Altered cochlear innervation in developing and mature naked and Damaraland mole rats

Compared to many other rodent species, naked mole rats (Heterocephalus glaber) have elevated auditory thresholds, poor frequency selectivity, and limited ability to localize sound. Because the cochlea is responsible for encoding and relaying auditory signals to the brain, we used immunofluorescence and quantitative image analysis to examine cochlear innervation in mature and developing naked mole rats compared to mice (Mus musculus), gerbils (Meriones unguiculatus), and Damaraland mole rats (Fukomys damarensis), another subterranean rodent. In comparison to mice and gerbils, we observed alterations in afferent and efferent innervation as well as their patterns of developmental refinement in naked and Damaraland mole rats. These alterations were, however, not always shared similarly between naked and Damaraland mole rats. Most conspicuously, in both naked and Damaraland mole rats, inner hair cell (IHC) afferent ribbon density was reduced, whereas outer hair cell afferent ribbon density was increased. Naked and Damaraland mole rats also showed reduced lateral and medial efferent terminal density. Developmentally, naked mole rats showed reduced and prolonged postnatal reorganization of afferent and efferent innervation. Damaraland mole rats showed no evidence of postnatal reorganization. Differences in cochlear innervation specifically between the two subterranean rodents and more broadly among rodents provides insight into the cochlear mechanisms that enhance frequency sensitivity and sound localization, maturation of the auditory system, and the evolutionary adaptations occurring in response to subterranean environments

Fructose-driven glycolysis supports anoxia resistance in the naked mole-rat

The African naked mole-rat’s ($\textit{Heterocephalus glaber}$) social and subterranean lifestyle generates a hypoxic niche. Under experimental conditions, naked mole-rats tolerate hours of extreme hypoxia and survive 18 minutes of total oxygen deprivation (anoxia) without apparent injury. During anoxia, the naked mole-rat switches to anaerobic metabolism fueled by fructose, which is actively accumulated and metabolized to lactate in the brain. Global expression of the GLUT5 fructose transporter and high levels of ketohexokinase were identified as molecular signatures of fructose metabolism. Fructose-driven glycolytic respiration in naked mole-rat tissues avoids feedback inhibition of glycolysis via phosphofructokinase, supporting viability. The metabolic rewiring of glycolysis can circumvent the normally lethal effects of oxygen deprivation, a mechanism that could be harnessed to minimize hypoxic damage in human disease.Work was supported aEuropean Research Council (294678), the Deutsche Forschungsgemeinschaft SFB 665 and Go865/9-1, NSF (grant #0744979 ), NIH (grants HL71626 and HL606

Cannabinoid Modulation of Development and Neuroprotection in African Naked Mole-Rats

Naked mole-rats display typical subterranean features, but also has unusual characteristics even among subterranean mammals due to unusually large colony sizes. The naked mole-rat provides insights into the complex interplay of evolutionary adaptations to the constraints of subterranean living, the hallmark of which is extreme hypoxia tolerance. Many of the known naked mole-rat adaptations are based in systems modulated by the endocannabinoid (eCB) system. This project characterized the three most relevant to naked mole-rat adaptations: neoteny, low oxygen tolerance, and attenuated hypercapnic pain response. I found that naked mole-rat expression of the endocannabinoid system is delayed in the hippocampus and prefrontal cortex. Additionally, the eCB system has developmentally dependent effects on behavior and physiology. I found novel effects in locomotion, synaptic facilitation, and synaptic plasticity. I also characterized the C-fiber pathway in naked mole-rats and investigating the attenuation of pain through the eCB system. I found that there is a functional purinergic response in naked mole-rats which cannabinoids attenuate, but not inflammatory response after activation. The final aim of the project was to assess the role of endocannabinoids in naked mole-rat hypoxia tolerance. In an analysis of naked mole-rat expression after 5 hours of hypoxia, there is a global decrease in multiple endocannabinoids which is not seen in mice. I looked at the effect of the eCB system on the hippocampus when exposed to anoxia. In immature animals, antagonizing the eCB system was protective. In another age-dependent response, adult animals responded equally to both agonism and antagonism of the eCB system to improve the time to depolarization. In both plasticity and anoxic protection, the adult response may be caused by a species dependent change in the balance of CB1r localization compared to excitatory and inhibitory synapses. Therefore, like other neonatally retained traits, the retention of an immature endocannabinoid system may be due to a need for adult naked mole-rats to mediate neuroprotection through the endocannabinoid system. This project identified multiple areas of interest for therapeutic potential in development, pain, and neuroprotection and reinforced the dynamic potential of the endocannabinoid system to modulate a vast array of functions

Gasotransmitter modulation of hypoglossal motoneuron activity

Obstructive sleep apnea (OSA) is characterized by sporadic collapse of the upper airway leading to periodic disruptions in breathing. Upper airway patency is governed by genioglossal nerve activity that originates from the hypoglossal motor nucleus. Mice with targeted deletion of the gene Hmox2, encoding the carbon monoxide (CO) producing enzyme, heme oxygenase-2 (HO-2), exhibit OSA, yet the contribution of central HO-2 dysregulation to the phenomenon is unknown. Using the rhythmic brainstem slice preparation that contains the preBötzinger complex (preBötC) and the hypoglossal nucleus, we tested the hypothesis that central HO-2 dysregulation weakens hypoglossal motoneuron output. Disrupting HO-2 activity increased the occurrence of subnetwork activity from the preBötC, which was associated with an increased irregularity of rhythmogenesis. These phenomena were also associated with the intermittent inability of the preBötC rhythm to drive output from the hypoglossal nucleus (i.e., transmission failures), and a reduction in the input-output relationship between the preBötC and the motor nucleus. HO-2 dysregulation reduced excitatory synaptic currents and intrinsic excitability in inspiratory hypoglossal neurons. Inhibiting activity of the CO-regulated H2S producing enzyme, cystathionine-g-lyase (CSE), reduced transmission failures in HO-2 null brainstem slices, which also normalized excitatory synaptic currents and intrinsic excitability of hypoglossal motoneurons. These findings demonstrate a hitherto uncharacterized modulation of hypoglossal activity through mutual interaction of HO‑2/CO and CSE/H2S, and support the potential importance of centrally‑derived gasotransmitter activity in regulating upper airway control

The naked mole-rat has a functional purinergic pain pathway despite having a non-functional peptidergic pain pathway.

Naked mole-rats (Heterocephalus glaber) have adaptations within their pain pathway that are beneficial to survival in large colonies within poorly ventilated burrow systems, with lower O2 and higher CO2 ambient levels than ground-level environments. These adaptations ultimately lead to a partial disruption of the C-fiber pain pathway, which enables naked mole-rats to not feel pain from the acidosis associated with CO2 accumulation. One hallmark of this disruption is that naked mole-rats do not express neuropeptides, such as Substance P and calcitonin gene-related peptide in their cutaneous C-fibers, effectively making the peptidergic pain pathway hypofunctional. One C-fiber pathway that remains unstudied in the naked mole-rat is the non-peptidergic, purinergic pathway, despite this being a key pathway for inflammatory pain. The current study aimed to establish the functionality of the purinergic pathway in naked mole-rats and the effectiveness of cannabinoids in attenuating pain through this pathway. Cannabinoids can manage chronic inflammatory pain in both humans and mouse models, and studies suggest a major downstream role for the purinergic receptor, P2X3, in this treatment. Here we used Ca2+-imaging of cultured dorsal root ganglion neurons and in vivo behavioral testing to demonstrate that the P2X3 pathway is functional in naked mole-rats. Additionally, formalin-induced inflammatory pain was reduced by the cannabinoid receptor agonist, WIN55 (inflammatory, but not acute phase) and the P2X3 receptor antagonist A-317491 (acute and inflammatory phases). This study establishes that the purinergic C-fiber pathway is present and functional in naked mole-rats and that cannabinoid-mediated analgesia occurs in this species.This work has been supported by the following: a grant from the National Science Foundation (1655494) and a UIC LAS Award for Faculty of Sciences to T.J.P., and a CRUK Grant (C56829/A22053) to ESS