Structural enrichment for laboratory mice: exploring the effects of novelty and complexity.

Providing structural enrichment is a widespread refinement method for laboratory rodents and other animals in captivity. So far, animal welfare research has mostly focused on the effect of increased complexity either by accumulating or combining different enrichment items. However, increasing complexity is not the only possibility to refine housing conditions. Another refinement option is to increase novelty by regularly exchanging known enrichment items with new ones. In the present study, we used pair-housed non-breeding female C57BL/6J and DBA/2N mice to investigate the effect of novelty when applying structural enrichment. We used a double cage system, in which one cage served as home cage and the other as extra cage. While the home cage was furnished in the same way for all mice, in the extra cage we either provided only space with no additional enrichment items (space), a fixed set of enrichment items (complexity), or a changing set of enrichment items (novelty). Over 5 weeks, we assessed spontaneous behaviors, body weight, and extra cage usage as indicators of welfare and preference. Our main results showed that mice with access to structurally enriched extra cages (complexity and novelty) spent more time in their extra cages and complexity mice had lower latencies to enter their extra cages than mice with access to the extra cages without any structural enrichment (space). This indicates that the mice preferred the structurally enriched extra cages over the structurally non-enriched space cages. We found only one statistically significant difference between the novelty and complexity condition: during week 3, novelty mice spent more time in their extra cages than complexity mice. Although we did not detect any other significant differences between the novelty and complexity condition in the present study, more research is required to further explore the potential benefits of novelty beyond complexity.

Not all mice are alike: Mixed-strain housing alters social behaviour.

The use of millions of mice in scientific studies worldwide emphasises the continuing need for a reduction of sample sizes, however, not at the expense of scientific validity. Split-plot designs have been suggested to enhance statistical power while allowing a reduction of animal numbers in comparison to traditional experimental designs. Recently, a promising approach of a split-plot design has been implemented and proven useful using mixed-strain housing of at least three different mouse strains. However, the impact of co-housing different strains of mice in one cage on animal welfare has still to be defined. This study aimed at comparing the effects of mixed-strain and same-strain housing of female C57BL/6J and DBA/2N mice on welfare and behaviour in two experimental phases. In a first phase, mice were housed in either mixed- or same-strain pairs. Home cage behaviour, activity rhythm, body weight, and faecal corticosterone metabolites were assessed. Furthermore, tests for anxiety-like and exploratory behaviour as well as spatial learning were performed. In a second phase, sociability was investigated in newly formed mixed-strain quartets. Mixed-strain housing did not induce alterations in anxiety, locomotion, learning, stereotypic behaviour, and stress hormone levels. However, changes in social behaviours and activity rhythm were observed. Increased agonistic and decreased socio-positive behaviours might point towards mild impacts on welfare in C57BL/6J mice under co-housing conditions. Altogether, scientific research may greatly benefit from co-housing mice of different strains within the same cages (e.g. for the realisation of a split-plot design), provided that strains are carefully selected for compatibility.

The neuroscience of positive emotions and affect: Implications for cultivating happiness and wellbeing.

This review paper provides an integrative account regarding neurophysiological correlates of positive emotions and affect that cumulatively contribute to the scaffolding for happiness and wellbeing in humans and other animals. This paper reviews the associations among neurotransmitters, hormones, brain networks, and cognitive functions in the context of positive emotions and affect. Consideration of lifespan developmental perspectives are incorporated, and we also examine the impact of healthy social relationships and environmental contexts on the modulation of positive emotions and affect. The neurophysiological processes that implement positive emotions are dynamic and modifiable, and meditative practices as well as flow states that change patterns of brain function and ultimately support wellbeing are also discussed. This review is part of "The Human Affectome Project" (http://neuroqualia.org/background.php), and in order to advance a primary aim of the Human Affectome Project, we also reviewed relevant linguistic dimensions and terminology that characterizes positive emotions and wellbeing. These linguistic dimensions are discussed within the context of the neuroscience literature with the overarching goal of generating novel recommendations for advancing neuroscience research on positive emotions and wellbeing.

Have I been here before? Complex interactions of age and test experience modulate the results of behavioural tests.

Recently, a discussion about the reproducibility of results from behavioural phenotyping experiments has emerged. A huge emphasis has therefore been put on the identification of those factors that might limit the reproducibility of behavioural data. As a comprehensive phenotypic characterisation can involve testing of the same animal repeatedly over a specific time period, the aim of the present study was to systematically investigate effects of two potentially confounding variables, age of the animals and test experience. For this purpose, the behaviour of 48 male C57BL/6 J mice of two different ages (9 vs. 13 weeks) was assessed in a battery of common behavioural tests measuring anxiety-like and exploratory behaviour (Elevated Plus Maze, Dark-Light test, Open Field test, Novel Cage test). While half of the mice of each age group was naïve to the test battery, the other half had experienced the same tests before. Besides main effects of both age and test experience on anxiety-like and exploratory behaviour, the analysis also revealed profound interactions between these factors. More precisely, an effect of age was apparent in experienced but not in naïve mice. Furthermore, the effect of previous test experience was more pronounced in older than in younger mice. These findings clearly demonstrate that experimental factors, such as age and test experience, can influence behavioural data not just additively, but also in a complex, interactive way. To provide robust and reproducible results, it is thus fundamental to consider such factors systematically in the study design.

Technology or ecology? New tools to assess cognitive judgement bias in mice.

Cognitive judgement bias tests have become important new tools for the assessment of animal emotions. They allow for the inference of an animal's emotional state based on ambiguous cue interpretations. As mice are the predominantly used animal model for cognitive and behavioural neuroscience, research in this field would considerably benefit from the development of suitable judgement bias tests for this species. Against this background, we aimed to implement two different active choice cognitive judgement bias paradigms for mice in a methodological study. For this purpose, two experiments were conducted: in experiment I, an automated, vision-based touchscreen technique was applied, allowing for the direct translation of tasks from rodents to humans and vice versa. Experiment II comprised a task relying on more ecologically relevant cues in form of tunnels of different lengths. While the touchscreen task was characterized by automation-related advantages such as the possibility to present many trials per session and a high convenience for the experimenter, the tunnel task was learned faster by the mice. In both tests, however, the response to the trained and ambiguous conditions resulted in a graded curve, the basic requirement for proving task validity. Thus, both the translational touchscreen task as well as the ecologically more relevant tunnel task could successfully be implemented and provide new tools for the future assessment of cognitive judgement biases in mice.

Brain serotonin deficiency affects female aggression.

The neurotransmitter serotonin plays a key role in the control of aggressive behaviour. While so far most studies have investigated variation in serotonin levels, a recently created tryptophan hydroxylase 2 (Tph2) knockout mouse model allows studying effects of complete brain serotonin deficiency. First studies revealed increased aggressiveness in homozygous Tph2 knockout mice in the context of a resident-intruder paradigm. Focussing on females, this study aimed to elucidate effects of serotonin deficiency on aggressive and non-aggressive social behaviours not in a test situation but a natural setting. For this purpose, female Tph2 wildtype (n = 40) and homozygous knockout mice (n = 40) were housed with a same-sex conspecific of either the same or the other genotype in large terraria. The main findings were: knockout females displayed untypically high levels of aggressive behaviour even after several days of co-housing. Notably, in response to aggressive knockout partners, they showed increased levels of defensive behaviours. While most studies on aggression in rodents have focussed on males, this study suggests a significant involvement of serotonin also in the control of female aggression. Future research will show, whether the observed behavioural effects are directly caused by the lack of serotonin or by potential compensatory mechanisms.

Varying Social Experiences in Adulthood Do Not Differentially Affect Anxiety-Like Behavior But Stress Hormone Levels.

Social experiences can have profound effects on an individual's level of anxiety. While various studies have addressed consequences of experiences of a specific type, e.g., social defeat, a recent study in mice investigated the impact of combinations of adverse and beneficial social experiences. Quite surprisingly, mice exposed to benefits during early life phases followed by escapable adversity in adulthood displayed lowest levels of anxiety, even compared to individuals having experienced throughout beneficial conditions. The present study aimed to elucidate whether this phenomenon is restricted to these specific life phases or whether it also exists when all these experiences are made in full adulthood. For this purpose, we compared anxiety-like behavior and stress response of adult male mice exposed to escapable social defeat following beneficial social experiences to that of mice exposed to either throughout adverse or throughout beneficial conditions. More precisely, we performed three established behavioral paradigms measuring anxiety-like behavior and assessed corticosterone metabolites non-invasively via feces sampling. Interestingly, we found no effects of social experience on anxiety-like behavior. In contrast to that, the animals' stress hormone levels were profoundly affected by current social conditions: escapable social defeat (adverse condition) led to an increase in corticosterone metabolite concentrations, whereas living with a female (beneficial condition) led to a decrease. Thus, on the one hand this study suggests the importance of the timing of social experience for affecting an individual's level of anxiety. On the other hand, it demonstrates that anxiety and stress hormone levels can be affected separately by social experience during adulthood.

What a difference a day makes-female behaviour is less predictable near ovulation.

'Animal personalities' have been shown to exist in many species. Yet, fluctuations in the stability of these inter-individual behavioural differences are not well understood. Against this background, we wondered whether behavioural consistency over time is affected by the reproductive cycle. Female mice were tested twice at an interval of eight weeks in four paradigms assessing social interest as well as anxiety-like behaviour and exploratory locomotion. Twenty-two individuals were tested repeatedly near ovulation, whereas another twenty-two were tested repeatedly in the non-receptive phase. While we found no major behavioural effects at the group level, the reproductive state indeed had profound effects on behavioural stability over time: social interest as well as anxiety-like behaviour proved to be significantly less predictable near ovulation. It is generally believed that phenotypic plasticity is limited due to the costs it brings about. In this context, our data indicate that females accept higher costs in phases directly related to fitness maximization.

A Time to Wean? Impact of Weaning Age on Anxiety-Like Behaviour and Stability of Behavioural Traits in Full Adulthood.

In mammals, weaning constitutes an important phase in the progression to adulthood. It comprises the termination of suckling and is characterized by several changes in the behaviour of both mother and offspring. Furthermore, numerous studies in rodents have shown that the time point of weaning shapes the behavioural profile of the young. Most of these studies, however, have focused on 'early weaning', while relatively little work has been done to study 'late weaning' effects. The aim of the present study was therefore to explore behavioural effects of 'late weaning', and furthermore to gain insights into modulating effects of weaning age on the consistency of behavioural expressions over time. In total, 25 male and 20 female C57BL/6J mice, weaned after three (W3) or four (W4) weeks of age, were subjected to a series of behavioural paradigms widely used to assess anxiety-like behaviour, exploratory locomotion, and nest building performance. Behavioural testing took place with the mice reaching an age of 20 weeks and was repeated eight weeks later to investigate the stability of behavioural expressions over time. At the group level, W4 mice behaved less anxious and more explorative than W3 animals in the Open Field and Novel Cage, while anxiety-like behaviour on the Elevated Plus Maze was modulated by a weaning-age-by-sex interaction. Furthermore, weaning age shaped the degree of behavioural stability over time in a sex-specific way. While W3 females and W4 males displayed a remarkable degree of behavioural stability over time, no such patterns were observed in W3 males and W4 females. Adding to the existing literature, we could thus confirm that effects of weaning age do indeed exist when prolonging this phase, and were furthermore able to provide first evidence for the impact of weaning age and sex on the consistency of behavioural expressions over time.

Benefits of a "vulnerability gene"? A study in serotonin transporter knockout mice.

Over the past years, certain "vulnerability genes" have been identified that play a key role in the development of mood and anxiety disorders. In particular, a low-expressing variant of the human serotonin transporter (5-HTT) gene has been described that renders individuals more susceptible to adverse experience and hence to the development of psychiatric diseases. However, some authors have recently argued that lower 5-HTT expression not only increases vulnerability to adverse experiences, but also enhances susceptibility to beneficial experiences, thus promoting phenotypic plasticity. The aim of the present study was to assess the effects of 5-HTT expression on susceptibility to beneficial experience in a hypothesis-driven experimental approach. Using a well-established rodent model for the human polymorphism, male heterozygous 5-HTT knockout (HET) and 5-HTT wildtype (WT) mice were either provided with the beneficial experience of cohabitation with a female (mating experience) or kept as naïve controls in single-housing conditions. Following the experimental treatment, they were tested for their anxiety-like behaviour and exploratory locomotion in three widely used behavioural tests. Interestingly, while cohabitation reduced anxiety-like behaviour and increased exploratory locomotion in the open field test in HET mice, it did not affect WT mice, pointing to a genotype-dependent susceptibility to the beneficial experience. Thus, our results might support the view of the low expressing version of the 5-HTT gene as a "plasticity" rather than a "vulnerability" variant.