[Relation between intactness and adenine nucleotide pattern of ejaculated bull spermatozoa]

The intactness of bull spermatozoa was determined by exclusion of external succinate. This was oxygraphically measured in the presence of cytochrome c after inhibition of the cell respiration with rotenone. An addition of cytochrome c was necessary for a reproducible respiration of damaged cells. The portion of intact cells determined in this way correlated with that obtained by the fluorescent dye primuline. The content of adenine nucleotides and the percentage of ATP of washed fresh spermatozoa depended on the intactness of the sperm sample. The adenine nucleotide pattern of spermatozoa after deep freezing corresponded to that of washed fresh ejaculates with a high percentage of damaged cells. The major part of ejaculates tested showed only a small increase of damaged cells during substrate-free incubations at 38 degrees C up to 4 h under aerobic and anaerobic conditions. Ejaculates which had shown a drastic increase of damaged cells were not recognizable by the initial portion of damaged cells. Also a lowered content of ATP in the cells was no indicator of an increased fragility of the cell membrane. A remarkable loss of total adenine nucleotides was observed under conditions of cold shock. It indicates that the changes in the sperm membrane induced by cold shock were qualitatively different in comparison with changes caused by storage and by a slow decrease in temperature.