Assessment of Progesterone-Induced Acrosome Reaction by Biotinylated Monoclonal Antibody Probes

Authors

  • Chi-Yu Gregory Lee,

    Corresponding author
    1. Andrology Laboratory, Department of Obstetrics and Gynecology, The University of British Columbia, Vancouver, BC V6T 2B5, Canada
      F107, UBC Hospital, 2211 Wesbrook Mall, Vancouver, BC, Canada V6T 2B5.
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  • Afsaneh Motamed Khorasani,

    1. Andrology Laboratory, Department of Obstetrics and Gynecology, The University of British Columbia, Vancouver, BC V6T 2B5, Canada
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  • Sonam Dorjee

    1. Andrology Laboratory, Department of Obstetrics and Gynecology, The University of British Columbia, Vancouver, BC V6T 2B5, Canada
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F107, UBC Hospital, 2211 Wesbrook Mall, Vancouver, BC, Canada V6T 2B5.

Abstract

PROBLEM: To develop an immunohistochemical assay for determination of acrosome-reacted human sperm and to study the effects of progesterone and cholesterol treatment on human sperm acrosome reaction.

METHOD OF STUDY: Three distinct anti-sperm monoclonal antibodies were biotinylated and used as probes for assessment of acrosome reaction in a 30-min immunohistochemical assay. Progesterone and/or cholesterol were added to sperm preparation to influence the acrosome reaction in different experimental conditions.

RESULTS: Percentages of acrosome-intact sperm decreased significantly during the 18-hr incubation. Acrosome reaction could be induced by progesterone as early as 2 hr after sperm incubation in human tubal fluid. The degree of progesterone-induced acrosome reaction was time dependent and the optimal effect was reached by adding 10 μg/ml progesterone for 30 min incubation. Progesterone-induced acrosome reaction was shown to be hormone-concentration dependent with 50% stimulation at 1 μg/ml. Cholesterol (1 μg/ml) was found to inhibit progesterone-induced acrosome reaction either by co-incubation with human sperm during capacitation, or by simultaneous incubation with progesterone during acrosome reaction induction.

CONCLUSIONS: Assessment of human sperm acrosomal status by avidin-biotin immunohistochemical assay can be a routine in clinical laboratories for male infertility services. Cholesterol can inhibit progesterone-induced acrosome reaction, possibly by its modifications of sperm plasma membrane and/or interference of progesterone binding to its surface receptors.

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