Mechanisms of cryoprotection in freezing tolerant animal systems

The theoretical mechanisms of freezing protection afforded by the natural occurrence of glycerol in an adult, freezing-tolerant insect have been considered in light of recent findings. While unequivocal identification of the specific site or sites of action of glycerol is yet obscured in a maze of interactions, it is apparent that a multicellular system (organism) that naturally possesses glycerol in high, nonlethal concentrations, that can maintain activity in the presence of tissue ice, that can survive frequent and prolonged freeze-thaw encounters, and finally, that can regulate cryoprotectant levels in the presence of changing environmental conditions should be a major focus of future studies.The carabid beetle, Pterostichus brevicornis has been found to regulate glycerol levels in response to fluctuating ambient temperatures even while frozen. Hemolymph freezing points and whole-body supercooling points correlated well with changes in glycerol. Freezing and supercooling points decreased 0.9 ° C per 4 g/100 ml increase in glycerol.An interpretation of the data accumulated on insect studies and integrated with data from other multicellular systems supports the theory that a single site of freezing concept as applied to considerations of cryoprotection and cryoinjury may not be realistic. Each level of events occurring during the freezing process in glycerolated and nonglycerolated animal systems has been discussed. There appears little room for speculative separation and isolation of the site of action for the events of freezing (and thawing) represent a continuum with changes in each parameter directly dependent upon the entire sequence.