This study evaluated DNA degradation in nuclei of muscle cells during ischemia in a rabbit limb amputation model. Seven New Zealand rabbits were used. The left rear limb of the animal was amputated. The quadriceps femoris muscle was biopsied, as well as the rabbit carotid arteries. Half of the tissue samples were preserved at a cold temperature (2-4 degrees C), and the other half were kept at room temperature (25 degrees C, global ischemia). Immediately and 4, 8, 12, 16, and 24 h after amputation, muscle tissue samples were stained with the Feulgen reaction and analyzed by an image-analysis system to detect DNA contents in muscle cell nuclei. Samples from carotid arteries were examined with a transmission electron microscope. The results showed that the DNA content in muscle cell nuclei was slightly decreased at 4 and 8 h of global ischemia. At 12 h after global ischemia, the DNA content was found to be significantly decreased (80.03% of baseline). The DNA content dropped to 53.93% of baseline at 16 h of global ischemia. However, in the muscle that was preserved at cold temperature, the DNA contents were only slightly changed along with the ischemia time at 24 h. The DNA changes were confirmed by transmission electron microscopic observations. The present findings could provide evidence for the study of revascularization of ischemic muscle.