Abstract
WE have examined the incorporation of radiocarbon from [14C] glucose, [2-14C] acetate and [1 : 4-14C2] succinate into the soluble metabolic intermediates of bull semen. Samples (50 µl.) of the semen, used within 6 h of collection, were mixed with 50 µl. of a solution containing 0.1 mM adenosine triphosphate, 2 mM magnesium sulphate, 10 mM potassium chloride and 1µc. of the labelled substrate dissolved in 0.1 M potassium phosphate buffer, pH. 7.4. The mixtures were incubated aerobically at 37° C for 30 min. At the end of the incubation period the enzyme systems were inactivated by the addition of 400 µl. of boiling ethanol and the mixtures centrifuged at 500g. The radioactive substances present in the supernatants were separated by two-dimensional paper chromatography, visualized by autoradiography and the carbon-14 measured by the techniques described by Smith and Moses1.
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Smith, M. J. H., and Moses, V., Biochem. J., 76, 579 (1960).
Redenz, E., Biochem. Z., 257, 234 (1933).
Mann, T., The Biochemistry of Semen (London: Methuen and Co., Ltd., 1954).
Bhargava, P. M., Bishop, M. W. H., and Work, T. S., Biochem. J., 73, 242 (1959).
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HINES, W., SMITH, M. Intermediary Metabolism in Bull Semen. Nature 199, 179 (1963). https://doi.org/10.1038/199179a0
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DOI: https://doi.org/10.1038/199179a0
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