Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, Ceará, Brazil.
Postgraduate Program in Veterinary Sciences, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, Ceará, Brazil.
Mol Reprod Dev. 2023 Dec;90(12):810-823. doi: 10.1002/mrd.23700. Epub 2023 Sep 6.
This study assessed the histones methylation profile (H3K4me3 and H3K9me3) in late preantral (PA) and early antral (EA) caprine follicles grown in vivo and in vitro, and the anethole effect during in vitro culture of PA follicles. Uncultured in vivo-grown follicles (PA, n = 64; EA, n = 73) were used as controls to assess the methylation profile and genes' expression related to apoptosis cascade (BAX, proapoptotic; BCL2, antiapoptotic), steroidogenesis (CYP17, CYP19A1), and demethylation (KDM1AX1, KDM1AX2, KDM3A). The isolated PA follicles (n = 174) were cultured in vitro for 6 days in α-MEM in either absence (control) or presence of anethole. After culture, EA follicles were evaluated for methylation, mRNA abundance, and morphometry. Follicle diameter increased after culture, regardless of treatment. The methylation profile and the mRNA abundance were similar between in vivo-grown PA and EA follicles. Anethole treatment led to higher H3K4me3 fluorescence intensity in EA follicles. The mRNA abundances of BAX, CYP17, and CYP19A1 were higher, and BCL2 and KDM3A were lower in in vitro-grown EA follicles than in vivo-grown follicles. In conclusion, in vitro follicle culture affected H3K4me3 fluorescence intensity, mRNA abundance of apoptotic genes, and steroidogenic and demethylase enzymes compared with in vivo-grown follicles.
本研究评估了体内和体外培养的晚期前腔卵泡（PA）和早期腔卵泡（EA）的组蛋白甲基化谱（H3K4me3 和 H3K9me3），以及茴香脑在 PA 卵泡体外培养中的作用。未培养的体内生长的卵泡（PA，n=64；EA，n=73）被用作对照，以评估与凋亡级联（BAX，促凋亡；BCL2，抗凋亡）、类固醇生成（CYP17、CYP19A1）和去甲基化（KDM1AX1、KDM1AX2、KDM3A）相关的甲基化谱和基因表达。分离的 PA 卵泡（n=174）在 α-MEM 中体外培养 6 天，要么在无处理（对照）的情况下，要么在茴香脑存在的情况下培养。培养后，评估 EA 卵泡的甲基化、mRNA 丰度和形态计量学。无论处理与否，卵泡直径在培养后均增加。体内生长的 PA 和 EA 卵泡的甲基化谱和 mRNA 丰度相似。茴香脑处理导致 EA 卵泡中的 H3K4me3 荧光强度增加。与体内生长的卵泡相比，体外生长的 EA 卵泡中 BAX、CYP17 和 CYP19A1 的 mRNA 丰度更高，BCL2 和 KDM3A 的 mRNA 丰度更低。总之，与体内生长的卵泡相比，体外卵泡培养影响 H3K4me3 荧光强度、凋亡基因以及类固醇生成和去甲基化酶的 mRNA 丰度。
