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Related Experiment Videos

Surface-active phospholipase A2 in mouse spermatozoa.

J K Thakkar, J East, D Seyler

    Biochimica Et Biophysica Acta
    |November 1, 1983
    PubMed
    Summary
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    A calcium-dependent phospholipase A2 enzyme in mouse sperm membranes is characterized. This enzyme is crucial for fertilization, showing increased activity during the acrosome reaction and requiring calcium for function.

    Area of Science:

    • Biochemistry
    • Cell Biology
    • Reproductive Biology

    Background:

    • Sperm function during fertilization involves complex membrane events.
    • Phospholipase A2 enzymes are implicated in membrane remodeling and signaling.

    Purpose of the Study:

    • To characterize a calcium-dependent phospholipase A2 (PLA2) associated with mouse sperm membranes.
    • To investigate the enzyme's localization, activity, and potential role in fertilization.

    Main Methods:

    • Enzyme activity assays using labeled substrates and sperm sonicates.
    • Centrifugation to determine enzyme localization.
    • Ionophore-induced acrosome reaction to assess activity changes.
    • Inhibition studies with various cations and pharmacological agents.

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    Main Results:

    • A membrane-associated PLA2 was identified in mouse sperm, with optimal activity at pH 8.0.
    • Enzyme activity increased 200% during the ionophore-induced acrosome reaction.
    • The enzyme requires calcium (Ca2+) and is inhibited by zinc and gold ions.
    • Lysophospholipase activity was detected, but not phospholipase A1.
    • Indomethacin, sodium meclofenamate, and mepacrine inhibited the enzyme.

    Conclusions:

    • Mouse sperm possess a membrane-bound, calcium-dependent phospholipase A2.
    • This enzyme is activated during the acrosome reaction and may be involved in membrane fusion events during fertilization.