When guinea-pig spermatozoa were suspended in a minimal culture medium (MCM-PL), 2-deoxy-D-glucose (100 microM) and 2-amino-2-deoxy-D-glucose (100 microM) were potent inhibitors of the acrosome reaction without affecting the sperm motility, whereas the N-acetyl derivative 2-acetamido-2-deoxy-D-glucose (2 mM) had no inhibitory effect. The addition of D-glucose (2 mM) partly inhibited the percentage acrosome reaction of spermatozoa suspended in Medium MCM-PL, but DL-alpha-glycerophosphate (2 mM) and myo-inositol (2 mM) had no effect. In addition, DL-alpha-glycerophosphate (2 mM) did not overcome the inhibitory effect of 2-deoxy-D-glucose on the sperm acrosome reaction. The inhibitory action of 2-deoxy-D-glucose (100 microM) on the sperm acrosome reaction assessed after a 3-h incubation was irreversible and was only completely effective if the sugar was added within 30 min of the start of incubation. When spermatozoa suspended in Medium MCM-PL were treated with 2-deoxy-D-glucose (100-200 microM) for an extended incubation up to 6 h, the inhibitory effect of 2-deoxy-D-glucose was partly overcome. Spermatozoa treated with 2-deoxy-D-glucose had significantly reduced concentrations of ATP after incubation for 2 h in Ca2+-free media, compared with the ATP concentrations of spermatozoa preincubated for 2 h in Ca2+-free media that supported acrosome reactions. The addition of Ca2+ (5 mM) caused a rapid decrease in ATP concentrations of spermatozoa suspended in Medium MCM-PL, while the addition of the monovalent ionophore monensin (50 microM) and Ca2+ stimulated sperm acrosome reactions as well as an additional decline in the sperm ATP concentrations. However, monensin (50 microM) in the absence of Ca2+ caused only a slight decline in the sperm ATP concentrations over the 15-min incubation period. The depletion of the sperm ATP concentrations by 2-deoxy-D-glucose may retard completion of the capacitation process and the resultant acrosome reaction.

Guinea pig spermatozoa were collected from the caudae epididymides in various isotonic solutions and the intracellular sodium and potassium content was determined by atomic absorption spectroscopy. The sperm intracellular Na and K content was found to be influenced by large variations in the extracellular concentrations of these ions. Treatment of spermatozoa suspended in a saline‐based solution with the monovalent ionophores monensin or nigericin caused an approximate 2‐fold increase in the intracellular Na content and a 3–6 fold decrease in the intracellular K content. Incubation of the spermatozoa in a K+‐free minimal culture medium (MCM‐PL) at a pH of 7.6 or 8.3 for 2 hr caused an approximate 2‐fold increase in the sperm intracellular Na content and a 5‐fold decrease in the intracellular K content. The motile spermatozoa incubated for 2 hr at pH 7.6 showed less than 5% acrosome reactions, compared with 30–40% acrosome reactions after incubation at pH 8.3, in response to the addition of 5 mM Ca2+. Changes in the sperm intracellular elemental composition during culture in vitro, which may lead to an acrosome reaction, are discussed.