Arctic Ground Squirrels Research Articles

Effects of Hyperoxia on Brain Tissue Oxygen Tension in Non-Sedated, Non- Anesthetized Arctic Ground Squirrels: An Animal Model of Hyperoxic Stress

Arctic Ground Squirrels (AGS) are classic hibernators known for their tolerance to hypoxia. AGS have been studied as a model of hypoxia with potential as a medical research model. Problem statement: Their unique resistance to the stressors of low oxygen led us to hypothesize that AGS might also be adaptable to hyperoxia. Approach: This study examined the physiological pattern associated with hyperoxia in response to brain tissue oxygen partial pressure (PtO2), brain temperature (Tbrain), global oxygen consumption (VO2) and respiratory frequency (fR) using non-sedated and nonanesthetized Arctic Ground Squirrels (AGS) and rats. Results: We found that 1) 100% inspired oxygen (FiO2) increased the baseline values of brain PtO2 significantly in both summer euthermic AGS (24.4 ± 3.6-87.3 ± 3.6 mmHg, n=6) and in rats (18.2 ± 5.2-73.3 ± 5.2 mmHg, n = 3); PtO2 was significantly higher in AGS than in rats during hyperoxic exposure; 2) hyperoxic exposure had no effect on brain temperature in either AGS or rats, with the brain temperatures maintaining constancy before, during and after 100% O2 exposure; 3) systemic metabolic rates increased significantly during hyperoxic exposure in both euthermic AGS and rats; moreover, VO2 were significantly lower in AGS than in rats during hyperoxic exposure; 4) the respiratory rates for rats were maintained before, during and after 100% O2 exposure, while the respiratory responding patterns to hyperoxic exposure changed after exposure in AGS. AGS fR was significantly lower after hyperoxic exposure than before the exposure. Conclusion: These results suggest that hyperoxic ventilation induced PtO2 and VO2 differences between AGS and rats and led to altered respiratory patterns between these species. AGS and the rat serves as an excellent comparative model for hypoxic and hyperoxic stress studies of the brain.

Phenological variation in annual timing of hibernation and breeding in nearby populations of Arctic ground squirrels

Ecologists need an empirical understanding of physiological and behavioural adjustments that animals can make in response to seasonal and long-term variations in environmental conditions. Because many species experience trade-offs between timing and duration of one seasonal event versus another and because interacting species may also shift phenologies at different rates, it is possible that, in aggregate, phenological shifts could result in mismatches that disrupt ecological communities. We investigated the timing of seasonal events over 14 years in two Arctic ground squirrel populations living 20 km apart in Northern Alaska. At Atigun River, snow melt occurred 27 days earlier and snow cover began 17 days later than at Toolik Lake. This spatial differential was reflected in significant variation in the timing of most seasonal events in ground squirrels living at the two sites. Although reproductive males ended seasonal torpor on the same date at both sites, Atigun males emerged from hibernation 9 days earlier and entered hibernation 5 days later than Toolik males. Atigun females emerged and bred 13 days earlier and entered hibernation 9 days earlier than those at Toolik. We propose that this variation in phenology over a small spatial scale is likely generated by plasticity of physiological mechanisms that may also provide individuals the ability to respond to variation in environmental conditions over time.

Insular arctic ground squirrels (Spermophilus parryii) of the North Pacific: indigenous or exotic?

We lack critical information for oceanic archipelagos worldwide related to the origin and status of insular faunas. In southwestern Alaska, in particular, a need exists to tease apart whether specific insular populations are naturally occurring or are the result of exotic introductions by humans. We analyzed variation in mitochondrial sequences of the cytochrome-b gene and 8 nuclear microsatellite loci across 215 individuals representing 17 populations (12 insular) to refine our understanding of the history of the previously identified Southwest clade of arctic ground squirrels (Spermophilus parryii). We found significant geographic structure that suggests long-term isolation and diversification (Ushugat Island and Cold Bay), but we also documented closely related populations that are likely the result of human-mediated introductions. The latter instances (Kavalga and Unalaska islands) corroborate reports from early Alaska explorers. DOI: 10.1644/09-MAMM-A386.1.

Habitat selection by arctic ground squirrels (Spermophilus parryii)

Abstract Arctic ground squirrels (Spermophilus parryii) are abundant, colonial, noncyclical small mammals whose within-range distribution patterns have received little scientific attention. The distribution of arctic ground squirrels may drive the abundance and spatial arrangement of other arctic ecosystem components, because they serve as a prey item, plant predator, and ecological engineer. We modeled ground squirrel habitat selection using exponential resource selection functions. To determine whether the presence of adjacent squirrel burrows influenced our indices of habitat selection, we evaluated autocovariates in our models that accounted for the predicted relative probability of ground squirrel presence within neighborhoods of various sizes around each focal area. Our models demonstrated selection by ground squirrels for well-drained substrates and sloped or convex terrain, and against wet and hummocky terrain and areas of high greenness. At a large spatial scale highly selected habitat was predic...

Early Wisconsinan (MIS 4) Arctic ground squirrel middens and a squirrel-eye-view of the mammoth-steppe

Fossil arctic ground squirrel ( Spermophilus parryii) middens were recovered from ice-rich loess sediments in association with Sheep Creek-Klondike and Dominion Creek tephras (ca 80 ka) exposed in west-central Yukon. These middens provide plant and insect macrofossil evidence for a steppe-tundra ecosystem during the Early Wisconsinan (MIS 4) glacial interval. Midden plant and insect macrofossil data are compared with those previously published for Late Wisconsinan middens dating to ∼25–29 14C ka BP (MIS 3/2) from the region. Although multivariate statistical comparisons suggest differences between the relative abundances of plant macrofossils, the co-occurrence of steppe-tundra plants and insects (e.g., Elymus trachycaulus, Kobresia myosuroides, Artemisia frigida, Phlox hoodii, Connatichela artemisiae) provides evidence for successive reestablishment of the zonal steppe-tundra habitats during cold stages of the Late Pleistocene. Arctic ground squirrels were well adapted to the cold, arid climates, steppe-tundra vegetation and well-drained loessal soils that characterize cold stages of Late Pleistocene Beringia. These glacial conditions enabled arctic ground squirrel populations to expand their range to the interior regions of Alaska and Yukon, including the Klondike, where they are absent today. Arctic ground squirrels have endured numerous Quaternary climate oscillations by retracting populations to disjunct “interglacial refugia” during warm interglacial periods (e.g., south-facing steppe slopes, well-drained arctic and alpine tundra areas) and expanding their distribution across the mammoth-steppe biome during cold, arid glacial intervals.

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.

Shotgun Proteomics Analysis of Hibernating Arctic Ground Squirrels

Mammalian hibernation involves complex mechanisms of metabolic reprogramming and tissue protection. Previous gene expression studies of hibernation have mainly focused on changes at the mRNA level. Large scale proteomics studies on hibernation have lagged behind largely because of the lack of an adequate protein database specific for hibernating species. We constructed a ground squirrel protein database for protein identification and used a label-free shotgun proteomics approach to analyze protein expression throughout the torpor-arousal cycle during hibernation in arctic ground squirrels (Urocitellus parryii). We identified more than 3,000 unique proteins from livers of arctic ground squirrels. Among them, 517 proteins showed significant differential expression 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 months after hibernation had ended (non-hibernating). Consistent with changes at the mRNA level shown in a previous study on the same tissue samples, proteins involved in glycolysis and fatty acid synthesis were significantly underexpressed at the protein level in both late torpid and early aroused animals compared with non-hibernating animals, whereas proteins involved in fatty acid catabolism were significantly overexpressed. On the other hand, when we compared late torpid and early aroused animals, there were discrepancies between mRNA and protein levels for a large number of genes. Proteins involved in protein translation and degradation, mRNA processing, and oxidative phosphorylation were significantly overexpressed in early aroused animals compared with late torpid animals, whereas no significant changes at the mRNA levels between these stages had been observed. Our results suggest that there is substantial post-transcriptional regulation of proteins during torpor-arousal cycles of hibernation.

Evidence for Selective Caching by Arctic Ground Squirrels Living in Alpine Meadows in the Yukon

Male arctic ground squirrels (Spermophilus parryii) rely on food they cached the previous year for the energy they need to compete for mates each spring. We collected cheek-pouch contents of arctic ground squirrels trapped during three summers (2000–02) as an indication of what squirrels cached. Among adults, both males and females carried material in their cheek pouches, but males did so more frequently than females (4.4% vs. 0.6% of captures). Males carried material later in the summer than females, and also carried different material (seeds and rhizomes as opposed to nesting material). These differences probably reflect different purposes of cheek-pouch contents—females carried material for immediate use, whereas males carried food for caching. Only 24 of over 100 species of vascular plants growing at our alpine study site were carried, and presumably cached, by male arctic ground squirrels. The seeds or rhizomes of one species, Polygonum viviparum, were found in over 90% of cheek-pouch contents examined, even though that species grew at relatively low density and was no more common than another species in the same genus (Polygonum bistorta) that was never found in cheek-pouch contents. Collectively, this evidence indicates that males are highly selective in what species they cache. Many of the species carried by arctic ground squirrels in this study have also been found in Pleistocene fossil caches from central Yukon, indicating that food preferences of this species may have remained stable over time.

Ground Squirrel: The Mysterious Rodent of Kodiak

Chirikof Island is well separated from the Alaska Peninsula and other landmasses. Thus its Arctic ground squirrels evidently were introduced by humans, as were all other land mammals on the island. It is unlikely that fur companies introduced ground squirrels. Rather, the animals appear to have been released there by Natives, particularly the Alutiiq of Kodiak. Radiocarbon dating of two archaeological habitation sites from which ground squirrel bones have been recovered confirms that the context of the bones is prehistoric in age and predates Russian settlement in the area by two centuries in one case, and probably by more than a millennium in the other case. Additionally, the time line for contact for the Semidi Islands and for Marmot Island (near Afognak Island) renders it likely that ground squirrels found on those islands were also introduced in pre-contact times.

Odontometric Patterns in the Radiation of Extant Ground-dwelling Squirrels within Marmotini (Sciuridae: Xerini)

I document odontometric variation across ground-dwelling squirrels of the Holarctic tribe Marmotini. Dental size, which correlates well with published average body mass values across species, accounts for most odontometric variation across the clade. Dental shape variation primarily reflects relative size of P3 (upper cheek teeth) and relative width of p4–m1 + length of m3 (lower cheek teeth). Shape variables and relative tooth crown height covary significantly across species, suggesting a common functional complex or shared genetic control. When dental morphology is mapped on published DNA-based phylogenies, Sciurotamias (Chinese rock squirrels), Ammospermophilus (antelope squirrels), and basal subgenera within Spermophilus (ground squirrels [Callospermophilus and Otospermophilus]) are shown to retain inferred primitive dental morphology—small to moderate dental size, relatively small P3, relatively narrow p4–m1 + shortened m3, and relatively low toothcrown height. Other clades depart from this morphotype in size (very small in Tamias [chipmunks] and very large in Marmota [marmots]), tooth shape (especially 2 clades representing Eurasian and North American subgenus Spermophilus), or in both attributes (notably Cynomys [prairie dogs]), with frequent homoplasy. A plot of odontometric distance against published estimates of divergence time between sister clades suggests a roughly ‘‘clocklike’’ accumulation of odontometric change through time but highlights episodes of rapid odontometric evolution during the origins of Marmota, Cynomys, and Spermophilus parryii (arctic ground squirrel).

Protein kinase C epsilon activation delays neuronal depolarization during cardiac arrest in the euthermic arctic ground squirrel

During the pre-hibernation season, arctic ground squirrels (AGS) can tolerate 8 min of asphyxial cardiac arrest (CA) without detectable brain pathology. Better understanding of the mechanisms regulating innate ischemia tolerance in AGS has the potential to facilitate the development of novel prophylactic agents to induce ischemic tolerance in patients at risk of stroke or CA. We hypothesized that neuroprotection in AGS involves robust maintenance of ion homeostasis similar to anoxia-tolerant turtles. Ion homeostasis was assessed by monitoring ischemic depolarization (ID) in cerebral cortex during CA in vivo and during oxygen glucose deprivation in vitro in acutely prepared hippocampal slices. In both models, the onset of ID was significantly delayed in AGS compared with rats. The epsilon protein kinase C (epsilonPKC) is a key mediator of neuroprotection and inhibits both Na+/K+-ATPase and voltage-gated sodium channels, primary mediators of the collapse of ion homeostasis during ischemia. The selective peptide inhibitor of epsilonPKC (epsilonV1-2) shortened the time to ID in brain slices from AGS but not in rats despite evidence that epsilonV1-2 decreased activation of epsilonPKC in brain slices from both rats and AGS. These results support the hypothesis that epsilonPKC activation delays the collapse of ion homeostasis during ischemia in AGS.

Adaptive response of brain tissue oxygenation to environmental hypoxia in non-sedated, non-anesthetized arctic ground squirrels

The present study examined the physiological mechanisms of the responses of brain tissue oxygen partial pressure (P tO 2), brain temperature ( T brain), global oxygen consumption (V̇ O2), and respiratory frequency ( f R) to hypoxia in non-sedated and non-anesthetized arctic ground squirrels ( Spermophilus parryii, AGS) and rats. We found that (1) in contrast to oxygen partial pressure in blood (P aO 2), the baseline value of P tO 2 in summer euthermic AGS is significantly higher than in rats; (2) both P tO 2 and P aO 2 are dramatically reduced by inspired 8% O 2 in AGS and rats, but AGS have a greater capacity in P tO 2 to cope with environmental hypoxia; (3) metabolic rate before, during, and after hypoxic exposure is consistently lower in AGS than in rats; (4) the respiratory responding patterns to hypoxia in the two species differ in that f R decreases in AGS but increases in rats. These results suggest that (1) AGS have special mechanisms to maintain higher P tO 2 and lower P aO 2, and these levels in AGS represent a typical pattern of adaptation of heterothermic species to and a brain protection from hypoxia; (2) AGS brain responds to hypoxia through greater decreases in P tO 2 and decreased f R and ventilation. In contrast, rat brain responds to hypoxia by less reduction in P tO 2 and increased f R and ventilation.

Energetics of arousal episodes in hibernating arctic ground squirrels

Arctic ground squirrels overwintering in northern Alaska experience average soil temperature of -10 degrees C. To examine energetic costs of arousing from hibernation under arctic compared to temperate conditions, captive ground squirrels were maintained in ambient temperatures (T(a)) of 2, -5 and -12 degrees C. Rates of oxygen consumption and carbon dioxide production were used to estimate metabolic rate and fuel use during the three phases of arousal episodes: rewarming, euthermia, and recooling. Respiratory quotient comparisons suggest exclusive use of lipid during rewarming and mixed fuel use during euthermia. Animals rewarming from torpor at T(a) -12 degrees C took longer, consumed more oxygen, and attained higher peak rates of oxygen consumption when compared to 2 degrees C. T(a) had no significant effect on cost or duration of the euthermic phase. Animals recooled faster at -12 degrees C than at 2 degrees C, but total oxygen consumption was not different. T(a) had no significant effect on the total cost of arousal episodes when all three phases are included. Arousal episodes account for 86% of estimated costs of a complete hibernation cycle including torpor when at 2 degrees C and only 23% at -12 degrees C. Thus, due to the higher costs of steady-state metabolism during torpor, proportional metabolic costs of arousal episodes at T(a) characteristic of the Arctic are diminished compared to relative costs of arousals in more temperate conditions.

Physiological oxidative stress after arousal from hibernation in Arctic ground squirrel

Hibernation in Arctic ground squirrels (AGS), Spermophilus parryii, is characterized by a profound decrease in oxygen consumption and metabolic demand during torpor that is punctuated by periodic rewarming episodes, during which oxygen consumption increases dramatically. The extreme physiology of torpor or the surge in oxygen consumption during arousal may increase production of reactive oxygen species, making hibernation an injurious process for AGS. To determine if AGS tissues experience cellular stress during rewarming, we measured carbonyl proteins, lipid peroxide end products and percent oxidized glutathione in brown adipose tissue (BAT) and liver of torpid, hibernating (hAGS), late arousal (laAGS), and cold-adapted, euthermic AGS (eAGS). In BAT carbonyl proteins and lipid peroxide end products were higher in eAGS and laAGS than in hAGS. By contrast, in liver, no significant difference in carbonyl proteins was observed. In another group of animals, comparison of carbonyl proteins and percent oxidized glutathione in frontal cortex, liver, and BAT of eAGS and hAGS showed no evidence of oxidative stress associated with torpor. These results indicate that increased thermogenesis associated with arousal AGS results in tissue specific oxidative stress in BAT but not in liver. Moreover, torpor per se is largely devoid of oxidative stress, likely due to suppression of oxidative metabolism.

PHF-like tau phosphorylation in mammalian hibernation is not associated with p25-formation

In Alzheimer's disease and related disorders, hyperphosphorylation of tau is associated with an increased activity of cyclin dependent kinase 5 (cdk5). Elevated cdk5 activity is thought to be due to the formation of p25 and thereby represents a critical element in the dysregulation of tau phosphorylation under pathological conditions. However, there is still a controversy regarding the correlation of p25 generation and tau pathology. Recently, we demonstrated physiological, paired helical filament-like tau phosphorylation that reversibly occurs in hibernating mammals. Here we used this model to test whether the tau phosphorylation in hibernation is associated with the formation of p25. Analysing brain material of arctic ground squirrels and Syrian hamsters we found no evidence for a hibernation dependent generation of p25. Hence, we suppose that phosphorylation of tau does not require the formation of p25. Instead we suggest that the truncation of p35 to p25 represents a characteristic of pathological alterations and may contribute to aggregation and deposition of hyperphosphorylated tau.

Impact of live trapping on stress profiles of Richardson’s ground squirrel ( Spermophilus richardsonii)

Researching the physiological ecology of natural populations requires an understanding of the impact of capture-induced stress because of its numerous effects on physiological processes. In many cases, initial blood samples to which comparisons are made are obtained well after capture and may differ markedly from free-ranging conditions. We examined the extent to which stress profiles of male Richardson’s ground squirrels ( Spermophilus richardsonii) were affected by short-term responses to live trapping. We compared stress profiles of true base animals (blood samples obtained <3 min of capture) with those of nominal base animals (blood samples obtained >1 h after capture). Total cortisol increased almost 40% whereas our measure of corticosteroid binding globulin (CBG) decreased by 21%, resulting in a two-fold increase in free cortisol levels in nominal base animals compared with true base animals. Capture caused androgen concentrations to fall to almost half of those of true base animals. Energy mobilization increased markedly (22% in glucose and 221% in free fatty acids). Although white blood cell counts did not change, the number of neutrophils was 48% higher in true base animals. There were no changes in hematocrit or lymphocyte counts. Although most of the changes were predictable, the changes in CBG and androgens were unexpected based on previous work on closely related Arctic ground squirrels ( Spermophilus parryii). Our results emphasize the value of obtaining true base measurements whenever possible in order to assess the directions and degree of bias introduced by trapping.

Modulation of gene expression in hibernating arctic ground squirrels

We performed a broadscale screening of differential gene expression using both high-throughput bead-array technology and real-time PCR assay in brown adipose tissue, liver, heart, hypothalamus, and skeletal muscle in hibernating arctic ground squirrels, comparing animals sampled after two durations of steady-state torpor, during two stages of spontaneous arousal episodes, and in animals after they ended hibernation. Significant seasonal and torpor-arousal cycle differences of gene expression were detected in genes involved in glycolysis, fatty acid metabolism, gluconeogenesis, amino acid metabolism, molecular transport, detoxification, cardiac contractility, circadian rhythm, cell growth and apoptosis, muscle dystrophy, and RNA and protein protection. We observed, for the first time, complex modulation of gene expression during multiple stages of torpor-arousal cycles. The mRNA levels of certain metabolic genes drop significantly during the transition from late torpor to early arousal, perhaps due to the rapid turnover of mRNA transcripts resulting from the translational demands during thermogenesis in early arousal, whereas the mRNA levels of genes related to circadian rhythm, cell growth, and apoptosis rise significantly in the early or late arousal phases during torpor-arousal cycle, suggesting the resumption of circadian rhythm and cell cycle during arousal.

Simultaneous measurement of brain tissue oxygen partial pressure, temperature, and global oxygen consumption during hibernation, arousal, and euthermy in non-sedated and non-anesthetized Arctic ground squirrels

This study reports an online temperature correction method for determining tissue oxygen partial pressure ( P t O 2 ) in the striatum and a novel simultaneous measurement of brain P t O 2 and temperature ( T brain) in conjunction with global oxygen consumption ( V O 2 ) in non-sedated and non-anesthetized freely moving Arctic ground squirrels (AGS, Spermophilus parryii). This method fills an important research gap—the lack of a suitable method for physiologic studies of tissue P O 2 in hibernating or other cool-blooded species. P t O 2 in AGS brain during euthermy (21.22 ± 2.06 mmHg) is significantly higher ( P = 0.016) than during hibernation (13.21 ± 0.46 mmHg) suggests brain oxygenation in the striatum is normoxic during euthermy and hypoxic during hibernation. These results in P t O 2 are different from blood oxygen partial pressure ( P a O 2 ) in AGS, which are significantly lower during euthermy than during hibernation and are actually hypoxic during euthermy and normoxic during hibernation in our previous study. This intriguing difference between the P O 2 of brain tissue and blood during these two physiological states suggests that regional mechanisms in the brain play a role in maintaining tissue oxygenation and protect against hypoxia during hibernation.

Dietary Fatty Acid Composition and the Hibernation Patterns in Free‐Ranging Arctic Ground Squirrels

Laboratory experiments have demonstrated that the amount of polyunsaturated fatty acids (PUFAs) in the diet before hibernation influences patterns of mammalian torpor. The hibernation ability of ground squirrels is greatest (longest torpor bouts, greatest number of animals entering torpor) when the PUFA content of their fall diets is 33-74 mg/g, under laboratory conditions. The extent to which natural fall diets both (a) vary in PUFA content and (b) influence the torpor patterns of free-ranging populations of hibernating mammals is unknown, however. We conducted a 3-yr study on the diet PUFA contents and subsequent hibernation patterns of free-ranging arctic ground squirrels (Spermophilus parryii) in the Brooks Range of Alaska. We found that the PUFA contents of fall diets varied more than threefold among individuals. Our study also revealed that arctic ground squirrels that consumed a moderate-PUFA (33-74 mg/g) diet had (a) longer torpor bouts, (b) fewer arousals from torpor, (c) shorter arousal periods, (d) more days in torpor, and (e) greater probability of persisting in the population than those that consumed a high-PUFA (>74 mg/g) diet during the fall. No animals were demonstrated to have consumed a diet representing low-PUFA (<33 mg/g) values. Our study is therefore the first to demonstrate that estimated dietary PUFA levels of a free-ranging hibernator influence subsequent torpor patterns.

Arctic Ground Squirrel (Spermophilus Parryii) Hippocampal Neurons Tolerate Prolonged Oxygen—Glucose Deprivation and Maintain Baseline ERK1/2 and JNK Activation Despite Drastic ATP Loss

Oxygen-glucose deprivation (OGD) initiates a cascade of intracellular responses that culminates in cell death in sensitive species. Neurons from Arctic ground squirrels (AGS), a hibernating species, tolerate OGD in vitro and global ischemia in vivo independent of temperature or torpor. Regulation of energy stores and activation of mitogen-activated protein kinase (MAPK) signaling pathways can regulate neuronal survival. We used acute hippocampal slices to investigate the role of ATP stores and extracellular signal-regulated kinase (ERK)1/2 and Jun NH(2)-terminal kinase (JNK) MAPKs in promoting survival. Acute hippocampal slices from AGS tolerated 30 mins of OGD and showed a small but significant increase in cell death with 2 h OGD at 37 degrees C. This tolerance is independent of hibernation state or season. Neurons from AGS survive OGD despite rapid ATP depletion by 3 mins in interbout euthermic AGS and 10 mins in hibernating AGS. Oxygen-glucose deprivation does not induce JNK activation in AGS and baseline ERK1/2 and JNK activation is maintained even after drastic depletion of ATP. Surprisingly, inhibition of ERK1/2 or JNK during OGD had no effect on survival, whereas inhibition of JNK increased cell death during normoxia. Thus, protective mechanisms promoting tolerance to OGD by AGS are downstream from ATP loss and are independent of hibernation state or season. Journal of Cerebral Blood Flow & Metabolism (2008) 28, 1307-1319; doi:10.1038/jcbfm.2008.20; published online 9 April 2008.