Metabolic characteristics of overwintering by the high-altitude dwelling Xizang plateau frog, Nanorana parkeri.

The Xizang plateau frog, Nanorana parkeri, has the highest altitudinal distribution of all frogs in the world and survives the cold of winter without feeding by entering into a hibernating state. However, little attention has been paid to its physiological and biochemical characteristics that support overwintering underwater in small ponds. Here, we measured metabolic rate and heart rate, and collected liver and muscle samples from N. parkeri in summer and winter for analysis of mitochondrial respiration rate, and activities and relative mRNA transcript expression of metabolic enzymes. Compared with summer-collected frogs, both resting metabolic rate and heart rate were significantly reduced in winter-collected frogs. Both state 3 and state 4 respiration of liver mitochondria were also significantly reduced in winter but muscle mitochondria showed a decline only in state 3 respiration in winter. The activities and corresponding mRNA expression of cytochrome c oxidase showed a marked decline in winter, whereas the activities and corresponding mRNA expression of lactate dehydrogenase increased in winter-collected frogs, compared to summer. The thermal sensitivity (Q values) for state 3 respiration rate by liver mitochondria, and activities of lactate dehydrogenase, and cytochrome c oxidase all increased in winter-collected frogs, compared with summer frogs, suggesting that overwintering frogs were more sensitive to changes in external temperature. Enzyme changes mainly result from lower overall quantities of these enzymes as well as post-translational modifications. We conclude that overwintering N. parkeri exhibit a seasonal, temperature-independent suppression of metabolism that is mediated at multiple levels: physiological, mitochondrial, gene expression and enzyme activity levels.