Matsuyama Shuichi, Nakamura Sho, Minabe Shiori, Sakatani Miki, Takenouchi Naoki, Sasaki Takuya, Inoue Yuki, Iwata Hisataka, Kimura Koji
Division of Animal Feeding and Management Research, Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization (NARO), Nasushiobara, Japan.
Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan.
Mol Reprod Dev. 2023 Mar;90(3):141-152. doi: 10.1002/mrd.23670. Epub 2023 Jan 16.
To investigate possible causes of reproductive failure, we conducted global endometrial gene expression analyses in fertile and subfertile cows. Ingenuity pathway analysis showed that RICTOR and SIRT3 are significant upstream regulators for highly expressed genes in fertile cows, and are predicted to be activated upstream regulators of normal mitochondrial respiration. Canonical pathway analysis revealed that these highly expressed genes are involved in the activation of mitochondrial oxidative phosphorylation. Therefore, in subfertile cows, the inactivation of RICTOR and SIRT3 may correlate with decreased capacity of mitochondrial respiration. Furthermore, the expression levels of most mitochondrial DNA genes and nuclear genes encoding mitochondrial proteins were higher in subfertile cows. The mitochondrial DNA copy number was significantly higher in the endometrium of subfertile cows, whereas the ATP content did not differ between fertile and subfertile cows. Quantitative reverse transcription-PCR analysis demonstrated that the expression of PGC1a, TFAM, MFN1, FIS1, and BCL2L13 were significantly lower in subfertile cows. In addition, transmission electron microscopy images showed mitochondrial swelling in the endometrial cells of the subfertile cow. These results suggest that poor-quality mitochondria accumulate in the endometrium owing to a reduced capacity for mitochondrial biogenesis, fusion, fission, and degradation in subfertile cows, and may contribute to infertility.
为了研究生殖失败的可能原因，我们对可育和不育奶牛的子宫内膜进行了全基因组表达分析。 Ingenuity通路分析表明，RICTOR和SIRT3是可育奶牛中高表达基因的重要上游调节因子，预计是正常线粒体呼吸的上游激活调节因子。 经典通路分析显示，这些高表达基因参与线粒体氧化磷酸化的激活。 因此，在不育奶牛中，RICTOR和SIRT3的失活可能与线粒体呼吸能力下降有关。 此外，大多数线粒体DNA基因和编码线粒体蛋白的核基因的表达水平在不育奶牛中较高。 不育奶牛子宫内膜中的线粒体DNA拷贝数显著更高，而可育和不育奶牛之间的ATP含量没有差异。 定量逆转录-PCR分析表明，不育奶牛中PGC1a、TFAM、MFN1、FIS1和BCL2L13的表达显著降低。 此外，透射电子显微镜图像显示不育奶牛子宫内膜细胞中的线粒体肿胀。 这些结果表明，由于不育奶牛中线粒体生物发生、融合、裂变和降解能力降低，低质量的线粒体在子宫内膜中积累，可能导致不育。
