The Intracellular Accumulation of Metabolites by Ram Spermatozoa During Incubation in the Presence of Exogenous Fructose

Abstract

A method is described by which sufficient spermatozoa can be separated rapidly from their incubation medium to assay individual metabolites accumulating from [U�14C]fructose. Acetate and lactate made up 60-80% of the substrate carbon accumulated within the cell. Acetate was the main intracellular metabolite and its concentration in the cell was 10-20 times its extracellular level. The intra-cellular accumulation of lactate varied with the period of incubation but its concen-tration in the cell was always below that in the medium. The remaining substrate carbon in the cell was made up of fructose and at least three other unidentified compounds. No accumulation of substrate carbon was detected in citric acid cycle intermediates.