The ovaries of 501 female eastern Atlantic bluefin tuna (Thunnus thynnus Linnaeus, 1758) captured in the Mediterranean Sea from May to September between 1998 and 2004 were analysed histologically. Body size at median sexual maturity (L-50) was 103.6 cm, fork length (FL), while 100% maturity was reached above 135 cm FL. The age analysis, based on the count of the translucent zones of the first spiniform ray of the first dorsal fin, showed that most of the specimens with FL = L-50 were 3 years old while 100% maturity was reached between 4 to 5 years. The reported evidence indicates that for the eastern Atlantic bluefin tuna stock, the size and age of first sexual maturity of females was lower than in the western Atlantic stock.\u
The histological analysis of eastern Atlantic bluefin tuna Thunnus thynnus ovaries caught from February to September 1999-2000, made it possible to distinguish the presence of seven oocyte developmental stages and allowed the characterization of six time-dependent ovary maturity stages. The ovaries of mature (fork length, L F ≥ 110 cm) bluefin tuna were non-active from August (spent period) to March (quiescent period) when they contained only perinucleolarstage oocytes. Ovary development started in April to early May (recrudescent period) with the appearance of oocytes at the lipid stage. Vitellogenesis appeared in mid-May (ripening period) and post-vitellogenesis occurred in late May to mid-June (pre-spawning period). In late June to early July, hydrated oocytes, a sign of imminent spawning, were found only in specimens caught in Balearic waters. Females ranging between 100 and 110 cm L F, captured during the recrudescent and ripening periods, had the largest oocytes at the lipid stage, most of which were degenerating. An extensive vitellogenic atresia was observed in the ovaries of five females caught during the spawning period in non-spawning areas
Oviductal environment affects preparation of gametes for fertilization, fertilization itself, and subsequent embryonic development. The aim of this study was to evaluate the effect of oviductal fluid and the possible involvement of deleted in malignant brain tumor 1 (DMBT1) on IVF in porcine and equine species that represent divergent IVF models. We first performed IVF after pre-incubation of oocytes with or without oviductal fluid supplemented or not with antibodies directed against DMBT1. We showed that oviductal fluid induces an increase in the monospermic fertilization rate and that this effect is canceled by the addition of antibodies, in both porcine and equine species. Moreover, pre-incubation of oocytes with recombinant DMBT1 induces an increase in the monospermic fertilization rate in the pig, confirming an involvement of DMBT1 in the fertilization process. The presence of DMBT1 in the oviduct at different stages of the estrus cycle was shown by western blot and confirmed by immunohistochemical analysis of ampulla and isthmus regions. The presence of DMBT1 in cumulus-oocyte complexes was shown by western blot analysis, and the localization of DMBT1 in the zona pellucida and cytoplasm of equine and porcine oocytes was observed using immunofluorescence analysis and confocal microscopy. Moreover, we showed an interaction between DMBT1 and porcine spermatozoa using surface plasmon resonance studies. Finally, a bioinformatic and phylogenetic analysis allowed us to identify the DMBT1 protein as well as a DMBT1-like protein in several mammals. Our results strongly suggest an important role of DMBT1 in the process of fertilization.
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