Biology of Reproduction, Vol 12, 545-551, Copyright © 1975 by Society for the Study of Reproduction

Tritium Release from [2-³H]D-Glucose as a Monitor of Glucose Consumption by Bovine Sperm

¹ Pennsylvania State University, Department of Biochemistry, University Park, Pennsylvania 16802

The rate of ³HOH release from [2-³H]glucose was shown to be a valid measure of the rate of glucose consumption by testicular and ejaculated bovine sperm. The sensitivity of the tritium release method permits precise estimation of glucose consumption rates under conditions where only a small fraction of the total glucose is consumed whereas the classical glucose oxidase method is rather imprecise under these conditions. Ejaculated sperm contain sufficient quantities of hexokinase, phosphoglucoisomerase and aldolase to account for the observed whole cell glycolytic rates. The enzymes phosphoglucomutase, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase were not detected in testicular or ejaculated bovine sperm. Thus, no correction for pentose cycle activity is required and the rate of glucose consumption is identical with flow of glucose through the Embden-Meyerhof glycolytic pathway. Rates of glucose consumption and tritium release were not identical. A molar ratio for glucose consumed to ³HOH released of 2.25 was determined for bovine sperm cells over a wide range of glucose consumption rates. Isotope discrimination and extent of tritium exchange with the medium account for this difference.

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