More exploratory house mice judge an ambiguous situation more negatively.

Exploration tendency, one of the most investigated animal personality traits, may be driven by either positive (when seeking interesting information) or negative (to reduce the uncertainty of the environment) affective/emotional profiles. To disentangle the valence of the affective state associated with exploration trait, we applied a judgment bias test to evaluate the animals' responses in an ambiguous situation, allowing an assessment of their affective state or mood. Experiments were carried out in male house mice (Mus musculus) of wild origin. Individual differences in exploration tendency were assessed by repeated open field and novel object tests. To evaluate the animals' judgment bias, we trained the subjects for 8 days in a 3-arm maze to discriminate between two extreme locations (outer arms: either positively reinforced with sugary water or less-positively reinforced with plain water), in terms of a shorter latency to approach the positively reinforced arm. After this learning criterion was reached, we repeatedly tested their responses to an ambiguous location (intermediate arm). The latencies to approach and consume the ambiguous reward were highly repeatable over the 3 days of testing; hence individuals expressed a stable judgment bias. Most importantly, more exploratory animals showed a more negative judgment bias, which supports the hypothesis that a higher exploration tendency was associated with a negative affective state. Further studies should investigate whether exploration in different situations might be due to distinct affective states.

Necessary Conditions for Social Transmission of Food Preference Through Feces in the House Mouse, Mus musculus domesticus.

Mice can obtain information about a new food source through olfactory cues of conspecifics and consequently develop an attraction for this diet. The social transmission of food preference (STFP) takes place directly, during an encounter with a conspecific or indirectly, via feces. In indirect STFP, the digestive process can degrade odorant compounds characterizing the food, impairing the matching between feces and food. In a previous study, indirect STFP was efficient when the information support was a composite odorant. We, thus, hypothesized that the acquisition of indirect STFP depends on the multiplicity of the odorant compounds present in diets. Tested in female house mice (Mus musculus domesticus) our results showed that a single odorant compound as information support was not sufficient to induce an indirect STFP. Chemical analysis did not reveal the presence of the compounds in feces suggesting that the degradation of diet cues during the digestive process prevented the pairing between feces and food. By using a process that limits the degradation of molecules, we performed indirect STFP when the pertinent information was represented by a single odorant compound and multiple odorant compounds. Unlike with multiple odorant compounds, our results did not show a clear indirect STFP with single odorant compound, despite their presence in feces confirmed by chemical analysis. We conclude that constraints associated to indirect STFP can be removed by the multiplicity of information characterizing the diet both by reducing the degradation risk during the digestive process and by allowing an accurate assessment of diet consumed by the conspecific.

Isolation calls in house mouse pups: Individual consistency across time and situations.

Isolation calls are emitted by the offspring of many mammalian species when separated from caregivers and siblings. Some studies indicate that isolation call rates constitute a consistent individual trait; others show that the young adjust their vocalization rate to the current situation. We studied this in the house mouse (Mus musculus domesticus) by exploring individual consistencies in pup isolation call rates and their potential modulation in different social situations. We carried out experiments including three treatments (repeated measurements) during consecutive days, all starting with an initial isolation of the pup, followed by (a) a reunion with mother and littermates and a second isolation hereafter, (b) the confrontation of isolated pup with cues of its own nest or (c) with cues of an unfamiliar adult male. The first treatment induced a significant increase, while the others induced significant decreases in pup isolation call rates. Pups showed consistent individual differences in initial call rates across the three days of testing (postnatal days 9-11), which were significantly associated with individual differences in call rates during the different treatments. We conclude that pup isolation calls represent a consistent, trait-like behaviour in the house mouse, which can also express flexibility in response to social cues.

Individual differences in early body mass affect thermogenic performance and sibling interactions in litter huddles of the house mouse.

We asked whether within-litter differences in early body mass are associated with differences in house mouse pups' thermogenic performance and whether such variation predicts individual differences in competitive interactions for thermally more advantageous positions in the huddle. We explored pups' thermogenic performance in isolation by measuring changes in (maximal) peripheral body temperatures during a 5-min thermal challenge using infrared thermography. Changes in peripheral body temperature were significantly explained by individual differences in body mass within a litter; relatively lighter individuals showed an overall quicker temperature decrease leading to lower body temperatures toward the end of the thermal challenge compared to heavier littermates. Within the litter huddle, relatively lighter pups with a lower thermogenic performance showed consistently more rooting and climbing behavior, apparently to reach the thermally advantageous center of the huddle. This suggests that within-litter variation in starting body mass affects the pups' thermal and behavioral responses to environmental challenges.

Efficiency of delayed reproduction in Mus spicilegus.

To cope with seasonally varying ecological constraints, some mammals temporally suppress breeding or delay their first reproduction. In field conditions, mound-building mice (Mus spicilegus) born in spring begin to reproduce when 2-3 months old, whereas individuals born at the end of summer delay their first reproduction for 6-8 months until the following spring. In order to test age effects on reproductive performance in M. spicilegus, sexually naïve mice were paired when 2-3 months old or at 6-8 months of age, and surveyed for reproduction. We show here that under laboratory conditions the aging of these mice does not impair their reproductive efficiency. Thus, the hypothesis of a lower reproductive potential in these relatively aged females seems to be contradicted. More surprisingly, the latency from pairing to the first reproduction was greater in the 2-3-month-old adults than in the delayed reproducers (6-8-month-old mice). Mound-building mice that are old enough to have overwintered do not suffer significant reproductive declines, but appear to reproduce as well and more quickly than younger first-time breeders.

Patterns and predictors of inter-litter differences in rabbit pup locomotor activity, based on an automatized quantification method.

Individual-level sibling interactions in the litter huddle have been studied extensively, especially in the domestic rabbit (Oryctolagus cuniculus). However, little is known about inter-litter differences in pup activity patterns during early postnatal life, in particular regarding the drivers of such variation. In our study on 2-3-day-old rabbit pups, we predicted lower locomotor activity in litters with lower mean body masses on the day of birth (starting body mass) and with lower daily milk intake per pup, possibly constituting a behavioral strategy of pups to cope with associated energetic constraints. For an automatized assessment of pup locomotor activity in the litter huddle, we successfully developed and validated a method based on the quantification of dissimilarities between consecutive frames of video footage. Using this method, we could confirm a U-shaped time course of litter-level locomotor activity, with maximum values shortly before and after the once-daily nursing typical for the rabbit. As predicted, between-litter variation in mean starting body mass and in daily milk intake affected the degree of locomotor activity in the litter huddle, in an interactive way. That is, in litters with heavier starting body masses, pup locomotor activity was greater in pups with an initially higher milk intake, suggesting that only pups with better body condition and a higher energy intake could afford higher levels of activity. This interaction was exclusively apparent during the middle phase of the 24 h inter-nursing interval, when litter activity was low. Shortly before nursing, when pups show higher levels of locomotor behavior in anticipation of the mother's arrival, and shortly after nursing when the pups were more active possibly due to adjustments of their positions in the huddle, activity levels were decoupled from pups' starting body mass and previous milk intake. Our findings highlight the importance of pup body mass and daily energy intake, two parameters known to be related to maternal characteristics, in shaping inter-litter differences in pup locomotor activity.

Early life parameters and personality affect oxidative status during adulthood in an altricial rodent.

It is increasingly recognized that alterations of the cellular oxidative status might be an important cost underlying challenging early life conditions. For example, an increased litter size can impose challenges as the offspring will face increased competition for maternal resources. Within a litter, individuals with relatively higher starting mass typically show higher growth rates, which can lead to increased oxidative damage. We investigated the long-term consequences of these early life parameters on the oxidative status in mature mound-building mice (Mus spicilegus). Individual differences in the animals' exploration tendency were assessed by repeated open field and novel object tests. We predicted less exploratory phenotypes, which typically show a higher stress responsiveness, to be particularly susceptible to possible effects of these early life parameters on oxidative status. We quantified oxidative damage of DNA (8-hydroxy-2'-deoxyguanosine levels, 8-OHdG) and proteins (protein carbonyl content, PCC), and activities of the antioxidants catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD) in liver and skeletal muscle tissue. 8-OHdG levels were positively associated with CAT and SOD in both tissues, indicating that increased oxidative DNA damage was associated with an upregulation of antioxidant production. Hepatic DNA damage after maturity was increased in animals from larger litters. In less exploratory animals, DNA damage and the activity of CAT and SOD in the muscle were increased, but only in individuals with higher relative starting mass (measured on postnatal day 9). This interaction may be explained by the typically higher adrenocortical activity in less exploratory phenotypes and by the higher growth in relatively heavier pups, two factors known to increase oxidative stress. These findings contribute to enlightening the complex interplay between early life conditions, personality, and oxidative status.

Is there a bias in spatial maze judgment bias tests? Individual differences in subjects' novelty response can affect test results.

Judgment bias tests have become an important tool in the assessment of animals' affective states. Subjects are first trained to discriminate between two cues associated with a positive and a less-positive outcome. After successful training, they are confronted with an ambiguous cue, and responses are used for judgment bias assessment. In spatial settings, ambiguous cue presentation is typically linked with novelty, i.e. to yet unexplored areas or areas to which the animal has a low degree of habituation. We hypothesized that in such settings, responses to ambiguity might be biased by the animals' perception of novelty. We conducted judgment bias tests in mound-building mice phenotyped for their exploration tendency. After subjects had learned to distinguish between the positively and less-positively rewarded arms of a maze, a new ambiguous middle-arm was introduced. During the first test trial, more exploratory, less neophobic individuals displayed higher bidirectional locomotion in the ambiguous arm, indicating intensive exploration. Although this resulted in longer latencies to the reward in more exploratory animals, we conclude that this did not reflect a 'more pessimistic judgment of ambiguity'. Indeed, during the following two trials, with increasing habituation to the ambiguous arm, the direction of the association was inversed compared to the first trial, as more exploratory individuals showed relatively shorter approach latencies. We suggest that in spatial test settings associating the ambiguous cue to novel areas, results can be confounded by subjects' personality-dependent motivational conflict between exploration and reaching the reward. Findings obtained under such conditions should be interpreted with care.

Transmission of food preference via faeces in male and female house mice: Who is a good provider of food cues?

The social transmission of food preference (STFP) is a phenomenon that allows rodents to use food information perceived from their conspecifics to guide their own food choices. This social information can be collected via olfactory cues, during direct social interactions, or indirectly, via faeces left in the environment by individuals. Although reducing the risks associated with a social confrontation, faeces also convey different types of information about traits and states of individuals which could affect the indirect STFP. Here, we evaluated in the house mouse, Mus musculus domesticus, which conspecifics could be good providers of food cues for indirect STFP. Our results indicated that female mice acquire an indirect STFP from faeces of adult females, familiar or not and from faeces of adult unfamiliar males. On the contrary, males do not establish an indirect STFP from faeces of males, whether they are familiar or not, nor from those of unfamiliar females. Indirect STFP was however effective in males when the faeces were those of an unfamiliar juvenile male. A prior habituation to the odour of an unfamiliar female allowed the establishment of indirect STFP in males. Conversely, the presence of faeces of another adult male during the presentation of faeces of an accustomed female precluded the acquisition of indirect STFP in males. This study suggested that in the context of the indirect STFP, females prioritize socio-olfactory information relative to food, whereas food cues were not priority information for males. Under these conditions, females appear to be the best vectors for disseminating food information within the population. These results are discussed according to the socio-spatial organization of the species.

Comparison of urinary scents of two related mouse species, Mus spicilegus and Mus domesticus.

Whereas the house mouse (Mus domesticus) has been studied extensively in terms of physiology/behavior and pheromonal attributes, the evolutionarily related mound-building mouse (Mus spicilegus) has received attention only recently due to its divergent behavioral traits related to olfaction. To date, no chemical studies on urinary volatile compounds have been performed on M. spicilegus. The rationale for our investigations was to determine if there are differences in urinary volatiles of intact and castrated M. spicilegus males and to explore further whether this species could utilize the same or structurally similar pheromones as the male house mouse, M. domesticus. The use of capillary gas chromatography/mass spectrometry (GC-MS) together with sorptive stir bar extraction sampling enabled quantitative comparisons between the intact and castrated M. spicilegus urinary profiles. Additionally, through GC-MS and atomic emission (sulfur-selective) detection, we identified qualitative molecular differences between intact M. spicilegus and M. domesticus. A series of volatile and odoriferous lactones and the presence of coumarin were the unique features of M. spicilegus, as was the notable absence of 2-sec-butyl-4,5-dihydrothiazole (a prominent M. domesticus male pheromone) and other sulfur-containing compounds. Castration of M. spicilegus males eliminated several substances, including delta-hexalactone and gamma-octalactone, and substantially decreased additional compounds, suggesting their possible role in chemical communication. Some other M. domesticus pheromone components were also found in M. spicilegus urine. These comparative chemical analyses support the notion of metabolic similarities as well as the uniqueness of some volatiles for M. spicilegus, which may have a distinct physiological function in reproduction and behavior.

Differences between fast and slow explorers in short-term tail temperature responses to handling in a rodent of wild origin.

Animals of different behavioral types typically show associated differences in their physiological stress response, including differential reactivity of the sympathetic nervous system. Infrared thermography offers the possibility to explore this link in a non-invasive way via the quantification of fine-scale changes in peripheral body temperature due to changes in cutaneous blood flow. We used this technique to investigate the association between exploration tendency, a behavioral trait frequently used to phenotype mammals and birds, and short-term thermal responses to challenge in a small rodent of wild origin, the mound-building mouse (Mus spicilegus). We applied a brief handling procedure consisting in the transfer of subjects into a small arena. This procedure led to a significant increase in subjects' maximum peripheral body temperature (mainly reflecting the temperature of the eyes) and to significant decreases in maximum temperatures at different positions on the tail. Maximum peripheral body and tail temperatures showed significant individual-level consistencies in response to repeated applications of the handling procedure, suggesting stable individual differences in the animals' sympathetic activity. We then compared the thermal responses to handling between 'fast' and 'slow' explorers, who were phenotyped through repeated open field and novel object tests. Fast explorers showed significantly lower tail temperatures than slow explorers shortly after handling, suggesting a stronger sympathetic reactivity in the former. Comparisons within sibling groups kept in different cages showed that the differences between explorer types were particularly pronounced during the first minute after handling, and increased in magnitude along the first millimeters distal to the tail base.

Transmission of food preference between unfamiliar house mice (Mus musculus domesticus) is dependent on social context.

Rodents obtain information about a new food source through olfactory cues of conspecifics and consequently develop an attraction for this diet. Generally, physical contact between an observer and a demonstrator that has recently consumed a novel food item is required to allow the social transmission of food preference (STFP). However, in natural populations of house mice, social encounters between unfamiliar individuals usually turn into a fight. Thus, social intolerance between the individuals involved could prevent STFP. It has been shown that the feces of rodents can act as a social stimulus and promote STFP in mice, which could reduce the social constraints associated with an encounter. Here, we examined the acquisition of the STFP in female house mice of wild origin (Mus musculus domesticus) after a direct encounter with a familiar and unfamiliar female, and after the presentation of olfactory marks of an unfamiliar female. Unlike in encounters between familiar females, our results did not provide significant support for the existence of STFP after encounters between two unfamiliar females, independently of the occurrence or absence of offensive agonistic behavior. However, STFP through olfactory marks of an unfamiliar female was effective. We suggest that the social context might strongly impair direct STFP, not necessarily via the unfamiliarity of the information provider but rather via its physical presence. (PsycINFO Database Record

Evaluation of the social bond: a new method tested in Mus spicilegus.

Innovative and fruitful studies of social bonds have been developed in recent years, although the methods used to establish the existence of a social bond between two individuals have not evolved significantly. Two types of paradigms have been currently used: the separation-reunion paradigm, which evaluates the distress caused by the disruption of the social bond, and choice paradigms, which test the specificity of the bond to a given individual. We have developed a new paradigm based on the idea that the cost an individual was ready to pay in order to gain access to a conspecific depended on the strength of the social bond between the two individuals. To test our paradigm we used mound-building mice, Mus spicilegus that present, in both males and females, a level of tolerance that differs greatly according to the degree of familiarity between the individuals. Our new method for testing social bond revealed unsuspected differences between males and females. Our results suggested that, at least in Mus spicilegus, strong social bonds were not necessary to the development of a high level of tolerance between individuals.

Paternal care in the mound-building mouse reduces inter-litter intervals.

In many rodent species males display paternal behaviour. The primary reported effect of this paternal care is to increase pup survival. In mammal females, pregnancy and lactation are energetically demanding, especially when they are concurrent in post-partum reproduction. To face this energy requirement, females generally lengthen the duration of their post-partum pregnancy. In the present study we tested whether paternal care could affect this duration in the monogamous mound building mouse Mus spicilegus. In this species, females have a short reproductive life that does not exceed 4 months. Reduction of inter-delivery latencies would then be an efficient way to increase reproductive success. In a male removal experiment, we showed that inter-delivery latency was shortened by male presence. Moreover, behavioural estimations of paternal involvement were correlated with inter-delivery latency. The longer the male spent inside the nest the shorter the inter-delivery latency. In the mound-building mouse, the female might be able to monopolise the parental care of a single male, which could be important for the evolution of monogamy. The characteristics of first reproduction as compared to post-partum reproduction suggest that it may contribute to the formation of a strong and exclusive social bond between the reproductive partners.

Body mass modulates huddling dynamics and body temperature profiles in rabbit pups.

Altricial mammals typically lack the physiological capacity to thermoregulate independently during the early postnatal period, and in litter-bearing species the young benefit strongly from huddling together with their litter siblings. Such litter huddles are highly dynamic systems, often characterized by competition for energetically favorable, central positions. In the present study, carried out in domestic rabbits Oryctolagus cuniculus, we asked whether individual differences in body mass affect changes in body temperature during changes in the position within the huddle. We predicted that pups with relatively lower body mass should be more affected by such changes arising from huddle dynamics in comparison to heavier ones. Changes in pups' maximum body surface temperature (determined by infrared thermography) were significantly affected by changes in the number of their neighbors in the litter huddle, and indeed these temperature changes largely depended on the pups' body mass relative to their litter siblings. Lighter pups showed significant increases in their maximum body surface temperature when their number of huddling partners increased by one or two siblings whereas pups with intermediate or heavier body mass did not show such significant increases in maximum body temperature when experiencing such changes. A similar pattern was found with respect to average body surface temperature. This strong link between changes in the number of huddling partners and body surface temperature in lighter pups might, on the one hand, arise from a higher vulnerability of such pups due to their less favorable body surface area-to-volume ratio. On the other hand, as lighter pups generally had fewer neighbors than heavier ones and thus typically a comparatively smaller body surface in contact with siblings, they potentially had more to gain from increasing their number of neighbors. The present findings might help to understand how individual differences in body mass within a litter lead to the emergence of individual differences in sibling interactions during early postnatal life in different species of altricial and litter-bearing mammals.

Assessment of anxiety in open field and elevated plus maze using infrared thermography.

Due to their direct inaccessibility, affective states are classically assessed by gathering concomitant physiological and behavioral measures. Although such a dual approach to assess emotional states is frequently used in different species including humans, the invasiveness of procedures for physiological recordings particularly in smaller-sized animals strongly restricts their application. We used infrared thermography, a non-invasive method, to assess physiological arousal during open field and elevated plus maze tests in mice. By measuring changes in surface temperature indicative of the animals' emotional response, we aimed to improve the inherently limited and still controversial information provided by behavioral parameters commonly used in these tests. Our results showed significant and consistent thermal responses during both tests, in accordance with classical physiological responses occurring in stressful situations. Besides, we found correlations between these thermal responses and the occurrence of anxiety-related behaviors. Furthermore, initial temperatures measured at the start of each procedure (open field, elevated plus maze), which can be interpreted as a measure of the animals' initial physiological arousal, predicted the levels of activity and of anxiety-related behaviors displayed during the tests. Our results stress the strong link between physiological correlates of emotions and behaviors expressed during unconditioned fear tests.

Deficit in reproduction in polygynously mated females of the monogamous mound-building mouse Mus spicilegus.

In monogamous mound-building mice Mus spicilegus, facultative polygyny was observed in April, at the beginning of the reproductive season. In order to evaluate the cost of polygyny, we compared, under laboratory conditions, the reproductive success of trios and pairs composed of sister females mated with unrelated brother males. Females in trios were able to tolerate each other during the reproductive period and to reproduce with the same male under spatial constraints. Nevertheless, polygyny had a strong negative effect on the reproductive success of the females. The average number of young per litter was smaller in trio females than in paired females, whereas the interval between two successive litters was higher. As a result, the number of litters and the number of young per time unit were smaller in each of the trio females compared with paired females. Agonistic behaviour being absent in trios, our results strongly suggest that living in trios led to pheromonal production that affected the female physiological state and reproduction. Males of trios did not obtain a better reproductive success than males in pairs.

Correlates between ear postures and emotional reactivity in a wild type mouse species.

Facial expressions have been extensively used to assess emotions in humans and thus could be extended to other species that also display facial movements. In mice both painful and fearful situations have been associated with particular shifts in facial expressions. Like other species, mice frequently show a great inter-individual variability when exposed to emotional situations, but so far no study has been conducted to investigate if facial expressions are related to these differences. The aim of this study is to explore if mice of wild origin (Mus spicilegus) express different facial expressions when confronted to novelty and to relate these mimics to their emotional reactivity profile. We used individual exploration scores in a novel odour test and in the elevated plus maze test as proxy measures of individual emotional reactivity. Our results showed that exploration scores in both tests were positively correlated, and both were related to the ear postures expressed by the individuals during their first exploration of the novel odour. This single component of facial expression was in fact a good indicator of inter-individual differences expressed in these two different tests suggesting a strong link between this marker and the individual emotional reactivity. These results highlight the great potential of facial expressions to assess emotional states in animals.

Social regulation of reproduction in the female mound-builder mouse (Mus spicilegus).

Social environment influences the reproductive physiology and sexual behaviour of the female house mouse Mus musculus. An all-female environment tends to suppress the oestrous cycles, whereas the presence of a male induces and synchronises sexual receptivity. However, reproductive responses to social environment may differ among the various species of rodents. In mound-builder female mice, Mus spicilegus, periods of sexual receptivity are interrupted by periods where adult females display a vaginal closure. We investigated the influence of different social environments on the vaginal opening and oestrous state of adult female M. spicilegus. Result showed that when females were grouped their vaginas were generally closed but that vaginal opening occurred when they were isolated or housed with a sexual partner. Females became sexually receptive when housed with a male, but when isolated their cervical smears did not reach characteristics of the oestrus. In female M. spicilegus, male presence thus has a stimulating effect on oestrous induction. Furthermore, cohabitation with females has an inhibiting effect on vaginal opening.

Seasonal reproduction and delayed sexual maturity in mound-building mice Mus spicilegus.

In the mound-building mouse, Mus spicilegus, which is found from Central to Eastern Europe, reproduction is seasonal and limited to spring and summer. In autumn, the mice build voluminous mounds composed of vegetable matter covered with earth, where juvenile animals (autumnal individuals) over-winter in groups without reproducing. Autumnal animals delay reproduction until the next spring when they are 6 months old. The influence and interactions of environmental (short light period and cold temperature (C conditions) compared with long light period and temperate temperature (T conditions)) and social factors (lack of odours from breeding adults (NB conditions) compared with presence of odours from breeding adults (B conditions)) on reproduction and sexual maturation were studied. Forty groups of three autumnal individuals (two males and a female or two females and a male) were placed in four experimental conditions (CB, CNB, TB and TNB), corresponding to interactions between environmental and social factors (n = 10 groups for each condition). Of the 40 groups only one initiated reproduction during the 18 weeks of cohabitation. Subsequently, animals were separated and isolated for 1 month and then paired with unfamiliar partners. Reproduction was monitored for an additional month, and 24 out of 39 females reproduced. In addition, of eight reproducing pairs placed in C conditions and 10 reproducing pairs maintained in T conditions, all but one pair continued reproduction. It was concluded that the delay in reproduction observed in autumnal individuals was the result of the social effects of living in groups as opposed to the environmental conditions of winter.