Bovine semen was analyzed to establish the source of glutamic oxaloacetic transaminase (GOT) and to determine the feasibility of using GOT release from the cell for determining semen quality. Experiments comparing the concentration of GOT in extracellular media after spermatozoa had been subjected to minimal and maximal stress and another measuring the amount of GOT in semen before and after a bilateral corpus epididymectomy led to the conclusion that GOT is primarily associated with the spermatozoa. The correlation between the releasable amount of GOT and sperm concentration was r = .80 (p ≤ .01). From a second group of experiments it was concluded that GOT release from the sperm cell was influenced by centrifuging at various gravitational forces and by freezing with different concentrations of glycerol or ethylene glycol. Freezing spermatozoa after different equilibration periods at 5° or thawing at different temperatures did not significantly alter GOT release. Another experiment involving 98 samples of semen from 35 Holstein bulls showed a significant correlation (r = .21, p ≤ .05) between GOT/109 spermatozoa and fertility. A significant correlation (r = .25, p ≤ .05) was also found between fertility and the amount of enzyme left in the sperm cell after normal freezing, indicating that the amount of enzyme left in the cell is important for fertility.