Model systems for regeneration: the spiny mouse, Acomys cahirinus.

The spiny mouse, Acomys spp., is a recently described model organism for regeneration studies. For a mammal, it displays surprising powers of regeneration because it does not fibrose (i.e. scar) in response to tissue injury as most other mammals, including humans, do. In this Primer article, we review these regenerative abilities, highlighting the phylogenetic position of the spiny mouse relative to other rodents. We also briefly describe the Acomys tissues that have been used for regeneration studies and the common features of their regeneration compared with the typical mammalian response. Finally, we discuss the contribution that Acomys has made in understanding the general principles of regeneration and elaborate hypotheses as to why this mammal is successful at regenerating.

Maternal separation followed by isolation-housing differentially affects prepulse inhibition of the acoustic startle response in C57BL/6 mice.

Exposure to chronic stress is associated with an increased incidence of neuropsychiatric dysfunction. The current study evaluated two competing hypotheses, the cumulative stress and the match/mismatch hypothesis of neuropsychiatric dysfunction, using two paradigms relating to exposure to "stress": pre-weaning maternal separation and post-weaning isolation-housing. C57BL/6 offspring were reared under four conditions: typical animal facility rearing (AFR, control), early handling (EH, daily 15 min separation from dam), maternal separation (MS, daily 4 hr separation from dam), and maternal and peer separation (MPS, daily 4 hr separation from dam and from littermates). After weaning, mice were either housed socially (2-3/cage) or in isolation (1/cage) and then tested for prepulse inhibition in adulthood. Isolation-housed MPS subjects displayed greater deficits in prepulse inhibition relative to socially-housed MPS subjects while socially-housed AFR subjects displayed greater deficits in prepulse inhibition relative to isolation-housed AFR subjects. The results indicate that these treatment conditions represent a potentially valuable model for evaluating the match/mismatch hypothesis in regards to neuropsychiatric dysfunction.

Older spiny mice (Acomys cahirinus) have delayed and spatially heterogenous ear wound regeneration.

The African spiny mouse (Acomys cahirinus) is a unique mammalian model of tissue regeneration, regenerating 4 mm ear-hole punches with cartilage, adipocytes, hair follicles, and muscle. However, the time to regenerate ear tissue varies from 20 to 90 days and muscle regeneration is inconsistent. Some report that older spiny mice have delayed regeneration without investigation on the regenerative capacity of muscle. We thought that delayed regeneration and inconsistent muscle regeneration could be linked via age-related nerve degeneration. While the current study found that spiny mice aged 6-9 months had delayed regeneration compared to 3-4 month-old spiny mice, the capacity of muscle regeneration was unrelated to age, and there was little evidence for age-related nerve degeneration. Instead, the regeneration of muscle, cartilage and adipocytes was spatially heterogeneous, declining in amount from the proximal to distal region of the regenerated tissue. Also, cartilage regeneration in the distal region was decreased in ≥22-month-old Acomys and adipocyte regeneration was decreased in those older than 6 months, compared to 3-4 month olds. While the underlying mechanisms for delayed and spatially heterogenous regeneration remain unclear, age and the spatial region of the regenerated tissue should be considered in experimental designs with spiny mice.

Bite Wounds and Dominance Structures in Male and Female African Spiny Mice (Acomys cahirinus): Implications for Animal Welfare and the Generalizability of Experimental Results.

Bite wounds due to aggression in male laboratory mice (Mus musculus) are a major welfare concern, often leading to attrition, chronic activation of the innate immune system, and significant impacts on the experimental results derived from the use of these animals as models. Bite wounding within the home-cage of spiny mice (Acomys cahirinus)-a valuable research model for wound healing and menstruation-is poorly characterized. While we have anecdotally observed frequent bite wounding in Acomys, the frequency of aggression within the home-cage, the severity of the bite wounds, and the types of dominance structures remain unstudied. Here, we report that 46% of Acomys cages in our colony had at least one bite wound over the course of a year and that same-sex pairs fought in the home-cage 10% of the time during their dark/active phase. Both sexes inflicted wounds and frequently engaged in agonistic behaviors, even with stable dominance structures. We found that females inflicted less severe bite wounds in same-sex housing. Also, aged females in same-sex pairs were never observed fighting, and no bite wounds were observed in aged Acomys. These results suggest that we should consider whether bite wounding negatively impacts our experimental results since physical trauma is known to alter menstrual cycling and healing.

Spiny mouse (Acomys): an emerging research organism for regenerative medicine with applications beyond the skin.

The spiny mouse (Acomys species) has emerged as an exciting research organism due to its remarkable ability to undergo scarless regeneration of skin wounds and ear punches. Excitingly, Acomys species demonstrate scar-free healing in a wide-range of tissues beyond the skin. In this perspective article, we discuss published findings from a variety of tissues to highlight how this emerging research organism could shed light on numerous clinically relevant human diseases. We also discuss the challenges of working with this emerging research organism and suggest strategies for future Acomys-inspired research.

A Systematic Review and Meta-Analysis of the Relationship Between Social Dominance Status and Common Behavioral Phenotypes in Male Laboratory Mice.

Background: Social dominance status (e.g., dominant or subordinate) is often associated with individual differences in behavior and physiology but is largely neglected in experimental designs and statistical analysis plans in biomedical animal research. In fact, the extent to which social dominance status affects common experimental outcomes is virtually unknown. Given the pervasive use of laboratory mice and culminating evidence of issues with reproducibility, understanding the role of social dominance status on common behavioral measures used in research may be of paramount importance. Methods: To determine whether social dominance status-one facet of the social environment-contributes in a systematic way to standard measures of behavior in biomedical science, we conducted a systematic review of the existing literature searching the databases of PubMed, Embase, and Web of Science. Experiments were divided into several domains of behavior: exploration, anxiety, learned helplessness, cognition, social, and sensory behavior. Meta-analyses between experiments were conducted for the open field, elevated plus-maze, and Porsolt forced swim test. Results: Of the 696 publications identified, a total of 55 experiments from 20 published studies met our pre-specified criteria. Study characteristics and reported results were highly heterogeneous across studies. A systematic review and meta-analyses, where possible, with these studies revealed little evidence for systematic phenotypic differences between dominant and subordinate male mice. Conclusion: This finding contradicts the notion that social dominance status impacts behavior in significant ways, although the lack of an observed relationship may be attributable to study heterogeneity concerning strain, group-size, age, housing and husbandry conditions, and dominance assessment method. Therefore, further research considering these secondary sources of variation may be necessary to determine if social dominance generally impacts treatment effects in substantive ways.

Social dominance hierarchy type and rank contribute to phenotypic variation within cages of laboratory mice.

A tacit assumption in laboratory animal research is that animals housed within the same cage or pen are phenotypically more similar than animals from different cages or pens, due to their shared housing environment. This assumption drives experimental design, randomization schemes, and statistical analysis plans, while neglecting social context. Here, we examined whether a domain of social context-social dominance-accounted for more phenotypic variation in mice than cage-identity. First, we determined that cages of mice could be categorized into one of three dominance hierarchies with varying degrees of dominance behavior between cage-mates, and low levels of agonistic behavior in the home-cage. Most groups formed dynamic hierarchies with unclear ranks, contrasting with recent accounts of stable transitive hierarchies in groups of mice. Next, we measured some phenotypic traits, and found that social dominance (i.e. dominance hierarchy type and degree of dominance behavior) consistently accounted for some phenotypic variation in all outcome measures, while cage-identity accounted for phenotypic variation in some measures but virtually no variation in others. These findings highlight the importance of considering biologically relevant factors, such as social dominance, in experimental designs and statistical plans.

Phenotypic variability between Social Dominance Ranks in laboratory mice.

The laboratory mouse is the most prevalent animal used in experimental procedures in the biomedical and behavioural sciences. Yet, many scientists fail to consider the animals' social context. Within a cage, mice may differ in their behaviour and physiology depending on their dominance relationships. Therefore, dominance relationships may be a confounding factor in animal experiments. The current study housed male and female C57BL/6ByJ mice in same-sex groups of 5 in standard laboratory conditions and investigated whether dominance hierarchies were present and stable across three weeks, and whether mice of different dominance ranks varied consistently in behaviour and physiology. We found that dominance ranks of most mice changed with time, but were most stable between the 2nd and 3rd week of testing. Phenotypic measures were also highly variable, and we found no relation between dominance rank and phenotype. Further, we found limited evidence that variation in measures of phenotype was associated with cage assignment for either males or females. Taken together, these findings do not lend support to the general assumption that individual variation among mice is larger between cages than within cages.

Evaluation of the effects of space allowance on measures of animal welfare in laboratory mice.

We studied how space allowance affects measures of animal welfare in mice by systematically varying group size and cage type across three levels each in both males and females of two strains of mice (C57BL/6ByJ and BALB/cByJ; n = 216 cages, a total of 1152 mice). This allowed us to disentangle the effects of total floor area, group size, stocking density, and individual space allocation on a broad range of measures of welfare, including growth (food and water intake, body mass); stress physiology (glucocorticoid metabolites in faecal boli); emotionality (open field behaviour); brain function (recurrent perseveration in a two-choice guessing task); and home-cage behaviour (activity, stereotypic behaviour). While increasing group size was associated with a decrease in food and water intake in general, and more specifically with increased attrition due to escalated aggression in male BALB mice, no other consistent effects of any aspect of space allowance were found with respect to the measures studied here. Our results indicate that within the range of conditions commonly found in laboratory mouse housing, space allowance as such has little impact on measures of welfare, except for group size which may be a risk factor for escalating aggression in males of some strains.

Effects of Cage Enrichment on Behavior, Welfare and Outcome Variability in Female Mice.

The manner in which laboratory rodents are housed is driven by economics (minimal use of space and resources), ergonomics (ease of handling and visibility of animals), hygiene, and standardization (reduction of variation). This has resulted in housing conditions that lack sensory and motor stimulation and restrict the expression of species-typical behavior. In mice, such housing conditions have been associated with indicators of impaired welfare, including abnormal repetitive behavior (stereotypies, compulsive behavior), enhanced anxiety and stress reactivity, and thermal stress. However, due to concerns that more complex environmental conditions might increase variation in experimental results, there has been considerable resistance to the implementation of environmental enrichment beyond the provision of nesting material. Here, using 96 C57BL/6 and SWISS female mice, respectively, we systematically varied environmental enrichment across four levels spanning the range of common enrichment strategies: (1) bedding alone; (2) bedding + nesting material; (3) deeper bedding + nesting material + shelter + increased vertical space; and (4) semi-naturalistic conditions, including weekly changes of enrichment items. We studied how these different forms of environmental enrichment affected measures of animal welfare, including home-cage behavior (time-budget and stereotypic behavior), anxiety (open field behavior, elevated plus-maze behavior), growth (food and water intake, body mass), stress physiology (glucocorticoid metabolites in fecal boluses and adrenal mass), brain function (recurrent perseveration in a two-choice guessing task) and emotional valence (judgment bias). Our results highlight the difficulty in making general recommendations across common strains of mice and for selecting enrichment strategies within specific strains. Overall, the greatest benefit was observed in animals housed with the greatest degree of enrichment. Thus, in the super-enriched housing condition, stereotypic behavior, behavioral measures of anxiety, growth and stress physiology varied in a manner consistent with improved animal welfare compared to the other housing conditions with less enrichment. Similar to other studies, we found no evidence, in the measures assessed here, that environmental enrichment increased variation in experimental results.