Decrease in nuclear Feulgen-positive material (DNA) upon aging in in vitro storage of bovine spermatozoa.

The Feulgen-DNA content of sperm cells from 5 bulls was studied by means of microspectrophotometry after storage at 5 degrees C for 2, 3, 5, and 10 days in a yolk-citrate diluent permitting slow aerobic metabolism. A subsample of sperm cells from each bull was subjected to the Feulgen technique on each of the storage days selected. The cells sampled on each of these days received a standard 12 minute, 60 degrees C hydrolysis. Absorption measurements at 546 mmicroof the individual cells indicated a marked progressive decrease in the Feulgen-DNA content of the stored spermatozoa. The loss of 30 per cent of the initial DNA at the end of 5 days' storage was highly significant statistically. This decrease approximately parallels the known decrease in fertility of stored sperm cells, as well as the increase in apparent embryonic mortality resulting from the use of similarly aged spermatozoa for artificial insemination.