Lectin-Binding Sites on Human Sperm During Acrosome Reaction: Modifications Judged by Electron Microscopy/Flow Cytometry

Abstract

Biochemical surface modifications occur during the capacitation and acrosome reaction of human sperm and among those, variations in the expression of carbohydrates moieties. A sequential study was performed with electronic microscopy and flow cytometry techniques, where the binding of 4 lectins was assessed on normal human sperm samples during in vitro induction of the acrosome reaction with calcium ionophore A-23187. Triticum vulgaris agglutinin (WGA) was shown to bind strongly the whole surface of sperm before induction of the acrosome reaction, and in lesser amounts after incubation with calcium ionophore. Arachis hypogea agglutinin (PNA) and mostly Concanavalia ensiformis agglutinin (Con-A) and Ulex europaeus agglutinin (UEA-I) binding evolved in an opposite pattern with an increase of the labeling parallel to that of GB24 antibody binding. Electron microscopy showed that the fluorescence patterns observed correlated with increased access to the inner membrane of the acrosome. This was significant 60 min after the induction of acrosome reaction. Lectin binding could be a useful tool to examine the ability of sperm samples to undergo the acrosome reaction.

Key Words:

• acrosome
• electron microscopy
• flow cytometry
• GB24
• human sperm
• ionophore
• lectins