Increased In Vitro Binding of Fresh and Frozen–Thawed Human Sperm Exposed to a Synthetic Peptide

Abstract

Prosaposin is a well-characterized, approximately 68-kDa protein found in many tissues and as a normal component of human semen. A fragment of prosaposin apparently is involved in primary sperm-egg binding. We hypothesized that binding of sperm from some men to egg investments would be increased by in vitro exposure of their sperm to a synthetic fragment of human prosaposin (FertPlus peptide). Hence, we evaluated samples of washed fresh or frozen-thawed human sperm after a 10-minute exposure to synthetic FertPlus peptide at 0 (control), 80, 160, 320, 640, or 1280 pM, followed by 1:50 dilution for evaluation of binding. The criterion of response was mean percentage of sperm bound to a substrate prepared from chicken egg membranes after sperm were incubated for 60 minutes at 37 degrees C in substrate-coated wells of a sperm-binding assay plate. For each seminal sample, data were normalized against the percentage of sperm bound for control aliquots, providing values for relative binding. With fresh sperm, relative binding was increased (P < 0.01) by exposure of sperm to peptide, and the effect was especially obvious at 1280 pM. Higher doses were not tested. Collectively at three study sites, exposure of fresh sperm to 1280 pM peptide substantially increased (above 99% confidence interval; on the basis of duplicate control samples) percentage of sperm bound for 25 of 74 (34%) samples. For frozen-thawed sperm, exposure to 1280 pM peptide increased binding for 29 of 65 (45%) samples. We concluded that for >30% of men, exposure of their sperm to this synthetic fragment of prosaposin at 1280 pM increased binding of sperm to an egg membrane substrate similar to that offered by the zona pellucida.