Institute of Veterinary Anatomy, Vetsuisse Faculty Zurich, University of Zurich, Lindau, ZH, 8315, Switzerland.
Department of Animal Reproduction, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA-CSIC), Madrid, Spain.
BMC Genomics. 2024 May 27;25(1):520. doi: 10.1186/s12864-024-10429-5.
Increasing evidence points to an active role of oviductal extracellular vesicles (oEVs) in the early embryo-maternal dialogue. However, it remains unclear whether oEVs contribute to the recognition of the presence of embryos and their quality in the oviduct. Hence, we examined whether the molecular cargo of oEVs secreted by bovine oviduct epithelial cells (BOEC) differs depending on the presence of good (≥ 8 cells, G) or poor (< 8 cells, P) quality embryos. In addition, differences in RNA profiles between G and P embryos were analyzed in attempt to distinguish oEVs and embryonic EVs cargos.
For this purpose, primary BOEC were co-cultured with in vitro produced embryos (IVP) 53 h post fertilization as follows: BOEC with G embryos (BGE); BOEC with P embryos (BPE); G embryos alone (GE); P embryos alone (PE); BOEC alone (B) and medium control (M). After 24 h of co-culture, conditioned media were collected from all groups and EVs were isolated and characterized. MicroRNA profiling of EVs and embryos was performed by small RNA-sequencing.
In EVs, 84 miRNAs were identified, with 8 differentially abundant (DA) miRNAs for BGE vs. B and 4 for BPE vs. B (P-value < 0.01). In embryos, 187 miRNAs were identified, with 12 DA miRNAs for BGE vs. BPE, 3 for G vs. P, 8 for BGE vs. GE, and 11 for BPE vs. PE (P-value < 0.01).
These results indicated that oEVs are involved in the oviductal-embryo recognition and pointed to specific miRNAs with signaling and supporting roles during early embryo development.
越来越多的证据表明,输卵管细胞外囊泡(oEVs)在早期胚胎-母体对话中发挥着积极作用。然而,oEVs 是否有助于识别输卵管中胚胎的存在及其质量仍不清楚。因此,我们研究了牛输卵管上皮细胞(BOEC)分泌的 oEVs 的分子货物是否因优质(≥8 细胞,G)或劣质(<8 细胞,P)胚胎的存在而有所不同。此外,还分析了 G 和 P 胚胎之间的 RNA 谱差异,试图区分 oEVs 和胚胎 EVs 货物。
为此,将原代 BOEC 与体外受精后 53 小时的 IVP 胚胎共培养,具体如下:与 G 胚胎共培养的 BOEC(BGE);与 P 胚胎共培养的 BOEC(BPE);G 胚胎单独培养(GE);P 胚胎单独培养(PE);BOEC 单独培养(B)和培养基对照(M)。共培养 24 小时后,从所有组收集条件培养基,并分离和鉴定 EVs。通过小 RNA-seq 对 EVs 和胚胎进行 miRNA 谱分析。
在 EVs 中,鉴定出 84 个 miRNA,其中 8 个差异丰度 miRNA(BGE 与 B 相比)和 4 个差异丰度 miRNA(BPE 与 B 相比)(P 值 < 0.01)。在胚胎中,鉴定出 187 个 miRNA,其中 12 个差异丰度 miRNA(BGE 与 BPE 相比),3 个差异丰度 miRNA(G 与 P 相比),8 个差异丰度 miRNA(BGE 与 GE 相比)和 11 个差异丰度 miRNA(BPE 与 PE 相比)(P 值 < 0.01)。
这些结果表明,oEVs 参与了输卵管-胚胎的识别,并指出了在早期胚胎发育过程中具有信号和支持作用的特定 miRNA。