Animal Sciences Research Center, University of Missouri, Columbia, MO 65211, USA.
National Agricultural Research Center for Kyushu Okinawa Region, National Agriculture and Food Research Organization, Koshi, Kumamoto 861-1192, Japan.
Genes (Basel). 2022 Feb 10;13(2):324. doi: 10.3390/genes13020324.
Recovery of bovine oocytes using the ovum pick-up (OPU) technique offers the advantage of rapid genetic improvement through propagation of desired genes from animals with high genetic qualities. However, the developmental competence of OPU-derived immature oocytes remains relatively poor. We previously found that cathepsin B gene expression and activity are increased in poor quality oocytes and embryos compared to good quality ones. In this study, we investigated the effect of E-64 (cathepsin B inhibitor) supplementation during in vitro maturation (IVM) on the developmental competence of OPU-derived immature oocytes and the quality of the produced blastocysts. Our results showed that supplementation of IVM medium with E-64 significantly improved the developmental competence of OPU-derived immature oocytes as evidenced by the significant increase of the blastocyst rate. Importantly, the presence of E-64 during IVM also significantly improved blastocyst quality by increasing the total cell number and decreasing the percentage of TUNEL positive cells. These results indicate that E-64 supplementation during IVM is a promising tool to improve the efficiency of OPU-IVF program by improving the developmental competence of OPU-derived immature oocytes.
使用卵母细胞抽吸 (OPU) 技术回收牛卵母细胞具有通过从具有高遗传品质的动物中传播所需基因来快速进行遗传改良的优势。然而，OPU 来源的不成熟卵母细胞的发育能力仍然相对较差。我们之前发现，与高质量的卵母细胞和胚胎相比，CATHEPSIN B 基因的表达和活性在质量较差的卵母细胞和胚胎中增加。在这项研究中，我们研究了在体外成熟 (IVM) 期间补充 E-64（CATHEPSIN B 抑制剂）对 OPU 来源的不成熟卵母细胞的发育能力和产生的囊胚质量的影响。我们的结果表明，在 IVM 培养基中补充 E-64 可显著提高 OPU 来源的不成熟卵母细胞的发育能力，囊胚率显著增加。重要的是，IVM 期间 E-64 的存在也通过增加总细胞数和降低 TUNEL 阳性细胞的百分比显著改善了囊胚质量。这些结果表明，在 IVM 期间补充 E-64 是一种很有前途的工具，可以通过提高 OPU 来源的不成熟卵母细胞的发育能力来提高 OPU-IVF 方案的效率。
