Recovery from repeated stressors: Physiology and behavior are affected on different timescales in house sparrows.

For decades, researchers across disciplines have been captivated by classifying, diagnosing, and avoiding the consequences of chronic stress. Despite the vast body of literature this has generated, we still lack the ability to predict which individuals or populations may be susceptible to stress-related pathologies. One critical unanswered question is whether the impacts of repeated stressors are reversible, or if instead they permanently alter an individual. In this study, we exposed house sparrows (Passer domesticus) to 6 days of random, repeated stressors, permitted them 0, 1, 3, or 6 days to recover, and then assessed changes in their body mass, hypothalamic-pituitaryadrenal (HPA) axis (baseline, stress-induced corticosterone, negative feedback strength), immune function, uric acid concentrations, DNA damage levels, and perch hopping activity. Body mass did not vary between groups after recovery. We found that the HPA axis and perch hopping were not significantly impacted by the 6 days of stressors, but that uric acid and DNA damage increased. Short recovery periods tended to negatively affect the HPA axis and reduced uric acid levels, but these were reversed with longer recovery periods. Following the recovery periods, the birds experienced an additional 6 days of random stressors and their responses were assessed again. All recovery times reduced perch hopping and immune function, but paradoxically, DNA damage was highest in the birds that had the longest amount of time to recover. These results show that recovery time affects responses to subsequent chronic stress in complex ways, and highlight the importance of multimodal, interdisciplinary approaches to studying stress physiology.

Recovery periods during repeated stress impact corticosterone and behavioral responses differently in house sparrows.

A number of studies have shown that chronic stress can negatively impact both physiology and behavior in a variety of organisms. What has yet to be extensively explored is whether these changes permanently alter an animal's functioning, or if they can be reversed. In this study, we used wild-caught house sparrows (Passer domesticus) to assess how recovery periods influence the physiological and behavioral impacts of an initial four days and subsequent four days of repeated stressors. Birds were randomly assigned to a recovery group and either experienced 0, 24, or 72 h of recovery between the two sets of stressors (cage rolling and cage tapping). We measured the regulation of the hypothalamic pituitary adrenal (HPA) axis by quantifying baseline and stress-induced corticosterone as well as negative feedback strength. We also assessed behavior using neophobia trials to measure how birds altered their approach towards novel objects and their overall activity. Both behavior and corticosterone responses were assessed before the experiment, after the recovery time, and following the final 4 days of stressors. We found that birds that experienced 24 h of recovery had reduced stress-induced corticosterone, but enhanced negative feedback relative to the pre-experiment sample. Additionally, 4 days of stressors was enough to significantly reduce approach latency towards novel objects; however, pre-experiment levels returned with longer periods of recovery. Finally, recovery time did not significantly influence responses to the second 4 days of stressors. Our results indicate that brief recovery periods partially ameliorate the hormonal and behavioral effects of repeated stress.

Cellular NAD replenishment confers marked neuroprotection against ischemic cell death: role of enhanced DNA repair.

BACKGROUND AND PURPOSE: NAD(+) is an essential cofactor for cellular energy production and participates in various signaling pathways that have an impact on cell survival. After cerebral ischemia, oxidative DNA lesions accumulate in neurons because of increased attacks by ROS and diminished DNA repair activity, leading to PARP-1 activation, NAD(+) depletion, and cell death. The objective of this study was to determine the neuroprotective effects of NAD(+) repletion against ischemic injury and the underlying mechanism. METHODS: In vitro ischemic injury was induced in rat primary neuronal cultures by oxygen-glucose deprivation (OGD) for 1 to 2 hours. NAD(+) was replenished by adding NAD(+) directly to the culture medium before or after OGD. Cell viability, oxidative DNA damage, and DNA base-excision repair (BER) activity were measured quantitatively up to 72 hours after OGD with or without NAD(+) repletion. Knockdown of BER enzymes was achieved in cultures using AAV-mediated transfection of shRNA. RESULTS: Direct NAD(+) repletion in neurons either before or after OGD markedly reduced cell death and OGD-induced accumulation of DNA damage (AP sites, single and double strand breaks) in a concentration- and time-dependent manner. NAD(+) repletion restored nDNA repair activity by inhibiting serine-specific phosphorylation of the essential BER enzymes AP endonuclease and DNA polymerase-beta. Knocking down AP endonuclease expression significantly reduced the prosurvival effect of NAD(+) repletion. CONCLUSIONS: Cellular NAD(+) replenishment is a novel and potent approach to reduce ischemic injury in neuronal cultures. Restoration of DNA repair activity via the BER pathway is a key signaling event mediating the neuroprotective effect of NAD(+) replenishment.