Behavioural Brain Research

Reports of honeybees demonstrating abstract concepts like sameness and difference marked a pivotal development in comparative psychology. Subsequent studies expanded the scope of concept learning in honeybee cognition, yet most evidence relies on a single method: the delayed-matching-to-sample task using a Y-maze. Whether this setup is uniquely effective or if alternative approaches could yield similar results remains unresolved. Additionally, the failure of bumblebees (Bombus spp.) to complete this task, despite honeybees demonstrating success, remains unexplained. This study compared the performance of honeybees (Apis mellifera) and bumblebees (Bombus terrestris) across matching-to-sample tasks with varying degrees of physical continuity between sample and target stimuli. The objectives were twofold: to evaluate an alternative method for assessing concept learning in both species and to investigate potential species differences in such tasks. Contrary to prior findings, neither species succeeded at the reported proficiency levels in simultaneous matching-to-sample tasks. Moreover, bumblebees outperformed honeybees in one task. These results are consistent with an explanation based on species-specific differences in visual attention mechanisms, and underscore the need for further research on concept learning in social bees.

Physical activity offers myriad benefits to health and well-being, in humans and other animals as well. In rodents, voluntary wheel running can attenuate the effects of both physical and social stressors on rodent social behavior. Whether wheel running affects rodent social behaviors per se remains less well understood. We conducted the current study to test whether home cage access to running wheels impacts the social behaviors of adult, group-housed C57BL/6J female mice during same-sex interactions with novel females. Group-housed females were either given continuous home cage running wheel access or a standard paper hut starting at weaning, and as adults, social behaviors were measured during interactions with novel females. In two cohorts, we found that 5 weeks of running wheel access during adolescence reduced the time that subject females spent investigating a novel female and also tended to reduce total ultrasonic vocalizations produced during interactions. These effects were not reversed by a 2-week period of running wheel removal but were recapitulated in a different cohort by 2 weeks of running wheel access in adulthood. Unexpectedly, we found that these effects on female social behavior were not due to wheel running per se, because females raised from weaning with immobile running wheels also showed low rates of social behaviors during same-sex interactions in adulthood. Overall, we find that the presence of a running wheel in the home cage has an enduring inhibitory influence on female social behavior during same-sex interactions, a finding that has implications for the design of studies that include same-sex interactions between female mice.

Sexual motivation is a complex concept involving both the initial drive to begin mating and the motivation to sustain copulation. Disruptions in sexual motivation are often observed in psychiatric disorders. This study proposes that sexual motivation can be divided into two distinct components: sexual incentive motivation and the drive to sustain copulation. To investigate this, we utilized the Motivation to Continue Copulation (MCC) test, which measures effort (nose pokes) to gain access to a sexual reward, and compared it with the Sexual Incentive Motivation (SIM) test and standard copulation tests. Male and female rats were trained on a fixed ratio (FR) 1 schedule using cheese rewards before transitioning to sexual rewards. After six FR1 sessions, the effort required increased to FR5 and progressive ratio (PR) schedules. Results revealed that sexual incentive motivation, measured by the SIM test, was higher in males after sexual experience, while females maintained consistent levels. In the MCC test, both males and females exhibited increased motivation to continue copulation with experience, but the motivation declined in the 2nd ejaculatory series. These findings demonstrate that sexual motivation comprises distinct components. The MCC test effectively measures the drive to sustain copulation, while the SIM test assesses incentive motivation. This distinction is crucial for advancing behavioral neuroscience and understanding sexual dysfunction in psychiatric conditions.

The midsession reversal (MSR) task is frequently used to study behavioral flexibility and decision strategies in animals. In a typical version of the task, subjects complete 80 trials in which they choose between two simultaneously presented stimuli, S1 and S2. During the first 40 trials, responses to S1 are reinforced, whereas responses to S2 are not. The contingencies then reverse without warning: From trial 41 to 80, only responses to S2 are reinforced. In birds, performance in this task is often characterized by anticipatory and perseverative errors around the reversal point, suggesting a reliance on elapsed time since the session began. In contrast, rats tested in operant conditioning chambers typically show near-optimal performance with few errors, a pattern often interpreted as evidence that rats rely primarily on local reinforcement cues rather than temporal information. The present study investigated whether rats exclusively rely on local cues in the MSR task or whether temporal information also contributes to the decision process. Two groups of rats were trained with different intertrial intervals (ITIs; 5 s or 10 s) while the reversal point remained fixed at Trial 41. During acquisition, both groups diplayed similar learning rates and near-optimal steady-state performance with minimal anticipatory or perseverative errors. However, when the ITI was manipulated in probe sessions, systematic shifts in switching behavior emerged. Rats adjusted their choices according to the temporal midpoint experienced during training rather than the nominal trial number of the reversal. These results suggest that rats rely on a mixed strategy that integrates local reinforcement cues with global timing information. Temporal control may therefore be present even when it is not expressed during standard training conditions.

RationaleRecent research on habits and skills has produced a wave of new theories regarding the shift in control from medial to lateral regions of the dorsal striatum, and how these regions are implicated in the selected and executed of action sequences. ObjectivesTo examine the comparative effects of muscimol/baclofen inactivation and dopamine D1 and D2 receptor agents in the dorsomedial (DMS) and dorsolateral (DLS) striatum on the performance of skilled action sequences. MethodsInfusions were made in well-trained rats using the five-step nose poke task to isolate the effects on initiation, execution and termination components of skilled action sequences. ResultsDLS inactivation produced sequencing deficits like those observed with pre-training lesions, indicating that the DLS is critical for both the acquisition and performance of sequences. Behaviour was unchanged following DMS inactivation, consistent with models of DMS disengagement following training. Infusions of D1 and D2 antagonists did not alter behaviour, however the D2 receptor agonist quinpirole increased sequence errors at a low dose and reduced sequences at the high dose in the DLS. DLS manipulations impaired sequence initiation and termination as well as reward transitions, while the chunking ballistic response pattern was largely unaltered, indicating that between-but not within-sequence actions rely on the DLS. ConclusionsSkilled action sequencing, including chunk transitions was dependent on DLS and its modulation by D2 receptors, but not on DMS function. Using a novel sequencing task, these results support the dissociable and dopamine-dependent role of the dorsal striatum subregions in performing skilled motor actions.

Exercise is a positive health behaviour associated with improved mood. However, the mechanisms underlying the benefits of exercise on affective health are unclear, particularly with respect to type of exercise and sex. Chronic exercise decreases neuroinflammation, which is linked to improvements in mood and anxiety. However, exercise is also a physiological stressor that can transiently upregulate systemic inflammation, and its effects on neuroinflammation are not well understood. This study examined how acute and chronic exercise affect circulating and brain cytokine levels and anxiety-related behaviour in young healthy male and female mice. In Experiment 1, mice were placed on a treadmill for a two-hour bout of moderate exercise. Two hours after exercise, animals were either tested in the open field or euthanized for measurement of cytokines (IL-1{beta}, TNF, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, IFN-{gamma}, KC/GRO). In Experiment 2, mice underwent an 8-week moderate treadmill exercise paradigm followed by open field testing and tissue collection. Acute exercise decreased time spent in the centre of the open field in males only, suggesting increased anxiety-like behaviour in males. Acute exercise increased IL-6 and decreased TNF in serum, and increased amygdala principal component 1 (loading IL-12p70, IL-10, IFN-{gamma}, and TNF) in both sexes. Chronic exercise increased open field centre entries, increased IL-6 in the prefrontal cortex, decreased TNF in the dorsal hippocampus, and had minimal effects on circulating cytokines in both sexes. These results demonstrate that the effects of exercise on anxiety-related behaviour and cytokine levels depend on recurrence, tissue, and brain region. New & NoteworthyOur work highlights the contrast between anxiogenic and anxiolytic effects of acute versus chronic exercise, respectively, in healthy mice. Acute and chronic exercise differentially affected circulating and brain cytokines, providing insight into physiological adaptations to exercise. Both sexes demonstrated similar cytokine responses to exercise. These similarities are novel with respect to exercise research and noteworthy given sex differences in anxiety with respect to acute exercise.

Traumatic brain injuries (TBIs) result from impact to or rapid displacement of the brain and can lead to various neurological deficits involving working memory, decision-making, and anxiety. While large-scale effects of brain damage are well-described for more severe TBIs, less is known about the extent and duration of cognitive deficits at the mild level. Interval timing can provide a helpful window into cognition in mice and humans. Interval-timing behavior is impaired in a wide range of neuropsychiatric disease states, such as Parkinsons disease. Furthermore, novel object recognition (NOR) and the Barnes maze (BM) tests are valuable assays for evaluating spatial learning, working memory, and anxiety-like behavior in mice. Here, we employed a weight-drop model of mild TBI (mTBI) to investigate changes in internal cognitive states resulting from mTBI treatment. mTBI mice were not significantly impaired in either interval timing or NOR, but they demonstrated impaired spatial memory in the Barnes Maze. Interestingly, within-sex comparisons revealed impairments in male mTBI mice in the interval-timing task and the NOR, suggesting that male and female mice may be differently affected by mTBIs.

Cannabis contains many bioactive compounds, including {Delta}9-Tetrahydrocannabinol (THC) and cannabidiol (CBD), which influence behavior through complex pharmacological interactions with endogenous targets. This study examines whether CBD influences THC-induced changes in motor activity, hypothermia, and antinociception traits across different THC:CBD ratios, sexes, and genetic backgrounds. Traits were measured in C57BL/6J (B6) and DBA/2J (D2) mice of both sexes following baseline intraperitoneal (i.p.) injection of vehicle (VEH) and two consecutive daily doses of VEH or THC (10 mg/kg) alone or in combination with 0.56, 5, or 10 mg/kg CBD (THC:0.56CBD, THC:5CBD, or THC:10CBD, respectively). Motor activity and hypothermia were quantified daily from 0 to 120 min following injection and antinociception was measured daily at 60 min. We found that CBD alters THC-induced changes in motor activity and hypothermia as a function of day, dose, time, sex, and strain. In D2 females, CBD dose-dependently attenuated the hypolocomotor effects of THC immediately following acute injection and enhanced these effects later at 75 min. Following repeated exposure, CBD dose-dependently enhanced THC-induced hypolocomotion in B6 females at 75 min and in D2 males at 30 min while attenuating THC-induced hypolocomotion in D2 females immediately following injection. In D2 females, CBD dose-dependently attenuated THC-induced hypothermia at 15 min and enhanced hypothermia relative to THC at 30 min in D2 males following acute injection. After repeated exposure, CBD dose-dependently enhanced THC-induced hypothermia in B6 females at 15 min and in D2 males from 30 to 120 mins, while attenuating hypothermia in D2 females at 30 min. No significant effects of CBD on antinociception were observed. Our results indicate that CBD can modulate some THC-induced traits acutely and after repeated exposure. Regulation of THC-induced behavioral responses is dependent on CBD dose, genetic background, and sex. A candidate gene search using brain gene expression in recombinant inbred mice revealed greater genetic variation in ion channel genes relative to key metabolic genes, suggesting an underlying pharmacodynamic mechanism. Future research and validation of molecular mechanisms underlying these differences is expected to enhance our understanding of potential health risks and clinical relevance of cannabis and cannabinoid compounds containing THC and CBD.

Accurate and efficient memory processing is essential for survival. Recent work in human subjects and animal models has suggested that memory processing may differ in meaningful ways between males and females. In mice, contextual fear memory (CFM) encoding, consolidation, and recall have been well studied, and the mouse hippocampus and amygdala have been implicated in these processes. The present study addresses how the specific contribution of these brain regions to each stage CFM processing in female vs. male mice. We find that male and female mice show no differences in CFM recall, nor in sleep behavior in the hours following single-trial contextual fear conditioning (CFC), which is essential for CFM consolidation. However, females - but not males - show significantly increased expression of cFos in dorsal hippocampal CA1 and CA2 neurons during CFM encoding. On the other hand, males - but not females - show increased cFos expression among DG granule cells during CFM consolidation. These findings highlight the fact that the neurobiological underpinnings of memory processing may differ between males and females, even when recall performance is identical. Scope statementHistorically, research on the neurobiological basis of memory processing has been carried out mainly in male subjects. Thus, our understanding of these mechanisms is biased towards male brain neurophysiology. Recent studies have variously reported performance differences for episodic memory tasks, in which male subjects perform better, worse, or the same as females. Here, we find that male and female mice perform similarly on a well-studied experimental memory task but nonetheless have differences in the relative activity of different brain structures during sequential stages of memory processing. This emphasizes the importance of including both males and females in memory studies, due to potential sex differences in the neurobiological substrates of memory.

The purpose of this study was to investigate the influence of treadmill belt speed on mechanically-induced (tail-pinch) and pharmacologically-induced (quipazine, a 5HT2A agonist) stepping behavior in one-day old rats. On postnatal day 1 (P1), male rat pups were tested on one of four moving treadmill belt speeds. Stepping was induced using a tail-pinch and quipazine administration to examine real-time adaptations on belt speeds. For tail-pinch-induced stepping in the forelimbs and hindlimbs, there was a significant effect of time but not belt speed. Step cycle duration was significantly shorter for both forelimbs and hindlimbs on the fast belt speed compared to all other belt speeds. For the forelimbs, this effect was driven by changes in stance phase duration. Compared to control the speed, step area was significantly larger on medium and fast speeds for forelimbs and slow and fast speeds for hindlimbs. For quipazine-induced stepping, forelimbs and hindlimbs showed significantly more steps across slow, medium, and fast belt speeds compared to the control speed. The forelimbs showed significantly shorter step cycle durations on the fast belt speed compared to the control belt speed. Again, this difference was driven by changes in the stance phase. There were no significant differences in step cycle duration, stance and swing phase durations, or step area between speeds for the hindlimbs. Overall, we showed that both mechanical and pharmacological stimulation is reliable at inducing stepping on a moving treadmill belt in neonatal rats, and P1 rats show real-time adaptations in response to a moving treadmill belt.

Ultrasonic vocalizations (USVs) are widely used in rodent social communication, yet the functional significance of male-male vocal interactions in mice remains unclear. Here, we investigated whether USVs produced during specific social behaviors influence the behavior of conspecifics. Using playback experiments, we compared responses to vocalizations recorded during chasing and being chased in male-male interactions. We found that USVs emitted by chased intruders consistently elicited approach behavior in receiver mice, whereas those emitted by chasing individuals did not. Acoustic analyses revealed that these vocalizations differed in syllable composition, with intruder calls containing a higher proportion of upward frequency-modulated syllables and exhibiting higher mean frequencies. In addition, the temporal organization of syllables appeared to contribute to the behavioral response. Together, these results suggest that male mice respond selectively to certain USV patterns associated with specific social contexts, indicating that acoustic features and temporal structure may jointly influence social approach behavior in mice. HighlightsO_LIBehavioral context (chased vs. chasing) shapes the composition of USV syllable types C_LIO_LIMale mice selectively approach USVs from chased intruders, but not chasing residents C_LIO_LIThe approach response exhibits high temporal synchrony across individual receivers C_LIO_LITemporal organization of syllables modulates approach behavior based on acoustic features C_LI

BackgroundThe three-chamber test (3CT) is widely used to assess social behaviour in mice, based on the assumption that time spent near a conspecific reflects motivation for social contact. However, the design of the task constrains interpretation, as behaviour may reflect exploration, novelty seeking, or territorial investigation rather than affiliative social motivation. In addition, key biological factors such as sex differences and social hierarchy are often overlooked. AimsWe hypothesised that the 3CT overestimates sociability and used a direct-interaction phase to investigate motivation for affiliative social contact. We also integrated social status to determine if this modulated behavioural patterns and interacted with sex. MethodsAdult male and female C57BL/6 mice (n = 32) were tested in a standard 3CT, followed by removal of the cage barrier to permit direct contact. Behavioural parameters were quantified, and social status was determined using the tube test. ResultsMales exhibited higher social interest index scores than females. Once the barrier was removed, both sexes displayed a negative direct sociability index, indicating greater environmental exploration than social engagement. Correlation analysis revealed no association between indirect and direct measures. Sex differences emerged primarily among submissive mice, with submissive males showing greater social investigation than submissive females. ConclusionThese findings suggest that standard 3CT indices reflect exploratory rather than affiliative social motivation. The modified paradigm incorporating direct interaction provides a more realistic assessment of social behaviour and challenges assumptions about intrinsic sociability in mice.

Sleep plays a key role in both physical recovery and welfare. However, sleep patterns remain poorly documented in animals, particularly in athletic horses. This study aimed to provide a detailed description of sleep quantity and quality in training Thoroughbred racehorses and to investigate their relationships with age, abnormal behaviours, and cortisol. Thirteen Thoroughbreds (2-7 years old) were continuously monitored in their home environment over three consecutive days. An ethogram was used to quantify the two main phases of sleep: Non Rapid Eye Movement sleep (NREM) and Rapid Eye Movement sleep (REM), as well as sleep interruptions (from day 1 at 12:00 a.m. to day 3 at 12:00 a.m.). Sleep Quality Indices (SQI), defined as the quantity of sleep divided by the number of sleep interruptions (SI), were calculated. Behavioural observations of four indicators of poor welfare (alertness, stereotypies, inactivity, aggressiveness towards humans) were performed using scan sampling, and salivary cortisol was measured each morning. Linear models were used to assess the links between sleep quantity and quality, age, mean cortisol, and abnormal behaviours. Sleep quantity was significantly associated with age: positively for total NREM sleep (ANOVA: {chi}{superscript 2} = 5.26, p < 0.05) and, negatively for total REM sleep (ANOVA: {chi}{superscript 2} = 4.46, p <0.05) and total recumbency duration (ANOVA: {chi}{superscript 2} = 5.68, p < 0.05), suggesting an age-related shift favouring NREM over REM. Morning cortisol concentrations and the frequency of abnormal behaviours were significantly higher in horses with lower sleep quality (cortisol: Total SQI, ANOVA: F = 5.26, p < 0.05; Combined SQI, ANOVA: F = 5.40, p < 0.05; abnormal behaviours: Total SQI, ANOVA: F = 4.07, p = 0.074), pointing to a potential link between stress or altered welfare and poorer sleep quality. These findings suggest that, whilst the type and duration of equine sleep may be mainly affected byage, sleep quality is associated with both cortisol levels and the expression of abnormal behaviours, indicating that poor sleep quality may be linked to poor welfare in this population of horses. Thus, sleep appears to be closely linked with racehorse welfare, highlighting the need for further investigation into how it is influenced by factors such as husbandry, training load, recovery, and performance.

Learning dance of a motor sequence involves the coordination of both oculomotor and manual motor systems through the practiced repetition of a fixed sequence of actions, resulting in automatized execution of movement through habit learning. This study aims to address whether a sequence-based learning paradigm centered on the visual-motor system can feasibly be measured while listening to music (Bar and DeSouza 2016). It aims to develop a new visual-motor-based learning paradigm with music, potentially promoting neuroplasticity and creating new interventional tools, building upon prior research that shows behavioural and putative neural changes following dance-based neurorehabilitation in people with Parkinson's disease (Bearss et al. 2024). Eye movements of 10 participants (8 female, 2 male) were tracked using the Eyelink 1000 Plus system during a 68-second eye-dance sequence. The experiment consisted of a learning phase, where participants observed the sequence five times with 30-second breaks, and a performance phase, where they performed the sequence five times from memory on a grey screen without visual cues. Music was incorporated into both phases to aid memorization of the 4 spatial locations. After each performance, the participant was shown a visual reinforcer and asked for their thoughts on how well they executed the dance. A visual reinforcer flashes one of three different colours: red, yellow, or green. Each colour corresponds to how many steps in the dance a participant performed correctly, with key points being: under one third, between one to two thirds, and over two thirds of total steps correct. Participants were scored based on timing of the steps as well for exact (1.00), good (0.66), slightly off (0.33) or missed (0) steps. Data was analyzed using R4.3.1, MATLAB, and Experiment Builder: Data Viewer software. Results showed a significant improvement in performance accuracy between the first session (g1; M = 40%, SD = 7.2%) and the last session (g5; M = 69.7%, SD = 22.8%). A repeated-measures ANOVA revealed a significant main effect of session on performance accuracy, F(4, 36) = 6.99, p < 0.001, 2G = 0.26, indicating that accuracy significantly improved over sessions. Post-hoc Bonferroni comparisons showed that accuracy in later sessions was significantly higher than earlier sessions, suggesting a defined learning curve and consolidation of performance pattern across repeated practice. Similarly, there was significant improvement in timing accuracy between the first session g1; M = 0.29, SD = 0.06) and the fifth session (g5; M = 0.46, SD = 0.12). A repeated-measures ANOVA revealed a significant main effect of session on timing precision, F(4, 36) = 11.67, p < 0.001, 2G = 0.25, indicating significant improvements in temporal control and coordination over sessions. Post-hoc Bonferroni comparisons showed that timing precision significantly improved between early and late sessions (e.g, g1-g4, p <0.01; g1-g5, p < 0.001), suggesting a defined learning curve and increase in precision across repeated practice. These findings suggest that visual-motor-based interventions have the potential to enhance motor and non-motor symptoms like depression and anxiety for neurodegenerative diseases such as Parkinson's Disorder (PD). The results provide a foundation for developing targeted therapies that integrate learning paradigms to improve functional outcomes, warranting further exploration of their long-term efficacy.

The misuse of opioid medications is a significant health issue in the United States. Very few studies have investigated the effect of opioids on perineuronal nets (PNNs), scaffold-like structures that surround neurons and are involved in the regulation of plasticity-dependent mechanisms such as development, learning and memory, and acquisition of addiction-like phenotypes. Regulation of PNNs in the orbitofrontal cortex (OFC) during periods of drug intoxication or withdrawal is widely unknown. In this study, male Wistar rats were injected with fentanyl (0.125 mg/kg, s.c.) or 0.9% saline twice daily for 7 days and once on day 8 (7continuous days following by 3 days of abstinence) or twice daily for 15 days (5 continuous days followed by 2 days of abstinence for more than 3 weeks) and twice on day 16. Antinociception was evaluated using the tail immersion test immediately before and 30 minutes after injections. Whole-brain coronal slices were collected, and immunohistochemistry was used to identify Wisteria Floribunda Agglutinin (WFA)-positive PNNs and parvalbumin (PV)-expressing cells. Results confirmed that repeated fentanyl injections induced tolerance to the antinociceptive effects, which normalized following acute abstinence periods. WFA intensity decreased following 8 days of injections. Analyses confirmed significant correlations between PV+ density and tail withdrawal latency following 8 days of fentanyl injections. These data confirm that repeated fentanyl injections modulate both WFA+ and PV+ expression in the rodent brain and antinociceptive tolerance in a duration-dependent manner. Overall, these data suggest that perineuronal nets may mediate opioid-induced behavioral effects, such as antinociceptive tolerance, following repeated administration and abstinence in rats.

Sex differences in responses to chronic stress are implicated in the higher prevalence of major depression and PTSD in females. Evidence of sex differences in endocannabinoid (eCB) physiology suggests that eCB signaling contributes to sexual disparities in fear conditioning and extinction. In adolescent male Sprague-Dawley rats, exposure to chronic-mild-unpredictable stress (CMS) resulted in enhanced trace-fear conditioning that was reversed by CB1 activation (Reich et al, 2013). In the present study, we assessed the effects of CMS and CB1 activation on hippocampal-dependent trace and contextual fear conditioning in adolescent female Sprague-Dawley rats. CMS exposure enhanced trace freezing behavior during memory recall compared to non-stress controls. This effect was not observed in contextually conditioned females. The CB1 receptor agonist, ACEA (0.1 mg/kg), administered prior to trace memory recall, but not prior to acquisition, significantly decreased freezing in both stress and non-stress females. ACEA significantly reduced baseline freezing behavior during trace memory recall in both stress and non-stress rats, however ACEA either 1) did not affect or 2) impaired short and long-term extinction in stress and non-stress females. In contextually conditioned females, ACEA decreased freezing during memory recall, although the effect was more robust in stress rats. ACEA impaired long-term contextual extinction in stress females while facilitating this in non-stress controls. However, ACEA had no effect or impaired short-term contextual extinction in both stress and non-stress groups. The results demonstrate that CMS enhances hippocampal-dependent episodic fear memories but has limited effects on contextual fear conditioning in female rats. These findings have implications in the use of medical cannabinoid treatment of disorders such as PTSD, as well as recreational cannabis use in adolescent/young adult females.

Changes within neurotransmitter systems are associated with variation in anxiety-related behavior. The adenosine signaling pathway has been associated with anxiety and caffeine has been utilized as a modulator. However, studies have not considered the impact of an individuals stress coping style (e.g. proactive, reactive) and corresponding differences in neuromolecular signaling that can influence the behavioral responses. To assess the role of adenosine signaling, we acutely treated reactive and proactive zebrafish with 50 mg/L caffeine and evaluated anxiety-like behavior using a novel tank diving test (NTDT). We then quantified whole-brain gene expression of genes representing distinct parts of the adenosine signaling pathway: adenosine receptors A1B, A2Aa, A2Ab, and A2B (adora1b, adora2aa, adora2ab, and adora2b, respectively) and enzymes adenosine deaminase (ada) and ecto-5-nucleotidase (nt5e). We found significant main effects of coping style, sex, treatment, and coping style by sex by treatment interaction effect on stress behaviors. Specifically, compared to controls, caffeine reduced stress behavior in only reactive males. We also observed significant differential baseline gene expression within the adenosine signaling pathway between the reactive and proactive strains, where reactive zebrafish expressed higher levels of adenosine receptors A1B, A2Ab, A2B, and adenosine deaminase and lower levels of adenosine receptor A2Aa than proactive zebrafish. These findings indicate that variation in adenosine signaling between the stress coping styles and sexes may be contributing to differences in anxiety-related behavior.

When novel nutrient-rich food sources become available to species sharing the same natural habitat, interspecies competition may arise, yielding insights into the ecological and social dynamics of the observed species. Here, we investigated food consumption patterns, and consequent social interactions, by two sympatric species of mice in response to a novel nutrient-rich food source. By deploying, in the mices natural habitat, baited video-monitored chambers, we collected, over a 5-month period, 1805 observations of food visiting by Apodemus agrarius and Apodemus flavicollis. We also documented interspecific encounters, with 86.7% of the cases showing agonism. In these interspecies agonistic encounters, A. flavicollis was always the initiator of agonism, attacking within 2 sec in 92.3% of the cases, and being dominant over A. agrarius in 84.6%. Analysis of food visiting behavior revealed that, initially, both species preferred nocturnality. However, after the interspecies fights, A. agrarius switched its temporal preference to diurnality, leading to temporal niche segregation between the two species and a significant reduction of interspecies encounters. Moreover, A. agrarius demonstrated hour-specific avoidance of A. flavicollis, visiting significantly less in hours with A. flavicollis compared to hours without. Through temporal niche switching, A. agrarius managed to access the food source safely, without fights. In contrast, A. flavicollis remained consistently nocturnal across the entire study. Notably, our study presents the first 24h foraging actogram for free-living rodents. Moreover, while rodent interspecific competition is a well-known phenomenon, most of what we know about it comes from indirect observations. Direct observations of rodent interspecific interactions in nature are rare. Our work is the first direct (video-monitored) observation of temporal switch-inducing interspecies interactions in nature. As free-living rodents are currently considered a major model system for the study of interspecific competition, these results may offer precious insights for a better understanding of social dynamics, especially in asymmetric relationships. Furthermore, our findings highlight the significance of considering temporal dynamics in studies of interspecific interactions.

Negative cognitive biases in depression are more pronounced in females than in males. This sex difference emerges during adolescence, a sensitive developmental stage when chronic stress exposure increases the risk of depression in adulthood. The neurobiology linking adolescent stress to sex-specific cognitive bias and resting-state network reorganization in adults remain poorly understood. The study aimed to investigate the longitudinal effects of chronic restraint stress (CRS) during adolescence on cognitive bias and functional connectome in emerging adulthood. 28 Wistar rats (sex-balanced; aged five weeks on arrival) were trained on a judgment bias task with distinct tactile cues signalling differential rewards. Cognitive bias was quantified from responses to ambiguous probe trials. Following training, animals were randomly and equally assigned to CRS or control groups (sex-balanced). Offline resting-state functional MRI scans were conducted at adolescent baseline (pre-CRS) and again in adulthood (post-CRS), followed by probe trials to assess neural and behavioural changes. Following CRS, females showed a greater tendency to shift toward negative bias than males (ratio of odds ratio=3.67). Furthermore, CRS significantly reduced functional connectivity between the left cerebellar-auditory and hypothalamic-thalamic networks only in females. Repeated-measures correlation between cognitive bias and network connectivity were not statistically significant across sex-by-group strata, potentially due to offline imaging and small sample size. However, intra-individual association revealed sex-specific trends, with CRS females showing moderately positive correlations and CRS males exhibiting a weak negative association. The results could inform stratified connectome-based interventions targeting adolescent stress exposures to potentially reduce the risk of adult depression. Six keywords: Resting-State Functional MRI, Chronic Restraint Stress, Judgement Bias, Open Field Test, Sex Differences

Background and aimsPerseverative behaviours are commonly assessed using operant paradigms in which rodents work for drugs or food under physiological deprivation, limiting translational relevance to some behavioural addictions. Here we validated an operant paradigm in which the acquired behaviour is driven neither by physiological needs nor hedonic responses. MethodsMice were trained to lever-press for green light. Exp.1 used a within-subjects design to examine lever discrimination and whether responding could be "satiated" by light preexposure. Exp.2 examined instrumental contingency using a between-subjects design, with light delivery equated between contingent and non-contingent groups. Exp.3 replaced green light with dim red light producing less retinal photoreceptor excitation but comparable heat to assess non-photic cues. Exp.4 examined whether green light could affect food seeking different motivational states. ResultsIn Exp.1, green light supported lever discrimination. Among high responders, the satiation effect was modest (<15% reduction) and did not deter lever pressing. In Exp.2, instrumental contingency promoted response acquisition whereas random light delivery did not. In Exp.3, dim red light failed to sustain behaviour, producing [~]50% response decrement. In Exp.4, light potentiated food seeking under ad libitum feeding. Discussion and conclusionsResponse-contingent light serves as a reward to establish operant responding, which cannot be explained by alerting effects or thermal cues. Our study bridges the gap between animal models and findings from humans that coloured light may exacerbate smartphone use and that light therapy may reshape reward circuits in individuals with Internet gaming disorder symptoms [Li et al. (2026) Advanced Science 13:e14044].