Genomic analysis of miRNAs in an extreme mammalian hibernator, the Arctic ground squirrel

MicroRNAs (miRNAs) are 19- to 25-nucleotide-long small and noncoding RNAs now well-known for their regulatory roles in gene expression through posttranscriptional and translational controls. Mammalian hibernation is a physiological process involving profound changes in set-points for food consumption, body mass and growth, body temperature, and metabolic rate in which miRNAs may play important regulatory roles. In an initial study, we analyzed miRNAs in the liver of an extreme hibernating species, the Arctic ground squirrel ( Spermophilus parryii ), using massively parallel Illumina sequencing technology. We identified >200 ground squirrel miRNAs, including 18 novel miRNAs specific to ground squirrel and mir-506 that is fast evolving in the ground squirrel lineage. Comparing animals sampled after at least 8 days of continuous torpor (late torpid), within 5 h of a spontaneous arousal episode (early aroused), and 1–2 mo after hibernation had ended (nonhibernating), we identified differentially expressed miRNAs during hibernation, which are also compared with the results from two other miRNA profiling methods: Agilent miRNA microarray and real-time PCR. Among the most significant miRNAs, miR-320 and miR-378 were significantly underexpressed during both stages of hibernation compared with nonhibernating animals, whereas miR-486 and miR-451 were overexpressed in late torpor but returned in early arousal to the levels similar to those in nonhibernating animals. Analyses of their putative target genes suggest that these miRNAs could play an important role in suppressing tumor progression and cell growth during hibernation. High-throughput sequencing data and microarray data have been submitted to GEO database with accession: GSE19808 .

Absence of cellular stress in brain after hypoxia induced by arousal from hibernation in Arctic ground squirrels

Although hypoxia tolerance in heterothermic mammals is well established, it is unclear whether the adaptive significance stems from hypoxia or other cellular challenge associated with euthermy, hibernation, or arousal. In the present study, blood gases, hemoglobin O2 saturation (So2), and indexes of cellular and physiological stress were measured during hibernation and euthermy and after arousal thermogenesis. Results show that arterial O2 tension (PaO2) and So2 are severely diminished during arousal and that hypoxia-inducible factor (HIF)-1α accumulates in brain. Despite evidence of hypoxia, neither cellular nor oxidative stress, as indicated by inducible nitric oxide synthase (iNOS) levels and oxidative modification of biomolecules, was observed during late arousal from hibernation. Compared with rats, hibernating Arctic ground squirrels ( Spermophilus parryii) are well oxygenated with no evidence of cellular stress, inflammatory response, neuronal pathology, or oxidative modification following the period of high metabolic demand necessary for arousal. In contrast, euthermic Arctic ground squirrels experience mild, chronic hypoxia with low So2 and accumulation of HIF-1α and iNOS and demonstrate the greatest degree of cellular stress in brain. These results suggest that Arctic ground squirrels experience and tolerate endogenous hypoxia during euthermy and arousal.

Ascorbate dynamics and oxygen consumption during arousal from hibernation in Arctic ground squirrels

During hibernation in Arctic ground squirrels ( Spermophilus parryii), O2 consumption and plasma leukocyte counts decrease by >90%, whereas plasma concentrations of the antioxidant ascorbate increase fourfold. During rewarming, O2 consumption increases profoundly and plasma ascorbate and leukocyte counts return to normal. Here we investigated the dynamic interrelationships among these changes. Plasma ascorbate and uric acid (urate) concentrations were determined by HPLC from blood samples collected at ∼15-min intervals via arterial catheter; leukocyte count and hematocrit were also determined. Body temperature, O2 consumption, and electromyographic activity were recorded continuously. Ascorbate, urate, and glutathione contents in body and brain samples were determined during hibernation and after arousal. During rewarming, the maximum rate of plasma ascorbate decrease occurred at the time of peak O2 consumption and peak plasma urate production. The ascorbate decrease did not correlate with mouth or abdominal temperature; uptake into leukocytes could account for only a small percentage. By contrast, liver and spleen ascorbate levels increased significantly after arousal, which could more than account for ascorbate clearance from plasma. Brain ascorbate levels remained constant. These data suggest that elevated concentrations of ascorbate {[Asc]} in plasma {[Asc]p} provide an antioxidant source that is redistributed to tissues during the metabolic stress that accompanies arousal.

The influence of herbivores and neighboring plants on risk of browsing: a case study using arctic lupine (Lupinus arcticus) and arctic ground squirrels (Spermophilus parryii plesius)

We examined how herbivore distribution and density, neighboring plant density and species composition, and individual plant morphology all influence the risk that individual arctic lupines (Lupinus arcticus) will be browsed by arctic ground squirrels (Spermophilus parryii plesius). Risk of being browsed was significantly influenced by the number of resident ground squirrels but not by overall squirrel density at a site. As the leaf density of neighboring conspecifics increased, risk of browsing to an individual lupine decreased except when palatable neighbors were also present. The presence of other palatable species increased the risk of browsing. Risk was highest when both lupine and other palatable neighbors were present. The presence of unpalatable neighbors reduced the risk of browsing of individual lupines. We discuss these results in the context of three hypotheses: (1) attractant decoy, (2) resource concentration, and (3) repellent plant. No single hypothesis accounts for our observations, but an interaction between herbivores, neighbors, and individual lupine morphology determined risk of browsing.