Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan.
Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan.
Theriogenology. 2024 Oct 15;228:30-36. doi: 10.1016/j.theriogenology.2024.07.021. Epub 2024 Jul 25.
Objectives of the current study were to examine the effects of exogenous expression of PGC-1α, which is a transcription factor responsive for controlling mitochondrial DNA (mtDNA) replication, mitochondria quantity control, mitochondrial biogenesis, and reactive oxygen species (ROS) maintenance, in porcine oocytes during in-vitro maturation (IVM) on the developmental competence, as well as mitochondrial quantity and function. Exogenous over-expression of PGC-1α by injection of the mRNA construct into oocytes 20 h after the start of IVM culture significantly increased the copy number of mtDNA in the oocytes, but reduced the incidences of oocytes matured to the metaphase-II stage after the IVM culture for totally 44 h and completely suppressed the early development in vitro to the blastocyst stage following parthenogenetic activation. The exogenous expression of PGC-1α also significantly induced spindle defects and chromosome misalignments. Furthermore, markedly higher ROS levels were observed in the PGC-1α-overexpressed mature oocytes, whereas mRNA level of SOD1, encoded for a ROS scavenging enzyme, was decreased. These results conclude that forced expression of PGC-1α successfully increase mtDNA copy number but led to increased ROS production, evidently by downregulation of SOD1 gene expression, inducement of spindle aberration/chromosomal misalignment, and consequently reduction in the meiotic and developmental competences of porcine oocytes.
本研究的目的是检验在猪卵母细胞体外成熟(IVM)期间,过表达转录因子 PGC-1α(其可控制线粒体 DNA(mtDNA)复制、线粒体数量控制、线粒体生物发生和活性氧(ROS)的维持)对卵母细胞的发育能力以及线粒体数量和功能的影响。在 IVM 培养开始后 20 小时,通过注射 mRNA 构建体将 PGC-1α 外源性过表达到卵母细胞中,显著增加了卵母细胞中 mtDNA 的拷贝数,但降低了 IVM 培养 44 小时后成熟为中期 II 阶段的卵母细胞的发生率,并完全抑制了孤雌激活后的早期体外发育至囊胚阶段。PGC-1α 的外源表达也显著诱导了纺锤体缺陷和染色体排列不齐。此外,在过表达 PGC-1α 的成熟卵母细胞中观察到明显更高的 ROS 水平,而编码 ROS 清除酶的 SOD1 mRNA 水平降低。这些结果表明,强制表达 PGC-1α 成功增加了 mtDNA 拷贝数,但导致 ROS 产生增加,显然是通过下调 SOD1 基因表达、诱导纺锤体异常/染色体排列不齐,从而降低了猪卵母细胞的减数分裂和发育能力。