Effects of acute heat exposure on oxidative stress and antioxidant defenses in overwintering frogs, Nanorana parkeri

Climate warming is intensifying on the Tibetan Plateau and poses a serious threat to amphibians that live there. Although hibernation physiology and ecology of Nanorana parkeri, a frog species native to the Tibetan plateau, has been well studied, little information is available about the physiological and biochemical responses to acute rising temperature. Here, we conducted an acute warming experiment comparing hibernating N. parkeri (acclimated at 4 °C) and frogs acutely warmed to 10 °C for 12 hours, comparing indicators of oxidative stress and antioxidant defense between the two groups. Acute warming led to a significant increase in the content of oxidized glutathione and the ratio of oxidized:reduced glutathione in liver and muscle, indicating that rapid warming causing oxidative stress could be a negative factor for frogs inhabiting the Tibetan plateau. Malondialdehyde content increased by 57% only in muscle but decreased significantly in three tissues. Protein carbonyl groups rose by 15% in brain, 28% in liver, and 21% in muscle after heat exposure but vitamin C content in heart decreased by 44%. Acute heat exposure also induced tissue-specific upregulation of antioxidant enzyme activities. Catalase activity increased by 27% in heat-exposed frogs, as compared to controls, and glutathione peroxidase activity rose by 12% in brain, 30% in liver, and 12% in muscle. Glutathione-S-transferase activity was also enhanced in heart and muscle of heat-exposed frogs. Acute warming also resulted in a rise in total antioxidant capacity in all tissues examined except kidney, relative to controls. In summary, our findings show that acute heat exposure to hibernating N. parkeri causes a tissue-specific increase in oxidative damage and antioxidant defenses, with skeletal muscle being the most affected tissue. These results reveal the physiological responses to acute temperature change in overwintering N. parkeri.