number of attacks by the resident mice against the intruder, mechanistically associated with an increase in glutamate levels and pGluN2BTyr1472 but not pGluN2ATyr1244, and GLT-1 downregulation. The abnormalities in mitochondrial Ca2+ influx, SDH, ΔѰm, and H2O2 implies in deficient energy support to the synaptic machinery. The ND/CEF group displayed a decreased aggressive behavior, normalization of glutamate and pGluN2BTyr1472levels, and mitochondrial function at synaptic terminals. In conclusion, the pharmacological modulation of GLT-1 highlights its relevance as an astrocytic target against highly impulsive and aggressive phenotypes.

Topiramate inhibits offensive aggression through targeting ventrolateral periaqueductal gray

Topiramate is an approved antiepileptic drug clinically used to treat epilepsy and prevent migraines. Currently, topiramate has been found to be effective in treating aggressive symptoms in neuropsychiatric patients. In preclinical studies, however, the effects and mechanisms of topiramate on offensive aggression are still largely uninvestigated. Our previous studies indicated that glutamatergic transmission in the ventrolateral periaqueductal gray (vlPAG) plays a crucial role in regulating elements of offensive aggressive behaviors. In the present work, we investigated the actions of topiramate on vlPAG glutamatergic transmission and aggressive behaviors in group-housed (GH) and socially isolated (SI) rats. The results suggested that a single injection of topiramate systemically and dose-dependently inhibited elements of offensive aggressive behaviors of both GH and SI rats in the resident-intruder test (RIT), with long-lasting effective time profiles in SI rats. Moreover, systemic single administration of topiramate reduced the frequency and amplitude of miniature excitatory postsynaptic currents (mEPSCs) in the vlPAG. Bath perfusion of topiramate directly decreased the frequency and amplitude of mEPSCs and shortened the amplitude of evoked excitatory postsynaptic currents (EPSCs) in the vlPAG. Furthermore, intra-vlPAG single microinjection of topiramate dose-dependently inhibited offensive aggressive behaviors in GH and SI rats in a time-dependent manner. Additionally, both systemic and local topiramate inhibited offensive aggressive behaviors in a (2R,6R)-hydroxynorketamine (HNK)-dependent rat model. In conclusion, the present results suggest that topiramate exerts anti-aggressive roles through its inhibitory actions on glutamatergic activities in the vlPAG. These preclinical results support topiramate as a candidate drug to treat patients with heightened offensive aggression.

Influence of Diazepino[1,2-a]benzimidazole derivative (DAB-19) on behavioral aspects of animals

Introduction: Diazepino[1,2-a]benzimidazole derivatives showed anxiolytic (EPM, L/D box, Vogel test), antidepres­sant (Tail Suspension test, Porsolt test), anticonvulsant (Pentylenetetrazole-Induced seizures) and analgesic (Tail-Flick, Hot Plate) actions, which were described earlier (Spasov et al. 2020). 11-(4-tert-butylbenzyl)-2,3,4,5-tetrahydro[1,3]diazepino[1,2-a]benzimidazole hydrobromide (compound DAB-19) has evident anxiolytic, antidepressant and anticon­vulsant effects. In the present study, compound DAB-19 was screened for its influence on animals` behavior patterns, such as aggression, obsessive-compulsive behavior, emotional lability, and unsociability.&#x0D; Materials and methods: 112 outbred white animals (rats and mice) were used. Compound DAB-19 (2.34 mg/kg, p.o.) and diazepam (2 mg/kg, p.o.) were injected to the treatment groups. Using an Open Field (OF) test, we evaluated a spontaneous motor activity, a search activity of mice and a degree of their emotionality. During a Marble Burying (MB) test, we monitored anxiolytic and anticompulsive effects of DAB-19. Using a Resident-intruder (RI) test, we evaluated a degree of aggression in the experimental animals, properties of their social interaction, as well as defensive and individual behavior.&#x0D; Results and discussion: Compound DAB-19 has positive influence on the search activity, mood stabilizing activity and antiaggressive actions. Administration of both DAB-19 (2.34 mg/kg, p.o.) and diazepam (2 mg/kg, p.o.) reduced anxiety-like behavior in OP, RI and MB tests, as indicated by a significantly increased number of entries to the center of OP; a decreased number of marbles buried in MB and reduced aggressive behavior in RI. It was stated that using DAB-19 leads to a decrease in affective reactions of the animals – aggression, obsessive-compulsive behavior, neurotic condition and emotional instability.&#x0D; Conclusions: The compound (DAB-19) produces anxiolytic-like effects, compared with those of diazepam, in various anxiety paradigms.

Pharmacological analysis of the effects of the neuropeptide Y antagonist BMS 193885 on the emotional, intraspecies behavior and reinforcing properties of ethanol in rats

Purpose. Our previously data on orexigenic peptides (orexin, ghrelin) showed antagonists of peptides receptors as correctors of the emotional-motivational and cognitive spheres. Currently, a close relationship between ghrelin and orexin with neuropetide Y has been shown in feeding and emotional behavior.&#x0D; The aim of this work was to analyze the effect of the NPY antagonist Y1R BMS 193885 on emotional and intraspecies behavior, as well as on the reinforcing properties of ethanol in rats.&#x0D; Methods. We used the open field test, elevated plus-maze, Porsolts forced swimming test, resident intruder test, conditional place preference (CPP). BMS 193885 1 mg/ml, 20 l intranasally did not cause an anxiogenic effect in the elevated plus-maze.&#x0D; Results. In the Porsolts test, there was also no increase in the level of depression. Moreover, there was a significant decrease in the number and time of dives, as an indirect indicator of a decrease in the level of depression. At the same time, in the resident intruder test were decreased protective behavior, as an indicator of a decrease in the stress of intraspecific interaction in the absence of aggression. Moreover, local movements were increased in the open field test as an indicator of the animals activity impaired by fear. BMS 193885 had no effect on the expression of the CPP of ethanol.&#x0D; Conclusion. Thus, it was previously shown that the BMS 193885 is a powerful, selective, brain-penetrating Y1 receptor antagonist, it reduces food intake and body weight in animal models of obesity both after acute and chronic administration. Our data indicate that the decrease in food intake is not associated with the level of anxiety, depression, or with a change in intraspecific interaction. It has been previously shown that NPY reduces alcohol consumption. Our data indicate that the Y1R antagonist of the neuropeptide Y BMS 193885 does not cause a change in the CPP of alcohol.

Bidirectional effects of Topiramate on anterior cingulate cortex region related aggressive behavior

Topiramate, an antiepileptic drug, has been found to be useful for the treatment of aggression in clinical populations as well as in animal models of aggression. However, increases in aggression were also observed under lower doses of Topiramate. Subsequently, Topiramate produced an inverted U-shaped dose response curve, with increases in aggression at low doses, whereas higher doses engendered anti-aggressive effects. In our previous study, we demonstrated that Topiramate modulates brain activity in the prefrontal areas involved in the modulation of the subcortical circuit mediating aggression, and we found indirect evidence that the anterior cingulate cortex (ACC) could be a key site where Topiramate may exert its dose-response effects on aggression. In this study, we performed site-directed injections of Topiramate (0.1 and 0.3 mM) into the ACC on offensive behaviors in isolation-inducedaggression paradigm. By using the resident-intruder test, we demonstrated that Topiramate microinfusion into the ACC at low doses produced increases in aggression, as evidenced by shorter attack latencies (p < 0.01) and increased attack duration (p < 0.01), without affecting the social behavior. In contrast, higher doses engendered anti-aggressive effects, by increasing the attack latencies (p < 0.001), decreasing attack number (p < 0.001) and decreasing attack duration (p < 0.001), as well as an increase in the social behavior. Our findings supported that Topiramate at higher concentrations appeared more efficient to decrease aggression in treated mice. Therefore, we suggest that the ACC is a key brain region in which Topiramate may exert its dose-response effects on aggressive and antisocial behaviors observed in populations with psychotic disorders.

Male-specific features are reduced in Mecp2-null mice: analyses of vasopressinergic innervation, pheromone production and social behaviour.

Deficits in arginine vasopressin (AVP) and oxytocin (OT), two neuropeptides closely implicated in the modulation of social behaviours, have been reported in some early developmental disorders and autism spectrum disorders. Mutations in the X-linked methyl-CpG-binding protein 2 (MECP2) gene are associated to Rett syndrome and other neuropsychiatric conditions. Thus, we first analysed AVP and OT expression in the brain of Mecp2-mutant mice by immunohistochemistry. Our results revealed no significant differences in these systems in young adult Mecp2-heterozygous females, as compared to WT littermates. By contrast, we found a significant reduction in the sexually dimorphic, testosterone-dependent, vasopressinergic innervation in several nuclei of the social brain network and oxytocinergic innervation in the lateral habenula of Mecp2-null males, as compared to WT littermates. Analysis of urinary production of pheromones shows that Mecp2-null males lack the testosterone-dependent pheromone darcin, strongly suggesting low levels of androgens in these males. In addition, resident-intruder tests revealed lack of aggressive behaviour in Mecp2-null males and decreased chemoinvestigation of the intruder. By contrast, Mecp2-null males exhibited enhanced social approach, as compared to WT animals, in a 3-chamber social interaction test. In summary, Mecp2-null males, which display internal testicles, display a significant reduction of some male-specific features, such as vasopressinergic innervation within the social brain network, male pheromone production and aggressive behaviour. Thus, atypical social behaviours in Mecp2-null males may be caused, at least in part, by the effect of lack of MeCP2 over sexual differentiation.

Adolescent oxytocin treatment affects resident behavior in aggressive but not tame adult rats

There are indications that exposing adolescent rodents to oxytocin (OT) promotes social activity and reduces anxiety in adulthood. Adult male gray rats selected for elimination and enhancement of the aggressive response to humans, when exposed to OT, showed divergent changes in the resident behavior towards the intruder. It could be assumed that adolescent administration of both OT and antagonist of OT receptor (OTR) would also have different long-term effects on resident behavior and startle reflex in adult aggressive and tame rats. The aim of this work is to study the long-term effects of adolescent administration of both OT and antagonist of OT receptor (OTR) on resident behavior and startle reflex in adult tame and aggressive male gray rats. Starting at the age of 28 days, the animals received nasal applications of 5 μL of oxytocin solution (1 μg / μL) or saline for 5 days (daily). At the age of two months, the acoustic startle amplitude was assessed in two series of 5 acoustic stimuli. The resident-intruder test was performed one week later. Antagonist of OT receptor l-368,899 was administered intraperitoneally (i.p.) once at a dose of 5 mg/kg at the age of 30–33 days. Subsequent startle reflex tests were performed 20 days later, at the age of 50–53 days. A week later, the resident-intruder test was performed on the same rats. The startle amplitude in aggressive rats of the control group (in two series of acoustic stimuli) and those having received saline (in the first series) was larger than in the corresponding tame groups. Oxytocin and saline solutions did not significantly affect the startle amplitude compared to control animals. After saline administration, the attack latency in tame rats was longer than in aggressive rats (P <0.05). Oxytocin treatment caused a prolongation of this period in aggressive males compared with control animals receiving saline solution (P <0.01). In addition, oxytocin administration in aggressive males caused an increase in the time of social behavior, which did not include aggressive and same-sex behavior, as compared with the corresponding control animals (P <0.05). Exogenous oxytocin receptor antagonist (l-368,899) did not affect the startle amplitude and behavior in the resident-intruder test in aggressive and tame male rats. Adolescent OT treatment causes a prolongation of both the attack latency and social behavior in the resident-intruder test in adult aggressive male rats, but does not affect these parameters in tame rats.

Social dysfunction in the neurodevelopmental model of schizophrenia in male and female rats: Behavioural and biochemical studies

Social dysfunction is among the core symptoms of schizophrenia. The neuropeptides oxytocin (OXT) and vasopressin (VP) are involved in the regulation of social behaviour and social cognition. There are indications that both of these neurotransmitter systems are altered in schizophrenia. Prenatal (embryonic day 17) exposure to the neurotoxin methylazoxymethanol acetate (MAM; 22 mg/kg) leads to a schizophrenia-like phenotype in rats and has been used as a model of schizophrenia symptoms. Here, we examined the social phenotype of MAM-exposed female and male rats and measured concentrations of OXT, VP and their specific receptors in various brain areas involved in the control of social behaviour.We report decreases in social behaviour and ultrasonic vocalisations (USVs) in the MAM rats during social encounters. Specifically, the duration of social interactions and number of corresponding USVs were reduced in this group. In the MAM rats, “positive” 50-kHz USVs were flatter, i.e., displayed lower bandwidth, a greater percentage of “short” calls and lower percentage of frequency-modulated calls. The MAM animals exhibited diminished interest towards social stimuli in olfactory preference tests. In the resident-intruder test, MAM exposure reduced dominance behaviour only in the males. We also report cognitive impairments, including reduced novel object recognition and cognitive inflexibility in the attentional set shifting test, and decreased OXT and OXT receptor concentrations in the prefrontal cortex and hypothalamus and VP and VP receptors in the hypothalamus in the MAM rats.Deficits in central OXT and VP systems may underlie abnormalities present in the MAM model of schizophrenia.

GluD1 knockout mice with a pure C57BL/6N background show impaired fear memory, social interaction, and enhanced depressive-like behavior.

The GluD1 gene is associated with susceptibility for schizophrenia, autism, depression, and bipolar disorder. However, the function of GluD1 and how it is involved in these conditions remain elusive. In this study, we generated a Grid1 gene-knockout (GluD1-KO) mouse line with a pure C57BL/6N genetic background and performed several behavioral analyses. Compared to a control group, GluD1-KO mice showed no significant anxiety-related behavioral differences, evaluated using behavior in an open field, elevated plus maze, a light-dark transition test, the resident-intruder test of aggression and sensorimotor gating evaluated by the prepulse inhibition test. However, GluD1-KO mice showed (1) higher locomotor activity in the open field, (2) decreased sociability and social novelty preference in the three-chambered social interaction test, (3) impaired memory in contextual, but not cued fear conditioning tests, and (4) enhanced depressive-like behavior in a forced swim test. Pharmacological studies revealed that enhanced depressive-like behavior in GluD1-KO mice was restored by the serotonin reuptake inhibitors imipramine and fluoxetine, but not the norepinephrine transporter inhibitor desipramine. In addition, biochemical analysis revealed no significant difference in protein expression levels, such as other glutamate receptors in the synaptosome and postsynaptic densities prepared from the frontal cortex and the hippocampus. These results suggest that GluD1 plays critical roles in fear memory, sociability, and depressive-like behavior.

Effect of a partial cage dividing enrichment on aggression-associated parameters in group-housed male C57BL/6NCrl mice

Group-housing is highly important for social animals. Group-housing of male mice in captivity though often leads to aggression with partially disastrous consequences for the animals as well as for the quality of experimental data. In this study we investigated the effect of a novel “cross-enrichment”, i.e. a colored partial cage divider, which is provided in transparent or black and which is partly separating the cage in four small areas. Group-housed male C57BL/6NCrl mice (three per cage) were maintained under either standard conditions (nestlet group) or enriched conditions (nestlet + cage divider in black [EB-group] or in transparent [ET-group]) for eight weeks. Several physiological parameters (body weight, blood glucose, stress induced hyperthermia, fecal corticosterone metabolites and organ weights) and behavioral tests (Nest test, Openfield/social Novel-Object, Dark-Light-Box, Hotplate and Resident-Intruder test) were measured/performed to determine enrichment-induced effects. In comparison to nestlet- and ET-group animals, EB-mice showed significant increased stress-associated parameters, i.e. in the blood glucose concentration. Furthermore, EB animals seemed to have enhanced emotional stress with a poorer outcome in the nest test and a higher amount of fecal boli at the end of the social Novel-Object test. Additionally, EB-mice behaved more aggressively towards conspecifics after cleaning cages. We conclude that the opacity of the tested partial cage dividers has a huge impact on aggressive behavior and therefore may lead to significant changes in behavioral and physical measures potentially altering research outcomes.

Effects of Testicular and Non-Testicular Testosterone on Territorial and Isolation-induced Aggressive Behavior of Male Layer Chicks.

Testosterone (T) is known to induce aggressive behavior, particularly in male animals. However, our recent results showed that a certain kind of aggressive behavior is T-independent; moreover, the role of T in chicken territorial and isolation-induced aggressive behavior has not yet been investigated. In addition, castration alone is insufficient to evaluate the role of T in aggressive behavior because we found that non-testicular T concentration, probably derived from the adrenal gland, in the blood of castrated chicks was low, but not zero. In the present study, therefore, the role of testicular T in chicken aggressive behavior was evaluated through castration, and the role of nontesticular T was assessed using the subcutaneous implantation of flutamide, a non-steroidal antiandrogen, in the castrated male layer chicks. Resident-intruder (R-I) and social interaction (SI) tests were used to quantitatively monitor territorial and isolation-induced aggressive behavior, respectively. Castration and drug implantation of the chicks were performed at 14 days of age. The R-I test was performed at 29 and 30 days of age, and the SI test was performed at 31 and 32 days of age. The total aggression frequencies (TAFs) and aggression establishment rate (AER) were used as indices of chick aggressive behavior. In the R-I test, castration significantly decreased the TAFs but the AER was not affected by castration or flutamide implantation. In the SI test, on the other hand, there were no significant differences in the TAFs, but the AER tended to increase in the intact chicks and decrease in the flutamide-implanted, castrated male chicks. These results suggest that the role of T in chicken aggression depends on the differences in social context of the behavior, and that both testicular and non-testicular T play an important role in the occurrence of isolation-induced aggression in male layer chicks.

Concomitant Evaluation of Heart Period and QT Interval Variability Spectral Markers to Typify Cardiac Control in Humans and Rats.

The variability of heart period, measured as the time distance between two consecutive QRS complexes from the electrocardiogram (RR), was exploited to infer cardiac vagal control, while the variability of the duration of the electrical activity of the heart, measured as the time interval from Q-wave onset to T-wave end (QT), was proposed as an indirect index of cardiac sympathetic modulation. This study tests the utility of the concomitant evaluation of RR variability (RRV) and QT variability (QTV) markers in typifying cardiac autonomic control of humans under different experimental conditions and of rat groups featuring documented differences in resting sympatho-vagal balance. We considered: (i) 23 healthy young subjects in resting supine position (REST) undergoing head-up tilt at 45° (T45) and 90° (T90) followed by recovery to the supine position; (ii) 9 Wistar (WI) and 14 wild-type Groningen (WT) rats in unstressed conditions, where the WT animals were classified as non-aggressive (non-AGG, n = 9) and aggressive (AGG, n = 5) according to the resident intruder test. In humans, spectral analysis of RRV and QTV was performed over a single stationary sequence of 250 consecutive values. In rats, spectral analysis was iterated over 10-min recordings with a frame length of 250 beats with 80% overlap and the median of the distribution of the spectral markers was extracted. Over RRV and QTV we computed the power in the low frequency (LF, from 0.04 to 0.15 Hz in humans and from 0.2 to 0.75 Hz in rats) band (LFRR and LFQT) and the power in the high frequency (HF, from 0.15 to 0.5 Hz in humans and from 0.75 to 2.5 Hz in rats) band (HFRR and HFQT). In humans the HFRR power was lower during T90 and higher during recovery compared to REST, while the LFQT power was higher during T90. In rats the HFRR power was lower in WT rats compared to WI rats and the LFQT power was higher in AGG than in non-AGG animals. We concluded that RRV and QTV provide complementary information in describing the functioning of vagal and sympathetic limbs of the autonomic nervous system in humans and rats.

P2X7 receptor knockout mice display less aggressive biting behaviour correlating with increased brain activation in the piriform cortex

P2X7 receptors are implicated in the pathophysiology of psychiatric conditions such as depression and bipolar disorder. P2X7 receptors regulate the release of pro-inflammatory cytokines from microglia, and gain-of-function P2X7 mutations may contribute to the neuroinflammation found in affective disorders. However, the role of this receptor in mediating other mental health conditions and aberrant behaviours requires further examination. The current study we investigated the effects of germline genetic deletion of P2xr7 on social and marble burying behaviours in mice throughout the critical adolescent developmental period. Marble burying behaviour is thought to provide a mouse model of obsessive-compulsive disorder (OCD). We also characterised the effects of P2rx7 deletion on aggressive attack behaviour in adult mice and subsequently quantifieded microglial cell densities and c-Fos expression, a marker of neuronal activation. P2rx7 knockout mice displayed reduced OCD-related marble burying behaviour which was most pronounced in late adolescence/early adulthood. P2rx7 knockout mice also exhibited reduced aggressive attack behaviours in adulthood in the resident-intruder test. Reduced aggression in P2xr7 knockout mice did not coincide with changes to microglial cell densities, however c-Fos expression was elevated in the piriform cortex of P2rx7 knockout mice compared to wildtype mice. This study suggests that the P2X7 receptor might serve as a novel target for serenic or anti−OCD therapeutics.

Early life stress induces submissive behavior in adult rats

Maternal-deprivation of rodent pups is a relevant model of extreme early-life stress that can be relevant to the understanding of long-term effects of war, migration, parental loss and displacement. Although even mild stress during infancy affects brain development and behavior, the current study focused on the effects of six hour daily maternal-separation, a model that reflects the severe distress often experienced in those circumstances. This study emphasizes the effect of maternal separation on social behavior in the context of a variety of factors that measure cognitive and emotional behavior which were subject to principle component analysis.Sprague-Dawley pups were separated from the dam for 6 h each day during the first 3-weeks of life and underwent a battery of behavioral tests at 3-months of age.We found that rodents exposed to postnatal maternal deprivation displayed submissive behavior in resident-intruder and dominant-submissive tests, as well as significantly more anxiety and anhedonia than control rats.The results of multivariate statistical analysis show that the dominant-submissive behavior correlates with depressive, anxiety and social behavior and can be predicted with an accuracy of 86.2%. The increased submissive behavior in male rats that had been subjected to severe postnatal stress suggests that exposure to stress during infancy and childhood could have long-term effects on social relationships. The mechanism of the long-term effects on depression, anxiety and submissive behavior requires further investigation.