Sugiyama Miyako, Kawahara-Miki Ryoka, Kawana Hirosuke, Shirasuna Koumei, Kuwayama Takehito, Iwata Hisataka
Department of Animal Science, Tokyo University of Agriculture, Kanagawa 243-0034, Japan.
J Reprod Dev. 2015;61(4):251-9. doi: 10.1262/jrd.2015-001. Epub 2015 Apr 12.
Mitochondrial numbers increase during oocyte growth. In this study, we collected oocytes and granulosa cell complexes (OGCs) from early antral follicles (EAFs) of aged cows (> 120 months of age) and examined the effects of resveratrol on mitochondrial generation, degradation, and quality in oocytes grown in vitro. We also examined the effects of resveratrol on gene expression of the granulosa cells. Resveratrol (20 µM) enhanced the expression of SIRT1 and induced autophagy in both granulosa cells and oocytes derived from aged cows. Culturing the OGCs with resveratrol increased mitochondrial DNA copy numbers in oocytes grown in vitro. Furthermore, resveratrol increased the ATP content in oocytes and improved the developmental ability of the oocytes to the blastocyst stage. Gene expression profiles in granulosa cells, as evaluated by next-generation sequencing technology, revealed that resveratrol enhanced the expression of EIF2-related genes but downregulated the expression of mammalian target of rapamycin (mTOR)-, inflammation-, and cholesterol homeostasis-related genes in granulosa cells. In conclusion, resveratrol affected both oocytes and granulosa cells derived from aged cows and improved the quality of oocytes grown in vitro through upregulation of mitochondrial biogenesis and degradation in growing oocytes and conditioning of granulosa cells.
在卵母细胞生长过程中，线粒体数量会增加。在本研究中，我们从老龄奶牛（>120月龄）的早期窦状卵泡（EAFs）中收集卵母细胞和颗粒细胞复合体（OGCs），并检测白藜芦醇对体外培养的卵母细胞中线粒体生成、降解及质量的影响。我们还检测了白藜芦醇对颗粒细胞基因表达的影响。白藜芦醇（20μM）增强了老龄奶牛来源的颗粒细胞和卵母细胞中SIRT1的表达并诱导了自噬。用白藜芦醇培养OGCs可增加体外培养卵母细胞中的线粒体DNA拷贝数。此外，白藜芦醇增加了卵母细胞中的ATP含量，并提高了卵母细胞发育至囊胚阶段的能力。通过下一代测序技术评估颗粒细胞中的基因表达谱，结果显示白藜芦醇增强了颗粒细胞中EIF2相关基因的表达，但下调了颗粒细胞中雷帕霉素靶蛋白（mTOR）、炎症及胆固醇稳态相关基因的表达。总之，白藜芦醇对老龄奶牛来源的卵母细胞和颗粒细胞均有影响，并通过上调生长中卵母细胞的线粒体生物发生和降解以及调节颗粒细胞来改善体外培养卵母细胞的质量。
