Tissue-resident macrophages specifically express Lactotransferrin and Vegfc during ear pinna regeneration in spiny mice.

The details of how macrophages control different healing trajectories (regeneration vs. scar formation) remain poorly defined. Spiny mice (Acomys spp.) can regenerate external ear pinnae tissue, whereas lab mice (Mus musculus) form scar tissue in response to an identical injury. Here, we used this dual species system to dissect macrophage phenotypes between healing modes. We identified secreted factors from activated Acomys macrophages that induce a pro-regenerative phenotype in fibroblasts from both species. Transcriptional profiling of Acomys macrophages and subsequent in vitro tests identified VEGFC, PDGFA, and Lactotransferrin (LTF) as potential pro-regenerative modulators. Examining macrophages in vivo, we found that Acomys-resident macrophages secreted VEGFC and LTF, whereas Mus macrophages do not. Lastly, we demonstrate the requirement for VEGFC during regeneration and find that interrupting lymphangiogenesis delays blastema and new tissue formation. Together, our results demonstrate that cell-autonomous mechanisms govern how macrophages react to the same stimuli to differentially produce factors that facilitate regeneration.

SAD rats: Effects of short photoperiod and carbohydrate consumption on sleep, liver steatosis, and the gut microbiome in diurnal grass rats.

Seasonal affective disorder (SAD) is a recurrent depression triggered by exposure to short photoperiods, with a subset of patients reporting hypersomnia, increased appetite, and carbohydrate craving. Dysfunction of the microbiota - gut - brain axis is frequently associated with depressive disorders, but its role in SAD is unknown. Nile grass rats (Arvicanthis niloticus) are potentially useful for exploring the pathophysiology of SAD, as they are diurnal and have been found to exhibit anhedonia and affective-like behavior in response to short photoperiods. Further, given grass rats have been found to spontaneously develop metabolic syndrome, they may be particularly susceptible to environmental triggers of metabolic dysbiosis. We conducted a 2 × 2 factorial design experiment to test the effects of short photoperiod (4 h:20 h Light:Dark (LD) vs. neutral 12:12 LD), access to a high concentration (8%) sucrose solution, and the interaction between the two, on activity, sleep, liver steatosis, and the gut microbiome of grass rats. We found that animals on short photoperiods maintained robust diel rhythms and similar subjective day lengths as controls in neutral photoperiods but showed disrupted activity and sleep patterns (i.e. a return to sleep after an initial bout of activity that occurs ~ 13 h before lights off). We found no evidence that photoperiod influenced sucrose consumption. By the end of the experiment, some grass rats were overweight and exhibited signs of non-alcoholic fatty liver disease (NAFLD) with micro- and macro-steatosis. However, neither photoperiod nor access to sucrose solution significantly affected the degree of liver steatosis. The gut microbiome of grass rats varied substantially among individuals, but most variation was attributable to parental effects and the microbiome was unaffected by photoperiod or access to sucrose. Our study indicates short photoperiod leads to disrupted activity and sleep in grass rats but does not impact sucrose consumption or exacerbate metabolic dysbiosis and NAFLD.

From grouping and cooperation to menstruation: Spiny mice (Acomys cahirinus) are an emerging mammalian model for sociality and beyond.

While spiny mice are primarily used as a model for Type II diabetes and for studying complex tissue regeneration, they are also an emerging model for a variety of studies examining hormones, behavior, and the brain. We began studying the spiny mouse to take advantage of their highly gregarious phenotype to examine how the brain facilitates large group-living. However, this unique rodent can be readily bred and maintained in the lab and can be used to ask a wide variety of scientific questions. In this brief communication we provide an overview of studies that have used spiny mice for exploring physiology and behavior. Additionally, we describe how the spiny mouse can serve as a useful model for researchers interested in studying precocial development, menstruation, cooperation, and various grouping behaviors. With increasingly available technological advancements for non-traditional organisms, spiny mice are well-positioned to become a valuable organism in the behavioral neuroscience community.

Matrisomal components involved in regenerative wound healing in axolotl and Acomys: implications for biomaterial development.

Achieving regeneration in humans has been a long-standing goal of many researchers. Whereas amphibians like the axolotl (Ambystoma mexicanum) are capable of regenerating whole organs and even limbs, most mammals heal their wounds via fibrotic scarring. Recently, the African spiny mouse (Acomys sp.) has been shown to be injury resistant and capable of regenerating several tissue types. A major focal point of research with Acomys has been the identification of drivers of regeneration. In this search, the matrisome components related to the extracellular matrix (ECM) are often overlooked. In this review, we compare Acomys and axolotl skin wound healing and blastema-mediated regeneration by examining their wound healing responses and comparing the expression pattern of matrisome genes, including glycosaminoglycan (GAG) related genes. The goal of this review is to identify matrisome genes that are upregulated during regeneration and could be potential candidates for inclusion in pro-regenerative biomaterials. Research papers describing transcriptomic or proteomic coverage of either skin regeneration or blastema formation in Acomys and axolotl were selected. Matrisome and GAG related genes were extracted from each dataset and the resulting lists of genes were compared. In our analysis, we found several genes that were consistently upregulated, suggesting possible involvement in regenerative processes. Most of the components have been implicated in regulation of cell behavior, extracellular matrix remodeling and wound healing. Incorporation of such pro-regenerative factors into biomaterials may help to shift pro-fibrotic processes to regenerative responses in treated wounds.

Generation and characterization of two immortalized dermal fibroblast cell lines from the spiny mouse (Acomys).

The spiny mouse (Acomys) is gaining popularity as a research organism due to its phenomenal regenerative capabilities. Acomys recovers from injuries to several organs without fibrosis. For example, Acomys heals full thickness skin injuries with rapid re-epithelialization of the wound and regeneration of hair follicles, sebaceous glands, erector pili muscles, adipocytes, and dermis without scarring. Understanding mechanisms of Acomys regeneration may uncover potential therapeutics for wound healing in humans. However, access to Acomys colonies is limited and primary fibroblasts can only be maintained in culture for a limited time. To address these obstacles, we generated immortalized Acomys dermal fibroblast cell lines using two methods: transfection with the SV40 large T antigen and spontaneous immortalization. The two cell lines (AcoSV40 and AcoSI-1) maintained the morphological and functional characteristics of primary Acomys fibroblasts, including maintenance of key fibroblast markers and ECM deposition. The availability of these cells will lower the barrier to working with Acomys as a model research organism, increasing the pace at which new discoveries to promote regeneration in humans can be made.

Comparative analysis of Acomys cahirinus and Mus musculus responses to genotoxicity, oxidative stress, and inflammation.

The Egyptian spiny mouse, Acomys cahirinus, is a recently described model organism for regeneration studies. It has surprising powers of regeneration with relatively fast repairing mechanisms and reduced inflammation form compared to other mammals. Although several studies have documented the exceptional capabilities of Acomys to regenerate different tissues after injury, its response to different cellular and genetic stresses is not yet investigated. Therefore, the current study aimed to investigate Acomys abilities to resist genotoxicity, oxidative stress and inflammation induced by acute and subacute treatments with lead acetate. Responses of Acomys were compared with those of the lab mouse (Mus musculus), which displays signatures of the "typical" mammalian response to various stressors. Cellular and genetic stresses were induced by using acute and subacute doses of Lead acetate (400 mg/kg and 50 mg/kg for 5 days, respectively). The assessment of genotoxicity was carried out by using comet assay, while oxidative stress was evaluated by measuring the biomarkers; MDA, GSH and antioxidant enzymes CAT and SOD. Moreover, inflammation was assessed by analyzing the expression of some inflammatory-regeneration-related genes: CXCL1, IL1-ß, and Notch 2 and immunohistochemical staining of TNF-α protein in brain tissue, in addition to histopathological examination of brain, liver, and kidneys. The obtained results revealed a unique resistance potency of Acomys to genotoxicity, oxidative stress, and inflammation in certain tissues in comparison to Mus. Altogether, the results revealed an adaptive and protective response to cellular and genetic stresses in Acomys.

Bioinspired Strategies for Wound Regeneration.

Regeneration allows animals to replace and restore injured tissues. Animal phyla have evolved different regenerative strategies to increase survival advantages. In contrast to the earlier principle that regeneration recapitulates development, recent studies indicate that wound healing in adult mammals is modified by the inflammatory response to injury, and biochemical signaling from immune and other cellular systems may modulate wound reparative responses to achieve successful tissue regeneration. Here we briefly survey different regenerative strategies used by animals across different phyla. We next focus on skin regeneration using the mouse wound-induced hair neogenesis model as an example to show the circumstances required to rebuild a new, morphogenetically competent field in the adult mammalian skin. Parallel investigations in African spiny mice (Acomys sp.) have further shown that skin rigidity can also modulate wound bed properties to facilitate de novo formation of skin appendages. These regenerating, periodically arranged hair primordia emerge using Turing activator/inhibitor principles with activities derived from sources that differ from those used in embryonic development, including the mechanical environment. Thus, a novel combination of biochemical, immunological, and mechanical signaling strategies can work together to achieve successful cutaneous regeneration in adult animals, potentially inspiring novel therapeutic strategies.

Mammalian organ regeneration in spiny mice.

Fibrosis-driven solid organ failure is a major world-wide health burden with few therapeutic options. Spiny mice (genus: Acomys) are terrestrial mammals that regenerate severe skin wounds without fibrotic scars to evade predators. Recent studies have shown that spiny mice also regenerate acute ischemic and traumatic injuries to kidney, heart, spinal cord, and skeletal muscle. A common feature of this evolved wound healing response is a lack of formation of fibrotic scar tissue that degrades organ function, inhibits regeneration, and leads to organ failure. Complex tissue regeneration is an extremely rare property among mammalian species. In this article, we discuss the evidence that Acomys represents an emerging model organism that offers a unique opportunity for the biomedical community to investigate and clinically translate molecular mechanisms of scarless wound healing and regeneration of organ function in a mammalian species.

Breathing patterns and CO2 production in adult spiny mice (Acomys cahirinus).

The spiny mouse (Acomys) is a precocial mammal with unique regenerative abilities. We used whole-body plethysmography to describe the breathing patterns and CO2 production (VCO2) of adult spiny mice (n = 10 male, 10 female) and C57BL/6 mice (n = 9 male, 11 female). During quiet breathing, female but not male spiny mice had lower tidal volumes and CO2 production vs. C57BL/6 mice. During extended hypoxia (30 min), male and female spiny mice decreased VCO2 and tidal volume to a greater degree than C57BL/6 mice. During an acute hypoxic-hypercapnic respiratory challenge (10% O2, 7% CO2), male and female spiny mice had blunted ventilatory responses as compared to C57BL/6 mice, primarily from a diminished increase in respiratory rate. These data establish a baseline for studies of respiratory physiology and neurobiology in spiny mice in the context of their remarkable regenerative capacity and their unique background of a desert dwelling species.

Wound healing and regeneration in spiny mice (Acomys cahirinus).

The winds of Patagonia are referred to by locals as "The Broom of God" because they sweep away the less fit species that cannot survive there. Fitness as an evolutionary trait has been considered as fundamental for many aspects of morphogenesis and behavior in metazoans. Yet, it has not received much attention in the area of wound healing, despite the obvious relevance of this polygenic trait to an organism's survival in nature. In this chapter, we review the evidence that the rodent species Acomys cahirinus is an emerging mammalian model system that has evolved a non-typical (for mammals) wound healing response that offers unique opportunities for the study of organ regeneration without fibrosis in an adult mammalian species.

Social recognition and short-term memory in two taxa of striped mouse with differing social systems.

The ability to distinguish between familiar and strange conspecifics is important in group-living animals and influences the types of interactions between conspecifics. Social systems differ in sister taxa of the striped mouse genus Rhabdomys originating from different environments. Xeric-adapted R. pumilio displays facultative group-living whereas the mesic-adapted R. d. chakae is solitary. We assessed social recognition and attraction to strangers in females of two populations each of R. pumilio and R. d. chakae by means of a social discrimination task. We used a three-chamber apparatus developed in an established protocol and measured the latency of test females to approach and the duration of their investigation of stimulus females. Differences in social recognition of and preference for unfamiliar conspecifics in group-living and solitary-living taxa occurred at the taxon-level, even though constituent populations occurring kilometers apart showed similar responses. Females differed in the latency (testing phase) and duration of investigation (familiarization and testing phases) inter-specifically but not intra-specifically. Female R. pumilio approached stimulus females faster than female R. d. chakae. Female R. pumilio also investigated stimulus females for longer, regardless of stimulus type compared to R. d. chakae, but both taxa spent more time investigating familiar females than novel females and approached the familiar females faster than novel females. Social recognition, short-term memory, and social preference do not appear to differ between closely related taxa and differences in behavior between the two taxa might be related to inherent personality and social proclivity.

Seasonal changes in problem-solving in wild African striped mice.

Innovative problem-solving ability is a predictor of whether animals can successfully cope with environmental changes. These environmental changes can test the limits of animals, for example when energy availability decreases seasonally and, hence, problem-solving performance decreases because less energy is available for cognitive processes. Here, we investigated: (1) how problem-solving performance changed between seasons that differed significantly in food availability; (2) whether these changes were related to environmentally induced physiological changes in blood glucose and ketone levels, indicators of energy availability; and (3) whether individual variation in problem-solving was related to sex differences. We studied 99 free-ranging African striped mice, Rhabdomys pumilio, in the Succulent Karoo, South Africa, 55 during the hot dry summer with low food availability and 44 during the cold wet winter with higher food availability. We measured their problem-solving abilities using a food extraction task and found no seasonal differences in problem-solving success. However, mice solved the problem faster in summer versus winter. In summer, food availability was reduced and blood ketones increased but there was no seasonal difference in blood glucose levels. There were no correlation between problem-solving performance and blood glucose or ketone levels. Overall, more males solved the task than females. It appears that in striped mice cognitive functions can be maintained under harsh environmental conditions.

Inoculation route-dependent Lassa virus dissemination and shedding dynamics in the natural reservoir - Mastomys natalensis.

Lassa virus (LASV), a Risk Group-4 zoonotic haemorrhagic fever virus, affects sub-Saharan African countries. Lassa fever, caused by LASV, results in thousands of annual deaths. Although decades have elapsed since the identification of the Natal multimammate mouse (Mastomys natalensis) as a natural reservoir of LASV, little effort has been made to characterize LASV infection in its reservoir. The natural route of infection and transmission of LASV within M. natalensis remains unknown, and the clinical impact of LASV in M. natalensis is mostly undescribed. Herein, using an outbred colony of M. natalensis, we investigate the replication and dissemination dynamics of LASV in this reservoir following various inoculation routes. Inoculation with LASV, regardless of route, resulted in a systemic infection and accumulation of abundant LASV-RNA in many tissues. LASV infection in the Natal multimammate mice was subclinical, however, clinical chemistry values were transiently altered and immune infiltrates were observed histologically in lungs, spleens and livers, indicating a minor disease with coordinated immune responses are elicited, controlling infection. Intranasal infection resulted in unique virus tissue dissemination dynamics and heightened LASV shedding, compared to subcutaneous inoculation. Our study provides important insights into LASV infection in its natural reservoir using a contemporary infection system, demonstrating that specific inoculation routes result in disparate dissemination outcomes, suggesting intranasal inoculation is important in the maintenance of LASV in the natural reservoir, and emphasizes that selection of the appropriate inoculation route is necessary to examine aspects of viral replication, transmission and responses to zoonotic viruses in their natural reservoirs.

Adaptations in Hippo-Yap signaling and myofibroblast fate underlie scar-free ear appendage wound healing in spiny mice.

Spiny mice (Acomys cahirinus) are terrestrial mammals that evolved unique scar-free regenerative wound-healing properties. Myofibroblasts (MFs) are the major scar-forming cell type in skin. We found that following traumatic injury to ear pinnae, MFs appeared rapidly in both Acomys and mouse yet persisted only in mouse. The timing of MF loss in Acomys correlated with wound closure, blastema differentiation, and nuclear localization of the Hippo pathway target protein Yap. Experiments in vitro revealed an accelerated PP2A-dependent dephosphorylation activity that maintained nuclear Yap in Acomys dermal fibroblasts (DFs) and was not detected in mouse or human DFs. Treatment of Acomys in vivo with the nuclear Yap-TEAD inhibitor verteporfin prolonged MF persistence and converted tissue regeneration to fibrosis. Forced Yap activity prevented and rescued TGF-ß1-induced human MF formation in vitro. These results suggest that Acomys evolved modifications of Yap activity and MF fate important for scar-free regenerative wound healing in vivo.

Establishment of a Genetically Confirmed Breeding Colony of Mastomys natalensis from Wild-Caught Founders from West Africa.

Mastomys natalensis are a ubiquitous and often dominant rodent across sub-Saharan Africa. Importantly, they are a natural reservoir for microbial pathogens including Lassa virus (LASV), the etiological agent of Lassa fever in humans. Lassa-infected rodents have been documented across West Africa and coincide with regions where annual outbreaks occur. Zoonotic transmission to humans most often occurs directly from infected rodents. Little is known about LASV infection kinetics and transmissibility in M.natalensis, primarily due to available animals. Here, we describe the establishment of a laboratory breeding colony of genetically confirmed M.natalensis from wild-captured rodents. This colony will provide a convenient source of animals to study LASV and other emerging pathogens that utilize M. natalensis in their enzootic lifecycles.

Sons benefit from paternal care in African striped mice.

Mammalian paternal care is rare and is often linked to enhanced fitness under particular ecological conditions. The proximate consequences of paternal care on offspring are lacking, however. Here, we tested whether levels of paternal care predict the behavioural, cognitive and physiological development of sons in the naturally paternal African striped mouse (Rhabdomys pumilio). We focused on sons raised in two treatments: biparental (both parents) or uniparental (mother alone) families. We recorded levels of interactions between pups with both parents, and later assessed the behaviour, cognition and physiology of sons at three developmental stages: juvenile, sub-adult and adult (sexual maturity). Sons from biparental families showed (a) reduced anxiety as juveniles; (b) greater exploration and social interaction at different stages; (c) better cognition; and (d) reduced corticosterone concentrations than sons from uniparental families. In contrast, sons from uniparental families showed greater levels of paternal care, although prolactin concentrations did not differ between treatments. Paternal care in striped mice enhances fitness of males. Here, we also show that sons benefit psychologically and physiologically through interactions with their fathers. However, sons also trade-off such benefits against their own paternal care behaviour, suggesting that fathers influence the development of their son's phenotype in complex ways.

Olfaction alters spatial memory strategy of scatter-hoarding animals.

Although it has been suggested that olfaction is closely interconnected with hippocampal systems, whether olfaction regulates spatial memory strategy remains never known. Furthermore, no study has examined how olfaction mediates spatial memory established on the external objects, for example, caches made by scatter-hoarding animals. Here, we experimentally induced nondestructive and reversible olfaction loss of a scatter-hoarding animal Leopoldamys edwardsi, to test whether and how olfaction regulates spatial memory to mediate cache recovery and pilferage. Our results showed that the normal L. edwardsi preferred to pilfer caches of others rather than to recover their own using accurate spatial memory (35.7% vs. 18.6%). Anosmic L. edwardsi preferred to recover the caches they made prior to olfaction loss rather than to pilfer from others relied on spatial memory (54.2% vs. 36.0%). However, L. edwardsi with anosmia showed no preference either to the caches they established after olfaction loss or caches made by others (25.8% vs. 29.1%). These collectively indicate that olfaction loss has a potential to affect new memory formation but not previously established spatial memory on caches. Our study first showed that olfaction modified spatial memory strategy in cache recovery and pilferage behaviors of scatter-hoarding animals. We suggest that future studies pay more attention to the evolution of olfaction and its relationship with spatial memory strategy.

Of city and village mice: behavioural adjustments of striped field mice to urban environments.

A fundamental question of current ecological research concerns the drives and limits of species responses to human-induced rapid environmental change (HIREC). Behavioural responses to HIREC are a key component because behaviour links individual responses to population and community changes. Ongoing fast urbanization provides an ideal setting to test the functional role of behaviour for responses to HIREC. Consistent behavioural differences between conspecifics (animal personality) may be important determinants or constraints of animals' adaptation to urban habitats. We tested whether urban and rural populations of small mammals differ in mean trait expression, flexibility and repeatability of behaviours associated to risk-taking and exploratory tendencies. Using a standardized behavioural test in the field, we quantified spatial exploration and boldness of striped field mice (Apodemus agrarius, n = 96) from nine sub-populations, presenting different levels of urbanisation and anthropogenic disturbance. The level of urbanisation positively correlated with boldness, spatial exploration and behavioural flexibility, with urban dwellers being bolder, more explorative and more flexible in some traits than rural conspecifics. Thus, individuals seem to distribute in a non-random way in response to human disturbance based on their behavioural characteristics. Animal personality might therefore play a key role in successful coping with the challenges of HIREC.

Neighborhood effects on the tannin-related foraging decisions of two rodent species under semi-natural conditions.

Neighborhood effects on seed predation and dispersal processes are usually seed-characteristic-dependent; however, how seeds with certain characteristics affect the foraging behavior of rodents in relation to other seeds nearby is unclear. Because large differences in seed characteristics between neighboring seeds may lead to significant differences in rodent foraging preferences, we hypothesized that neighborhood effects were more likely to be detected when paired seeds differed in seed characteristics. We investigated the foraging decisions of two rodent species, the red spiny rat Maxomys surifer and the Chinese white-bellied rat Niviventer confucianus, in semi-natural enclosures by presenting them with artificial seeds containing different levels of tannin (0, 3%, and 6% tannin). Both rodents showed similar preferences and preferentially consumed high-tannin seeds (6% tannin) and scatter hoarded low-tannin seeds (0 tannin). The scatter hoarding of low-tannin (0 tannin) and high-tannin (6% tannin) seeds was significantly higher when these seeds were neighboring higher-tannin seeds than when they neighbored lower-tannin seeds, whereas the scatter hoarding of intermediate-tannin seeds (3% tannin) varied little when they had different neighbors. High-tannin-seed (6% tannin) scatter hoarding was lowest when they neighbored low-tannin seeds (0 tannin), while low-tannin-seed (0 tannin) scatter hoarding was highest when they neighbored high-tannin seeds (6% tannin). Therefore, the seeds that the rodents scatter hoarded were next to (neighbored) seeds that they preferred to eat immediately, and vice versa. Our findings suggest that seed neighborhood effects affect rodent foraging behavior and the relationship between plants and rodents, and may have a profound effect on the regeneration and spatial structure of plant communities.

Regeneration in the spiny mouse, Acomys, a new mammalian model.

We describe the tissues and organs that show exceptional regenerative ability following injury in the spiny mouse, Acomys. The skin and ear regenerate: hair and its associated stem cell niches, sebaceous glands, dermis, adipocytes, cartilage, smooth muscle, and skeletal muscle. Internal tissues such as the heart, kidney, muscle, and spinal cord respond to damage by showing significantly reduced inflammation and improved regeneration responses. The reason for this improved ability may lie in the immune system which shows a blunted inflammatory response to injury compared to that of the typical mammal, but we also show that there are distinct biomechanical properties of Acomys tissues. Examining the regenerative behavior of closely related mammals may provide insights into the evolution of this remarkable property.