Stranger Mouse Research Articles

Abstract WMP75: Neurobehavioral Outcomes Following Left- and Right-Sided Intracerebral Hemorrhage in Mice

Background: Injury location is an established outcome predictor in intracerebral hemorrhage (ICH), with most studies focusing on outcomes relating to motor deficits. However, location may also affect other neurobehavioral deficits following ICH, such as mood and cognitive decline. To address this gap, we studied neurobehavioral outcomes following left and right-sided ICH in mice. Methods: Three-month-old male mice (C57BL/6) were given a left (n=5) or right-sided (n=5) ICH. In short, type VII collagenase was injected into the left or right striatum (0.0 mm anterior, +/- 2.3 mm lateral, and 3.6 mm deep from bregma). Naïve mice (n=5) were used as controls. Behavior was assessed at day 28 post-ICH using a video-recorded three-chamber sociability task in which one chamber had an enclosed male stranger mouse. Distance traveled, time in each chamber, and time exploring the stranger mouse were quantified using automated software (AnyMaze). Data were analyzed by one-way ANOVA and Tukey post-hoc tests. Results: ICH location had differential effects on behavioral outcomes. Compared to naïve and right-sided ICH mice, left-sided ICH mice traveled a greater overall distance exploring the chambers (both p<0.05), spent a greater overall time immobile in the stranger mouse zone (both p<0.001) and less time oriented towards the stranger mouse (both p<0.05). Furthermore, when comparing naive and left-sided ICH mice, left-sided mice had a higher frequency of investigations of the empty cage (p<0.05). No significant differences were found between the naïve and right-sided ICH mice. Conclusions: These results reveal differences in behavioral phenotypes associated with left- and right-sided ICH.

Metabolic, neuroendocrine and behavioral effects of social defeat in male and female mice using the chronic non-discriminatory social defeat stress model

Stress-related disorders predominately affect females, yet preclinical models of chronic stress exclusively use males especially in models where social stressors are studied. Here, we implemented a 21-day novel social defeat paradigm in which a female and male C57 intruder are simultaneously placed in the cage of a territorial, resident CD-1 male mouse, and the resident proceeds to attack both intruders. Mice were given access to a regular laboratory diet, high in carbohydrates, and a palatable diet, high in fat. Chronic social defeat stress using this paradigm resulted in increased caloric intake in male and female mice, with the effects being more pronounced in females. We observed sex differences in high fat diet intake in response to stress, which was correlated with higher levels of plasma ghrelin observed in female mice but not male mice. Furthermore, females exposed to chronic stress displayed changes in growth hormone secretatogue receptor (ghsr) and neuropeptide-y (npy) expression in the arcuate nucleus of the hypothalamus, potentially increasing ghrelin sensitivity and inducing changes in diet choice and caloric intake. Behavioral results show that females tended to spend more time interacting during the social interaction test, compared to males who displayed higher vigilance towards the stranger mouse. Overall, our results highlight unique neurometabolic alterations in female mice in response to stress that is not present in male mice and may be important for coping with chronic stress and sustaining reproductive function.

The anterior insular cortex processes social recognition memory

Impaired social abilities are characteristics of a variety of psychiatric disorders such as schizophrenia, autism spectrum disorder, and bipolar disorder. Studies consistently implicated the relationship between the anterior insular cortex (aIC) and social ability, however, how the aIC involves in processing specific subtypes of social ability was uninvestigated. We, therefore, investigated whether the absence or presence of the aIC affects the social behaviors of mice. We found that electrolytic lesions of the aIC specifically impaired mice’s ability to recognize a novel stranger mouse, while the sociability of the aIC-lesioned mice was intact. Interestingly, the aIC-lesioned mice were still distinguished between a mouse that had been housed together before the aIC lesion and a novel mouse, supporting that retrieval of social recognition memory may not involve the aIC. Additional behavioral tests revealed that this specific social ability impairment induced by the aIC lesion was not due to impairment in olfaction, learning and memory, locomotion, or anxiety levels. Together our data suggest that the aIC is specifically involved in processing social recognition memory, but not necessarily involved in retrieving it.

Does the stranger mouse strain matter to female BTBR mice?

Autism spectrum disorder (ASD) is characterized by deficits in social communication and repetitive behaviors/restricted interests. One mouse model of ASD is the BTBR T+Itprtf/J (BTBR) mice which display low levels of social behavior in several tests. The social approach test is used to examine the preference for social interaction between a stranger mouse or a novel object. While female BTBR mice have been used in the social approach test, no one has examined the degree to which the strain of the stranger mouse will affect social behavior. The current experiment tested female BTBR subject mice in the social approach test with stranger mice from different strains including the BTBR, 129S1/SvImJ (129), and C57BL/6J (B6) mice, of which the B6 mice are most social. The results show that female BTBR mice overall spent significantly more time in the stranger mouse chamber. However, further analysis revealed that the subject mice spent significantly more time in the stranger mouse chamber when the stranger was from the B6 strain, but not the BTBR or 129 strains. The BTBR female mice also sniffed the B6 and 129 stranger mice more than the novel object. This suggests that BTBR females are more social with mice that display high levels of social behavior, but less so with less social mice.

Social Relationship as a Factor for the Development of Stress Incubation in Adult Mice.

While stress reactions can emerge long after the triggering event, it remains elusive how they emerge after a protracted, seemingly stress-free period during which stress incubates. Here, we study the behavioral development in mice isolated after observing an aggressive encounter inflicted upon their pair-housed partners. We developed a spatially resolved fine-scale behavioral analysis and applied it to standard behavioral tests. It reveals that the seemingly sudden behavioral changes developed gradually. These behavioral changes were not observed if the aggressive encounter happened to a stranger mouse, suggesting that social bonding is a prerequisite for stress incubation in this paradigm. This finding was corroborated by hemisphere-specific morphological changes in cortex regions centering at the anterior cingulate cortex, a cognitive and emotional center. Our non-invasive analytical methods to capture informative behavioral details may have applications beyond laboratory animals.

Effects of a 33-ion sequential beam galactic cosmic ray analog on male mouse behavior and evaluation of CDDO-EA as a radiation countermeasure

In long-term spaceflight, astronauts will face unique cognitive loads and social challenges which will be complicated by communication delays with Earth. It is important to understand the central nervous system (CNS) effects of deep spaceflight and the associated unavoidable exposure to galactic cosmic radiation (GCR). Rodent studies show single- or simple-particle combination exposure alters CNS endpoints, including hippocampal-dependent behavior. An even better Earth-based simulation of GCR is now available, consisting of a 33-beam (33-GCR) exposure. However, the effect of whole-body 33-GCR exposure on rodent behavior is unknown, and no 33-GCR CNS countermeasures have been tested. Here astronaut-age-equivalent (6mo-old) C57BL/6J male mice were exposed to 33-GCR (75cGy, a Mars mission dose). Pre-/during/post-Sham or 33-GCR exposure, mice received a diet containing a 'vehicle' formulation alone or with the antioxidant/anti-inflammatory compound CDDO-EA as a potential countermeasure. Behavioral testing beginning 4mo post-irradiation suggested radiation and diet did not affect measures of exploration/anxiety-like behaviors (open field, elevated plus maze) or recognition of a novel object. However, in 3-Chamber Social Interaction (3-CSI), CDDO-EA/33-GCR mice failed to spend more time exploring a holder containing a novel mouse vs. a novel object (empty holder), suggesting sociability deficits. Also, Vehicle/33-GCR and CDDO-EA/Sham mice failed to discriminate between a novel stranger vs. familiarized stranger mouse, suggesting blunted preference for social novelty. CDDO-EA given pre-/during/post-irradiation did not attenuate the 33-GCR-induced blunting of preference for social novelty. Future elucidation of the mechanisms underlying 33-GCR-induced blunting of preference for social novelty will improve risk analysis for astronauts which may in-turn improve countermeasures.

A Temporal Activity of CA1 Neurons Underlying Short-Term Memory for Social Recognition Altered in PTEN Mouse Models of Autism Spectrum Disorder.

Memory-guided social recognition identifies someone from previous encounters or experiences, but the mechanisms of social memory remain unclear. Here, we find that a short-term memory from experiencing a stranger mouse lasting under 30 min interval is essential for subsequent social recognition in mice, but that interval prolonged to hours by replacing the stranger mouse with a familiar littermate. Optogenetic silencing of dorsal CA1 neuronal activity during trials or inter-trial intervals disrupted short-term memory-guided social recognition, without affecting the ability of being sociable or long-term memory-guided social recognition. Postnatal knockdown or knockout of autism spectrum disorder (ASD)-associated phosphatase and tensin homolog (PTEN) gene in dorsal hippocampal CA1 similarly impaired neuronal firing rate in vitro and altered firing pattern during social recognition. These PTEN mice showed deficits in social recognition with stranger mouse rather than littermate and exhibited impairment in T-maze spontaneous alternation task for testing short-term spatial memory. Thus, we suggest that a temporal activity of dorsal CA1 neurons may underlie formation of short-term memory to be critical for organizing subsequent social recognition but that is possibly disrupted in ASD.

Therapeutic Potential of Metformin to Ameliorate Behavioral Symptoms of Autism

Autism spectrum disorders (ASD) affect 1/39 boys and 1/144 girls and manifest in lifelong social interaction deficits and restrictive-repetitive traits of varying severity. Currently no treatments capable of both enhancing social interaction and curbing restrictive repetitive behaviors have been identified. In light of strikingly higher incidence of autism in males versus females, it is surprising that treatments for dysfunctions of steroid hormone metabolism have not been more extensively studied. Clinical evidence supports this approach, as in several studies prepubertal boys and adolescent girls with autism had more androgens and other steroids than age matched children without autism. The antidiabetic drug metformin was found to inhibit adrenergic steroid synthesis, particularly of androgens, through antagonism of melanocortin receptors. Metformin is already co-administered with antipsychotics to manage weight in some autism patients. The goal of this study was to establish in mice if an androgen inhibitor, metformin, can enhance social or reduce repetitive behavior with chronic use. BTBR T+Itpr3tf/J (BTBR) mice were used because this strain is a popular behavioral autism model with high testosterone, corticosterone and progesterone levels, and low serotonin (5-HT) transporter (SERT) density as compared to C57BL/6J (B6) mice, which were used as a strain control. Initially acute metformin treatment dose-dependently enhanced male BTBR mouse social interaction preference for a stranger mouse versus a novel object (p <0.05 different from control at 150 mg/kg dose, N= 9-11 mice), and had no adverse effects on B6 social behaviors. Metformin (150 mg/kg/d) was administered to 10 each adult male and female mice via drinking water for 28 days, yielding a steady state concentration in brain of 36 ± 12 ng/ml. A series of autism relevant behavior tests were performed to compare BTBR and B6 treatment responses. Three chamber social interaction preference tests revealed no effect of chronic metformin treatment on social preference in either strain by the measure of time spent in chambers. Yet BTBR male and female social dominance, measured by the tube test, increased slightly with chronic metformin treatment, but was still less than B6 mouse dominance, which was unaffected by chronic metformin treatment. Marble burying by mice over 30 min did not differ by strain, sex or metformin treatment. Cognitive flexibility in the water T-maze test, measured by number of errors made on day 1 of reversal learning, was unaffected by metformin treatment (F(1,100) = 0.02, p = 0.9). Males and females were pooled for this measure. [3H] histamine and [3H] metformin uptake were greater in hippocampal synaptosomes isolated from metformin treated male B6 mice than in untreated ones (N = 4). This finding may indicate chronic metformin treatment increases hippocampal expression of common transporters of these substrates such as organic cation transporters (OCTs), which also transport 5-HT and other monoamines. Overall, our findings indicate that metformin treatment can enhance social preference and dominance, and chronically it may enhance expression of OCTs.

Cranial irradiation impairs juvenile social memory and modulates hippocampal physiology

Cranial and craniospinal irradiation are the oldest central nervous system prophylaxis treatments considered for pediatric patients with acute lymphoblastic leukemia (ALL). However, survivors of childhood ALL that received cranial radiotherapy are at increased risk for deficits in neurocognitive skills. The continuous and dynamic response of normal tissue after irradiation has been identified as one of the causative factors for cognitive changes after cranial radiation therapy. The aim of our study was to investigate the radiation effects on social behavior and neuronal morphology in the hippocampus of adult mice. Twenty-oneday-old male C57BL/6 mice were irradiated with the small-animal radiation research platform (SARRP). Animals were given a single 10-Gy dose of radiation of X-ray cranial radiation. One month following irradiation, animals underwent behavioral testing in the Three-Chamber Sociability paradigm. Radiation affected social discrimination during the third stage eliciting an inability to discriminate between the familiar and stranger mouse, while sham successfully spent more time exploring the novel stranger. Proteomic analysis revealed dysregulation of metabolic and signaling pathways associated with neurocognitive dysfunction such as mitochondrial dysfunction, Rac 1 signaling, and synaptogenesis signaling. We observed significant decreases in mushroom spine density in the Cornu Ammonis 2 of the hippocampus, which is associated with sociability processing.

Environmental enrichment modulates affiliative and aggressive social behaviour in the neuroligin-3 R451C mouse model of autism spectrum disorder

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterised by impairments in social communication and the presence of restrictive and repetitive behaviours. A mouse model expressing an autism-associated R451C mutation in the gene encoding the synaptic adhesion protein neuroligin-3 (NL3) has been extensively characterised and shows altered behaviour relevant to core traits observed in ASD. Reported impairments in social behaviours in NL3R451C mice however remain controversial due to inconsistent findings in various assays across different laboratories. Such inconsistencies could plausibly be explained by an increased susceptibility of the NL3R451C mouse social phenotype to environmental modulation. To address this, NL3R451C mice were housed in standard or enriched housing from 4 weeks of age prior to behavioural testing. Enrichment rearing enhanced direct interactions with the stranger mouse in all mice in the three-chamber social interaction test however, NL3R451C mice did not show impairment in social interaction in the three-chamber test, in contrast with previous reports. Environmental enrichment enhanced aggressive behaviour in all mice, and did not specifically alter the heightened aggressive phenotype previously described in NL3R451C mice. Specific genotype effects of enrichment included reduced anxiety-like behaviour in WT mice, and lower locomotor activity levels in NL3 mice. While genotype-specific effects of enrichment were not seen on social behaviour, the general increase in affiliative social interaction and aggression seen in all mice, indicates that these behaviours, are vulnerable to change based on housing condition. Mouse models expressing ASD-associated mutations have great utility in elucidating the neurobiology underling development of core traits and it is crucial that efforts are focussed on those models exhibiting robust phenotypes. In light of the findings in the present study, we suggest approaches to improve replicability and reproducibility in mouse models of ASD.

Sex-Selective Effects on Behavior in a Mouse Model of Tuberous Sclerosis Complex.

Tuberous sclerosis complex (TSC) is an autosomal dominant genetic disorder that is caused by a mutation in either TSC1 or TSC2. TSC affects multiple systems of the body, and patients with TSC display a range of neurologic and behavioral manifestations including seizures, intellectual disability, autism spectrum disorders, attention deficit hyperactivity disorder, anxiety, and mood disorders. Whereas behavioral phenotypes of many mouse models have been studied, the effects of sex have, for the most part, not been explored. We studied adult male and female Tsc2 heterozygous and control mice to investigate the influence of sex and genotype on behavior. On a test of social preference, Tsc2 heterozygous mice, regardless of sex, demonstrated lower preference for the stranger mouse than control mice. In the open field, Tsc2 heterozygous males and control females habituated to the open field with decreasing anxiety-like behavior over time, whereas Tsc2 heterozygous females did not show habituation to the open field environment. We did not find any statistically significant effects of genotype on open field activity, learning and memory or motor function. Our results highlight phenotype differences in Tsc2 heterozygous mice, some of which are influenced by sex. A consideration of how sex influences the behavioral phenotypes of TSC is critical to develop a more complete understanding of the disorder and better target future pharmacological treatments.

Altered anxiety and social behaviors in a mouse model of Fragile X syndrome treated with hyperbaric oxygen therapy

Fragile X syndrome (FXS) is a common mental retardation syndrome. Anxiety and abnormal social behaviors are prominent features of FXS in humans. To better understand the effects of hyperbaric oxygen therapy (HBOT) on these behaviors, we analyzed anxiety-related and social behaviors in Fmr1 knockout mice treated by HBOT. In the open field test, HBOT group mice preferred the periphery to central areas and tended to run or walk along the wall. The results suggested that thigmotaxis was significantly increased in the HBOT group compared with the control group. In the elevated plus maze test, the percentage of distance traveled was significantly increased in the open arm and significantly decreased in the closed arm for HBOT group mice compared with control group mice. These results suggested that HBOT group mice displayed enhanced motor activity in the open arm and exhibited fewer anxiety-related behaviors. In the three-chambered social approach test, the HBOT group mice made more approaches to the wire cup containing an acquaintance mouse than control group mice in the sociability test and made more approaches to the wire cup containing a stranger mouse than control group mice in the social novelty preference test. The results suggested that HBOT group mice showed increased levels of social interaction and decreased “social anxiety” than the control group to partner mice in this test. Our findings indicated that HBOT resulted in altered anxiety and social behavior in Fmr1 knockout mice and could possibly be used as a treatment for FXS.

Social behaviour is altered in the insulin-regulated aminopeptidase knockout mouse

Oxytocin, and the closely related neuropeptide, vasopressin, are both known to modulate social behaviours. The pro-social effects of oxytocin are well-documented and have generated much interest into its suitability as a therapeutic for disorders characterised by social dysfunction. This study investigated the social phenotype of mice with a targeted deletion of the gene for insulin-regulated aminopeptidase, an enzyme involved in the degradation of oxytocin and vasopressin. In the 3-chamber sociability test, a genotype effect was observed and subsequent post hoc analysis revealed that male, but not female, insulin-regulated aminopeptidase knockout mice made significantly more approaches to the enclosure holding a stranger mouse than did wildtype mice (p = 0.0039). Male insulin-regulated aminopeptidase knockout mice also displayed decreased rearing (t = 2.309, df = 24, p = 0.0299) and locomotor activity (t = 2.134, df = 24, p = 0.043) in the open field test, suggestive of a reduced stress response to a novel environment. Our findings provide support for the role of insulin-regulated aminopeptidase in influencing social behaviour, possibly via modulation of oxytocin and vasopressin levels. The increase in social interaction observed in the male, but not female, insulin-regulated aminopeptidase knockout mice is in agreement with reports of sex differences in effects of oxytocin and vasopressin on social behaviours and should be explored further.

Inbred strain preference in the BTBR T+ Itpr3tf /J mouse model of autism spectrum disorder: Does the stranger mouse matter in social approach?

BTBR T+ Itpr3tf /J (BTBR) mice have been used as a model of autism spectrum disorder (ASD) due to their low levels of sociability and high levels of repetitive grooming. These experiments explored social behavior in the BTBR and C57BL/6J mice using variations of the three-chambered social approach test. In the first test, the subject mice had a choice between a stranger mouse of the same strain or from a strain with a different level of sociability. The BTBR male mice demonstrated a strong preference for the more social C57BL/6J stranger mouse, as did the C57BL/6J male mice, although more moderately with sniff time only. The C57BL/6J female mice showed a moderate preference, sniff time only, for the BTBR stranger mouse, whereas the BTBR female mice did not show a preference. The second experiment examined whether the subject mouse preferred a stranger mouse or bedding from the stranger mouse home cage. Male BTBR mice always preferred bedding, whereas the C57BL/6J male mice did not show a preference. Both BTBR and C57BL/6J female mice preferred bedding when the stranger mouse was a different strain but not when the stranger mouse was the same strain. Therefore, the stranger mouse strain seems to influence the preference of the female mice more than the male mice. The mice preferred spending time in the chamber with the social smell but not the actual stranger mouse although not always significantly. This suggests that contact with a stranger mouse is more stressful or anxiety provoking than the smell. Autism Res 2019, 12: 1184-1191. © 2019 International Society for Autism Research, Wiley Periodicals, Inc. LAY SUMMARY: BTBR T+ Itpr3tf /J (BTBR) mice have been used as a model of autism spectrum disorder (ASD) due to their low levels of sociability and high levels of repetitive grooming. These experiments explored social behavior in the BTBR and C57BL/6J mice using variations of the three-chambered social approach test. These experiments examined how the sociability level of the stranger mouse affected the subject mouse's preference and if social odor was preferable to a social situation in the BTBR mice. The BTBR male mice demonstrated a strong preference for the more social C57BL/6J stranger mouse, as did the C57BL/6J male mice. The C57BL/6J female mice showed a moderate preference for the BTBR stranger mouse, whereas the BTBR female mice did not show a preference for either stranger mouse. The second modification let the subject mouse have a choice between a stranger mouse or bedding. Male BTBR mice preferred bedding, regardless of the strain of the stranger mouse, whereas the C57BL/6J male mice did not show a preference. Both BTBR and C57BL/6J female mice preferred bedding when the stranger mouse was a different strain but showed no preference when the stranger mouse was from the same strain. The stranger mouse strain seems to influence the female mice more. Male BTBR mice preferred spending time in the chamber with the social smell but not the actual mouse, suggesting that actual contact with a stranger mouse is more stressful or anxiety provoking.

Differences in the expression of restricted repetitive behaviors in female and male BTBR T + tf/J mice

Autism spectrum disorder (ASD) is characterized by the expression of restricted repetitive behaviors (RRBs) and impairments in social recognition and communication. Epidemiological studies demonstrate males are three times more likely than females to be affected. Although this is the case, more recent studies suggest females may be underrepresented in these numbers due to standard clinical measures of RRBs and social behaviors. In addition, many studies examining mouse models of ASD exclude females due to the sex disparity in diagnoses. The present study examined how female and male BTBR T + Itpr3tf /J (BTBR) compare to control C57BL/6J mice on tests of RRBs (probabilistic reversal learning, repetitive grooming, spontaneous alternation, and marble burying) and social behaviors (three chambered social approach task). Utilizing a spatial reversal learning test with 80/20 probabilistic feedback, in which ASD individuals have exhibited deficits, we find that female BTBR mice do not show the same impairment found in male BTBR mice. Interestingly, control female C57BL/6J mice required more trials to reach criterion. Female BTBR mice expressed comparable rates of repetitive grooming, marble burying and spontaneous alternation compared to female C57BL/6J mice. Male BTBR mice expressed higher rates of grooming behavior and locomotor activity compared to male C57BL/6J mice, as found in previous studies. Similarly, male BTBR mice showed a reduction in both measures of social approach compared to controls. Both male and female BTBR mice showed a reduction in sniff time for the stranger mouse compared to controls. Together these findings demonstrate how female BTBR mice do not display the RRB profile expressed by male BTBR mice. Testing of repetitive behaviors in ASD needs to better reflect the sex differences in how RRBs manifest in females compared to their extensively researched male counterparts.

Developmental exposure to mercury chloride impairs social behavior in male offspring dependent on genetic background and maternal autoimmune environment

To date, the connection between inorganic mercury (Hg) and social behavior remains incompletely understood. The aim of this study was to investigate the influence of maternal autoimmunity by inorganic Hg (Hg2+) exposure on social behavior of offspring. Wild-type (WT) and immunoglobulin deficient (Ig−/−) B10.S dams fertilized by male WT B10.S or SJL mice were treated with 50 μM Hg chloride (HgCl2). Non-pregnant female WT B10.S mice were used to investigate factors regulating HgCl2-induced autoimmunity to brain. HgCl2 selectively impaired social behavior in male offspring, but not female offspring from WT B10.S dams × male SJL, in that only male offspring displayed reduced time distribution with the stranger mouse, decreased sniffing to the stranger mouse and increased self-grooming. HgCl2 did not disrupt social behavior of male or female offspring from WT B10.S dams × male WT B10.S or Ig−/− B10.S dams × male SJL. The offspring from WT and Ig−/− B10.S dams × male SJL had equivalent autoimmunity to brain antigens during HgCl2 exposure, indicating that maternal, but not offspring-derived anti-brain antibodies (Ab) impaired social behavior of the offspring. Non-pregnant WT B10.S mice treated with HgCl2 had increased anti-brain Ab dependent on increase in CD4 T cell activation and IFNγ signaling to macrophages. IFNγ interaction with macrophages drove B cells and plasma cells to produce IgG. Therefore, HgCl2 selectively impaired social behavior in males with certain genetic background via maternally derived anti-brain Ab production, thus providing a novel insight into our current understanding of Hg toxicity.

Deficiency of urokinase-type plasminogen activator and its receptor affects social behavior and increases seizure susceptibility

Extracellular proteolysis initiated by the binding of urokinase-type plasminogen activator (uPA) to its receptor (uPAR) regulates the development of inhibitory neuronal circuits in the cerebral cortex and tissue remodeling after epileptogenic brain injury. To study the function of different components of the uPA-uPAR system on behavior and epileptogenesis, and to complement our previous studies on naïve and injured mice deficient in the uPA-encoding gene Plau or the uPAR-encoding gene Plaur, we analyzed the behavioral phenotype, seizure susceptibility, and perineuronal nets surrounding parvalbumin-positive inhibitory interneurons in Plau and Plaur (double knockout dKO) mice. In a climbing test, dKO mice showed reduced interest towards the environment as compared with Wt mice (p < 0.01). In a social approach test, however, dKO mice spent more time than Wt mice exploring the compartment containing a stranger mouse than the empty compartment (p < 0.05). Moreover, in a social interaction test, dKO mice exhibited increased contact time (p < 0.01). Compared with Wt mice, the dKO mice also had a longer single contact duration (p < 0.001) with the stranger mouse. In the elevated plus-maze, grooming, and marble burying tests, the anxiety level of dKO mice did not differ from that of Wt mice. Rearing time in an exploratory activity test, and spatial learning and memory in the Morris swim navigation task were also comparable between dKO and Wt mice. In the pentylenetetrazol (PTZ) seizure-susceptibility test, dKO mice had a shorter latency to the first epileptiform spike (p = 0.0001) and a greater total number of spikes (p < 0.001) than Wt mice. The dKO genotype did not affect the number of cortical perineuronal nets. Our findings indicate that Plau/Plaur-deficiency leads to a more social phenotype toward other mice with diminished interest in the surrounding environment, and increased seizure susceptibility.

Influence of Sex on Cognition and Peripheral Neurovascular Function in Diabetic Mice.

Cognition impairment and peripheral neuropathy (DPN) are two major complications of diabetes. The aim of the present study is to investigate the effect of sex differences on cognition and DPN in diabetic mice. Male and female BKS.Cg-m+/+Leprdb/J (db/db) and db/m mice were used. At ages of 20 and 30 weeks, all animals were subjected to learning, memory and neurological function tests. Regional blood flow in footpad and sciatic nerves were measured using laser Doppler flowmetry. Our data showed that male db/db mice aged 20 weeks and 30 weeks spent significantly more time to locate the hidden platform in the correct quadrant and spent significantly less time exploring the cage with a new stranger mouse compared to aged-matched female db/db mice. Electrophysiological recordings showed that male db mice aged 30 weeks had significantly reduced motor and sensory nerve conduction velocity compared with females. Hot plate and tactile allodynia tests revealed that males exhibited significantly higher thermal and mechanical latency than females. Male db mice aged 30 weeks displayed significantly reduced blood perfusion in sciatic nerve and footpad tissues compared with females. In addition, compared with male and female non-diabetic db/m mice, db/db mice exhibited increased time spent on locating the hidden platform, decreased time spent on exploring the novel odor bead and an unfamiliar mouse, as well as showed significantly lower levels of blood flow, lower velocity of MCV and SCV, higher thermal and mechanical latencies. Blood glucose levels and body weight were not significantly different between male and female diabetic animals (age 30 weeks), but male db mice showed a higher serum total cholesterol content. Together, our data suggest that males develop a greater extent of diabetes-induced cognition deficits and peripheral neurovascular dysfunction than females.

Assessment of Topographic Memory in Mice in a Complex Environment Using the Hamlet Test.

Here we provide instructions to measure topographic memory in mice using the Hamlet test, a complex environment. The apparatus mimics a small hamlet with a central agora and five houses, which are functionalized since mice can drink, eat pellets, hide within a small maze, run in an activity wheel, or interact with a stranger mouse behind a grid. The houses are interconnected through a network of streets in a five-arm star shape, and a video tracking system takes information from the activity in each house or follows a single mouse by trajectometry. Training the mice in the Hamlet, in groups for several hours per day over several days or weeks, allows consolidation of topographic memory (i.e., route learning involving both allocentric and egocentric strategies). Analysis of topographic memory can be performed a posteriori in a probe test by depriving mice of water or food and measuring their ability to efficiently reach the "eat" or "drink" house. Control groups include mice tested in non-deprived condition and mice naïve to the Hamlet and tested in deprived or non-deprived conditions. The present article details the apparatus, procedures, and protocols that can be used to reliably habituate mice in this complex environment and measure topographic memory. © 2018 by John Wiley & Sons, Inc.

Topographical memory analyzed in mice using the Hamlet test, a novel complex maze.

The Hamlet test is an innovative device providing a complex environment for testing topographic memory in mice. Animals were trained in groups for weeks in a small village with a central agora, streets expanding from it towards five functionalized houses, where they can drink, eat, hide, run, interact with a stranger mouse. Memory was tested by depriving mice from water or food and analyzing their ability to locate the Drink/Eat house. Exploration and memory were analyzed in different strains, gender, and after different training periods and delays. After 2 weeks training, differences in exploration patterns were observed between strains, but not gender. Neuroanatomical structures activated by training, identified using FosB/ΔFosB immunolabelling, showed an involvement of the hippocampus-subiculum-parahippocampal gyrus axis and dopaminergic structures. Training increased hippocampal neurogenesis (cell proliferation and neuronal maturation) and modified the amnesic efficacy of muscarinic or nicotinic cholinergic antagonists. Moreover, topographical disorientation in Alzheimer's disease was addressed using intracerebroventricular injection of amyloid β25-35 peptide in trained mice. When retested after 7 days, Aβ25-35-treated mice showed memory impairment. The Hamlet test specifically allows analysis of topographical memory in mice, based on complex environment. It offers an innovative tool for various ethological or pharmacological research needs. For instance, it allowed to examine topographical disorientation, a warning sign in Alzheimer's disease.