Universidade Federal Fluminense (UFF), Faculdade de Medicina Veterinária, Vital Brazil Filho St., 64, 24230-340, Niterói, Rio de Janeiro, Brazil.
Universidade Federal Fluminense (UFF), Faculdade de Medicina Veterinária, Vital Brazil Filho St., 64, 24230-340, Niterói, Rio de Janeiro, Brazil.
Theriogenology. 2024 Jan 15;214:33-42. doi: 10.1016/j.theriogenology.2023.10.001. Epub 2023 Oct 5.
This study investigated the time course of lipid accumulation during IVM and assessed the role of lipid modulators added during IVM on lipid content, nuclear maturation, oxidative stress, mitochondrial activity, gene expression, and cryosurvival of cat oocytes. First, the lipid content of immature COCs was compared to those subjected to different IVM duration times (24, 28, and 32 h). Then, the lipid content was investigated after the use of different lipid modulators [conjugated linoleic acid (CLA), forskolin (FSK), l-carnitine (LC)]. Subsequently, both the CONTROL group and MIX 18 (CLA+FSK+LC) were compared regarding nuclear maturation, mitochondrial activity, reactive oxygen 19 species (ROS), and glutathione (GSH) levels, to the expression of SDHA, GDF9, BMP15, ZAR-1, 20 PRDX1, SIRT1, and SIRT3 genes (normalized by ACTB and YWHAZ genes); and to vitrification and 21 post-warming viability assessment. When not using any lipid modulator, an increase (P < 0.05) in lipid content could be observed after 28 h of IVM. The MIX group showed the greatest (P < 0.05) reduction in oocyte lipid content after 28 h of IVM. No difference (P > 0.05) was observed in the MII rate in the CONTROL (45%) and MIX (41%) groups and in mitochondrial activity ((1.00 ± 0.35 A U vs 1.19 ± 0.14 A U). Although ROS and GSH levels were higher (P < 0.05) in MIX than in CONTROL, the redox balance (ROS/GSH) was greater (P < 0.05) in the latter (C:1.00 ± 0.20 vs M:0.26 ± 0.06 A.U). The GDF9, HSP70, PRDX1, and SIRT1 transcripts were downregulated (P < 0.05) in MIX-oocytes, compared to the CONTROL. After vitrification, MIX (74%) presented a higher (P < 0.05) viability compared to control (53%). In conclusion, MIX can reduce the total lipid content and improve viability after cryopreservation, however, it seems to affect the oocyte metabolism in a way that still needs to be better understood in the cat biological model.
本研究调查了 IVM 过程中脂质积累的时间过程,并评估了在 IVM 期间添加脂质调节剂对卵母细胞脂质含量、核成熟、氧化应激、线粒体活性、基因表达和冷冻保存的影响。首先,比较了不成熟 COC 的脂质含量与经历不同 IVM 持续时间(24、28 和 32 小时)的 COC 的脂质含量。然后,研究了使用不同脂质调节剂[共轭亚油酸(CLA)、福司可林(FSK)、左旋肉碱(LC)]后的脂质含量。随后,将 CONTROL 组和 MIX 18(CLA+FSK+LC)进行比较,以评估核成熟度、线粒体活性、活性氧 19 种(ROS)和谷胱甘肽(GSH)水平、SDHA、GDF9、BMP15、ZAR-1、20 PRDX1、SIRT1 和 SIRT3 基因的表达(由 ACTB 和 YWHAZ 基因标准化);以及玻璃化和 21 次解冻后活力评估。当不使用任何脂质调节剂时,在 IVM 28 小时后可以观察到脂质含量增加(P<0.05)。MIX 组在 IVM 28 小时后表现出最大(P<0.05)的卵母细胞脂质含量降低。CONTROL(45%)和 MIX(41%)组的 MII 率和线粒体活性无差异(P>0.05)((1.00±0.35 A.U vs 1.19±0.14 A.U)。尽管 MIX 中的 ROS 和 GSH 水平较高(P<0.05),但后者的氧化还原平衡(ROS/GSH)更高(P<0.05)(C:1.00±0.20 vs M:0.26±0.06 A.U)。与 CONTROL 相比,MIX 卵母细胞中的 GDF9、HSP70、PRDX1 和 SIRT1 转录本下调(P<0.05)。玻璃化后,MIX(74%)的活力高于 CONTROL(53%)(P<0.05)。总之,MIX 可以降低总脂质含量并提高冷冻保存后的活力,但它似乎以一种仍需要在猫生物学模型中更好理解的方式影响卵母细胞代谢。