The naked mole-rat (Heterocephalus glaber) demonstrates a strict social and reproductive hierarchy. Oxytocin (OXT) is a peptide hormone that acts both peripherally and centrally in the regulation of a number of sexual and social behaviours. The main area of central production of this peptide is the paraventricular nucleus of the hypothalamus (PVN). The present study characterized differences that exist in OXT neurons in this region. Breeders and subordinates from established colonies were sacrificed and brains were processed for OXT-immunoreactivity. Four further groups of paired animals underwent various social and hormonal manipulations (opposite-sex paired, same sex-paired, opposite-sex/gonadectomised paired, opposite-sex/vasectomized paired) and were also used for analysis. Results showed that subordinate naked mole-rats had significantly more OXT-immunoreactive neurons in the PVN than either breeders or paired animals that had been gonadectomised. However, no differences were found on measures of OXT cell volume. Possible functional significance of these differences is discussed.

The naked mole-rat (Heterocephalus glaber) demonstrates a strict social and reproductive hierarchy. Oxytocin (OXT) is a peptide hormone that acts both peripherally and centrally in the regulation of a number of sexual and social behaviours. The main area of central production of this peptide is the paraventricular nucleus of the hypothalamus (PVN). The present study characterized differences that exist in OXT neurons in this region. Breeders and subordinates from established colonies were sacrificed and brains were processed for OXT-immunoreactivity. Four further groups of paired animals underwent various social and hormonal manipulations (opposite-sex paired, same sex-paired, opposite-sex/gonadectomised paired, opposite-sex/vasectomized paired) and were also used for analysis. Results showed that subordinate naked mole-rats had significantly more OXT-immunoreactive neurons in the PVN than either breeders or paired animals that had been gonadectomised. However, no differences were found on measures of OXT cell volume. Possible functional significance of these differences is discussed.

Subterranean rodents are interesting model group for studying of the circadian timing system due to their cryptic lifestyle and challenging living conditions. The filed data indicate that the locomotor activity of the silvery mole-rat (Heliophobius argenteocinereus) is influenced by temperature cycle, but the confirmation in laboratory study was to date missing.

Until recently, it was assumed that African mole-rats search for food randomly, as it is in concordance with Aridity food distribution hypothesis. However, recent studies indicate that some subterranean rodent species including mole-rats could be able to use plant chemicals (kairomones) for food localization. In my master thesis I conducted a battery of experiments on four mole-rat species to prove, that these species also posses the ability of kairomone guided foraging independently of their social organization.

Life underground is one of the most challenging tasks for animals. The subterranean environment is seasonally and diurnally stable and provides shelter from predators. On the other hand food is scarce, the cost of digging is very high and closed burrows create hypoxic and hypercapnic conditions. Thus the physiological and behavioral adaptations to save energy are very important for underground dwellers. Social thermoregulation is such behavioral adaptation. Additionally, it has been suggested that social animals suffer from ?isolation stress? and that they decrease their metabolic rates when other family members are present = socio-physiological effect. In this study I measured the resting metabolic rates (RMR) of social Mashona mole-rats (Fukomys darlingi) in isolated individuals, pairs and groups of three to eight individuals. Measurements were carried out at two ambient temperatures, in the thermoneutral zone (TNZ; 30°C) to test the presence of a socio-physiological effect and below their TNZ (20°C) to test the effect of social thermoregulation. A socio-physiological effect was distinctive neither in pairs nor in larger groups. At temperature below the TNZ the Mashona mole-rat saved 21% of its energetic expense in pairs due to social thermoregulation. With an increase in group size, energetic savings rose up to four animals. In larger groups, social thermoregulation did not influence the energetic expenditure, possibly because Mashona mole-rat´s families naturally contain around four to five adults.

Having an appropriate understanding of the behavioral and physiological impact of a disease requires thoroughly fielded tests and wide-ranging animal models to properly deduce generalized impacts of the disease under investigation. Through novel animal models, we acquire diverse insights into the mechanisms at play so that we may approach the problem at hand with fresh perspective and renewed vigor. Using the naked mole-rat (Heterocephalus glaber), a species becoming increasingly common in the medical sciences, my dissertation aimed to accomplish the aforementioned goals in two parts: 1) take the naked mole-rat through a battery of behavioral tasks to bolster our capabilities of using this animal model in medical research; and 2) introduce the naked mole-rat as an alternative model for the investigation of how loss of dental sensation alters behavior and neuroanatomy. Chapter two describes the many methods with which I tested the naked mole-rat across a variety of typical behavioral tasks for social dominance, learning and memory, anxiety, depression, and sociability. Included herein were tests to determine appropriate motivators for this subterranean species to perform such tasks. Additionally, due to the naked mole-rat’s unique disposition for biting, I measured their biting behaviors and compared them across rodent and other mammalian species. The results show that naked mole-rats exhibit large evolutionary divergence in their sensory capabilities and great consideration needs to be given to the proper behavioral tasks and subsequent evaluations of these behavioral paradigms. In chapter three, I evaluated the impact of tooth extraction on affective behaviors, learning and memory, and sociability. Chapter four sought to uncover underlying neuroplasticity associated with the behavioral tasks performed in chapter three. Additionally, in chapter four, I investigated plasticity of the traditional somatosensory pathway for tooth sensation. Tooth loss decreased neuronal density of contralateral ventral posteromedial nucleus of the thalamus and increased the neuronal density of the contralateral ventral dentate gyrus, indicating that tooth-loss induced neuronal plasticity may be more related to plasticity of pain circuitry or resultant from alterations in the muscles of mastication following tooth loss. Though no significant changes in the hippocampus arose from one year of living with no right incisor, I theorized as to which of the physiological idiosyncrasies exhibited by Heterocephalus glaber may have inhibited any observable plasticity due to tooth loss.

The seasonality of wild caught Cape mole-rats was investigated, focusing on endocrine, neuroendocrine and neuroanatomical parameters. The Cape mole-rat reproduces only during the winter rainfall period of the western Cape Province. Interestingly, the gonadal steroid hormones do not show any significant seasonal patterns in either males or females. In addition, the pituitary does not exhibit a reduced sensitivity to GnRH stimulation during the non-breeding season in either sex. Moreover, in the brain the distribution, number and size of GnRH perikarya do not differ with respect to season. Therefore, although only producing young during a specific period of the year, the Cape mole-rat does not appear to be physiologically repressed from reproducing throughout the year. The results of this study imply that the Cape mole-rat is an opportunistic breeder that is reproductively quiescent during the dry season as a consequence of ecological factors such as lack of favourable burrowing conditions and that when opportunities avail themselves following aseasonal rainfall, reproduction may occur. The seasonality and socially induced infertility of the Natal mole-rat was examined via endocrine, neuroendocrine and neuroanatomical investigations. In contrast with the phylogenetically closely related common mole-rat and highveld mole-rat, the Natal mole-rat does not appear to breed seasonally. Post-mortem examination of animals collected revealed pregnant reproductive females throughout the year. A similar scenario is reflected in gonadal steroid concentrations, with no major seasonal differences observed in either males or females. Likewise, pituitary sensitivity to GnRH does not change over season in males or females, and all neuroanatomical parameters investigated remained stable over season in both males and females. Natal mole-rats captured for this study occur in an area with a high annual rainfall, (almost double that of the common mole-rat habitat and one and a half times that of the highveld mole-rat) and the relatively higher rainfall and hence soil moisture content may provide the Natal mole-rat with increased foraging time a result of increased periods over which the soil is optimal for burrowing. Socially induced infertility in the Natal mole-rat appears to be enforced behaviourally as a result of active incest avoidance. The circulating testosterone concentrations were not significantly different in reproductive and non-reproductive males, and likewise oestrogen concentrations did not differ between reproductive and non-reproductive females. Progesterone concentrations were very low in subordinate females, but markedly higher in reproductive females. This is not surprising since the Natal mole-rat is an induced ovulator, thus coitus is required to induce the LH surge that stimulates ovulation. The sensitivity of the pituitary to a GnRH challenge did not differ between reproductive and non-reproductive animals from either sex. No neuroanatomical differences were observed between reproductive and non-reproductive animals of either sex, however, non-reproductive males and females had significantly higher amounts of GnRH in the median eminence, implying that GnRH is synthesized and transported to the median eminence but not released in the portal system to the anterior pituitary. These findings suggest that non-reproductive animals of both sexes are not sterile, but merely sexually quiescent. Copyright / Thesis (PhD)--University of Pretoria, 2008. / Zoology and Entomology / unrestricted

Background: A variety of microbial communities exist throughout the human and animal body. Genetics, environmental factors and long-term dietary habit contribute to shaping the composition of the gut microbiota. For this reason the study of the gut microbiota of a mammal exhibiting an extraordinary life span is of great importance. The naked mole-rat (Heterocephalus glaber) is a eusocial mammal known for its longevity and cancer resistance. Methods: Here we analyzed its gut microbiota by cultivating the bacteria under aerobic and anaerobic conditions and identifying their species by mass spectrometry. Results: Altogether, 29 species of microbes were identified, predominantly belonging to Firmicutes, and Bacteroidetes. The most frequent species were Bacillus megaterium (45.2 %), followed by Bacteroides thetaiotaomicron (19.4 %), Bacteroides ovatus, Staphylococcus sciuri and Paenibacillus spp., each with a frequency of 16.1 %. Conclusion: Overall, the gut of the naked mole-rat is colonized by diverse, but low numbers of cultivable microbes compared with humans and mice. The primary food plants of the rodents are rich in polyphenols and related compounds, possessing anti-microbial, anti-inflammatory, anti-oxidative as well as anti-cancer activity which may contribute to their exceptionally healthy life.

Background: The naked mole-rat (NMR) is a long-lived and cancer resistant species. Identification of potential anti-cancer and age related mechanisms is of great interest and makes this species eminent to investigate anti-cancer strategies and understand aging mechanisms. Since it is known that the NMR expresses higher liver mRNA-levels of alpha 2-macroglobulin than mice, nothing is known about its structure, functionality or expression level in the NMR compared to the human A2M. Results: Here we show a comprehensive analysis of NMR- and human plasma-A2M, showing a different prediction in glycosylation of NMR-A2M, which results in a higher molecular weight compared to human A2M. Additionally, we found a higher concentration of A2M (8.3±0.44 mg/mL vs. and 4.4±0.20 mg/mL) and a lower total plasma protein content (38.7±1.79 mg/mL vs. 61.7±3.20 mg/mL) in NMR compared to human. NMR-A2M can be transformed by methylamine and trypsin resulting in a conformational change similar to human A2M. NMRA2M is detectable by a polyclonal antibody against human A2M. Determination of tryptic and anti-tryptic activity of NMR and human plasma revealed a higher anti-tryptic activity of the NMR plasma. On the other hand, less proteolytic activity was found in NMR plasma compared to human plasma.

The naked mole-rat (Heterocephalus glaber) is one of the most social mammals on the planet. These animals live in underground colonies consisting of a breeding female (the “queen”), 1 to 3 breeding males and up to 300 nonreproductive “workers” organized in a dominance hierarchy. In addition to their eusociality, naked mole-rats have evolved many extreme biological characteristics including an exquisite sense of touch and insensitivity to certain types of pain. However, the overlap between the social and somatosensory abilities of naked mole-rats remains mysterious. In this work, we show that naked mole-rats exhibit caste-like behavioral signatures linked to eusociality, predominantly utilize snout-to-snout interactions in social behavior and possess a somatosensory profile to mechanical stimuli distinct from mice. In the first chapter of this work, we leveraged machine learning and molecular biology tools to create a behavioral atlas of naked mole-rat behavior. We first utilized a computational pipeline of pose-tracking using SLEAP and behavioral segmentation using keypoint-MoSeq to identify 20+ behavioral syllables. This showed that the queen naked mole-rat has a distinct behavioral phenotype from the workers, consisting of faster movements, less anxiety-like freezing, and less colony maintenance behaviors such as digging. We next showed that behavioral differences exist between the workers and that the dominance rank of a given individual could be predicted by its spontaneous behaviors in an open field. Relative rank differential appeared to have relevance for social behaviors: during a tube test for dominance, pairings of a high-ranked and low-ranked animal occurred more quickly and were won more frequently by the dominant animal. Snout touch played an integral role in these dominance tests (perhaps in the communication of individual ID information). This prompted us to investigate snout-to-snout interactions in the open field. We found that two familiar naked mole-rats from the same colony engaged in hundreds of snout interactions in a 10-minute period, and the number of interactions was nearly twice as high between two foreign animals. Follow-up experiments explored the molecular basis of this snout touch and showed that mechanosensory channels (e.g. Piezo2) are likely involved in social snout-to-snout interactions. Furthermore, trimming the sensory whiskers of naked mole-rats disrupts their ability to recognize conspecifics and alters their relative dominance relationships. These findings uncover face touch as a prominent social behavior in naked mole-rats that is intimately linked to social recognition. In the second chapter of this work, we more deeply investigated naked mole-rat somatosensation by using high-speed videography. We began by determining how naked mole-rats respond to both innocuous stimuli (cotton swab, dynamic brush) and noxious stimuli (light and heavy pinpricks) when given to the hindpaw. Compared to mice, naked mole-rats showed a distinct hindpaw phenotype, never responding to innocuous touch stimuli, responding to light pinprick, but rarely responding to heavy pinprick (normally the most noxious stimulus that elicits the strongest pain response in mice). Interestingly, naked mole-rats do respond to brush stimuli to the back skin with a dorsiflexion posture. These animals also exhibit an idiosyncratic withdrawal response to a brush applied to the snout skin which appears to be highly aversive. Interestingly, the velocity of this aversive snout withdrawal appears to be socially modulated and is decreased in the presence of another naked mole-rat in the testing chamber. This phenotype does not occur in mice and provides additional evidence that the naked mole-rat snout is not only extremely sensitive but plays a role in processing socially relevant information.