Abstract
TEST (TES (N-tris[hydroxymethyl]methyl-2-aminoethanesulfonic acid) and Tris) yolk buffer (TYB) has recently been shown to improve the binding capacity of spermatozoa to zona pellucidae. The present study had two objectives: 1) to elucidate which component(s) of TYB dominates this effect and 2) to define the responsible mechanism. Sperm samples obtained from subfertile men were incubated for 2 h in either TYB or media containing egg yolk lipoproteins or phospholipids. After incubation, sperm binding was tested by the hemizona assay. Yolk lipoprotein-treated spermatozoa bound hemizonae with efficiency equal to that of the spermatozoa incubated in control medium. Conversely, incubation of spermatozoa in media containing either TYB, yolk-phospholipids, or pure phosphatidylcholine (PC) resulted in a 2- to 3-fold increased binding capacity (p < 0.01). A close correlation was found between the effect of yolk-phospholipids and TYB on the binding capacity of the same sperm samples, compared to spermatozoa incubated in control medium. Incubation of spermatozoa in yolk phospholipid medium caused a dose-dependent increase of sperm binding capacity (p < 0.05). Treatment of sperm samples with 1 mg/ml or more of purified PC preparation also resulted in a reduction of the sperm cholesterol:phospholipid molar ratio. Significant correlations between the effects of the treatments on sperm cholesterol: phospholipid molar ratio and sperm binding were obtained with yolk-phospholipids (r = -0.55) or 1 mg/ml purified PC (r = -0.61). We conclude that 1) the enhanced binding capacity of human spermatozoa following TYB treatment is probably due to yolk-phospholipids, mainly egg yolk PC; and 2) it appears that the enhanced binding capacity of human spermatozoa following treatment with egg yolk-containing media may be a result of the reduction of the cholesterol:phospholipid molar ratio in the sperm cells.
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