The unfertilized eggs (UFE) of the solitary ascidian, Halocynthia roretzi, which are released naturally, are strictly self-sterile. However, ovarian eggs isolated after spawning, which are expected to develop into UFE on the following day, are self-fertile. Some exogenous proteases-trypsin, chymotrypsin, papain and elastase-induced self-sterility in the self-fertile ovarian eggs within an hour in vitro. The establishment of self-sterility by the exogenous protease did not require the synthesis of new protein, or the participation of follicle cells. Some of the ovarian eggs were able to differentiate into self-sterile eggs spontaneously in vitro. The protein synthesis inhibitors puromycin and cycloheximide had no effect on the spontaneous establishment of self-sterility. However, several protease inhibitors such as leupeptin, soybean trypsin inhibitor (SBTI) and antipain, did inhibit the spontaneous establishment of self-sterility. The possible participation of trypsin-like protease in the establishment of self-sterility in the ovary is discussed. Mol. Reprod. Dev. 52:99–106, 1999.
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