Faculty of Veterinary Medicine, Laboratory of Manipulation of Oocytes and Preantral Follicles (Lamofopa), State University of Ceara, Fortaleza, Ceará, Brazil.
Faculty of Veterinary Medicine, Laboratory of Ruminant Production and Nutrition (Lanuprumi), State University of Ceará, Fortaleza, Ceará, Brazil.
Mol Reprod Dev. 2020 Sep;87(9):966-977. doi: 10.1002/mrd.23410. Epub 2020 Aug 6.
This study aimed to evaluate the role of anethole during the in vitro culture of caprine early antral follicles. Early antral follicles were isolated from caprine ovaries and cultured for 18 days without (control) or with anethole (300 µg/ml). After culture, the cumulus-oocyte complexes were subjected to in vitro maturation, followed by parthenogenetic activation or in vitro fertilization (IVF) and embryo culture. Follicular walls were used for the quantification of messenger RNA (mRNA) of CYP19A1, CYP17, MMP-9, TIMP-2, Bax, and Bcl-2 genes, and culture medium was used for evaluation of ferric reducing antioxidant power (FRAP) and estradiol levels. After in vitro follicle culture (IVFC), anethole induced higher total antioxidant capacity, that is, it produced higher FRAP levels, reduced the Bax/Bcl-2 ratio, and increased the levels of mRNA for CYP19A1 and CYP17, which was associated with a greater estradiol production (p < .05). Also, anethole improved the ability of oocytes to resume meiosis and reach metaphase II stage, as well as yielded higher (p < .05) embryo production (e.g., morulas and blastocysts) in both parthenogenetic activation and IVF techniques. One pregnancy (Day 30) was obtained from IVFC with anethole. In conclusion, anethole promoted in vitro growth and maturation of goat early antral follicles and oocytes and enabled embryo production. Furthermore, this study reports, for the first time in goats, a pregnancy after IVF using oocytes originated from early antral follicles grown in vitro.
本研究旨在评估茴香脑在山羊早期腔前卵泡体外培养中的作用。早期腔前卵泡从山羊卵巢中分离出来，在无（对照）或有茴香脑（300μg/ml）的情况下培养 18 天。培养后，卵丘-卵母细胞复合物进行体外成熟，然后进行孤雌激活或体外受精（IVF）和胚胎培养。卵泡壁用于检测 CYP19A1、CYP17、MMP-9、TIMP-2、Bax 和 Bcl-2 基因的信使 RNA（mRNA），培养介质用于评估铁还原抗氧化能力（FRAP）和雌二醇水平。在体外卵泡培养（IVFC）后，茴香脑诱导更高的总抗氧化能力，即产生更高的 FRAP 水平，降低 Bax/Bcl-2 比值，并增加 CYP19A1 和 CYP17 的 mRNA 水平，这与更高的雌二醇产生有关（p<.05）。此外，茴香脑提高了卵母细胞恢复减数分裂并达到中期 II 阶段的能力，并在孤雌激活和 IVF 技术中产生了更高的（p<.05）胚胎产量（例如，桑葚胚和囊胚）。通过 IVFC 获得了一例妊娠（第 30 天）。综上所述，茴香脑促进了山羊早期腔前卵泡和卵母细胞的体外生长和成熟，并实现了胚胎的产生。此外，本研究首次在山羊中报道了使用体外培养的早期腔前卵泡来源的卵母细胞进行 IVF 后的妊娠。
