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Abstract：
Cumulus cell (CC) expansion is pivotal for oocyte maturation, during which CCs release factors that initiate paracrine signaling within the follicular fluid (FF). The FF is abundant in extracellular vesicles (EVs) that facilitate intercellular communication. Although bovine and murine EVs can control cumulus expansion, these effects have not been observed in equines. This study aimed to assess the impact of FF-derived EVs (ffEVs) on equine CC expansion, viability, and transcriptome. Cumulus-oocyte complexes (COCs) that underwent in vitro maturation (IVM) in the presence (200 µg protein/mL) or absence (control) of ffEVs were assessed for cumulus expansion and viability. CCs were isolated after 12 h of IVM, followed by RNA extraction, cDNA library generation, and subsequent transcriptome analysis using next-generation sequencing. Confocal microscopy images illustrated the internalization of labeled ffEVs by CCs. Supplementation with ffEVs significantly enhanced cumulus expansion in both compacted (Cp, p < 0.0001) and expanded (Ex, p < 0.05) COCs, while viability increased in Cp groups (p < 0.01), but decreased in Ex groups (p < 0.05), compared to the controls. Although transcriptome analysis revealed a subtle effect on CC RNA profiles, differentially expressed genes encompassed processes (e.g., MAPK and Wnt signaling) potentially crucial for cumulus properties and, consequently, oocyte maturation.
摘要：
卵丘细胞（CC）扩增是卵母细胞成熟的关键，在此期间，CCs释放启动卵泡液（FF）内旁分泌信号传导的因子。FF在促进细胞间通讯的细胞外囊泡(EV)中是丰富的。尽管牛和鼠电动汽车可以控制积云扩张，这些影响在马科动物中没有观察到。本研究旨在评估FF衍生电动汽车(ffEV)对马CC扩张的影响，生存能力，和转录组。在存在（200µg蛋白质/mL）或不存在（对照）的情况下进行体外成熟（IVM）的卵丘-卵母细胞复合物（COCs）评估了卵丘的扩张和生存力。在IVM12小时后分离CC，然后进行RNA提取,cDNA文库的产生，和随后使用下一代测序的转录组分析。共聚焦显微镜图像说明了CC对标记的ffEV的内化。补充ffEV显着增强了两个压实的积云扩张(Cp，p<0.0001)和扩展(Ex，p<0.05)COCs，而Cp组的活力增加（p<0.01），但Ex组下降(p<0.05)，与对照组相比。尽管转录组分析揭示了对CCRNA谱的微妙影响，差异表达的基因包括过程(例如，MAPK和Wnt信号）可能对积云特性至关重要，因此,卵母细胞成熟。
