Tokyo University of Agriculture, Atsugi City, Kanagawa, Japan.
Theriogenology. 2013 Mar 15;79(5):809-14. doi: 10.1016/j.theriogenology.2012.12.009. Epub 2013 Jan 22.
Antrum formation and estradiol (E2) secretion occur during early folliculogenesis. The objective was to determine the role of E2 in antrum formation of oocyte-granulosa cell complexes (OGCs) derived from porcine preantral follicles (PAFs). Supplementation of the culture medium with E2 (1 μg/mL) improved antrum formation of OGCs during 14 days of in vitro culture. Furthermore, adding 0.1 μg/mL androstenedione (a precursor of E2) to the medium also improved antrum formation. Concentration of E2 was higher in the medium of developmentally competent OGCs versus incompetent OGCs (8.5 vs. 3.5 ng/mL, P < 0.05). Fulvestrant (1 μg/mL), a competitive inhibitor of E2, completely inhibited antrum formation of OGCs that were cultured in medium containing either E2 (0.1 μg/mL) or androstenedione (0.1 μg/mL); however, increasing E2 to 1 μg/mL ameliorated the inhibitory effect. Conversely, in the case of early antral follicles, OGCs formed antrums without E2 supplementation. After E2 pretreatment, OGCs derived from PAFs formed antrums even when the OGCs were subsequently cultured in medium without E2. Furthermore, when OGCs derived from PAFs were cultured without E2 followed by an additional in vitro culture with E2, antrums were formed, albeit with the same period delay by the same pretreatment periods. In conclusion, E2 in the culture medium was indispensable for in vitro antrum formation of OGCs derived from PAFs; therefore, one of the roles of E2 is in the initiation of antrum formation.
腔的形成和雌二醇(E2)的分泌发生在卵泡早期发生。目的是确定 E2 在猪原始卵泡(PAFs)来源的卵母细胞-颗粒细胞复合体(OGCs)腔形成中的作用。在体外培养的 14 天中,培养基中添加 E2(1μg/mL)可改善 OGCs 的腔形成。此外,向培养基中添加 0.1μg/mL 雄烯二酮(E2 的前体)也可改善腔形成。在发育能力强的 OGCs 与发育能力弱的 OGCs 相比,培养基中 E2 的浓度更高(8.5 与 3.5ng/mL,P<0.05)。他莫昔芬(1μg/mL),E2 的竞争性抑制剂,完全抑制了在含有 E2(0.1μg/mL)或雄烯二酮(0.1μg/mL)的培养基中培养的 OGCs 的腔形成;然而,将 E2 增加到 1μg/mL 可改善抑制作用。相反,在早期腔卵泡的情况下,OGCs 在没有 E2 补充的情况下形成腔。E2 预处理后,即使随后在没有 E2 的培养基中培养,源自 PAF 的 OGCs 也会形成腔。此外,当源自 PAF 的 OGCs 在没有 E2 培养后再进行额外的 E2 体外培养时,也会形成腔,尽管相同的预处理期会导致相同的时期延迟。总之,培养基中的 E2 对于源自 PAF 的 OGCs 的体外腔形成是必不可少的;因此,E2 的作用之一是启动腔的形成。