RAM SPERMATOZOA STUDIED WITH TECHNIQUES FOR THE CYTOCHEMICAL DETECTION OF PHOSPHORYLASE AND SUCCINIC DEHYDROGENASE

Summary. Frozen ram spermatozoa gave a positive reaction for succinic dehydrogenase and a negative one for phosphorylase. An increase of the succinic dehydrogenase reaction after freezing is inter-preted as an augmentation of the cellular permeability. It is well known that ovine spermatozoa do not survive well at low tempera¬ tures. In order to study the effects of freezing upon the ovine spermatozoa we performed the reaction for succinic dehydrogenase after different times of freezing. Succinic dehydrogenase was chosen because it is involved in aerobic metabolism. Earlier papers about techniques for the demonstration of oxidative enzymes in intact cells include studies by Edwards & Valentine (1963), Balogh & Cohen (1964), Hrudka (1965) and Reissenweber (1966). In addition to succinic dehydrogenase, we also studied the enzyme phosphorylase. Semen was collected by artificial vagina, and volume, colour, motility, cell density and méthylène blue reduction time were estimated. We used only those samples that had normal values, and these were frozen at —79 ° C. Aliquots o