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Abstract：
In reproductive technologies, uncovering the molecular aspects of oocyte and embryo competence under different conditions is crucial for refining protocols and enhancing efficiency. RNA-seq generates high-throughput data and provides transcriptomes that can undergo additional computational analyses. This study presented the transcriptomic profiles of in vitro matured oocytes and blastocysts produced in vitro from buffalo crossbred (Bubalus bubalis), coupled with gene co-expression and module preservation analysis. Cumulus Oophorus Complexes, obtained from slaughterhouse-derived ovaries, were subjected to in vitro maturation to yield metaphase II oocytes (616) or followed in vitro fertilization and culture to yield blastocysts for sequencing (526). Oocyte maturation (72%, ±3.34 sd) and embryo development (21.3%, ±4.18 sd) rates were obtained from three in vitro embryo production routines following standard protocols. Sequencing of 410 metaphase II oocytes and 70 hatched blastocysts (grade 1 and 2) identified a total of 13,976 genes, with 62% being ubiquitously expressed (8,649). Among them, the differentially expressed genes (4,153) and the strongly variable genes with the higher expression (fold-change above 11) were highlighted in oocytes (BMP15, UCHL1, WEE1, NLRPs, KPNA7, ZP2, and ZP4) and blastocysts (APOA1, KRT18, ANXA2, S100A14, SLC34A2, PRSS8 and ANXA2) as representative indicators of molecular quality. Additionally, genes exclusively found in oocytes (224) and blastocysts (2,200) with specific biological functions were identified. Gene co-expression network and module preservation analysis revealed strong preservation of functional modules related to exosome components, steroid metabolism, cell proliferation, and morphogenesis. However, cell cycle and amino acid transport modules exhibited weak preservation, which may reflect differences in embryo development kinetics and the activation of cell signaling pathways between buffalo and bovine. This comprehensive transcriptomic profile serves as a valuable resource for assessing the molecular quality of buffalo oocytes and embryos in future in vitro embryo production assays.
摘要：
在生殖技术中，揭示不同条件下卵母细胞和胚胎能力的分子方面对于完善方案和提高效率至关重要。RNA-seq产生高通量数据，并提供可以进行额外计算分析的转录组。这项研究介绍了从水牛杂交（Bubalusbubalis）体外产生的体外成熟卵母细胞和胚泡的转录组学概况，结合基因共表达和模块保存分析。Oophorus积云复合物，从屠宰场衍生的卵巢获得，进行体外成熟以产生中期II卵母细胞(616)或随后进行体外受精和培养以产生用于测序的胚泡(526)。卵母细胞成熟（72%，±3.34sd)和胚胎发育(21.3%，按照标准方案，从三个体外胚胎生产程序中获得±4.18sd）速率。对410个中期II卵母细胞和70个孵化的囊胚（1级和2级）进行测序，共鉴定出13,976个基因，62%普遍表达(8,649)。其中,在卵母细胞中突出显示了差异表达基因（4,153）和具有较高表达的强可变基因（倍数变化高于11）（BMP15，UCHL1，WEE1，NLRP，KPNA7，ZP2和ZP4）和囊胚（APOA1，KRT18，ANXA2，S100A14，SLC34A2，PRSS8和ANXA2）为分子质量的代表性指标。此外,鉴定了仅在卵母细胞（224）和胚泡（2,200）中发现的具有特定生物学功能的基因。基因共表达网络和模块保存分析揭示了与外泌体组件相关的功能模块的强保存，类固醇代谢，细胞增殖，和形态发生。然而,细胞周期和氨基酸转运模块表现出弱保存，这可能反映了水牛和牛之间胚胎发育动力学和细胞信号通路激活的差异。这种全面的转录组学概况可作为未来体外胚胎生产测定中评估水牛卵母细胞和胚胎分子质量的宝贵资源。
