Response of Anatolian mountain frogs (Rana macrocnemis and Rana holtzi) to freezing, anoxia, and dehydration: Glucose as a cryoprotectant

Abstract

Cryoprotectants play an essential role in the survival of some amphibians in response to different stress conditions such as freezing, anoxia, and dehydration. Glucose is one of the cryoprotectants important for freeze-tolerant frogs. The aim of the present study was to investigate the survival strategies of Anatolian mountain frogs (Rana macrocnemis and Rana holtzi), which are terrestrial hibernators, by examining the changes in glucose and water content in some tissues at subzero temperatures. In the current study, animals were exposed to freezing (-2.5 °C), anoxia, and dehydration treatments. During these treatments, all frogs survived. The glucose levels in the plasma, liver, and skeletal muscle and the water content of the tissues were measured during the freezing, anoxia, and dehydration. Changes in body weight were also recorded in both species. During the freezing, a 3.3-fold increase was seen in the blood glucose level of R. macrocnemis (1.35 ± 0.25 to 4.45 ± 0.51 μmol mL^-1), whereas the blood glucose level of R. holtzi exhibited a 4.5-fold increase (1.90 ± 0.25 to 8.67 ± 2.22 μmol mL^-1). In the liver, a 6.7-fold increase was seen in the glucose level of R. macrocnemis (5.66 ± 0.15 to 38.27 ± 8.53 μmol g^-1) and the increase in R. holtzi was approximately 6.0-fold (2.25 ± 0.46 to 13.36 ± 1.32 μmol g^-1) during freezing. The liver glucose levels of both species also increased significantly in response to the anoxia and dehydration. In both species, the glucose levels of the skeletal muscle were found to be higher in dehydration than with freezing and anoxia. In conclusion, our results suggest that glucose may be identified as an important cryoprotectant that plays an important role in the survival of Anatolian mountain frogs during extreme conditions.

Keywords: Cryobiology; Freeze tolerance; Glucose levels; Rana holtzi; Rana macrocnemis.