Biology of Reproduction, Vol 13, 389-396, Copyright © 1975 by Society for the Study of Reproduction

Use of High Speed Dialysis to Prepare Bovine Sperm for MetaboliC Studies

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

Biochemical characteristics were compared for bovine sperm prepared by (a) simple dilution of semen with buffer, (b) dilution followed by centrifugation at 300g for 10 min and resuspension (no washing), (c) dilution followed by centrifugation and washing, and (d) dilution followed by high speed dialysis using a Dow Bio-Fiber 50 dialyzer. This sperm preparation has all high molecular weight (>5,000 daltons) components of semen, including the free floating cytoplasmic droplets. Parameters determined were sperm motility, intracellular ATP, ADP and AMP levels, energy charge, glucose consumption, pyruvate oxidation, total oxygen consumption and estimated endogenous oxidation rate in the presence of exogenous glucose. No difference in motility, adenine nucleotide levels or energy charge was noted when sperm prepared by each treatment were compared. All metabolic parameters of sperm prepared by methods b and d were identical to those for treatment a. Sperm prepared by treatment c use less endogenous substrate and were more dependent on consumption of exogenous glucose to satisfy energy requirements than sperm prepared by procedure d. Thus, high speed dialysis is a useful method of preparing sperm for metabolic studies which minimizes alteration of cellular function.