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Distribution of Radioactive Metabolites Between Bovine Spermatozoa and the Incubation Medium

Department of Dairy Science, Pennsylvania State University, University Park

ABSTRACT

Washed bovine spermatozoa were incubated with glutamate-U-¹⁴C, succinate-1,4- ¹⁴C, glucose-U-¹⁴C, and acetate-2-"C, and the distribution of radioactivity between sperm cells and incubation medium determined. Using density-gradient centrifugation through 11% lactose-1% formalin, only 1 to 2% of the recovered radioactivity from glutamate or succinate was in the sperm cell fraction; with glucose this figure was about 7%, but with acetate, 75%. Thus with some substrates, a reduction in interfering substances and consequent improvement in electrophoretic-chromatographic separation of metabolites can be obtained by removing the sperm and analyzing only the incubation medium.