Transcriptome profile analysis in bovine blastocysts developed in vivo after intrafollicular transfer of matured or immature oocytes. Transcriptome profile analysis in bovine blastocysts developed in vivo after intrafollicular transfer of matured or immature oocytes

This study aimed to investigate the transcriptome profile of bovine blastocysts produced in three different experimental conditions, namely I) VITRO (entire in vitro production of blastocysts), II) IFOT-M (Intrafollicular transfer of in vitro matured oocytes into preovulatory follicles) and III) IFOT-IM (Intrafollicular transfer of immature oocytes into dominant follicles) and ex vivo derived blastocysts (IV, VIVO) were used as a control. Accordingly, differentially expressed genes between in vitro and in vivo derived blastocysts which are involved in various fundamental roles including protein synthesis and protein degradation were identified. The study highlights insights about the attainment of embryonic developmental capacity and might be useful to improve embryo in vitro culture conditions. Overall design: Blastocysts produced in three different experimental conditions, namely I) VITRO (entire in vitro production of blastocysts), II) IFOT-M (Intrafollicular transfer of in vitro matured oocytes into preovulatory follicles) and III) IFOT-IM (Intrafollicular transfer of immature oocytes into dominant follicles) were compared with ex vivo derived blastocysts (IV, VIVO) serving as control. To determine the transcriptome profiles, pools of 15-20 blastocysts were analyzed by Affymetrix GeneChip® Bovine 1.0 ST array. Three independent biological replicates were investigated for each of the four experimental groups.

本研究旨在探究三种不同实验条件下制备的牛囊胚的转录组（transcriptome）谱，具体分组如下：Ⅰ) VITRO组（囊胚完全体外生产，entire in vitro production of blastocysts）、Ⅱ) IFOT-M组（体外成熟卵母细胞卵泡内移植至排卵前卵泡，Intrafollicular transfer of in vitro matured oocytes into preovulatory follicles）、Ⅲ) IFOT-IM组（未成熟卵母细胞卵泡内移植至优势卵泡，Intrafollicular transfer of immature oocytes into dominant follicles），并以离体来源囊胚（IV, VIVO）作为对照。据此，本研究鉴定出体外来源与体内来源囊胚间的差异表达基因，这些基因参与蛋白质合成与蛋白质降解等多种基础生物学过程。本研究揭示了胚胎发育能力获得的相关机制，可为优化胚胎体外培养体系提供理论支撑。实验整体设计：将上述三种实验条件下制备的牛囊胚与作为对照的离体来源囊胚（IV, VIVO）进行对比分析。为获取转录组谱，研究人员采用Affymetrix GeneChip® Bovine 1.0 ST芯片对15~20个囊胚的混合样本进行检测。四个实验组每组均设置三次独立生物学重复。