Characterization of gamma-glutamyltranspeptidase in the liver of the frog: 3. Response to freezing and thawing in the freeze-tolerant wood frog Rana sylvatica.

The freeze tolerant wood frog Rana sylvatica was studied to determine the impact of the freezing and thawing of this frog on the activity of gamma-glutamyltranspeptidase in the liver. On exposure to -2.5 degrees C, for 1, 12 and 24 h, frogs were found to be cool, covered with ice crystals and frozen, respectively. Thawing for 24 h at 4 degrees C recovered the frogs completely. A 45 per cent decrease in the liver weight: body weight ratio was notable after 1 h at -2.5 degrees C, suggestive of an early hepatic capacitance response. A glycemic response to freezing was observed: blood glucose levels exhibited a 55 per cent decrease after 1 h at -2.5 degrees C on cooling; a 10.5-fold increase after 12 h at -2.5 degrees C on the initiation of freezing; and a 22-fold increase after 24 h at -2.5 degrees C in the fully frozen state. Blood glucose levels remained elevated four-fold in the thawed state. Plasma insulin levels were increased twofold in the frozen state and 1.8-fold in the thawed state, while plasma ketone levels were increased 1.8-fold in the frozen state and 1.5-fold in the thawed state. Plasma total T3 levels were decreased by 22 per cent in the frozen state and normalized on thawing. In homogenates and plasma membranes isolated from the livers of Rana sylvatica, the activity of gamma-glutamyltrans-peptidase was found to be elevated at all stages of the freeze-thaw process. After 1, 12 and 24 h at -2.5 degrees C, activities were increased 2.5-, 2.3-, 2.4-fold respectively in the homogenates and 2.5-, 2.2-, 2.4-fold respectively in the plasma membranes. After thawing, activities were still increased 1.9-fold in both homogenates and plasma membranes. In homogenates prepared from the kidneys of Rana sylvatica, the activity of gamma-glutamyltranspeptidase was increased 1.4-fold after 1 h at -2.5 degrees C after which it returned to normal. The role of thyroid hormone in producing the increase in gamma-glutamyltranspeptidase in the liver of Rana sylvatica in response to freezing is discussed as is the significance of the enzyme increase in terms of hepatic cytoprotection and freeze tolerance.