By Jerry L. Hedrick (auth.), Jerry L. Hedrick (eds.)
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Extra info for The Molecular and Cellular Biology of Fertilization
Other mice, similarly immunized were mated continuously until obviously pregnant at which time the males were removed (11,13). 5 x 106/ml epididymal sperm from ZBW F1 males. Fertilization was determined at 5-6 h by the presence of two pronuclei and two polar bodies (11). To titrate the effect of monoclonal antibodies on fertilization, eggs in cumulus mass were incubated for 15 min with increasing concentrations of antibody (0-90 l~/ml) and washed three times in T6 medium before the addition of sperm.
30 A. MONROY This is most clearly shown by the futile heat production associated with a massive ATP breakdown and uncoupling of the sperm oxidative phosphorylation. Thus the so-called activation of the spermatozoon is actually a switching off of its metabolic machinery. THE ACTIVATION OF THE EGG The next question that I would like to address is whether the activation of the egg requires fusion with the spermatozoon or whether the processes leading to activation are started before actual fusion takes place.
This could then be split by a reducing agent into Fab1 fragments. The latter could then be oxidized to permanent Fab1 univalent fragments or combined with reduced Fab1 fragments with different specificity to produce "hybrid" bivalent, Fc-free antibodies (19). With this simple technology available we first showed (59,60) that treatment with Fab antisperm antibody rendered Arbacia punctulata sperm incapable of agglutinating with egg water and incapable of fertilizing eggs. It is of special interest that certain of the Fab antisera inhibited the fertilizing capacity of the sperm but did not inhibit fertilizin agglutination of the sperm.