A quantitative evaluation of the extent of inner mitochondrial membrane destruction after freezing-thawing based on functional studies.

An assay based on comparative investigation of ATPase and ATP synthetase changes in the activity in rat liver mitochondria after low temperature and uncoupler 2,4-dinitrophenol treatment is considered. By varying the activity of the respiratory chain, three extents of membrane cryoinjury could be distinguished by monitoring the changes in ATPase activity under different types of cryoinfluence. The first extent corresponds to a minimum membrane destruction, where the action of the respiratory chain compensates the changes in proton permeability and the cryotreatment does not change the ATPase activity. The second extent corresponds to changes in ion permeability which is partially compensated by the respiratory chain action. The third extent corresponds to a maximum membrane destruction and, therefore, maximum increase in permeability which is related to the irreversible stimulation of the ATPase activity and complete inhibition of the phosphorylation. In this study the extent of cryoinjury in mitochondrial preparations frozen and thawed at different rates was evaluated using this method.