Staining of the inner acrosomal membrane of human spermatozoa with concanavalin A lectin as an indicator of potential egg penetration ability

Carol Holden, A. O. Trounson

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19 Citations (Scopus)


Objective: To determine the sensitivity and functional significance of the fluorescein isothiocyanate concanavalin A (FITC-ConA) staining method of assessment of acrosomal status. Design: Treatments were assessed for their ability to induce human sperm acrosomal loss. Penetration of zona-free human eggs by treated spermatozoa was subsequently determined. Setting: Zona-free human eggs were obtained from the Infertility Medical Centre, Richmond, Victoria, Australia. Patients, Participants: None. Interventions: None. Main Outcome Measure: Acrosomal loss of human spermatozoa was determined at 4 hours and 10 hours of treatment incubation. Penetration of zona-free human eggs was assessed 16 hours after reinsemination. Results: Human spermatozoa incubated in a strontium- or lanthanum-based medium, or T6 + 10% maternal human serum (HS) supplemented with 12 mM 8-bromo cyclic guanosine 3,5'-monophosphate and 10 mM imidazole for a 4-hour period before transfer to fresh T6 + 10% HS for a further 6 hours, demonstrated a significant increase (P < 0.05) in acrosomal loss compared with T6 + 10% HS for a total 10-hour incubation. This increase in acrosomal loss with test treatments correlated with an increase in the development of pronuclei of zona-free human eggs (r = +0.98). Conclusions: The FITC-ConA staining procedure therefore reflects biological function as assessed by the penetration of zona-free human eggs and consequently provides a further research tool for the investigation of the human sperm acrosome reaction.

Original languageEnglish
Pages (from-to)967-974
Number of pages8
JournalFertility and sterility
Issue number5
Publication statusPublished - 1 Jan 1991
Externally publishedYes

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynaecology

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