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An in vitro penetration assay employing zona-free hamster eggs was used to study the effects of phospholipase A2, lysophosphatidyl choline, and fatty acid on the acrosome reaction of human spermatozoa. Human spermatozoa were preincubated for 4 hr in modified Biggers, Whitten, and Whittingham's medium (mBWW) containing a specific phospholipase A2 inhibitor, p-bromophenacyl bromide (p-BPB: 1 X 10(-5) - 1 X 10(-3) M), lysophosphatidyl choline (LC: 5-500 micrograms/ml), and arachidonic acid (AA: 5-500 micrograms/ml), prior to the addition of zona-free superovulated hamster eggs. Eggs were examined microscopically 2 or 4 hr later for evidence of swelling or decondensing sperm heads in the cytoplasm. Lysophosphatidyl choline increased penetration rates of spermatozoa from 43.2% (control) to 91.4% (LC: 50 micrograms/ml). Arachidonic acid also increased penetration rates from 51.6% (control) to 87.0% (AA: 5 micrograms/ml) and 80.5% (AA: 50 micrograms/ml). p-BPB decreased penetration rates from 90.6% (1% dimethyl sulfoxide) to 16.0% (1% dimethyl sulfoxide+p-BPB 1 X 10(-4) M). These results suggest that endogenous phospholipase A2 may break membrane phosphatidyl choline into lysophosphatidyl choline and fatty acid, when the acrosome reaction occurs.

Effects of phospholipase A2, lysophosphatidyl choline, and fatty acid on the acrosome reaction of human spermatozoa.