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In order to demonstrate whether ovoperoxidase hardens the zona of oocytes activated by incubating in M-S buffer supplemented with 20muM of Ca-ionophore A 23187, the effect of peroxidase inhibitors(250mM pheylhydrazine, 28mM sodilum sulfite, 350mM glycine ethyl ester and 50mM sodium azide), tyrosine analogue(12.5mM tyramine) and exogeneous peroxidase(50mutextrm{g}/ml horseradishperoxidase ; HRP) on zona hardening in ionorphore-treated oocytes were investigated. The results obtained from thses experiments were summarized as follows : 1. The zona solubility (t50) of ionophore-activated and DMSO-treated oocytes at 1, 2 and 3 hr of culture were 25.0, 31.6 and 40.6min., and 9.7, 10.8 and 15.5 min., respectively. The longest time required for zona lysis of ionophore activated oocytes at 1 hr after onset of ionophore treatment. The diferences int50 for zona was significantly greater as compared to DMSO-treated controls(P<0.01). 2. The inhibition rates of hardening in the oocytes treated with the phenylhydrazine, sodium sulfite, glycine ethyl ester and sodium azide, were 23.8, 61.9, 95.2 and 23.8%l, respectively, and the tyramine, was 14.3%. Several known peroxidase inhibitors and tyrosine analogue were blocked zona hardening in ionophore activated oocytes. 3. The treatment of exogeneous peroxidase promoted the zona hardening of activated oocytes but not unactivated oocytes. These resuls indicate that the ovoperoxidase apparently catalyzes the hardening of the zona following ionophore activation of mouse oocytes.
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