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Polyadenylation of tgo3�� mRNA occurred at the time TGVBD50?=?58.9?��?8.4 (n?=?3), coincident with that of cyclin B1 mRNA ( Figs. 6B, C), suggesting that the tgo3�� mRNA is translationally activated at a time similar to that of cyclin B1 mRNA. In contrast, polyadenylation of tg3�� mRNA was detected at the time TGVBD50?=?18.1?��?6.4 (n?=?3, p?BMS777607 oocytes 4?h after injection ( Fig.?7A and Table?2), in striking contrast to the tgo3�� mRNAs in transgenic oocytes, which localized to the animal pole ( Fig.?2B and Table?1). Dextran with a relative molecular mass equivalent to that of tgo3�� mRNA showed a similar distribution pattern after injection into oocytes ( Fig.?7B and Table?2), suggesting that the injected mRNA was not directed to localize; rather, it was passively diffused after injection. Translation of tgo3�� mRNA injected into the oocyte cytoplasm was initiated at timing much earlier than that of tgo3�� mRNA transcribed Evodiamine in the nucleus and transferred to the cytoplasm in transgenic zebrafish oocytes ( Fig.?5G). Among 4 oocytes Epigenetics Compound Library supplier examined, one was activated before MIH stimulation and the others were activated precociously after MIH stimulation ( Fig.?7C). These results clearly indicate that the translation of mRNAs produced in vitro and injected into oocytes was not precisely regulated in the oocytes. Therefore, nuclear events, such as transcription and splicing, are necessary to drive the specific localization in oocytes and to regulate the accurate timing of translational activation of cyclin B1 mRNA. In this study, we distinguished the role of cyclin B1 ORF from that of 3�� UTR in localization and translational control of the mRNA by using genetic analysis in combination with real-time imaging of the translation site and timing of temporally controlled mRNA in living cells. Our results demonstrated the following roles of the 3�� UTR: 1) directing mRNA localization to the animal pole of oocytes ( Fig.?3), 2) mediating translational repression in immature oocytes ( Fig.?4), and 3) promoting translational activation and cytoplasmic polyadenylation during oocyte maturation ( Fig.?4?and?Fig.?6). Furthermore, we demonstrated unexpected roles of the cyclin B1 ORF in anchoring the mRNA on the animal polar cytoplasm of full-grown oocytes ( Fig.?2?and?Fig.