Wang Ting, Na Jie
Center for Stem Cell Biology and Regenerative Medicine, School of Medicine, Tsinghua University, Beijing, China.
Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing, China.
Front Cell Dev Biol. 2021 Apr 15;9:648331. doi: 10.3389/fcell.2021.648331. eCollection 2021.
The nucleolus undergoes significant functional changes and plays important roles during mammalian oocyte meiotic maturation. Fibrillarin (FBL) is the component of nucleolar small nuclear ribonucleoprotein (snRNP) particle and localizes to the dense fibrillar component (DFC) of the nucleolus. We found that FBL-GFP displays an uneven and cloudy localization in the nucleolus of non-surrounded nucleolus (NSN) oocytes, while it distributes evenly and to a few bright dots in the surrounded nucleolus (SN) oocytes. Accordingly, NSN oocytes showed active nascent RNA transcription, while the SN group was transcriptionally quiescent. NSN geminal vesicles also contained more DNA damage marker γH2AX foci. Based on different FBL-GFP patterns in live oocytes, the ones with superior meiotic maturation potential can be identified. Global transcriptome profiling revealed a significant difference in single SN and NSN oocytes. Thus, FBL-GFP can serve as a marker for nucleolus activity, which also correlates with transcription activity and the quality of oocytes.
核仁在哺乳动物卵母细胞减数分裂成熟过程中经历显著的功能变化并发挥重要作用。纤维蛋白原(FBL)是核仁小核糖核蛋白(snRNP)颗粒的组成部分,定位于核仁的致密纤维成分(DFC)。我们发现,FBL-GFP在非包围核仁(NSN)卵母细胞的核仁中呈现不均匀且模糊的定位,而在包围核仁(SN)卵母细胞中则均匀分布并形成一些亮点。相应地,NSN卵母细胞显示出活跃的新生RNA转录,而SN组转录静止。NSN生发泡也含有更多的DNA损伤标记γH2AX焦点。基于活卵母细胞中不同的FBL-GFP模式,可以识别出具有更高减数分裂成熟潜力的卵母细胞。全局转录组分析揭示了单个SN和NSN卵母细胞之间存在显著差异。因此,FBL-GFP可作为核仁活性的标志物,这也与转录活性和卵母细胞质量相关。