Morphological and genomic shifts in mole-rat 'queens' increase fecundity but reduce skeletal integrity.

In some mammals and many social insects, highly cooperative societies are characterized by reproductive division of labor, in which breeders and nonbreeders become behaviorally and morphologically distinct. While differences in behavior and growth between breeders and nonbreeders have been extensively described, little is known of their molecular underpinnings. Here, we investigate the consequences of breeding for skeletal morphology and gene regulation in highly cooperative Damaraland mole-rats. By experimentally assigning breeding 'queen' status versus nonbreeder status to age-matched littermates, we confirm that queens experience vertebral growth that likely confers advantages to fecundity. However, they also upregulate bone resorption pathways and show reductions in femoral mass, which predicts increased vulnerability to fracture. Together, our results show that, as in eusocial insects, reproductive division of labor in mole-rats leads to gene regulatory rewiring and extensive morphological plasticity. However, in mole-rats, concentrated reproduction is also accompanied by costs to bone strength.

Some social animals are highly cooperative creatures that live in tight-knit colonies. Bees and ants are perhaps the most well-known examples of social insects, while Damaraland mole-rats and naked mole-rats, two rodent species found in southern and eastern Africa, are among the most cooperative mammal species. In these colony-forming animals, only one or a few females reproduce and these fertile females are frequently referred to as "queens". When an animal becomes a queen, her body shape can change dramatically to support the demands of high fertility and frequent reproduction. The molecular basis of such changes has been well-described in social insects. However, they are poorly understood in mammals. To address this knowledge gap, Johnston et al. studied how transitioning to queen status affects bone growth and structural integrity in Damaraland mole-rats, as well as body shape and size. The experiments compared non-breeding female mole-rats with other adult females recently paired with a male to become the sole breeder of a new colony. Johnston et al. also used bone-derived cells grown in the laboratory to assess underlying gene regulatory changes in new queen mole-rats. Johnston et al. showed that transitioning to the role of queen leads to a cascade of skeletal changes accompanied by shifts in the regulation of genetic pathways linked to bone growth. Queen mole-rats show accelerated growth in the spinal column of their lower back. These bones are called lumbar vertebrae and this likely allows them to have larger litters. However, queen mole-rats also lose bone growth potential in their leg bones and develop thinner thigh bones, which may increase the risk of bone fracture. Therefore, unlike highly social insects, mole-rats do not seem to have escaped the physical costs of intensive reproduction. This work adds to our understanding of the genes and physical traits that have evolved to support cooperative behaviour in social animals, including differences between insects and mammals. It also shows, with a striking example, how an animal's genome responds to social cues to produce a diverse range of traits that reflect their designated social role.

Cultural transmission of vocal dialect in the naked mole-rat.

Naked mole-rats (Heterocephalus glaber) form some of the most cooperative groups in the animal kingdom, living in multigenerational colonies under the control of a single breeding queen. Yet how they maintain this highly organized social structure is unknown. Here we show that the most common naked mole-rat vocalization, the soft chirp, is used to transmit information about group membership, creating distinctive colony dialects. Audio playback experiments demonstrate that individuals make preferential vocal responses to home colony dialects. Pups fostered in foreign colonies in early postnatal life learn the vocal dialect of their adoptive colonies, which suggests vertical transmission and flexibility of vocal signatures. Dialect integrity is partly controlled by the queen: Dialect cohesiveness decreases with queen loss and remerges only with the ascendance of a new queen.

Behavioural responses to environmental hypercapnia in two eusocial species of African mole rats.

Damaraland and naked mole rat are eusocial mammals that live in crowded burrows in which CO2 is elevated. These species are thought to be highly tolerant of CO2 but their behavioural responses to hypercapnia are poorly understood. We hypothesized that Damaraland and naked mole rats would exhibit blunted behavioural responses to hypercapnia and predicted that their activity levels would be unaffected at low to moderate (2-5%) CO2 but increased at > 7% CO2. To test this, we exposed Damaraland and naked mole rats to stepwise increases in environmental CO2 (0-10%) and measured activity, exploratory behaviour, and body temperature. Surprisingly, we found that both species exhibited no differences in movement velocity, distance travelled, zone transitions (exploration), or body temperature at any level of environmental hypercapnia. Conversely, when carbonic anhydrase was inhibited with acetazolamide (50 mg kg-1 intraperitonially, to increase whole-animal acidosis), exploration was significantly elevated relative to hypercapnic controls in both species at all levels of inhaled CO2, and naked mole rat body temperature decreased in > 7% CO2. We conclude that both species are largely non-responsive to environmental CO2, and that this tolerance may be dependent on bicarbonate buffering at the level of the kidney or within the blood.

Allo-parental care in Damaraland mole-rats is female biased and age dependent, though independent of testosterone levels.

In Damaraland mole-rats (Fukomys damarensis), non-breeding subordinates contribute to the care of offspring born to the breeding pair in their group by carrying and retrieving young to the nest. In social mole-rats and some cooperative breeders, dominant females show unusually high testosterone levels and it has been suggested that high testosterone levels may increase reproductive and aggressive behavior and reduce investment in allo-parental and parental care, generating age and state-dependent variation in behavior. Here we show that, in Damaraland mole-rats, allo-parental care in males and females is unaffected by experimental increases in testosterone levels. Pup carrying decreases with age of the non-breeding helper while the change in social status from non-breeder to breeder has contrasting effects in the two sexes. Female breeders were more likely than female non-breeders to carry pups but male breeders were less likely to carry pups than male non-breeders, increasing the sex bias in parental care compared to allo-parental care. Our results indicate that testosterone is unlikely to be an important regulator of allo-parental care in mole-rats.

Behavioural responses of naked mole rats to acute hypoxia and anoxia.

Naked mole rats (NMRs) are among the most hypoxia-tolerant mammals. Other species respond to hypoxia by either escaping the hypoxic environment or drastically decreasing behavioural activity and body temperature (Tb) to conserve energy. However, NMRs rarely leave their underground burrows, which are putatively hypoxic and thermally stable near the NMRs' preferred Tb Therefore, we asked whether NMRs are able to employ behavioural and thermoregulatory strategies in response to hypoxia despite their need to remain active and the minimal thermal scope in their burrows. We exposed NMRs to progressively deeper levels of hypoxia (from 21 to 0% O2) while measuring their behaviour and Tb Behavioural activity decreased 40-60% in hypoxia and Tb decreased slightly in moderate hypoxia (5-9%) and then further with deeper hypoxia (3% O2). However, even at 3% O2 NMRs remained somewhat active and warm, and continued to explore their environment. Remarkably, NMRs were active for greater than 90 s in acute anoxia and Tb and metabolic rate decreased rapidly. We conclude that NMRs are adapted to remain awake and functional even at the extremes of their hypoxia-tolerance. This adaptation likely reflects variable and challenging levels of environmental hypoxia in the natural habitat of this species.

Variation in growth of Damaraland mole-rats is explained by competition rather than by functional specialization for different tasks.

In some eusocial insect societies, adaptation to the division of labour results in multimodal size variation among workers. It has been suggested that variation in size and growth among non-breeders in naked and Damaraland mole-rats may similarly reflect functional divergence associated with different cooperative tasks. However, it is unclear whether individual growth rates are multimodally distributed (as would be expected if variation in growth is associated with specialization for different tasks) or whether variation in growth is unimodally distributed, and is related to differences in the social and physical environment (as would be predicted if there are individual differences in growth but no discrete differences in developmental pathways). Here, we show that growth trajectories of non-breeding Damaraland mole-rats vary widely, and that their distribution is unimodal, contrary to the suggestion that variation in growth is the result of differentiation into discrete castes. Though there is no evidence of discrete variation in growth, social factors appear to exert important effects on growth rates and age-specific size, which are both reduced in large social groups.

Sociality and the telencephalic distribution of corticotrophin-releasing factor, urocortin 3, and binding sites for CRF type 1 and type 2 receptors: A comparative study of eusocial naked mole-rats and solitary Cape mole-rats.

Various aspects of social behavior are influenced by the highly conserved corticotrophin-releasing factor (CRF) family of peptides and receptors in the mammalian telencephalon. This study has mapped and compared the telencephalic distribution of the CRF receptors, CRF1 and CRF2 , and two of their ligands, CRF and urocortin 3, respectively, in African mole-rat species with diametrically opposed social behavior. Naked mole-rats live in large eusocial colonies that are characterized by exceptional levels of social cohesion, tolerance, and cooperation in burrowing, foraging, defense, and alloparental care for the offspring of the single reproductive female. Cape mole-rats are solitary; they tolerate conspecifics only fleetingly during the breeding season. The telencephalic sites at which the level of CRF1 binding in naked mole-rats exceeds that in Cape mole-rats include the basolateral amygdaloid nucleus, hippocampal CA3 subfield, and dentate gyrus; in contrast, the level is greater in Cape mole-rats in the shell of the nucleus accumbens and medial habenular nucleus. For CRF2 binding, the sites with a greater level in naked mole-rats include the basolateral amygdaloid nucleus and dentate gyrus, but the septohippocampal nucleus, lateral septal nuclei, amygdalostriatal transition area, bed nucleus of the stria terminalis, and medial habenular nucleus display a greater level in Cape mole-rats. The results are discussed with reference to neuroanatomical and behavioral studies of various species, including monogamous and promiscuous voles. By analogy with findings in those species, we speculate that the abundance of CRF1 binding in the nucleus accumbens of Cape mole-rats reflects their lack of affiliative behavior.

Social condition and oxytocin neuron number in the hypothalamus of naked mole-rats (Heterocephalus glaber).

The naked mole-rat is a subterranean colonial rodent. In each colony, which can grow to as many as 300 individuals, there is only one female and 1-3 males that are reproductive and socially dominant. The remaining animals are reproductively suppressed subordinates that contribute to colony survival through their cooperative behaviors. Oxytocin is a peptide hormone that has shown relatively widespread effects on prosocial behaviors in other species. We examined whether social status affects the number of oxytocin-immunoreactive neurons in the paraventricular nucleus and the supraoptic nucleus by comparing dominant breeding animals to subordinate non-breeding workers from intact colonies. We also examined these regions in subordinate animals that had been removed from their colony and paired with an opposite- or same-sex conspecific for 6 months. Stereological analyses indicated that subordinates had significantly more oxytocin neurons in the paraventricular nucleus than breeders. Animals in both opposite- and same-sex pairs showed a decreased oxytocin neuron number compared to subordinates suggesting that status differences may be due to social condition rather than the reproductive activity of the animal per se. The effects of social status appear to be region specific as no group differences were found for oxytocin neuron number in the supraoptic nucleus. Given that subordinate naked mole-rats are kept reproductively suppressed through antagonism by the queen, we speculate that status differences are due either to oxytocin's anxiolytic properties to combat the stress of this antagonism or to its ability to promote the prosocial behaviors of subordinates.

Influence of spatial environment on maze learning in an African mole-rat.

In subterranean species where excavation is energetically expensive, efficient spatial navigation is vital to reducing the costs of locating important resources such as food and mates. While spatial navigational ability is positively correlated with sociality in subterranean mammals, we have a less clear understanding of the role of habitat complexity on navigational ability. We tested spatial navigational ability and memory in 12-18-month captive Natal mole-rats (Cryptomys hottentotus natalensis) maintained in a simple environment with no environmental enrichment and newly captured wild individuals from natural, complex burrow systems. In maze trials, mole-rats captured freshly from the wild made significantly fewer navigational errors, were more likely to successfully navigate the maze, travelled shorter distances and as a consequence, completed the maze in less time. Male mole-rats from both experimental treatments were more likely to complete the maze than females. Memory retention of the maze was tested on day two, seven, 30 and 60, respectively. The results were variable, although both groups showed a significant memory retention 60 days after testing. Our results highlight the potential importance of the environment (microhabitat complexity) on spatial cognitive performance in mole-rats.

Endocrine function and neurobiology of the longest-living rodent, the naked mole-rat.

Animals that have evolved exceptional capabilities, such as extraordinary longevity may reveal pertinent and potentially critical insights into biomedical research that are not readily apparent in standard laboratory animals. Naked mole-rats (Heterocephalus glaber; NMRs) are extremely long-lived (30 years) mouse-sized rodents. They clearly have evolved superior anti-aging mechanisms as evident by the markedly attenuated age-related decline in physiological function, sustained reproductive capacity and pronounced cancer resistance throughout their long-lives. These eusocial rodents, like the social insects, live in colonies with breeding restricted to one female and a few males. Subordinates are sexually monomorphic, yet retain the ability to become breeders, and can undergo growth surges and neural modifications at any time throughout their life. This plasticity in physiological and behavioral aspects may have contributed to their long-lives. Naked mole-rats show numerous adaptations to life underground including extreme tolerance of hypoxia, acid insensitivity, as well as independence of photoendocrine systems. Here we review what is known about their unique social structure, sensory systems, endocrinology and neurobiology, and highlight areas that may be pertinent to biogerontology.

Tolerance to unfamiliar conspecifics varies with social organization in female African mole-rats.

Intolerance to familiar conspecifics characterises solitary mole-rats and distinguishes them from social ones. However, no study has compared the patterns of tolerance to unfamiliar conspecifics. Theoretically, both solitary and social species should react similarly and show intolerance to unfamiliar same-sex conspecifics, unless the evolution of grouping has favoured higher tolerance to conspecifics among the social species. Our study compares tolerance to unfamiliar conspecifics in four African mole-rat species exhibiting varying degrees of sociality. Dyadic encounters between female unfamiliar conspecifics were performed in a neutral arena, and the assessment of social tolerance was based on both behavioural observations of amicable contact behaviour as opposed to aggression and avoidance behaviour and the assessment of a stress response to the encounter measured as an increase in plasma cortisol concentrations. Our results show that the two highly social species and the solitary one presented similar high levels of agonistic behaviours during encounters with unfamiliar conspecifics. Nevertheless, all three social species displayed social tolerance and did not show a stress arousal during encounters with unfamiliar conspecifics, a pattern that contrasted significantly with that evidenced in the solitary species. The results suggest that physiological and behavioural characteristics allow a higher tolerance to unfamiliar conspecifics in the social as opposed to the solitary mole-rat, and the adaptive value of these characteristics are discussed. Finally, we discuss why constraints on social tolerance may be an important limiting factor to take into account in theories concerning the evolution of grouping.

A subterranean mammal uses the magnetic compass for path integration.

Path integration allows animals to navigate without landmarks by continuously processing signals generated through locomotion. Insects such as bees and ants have evolved an accurate path integration system, assessing and coding rotations with the help of a general directional reference, the sun azimuth. In mammals, by contrast, this process can take place through purely idiothetic (mainly proprioceptive and vestibular) signals. However, without any stable external reference for measuring direction, path integration is highly affected by cumulative errors and thus has been considered so far as valid only for short-distance navigation. Here we show through two path integration experiments (homing and shortcut finding) that the blind mole rat assesses direction both through internal signals and by estimating its heading in relation to the earth's magnetic field. Further, it is shown that the greater the circumvolution and length of the traveled path, the more the animal relies on the geomagnetic field. This path integration system strongly reduces the accumulation of errors due to inaccuracies in the estimation of rotations and thus allows the mole rat to navigate efficiently in darkness, without the help of any landmark, over both short and long distances.

Eusociality in African mole-rats: new insights from patterns of genetic relatedness in the Damaraland mole-rat (Cryptomys damarensis).

After the discovery of eusociality in the naked mole-rat, it was proposed that inbreeding and high colony relatedness in this species were the major underlying factors driving cooperative breeding in African molerats. By contrast, field and laboratory studies of the eusocial Damaraland mole-rat (Cryptomys damarensis) have raised the possibility that this species is an obligate outbreeder, although the build-up of inbreeding over several generations could still occur. Using microsatellite markers, we show that most breeding pairs in wild colonies of the Damaraland mole-rat are indeed unrelated (R = 0.02 +/- 0.04) and that mean colony relatedness (R = 0.46 +/- 0.01), determined across 15 colonies from three separate populations, is little more than half that previously identified in naked mole-rats. This finding demonstrates that normal familial levels of relatedness are sufficient for the occurrence of eusociality in mammals. Variation in the mean colony relatedness among populations provides support both for the central role played by ecological constraints in cooperative breeding and for the suggestion that inbreeding in naked mole-rats is a response to extreme constraints on dispersal. Approaches that determine the relative importance of an array of extrinsic factors in driving social evolution in African mole-rats are now required.

Reproductive suppression in female Damaraland mole-rats Cryptomys damarensis: dominant control or self-restraint?

Colonies of Damaraland mole-rats Cryptomys damarensis exhibit a high reproductive skew. Typically one female breeds and the others are anovulatory. Two models, the dominant control model (DCM) and the self-restraint model (SRM), have been proposed to account for this reproductive suppression. The DCM proposes that suppression is under the control of the dominant breeder and is imposed by mechanisms such as aggression, pheromones and interference with copulation, whereas the SRM does not involve aggression directed towards non-breeders and may function in order to minimize inbreeding. We investigated potential proximate mechanisms involved in the suppression of females in a series of experiments. Socially induced stress through aggression did not appear to be responsible for anovulation. Nor did breeders actively interfere with subordinate copulation. Females were physiologically suppressed when housed in intact colonies. However, as predicted by the DCM, they did not become reproductively active when removed from the presence of breeders. We found no evidence that pheromonal cues block ovulation. We suggest that the SRM is the basic model found in the Damaraland mole-rat and that self-restraint functions in order to minimize inbreeding by restricting reproduction until an unrelated male is present. This would explain the rapid onset of reproductive activation in females when paired with an unrelated male, as demonstrated in this study.

Social stress in neighboring and encountering blind mole-rats (Spalax ehrenbergi).

Blind mole-rats (Spalax ehrenbergi) are solitary aggressive subterranean rodents. They inhabit individual territories, comprised of branched tunnels. Each such tunnel system is completely separate from that of any neighboring mole-rat. Although intraspecific encounters between neighbors are infrequent, when they do occur, they may result in the injury or death of one or both animals. Avoidance of encounters may be due to the awareness of a neighbor's whereabouts through scent-marking and/or seismic (vibratory) communication. The present study was intended to examine whether encounters between individual mole-rats result in physiological stress. Two experimental conditions were designed to simulate natural situations: a brief encounter between two neighboring mole-rats, taking place either once or several times and long-term residency of neighbors whose only contact was either vibratory or vibratory plus odor communication. Blood samples were taken before, during, and after encounters in the first experiment and at set intervals in the second. The blood variables measured were blood glucose levels (BGL) and neutrophil/lymphocyte ratio (N/L). Blood glucose levels and neutrophil/lymphocyte ratio ratios increased in both members of encountering pairs. Long-term residency with a neighbor resulted in the establishment of a dominant-subordinate relationship through vibratory communication only, with increased neutrophil/lymphocyte ratio ratio in the subordinate males. However, long-term residency of males exposed to both vibrations and odors of neighboring males resulted in the death of both individuals. It seems that brief direct encounters and long-term neighboring conditions without physical contact are sufficient to cause severe stress to mole-rats. It is possible that in the wild, in some situations in which neighboring mole-rats cannot avoid constant exposure to each other's vibratory and odor signals, the consequent extensive stress may result in death.

Hormonal and behavioural correlates of male dominance and reproductive status in captive colonies of the naked mole-rat, Heterocephalus glaber.

Naked mole-rat colonies are societies with a high reproductive skew, breeding being restricted to one dominant female (the 'queen') and 1-3 males. Other colony members of both sexes are reproductively suppressed. Experimental removal of breeding males allowed us to investigate the relationship between urinary testosterone and cortisol, dominance rank, and male reproductive status. Dominance rank was strongly correlated with body weight, age, and urinary testosterone titres in males. No relationship between urinary cortisol levels and male reproductive status or dominance was found. Breeding males were among the highest-ranking, heaviest and oldest males in their respective colonies, and were succeeded by other high-ranking, large, old colony males. In contrast to females, no evidence of competition over breeding status was observed among males. Male-male agonism was low both before and after removal of breeders and mate guarding was not observed. The lower reproductive skew for males compared with female skew or queen control over male reproduction may explain why males compete less strongly than females over breeding status after removal of same-sexed breeders.