Ferronato Giuliana de A, Rosa Paola M da S, Bridi Alessandra, Santos Angélica Camargo Dos, Nociti Ricardo P, Chiaratti Marcos Roberto, Perecin Felipe, Meirelles Flávio V, Sangalli Juliano R, Silveira Juliano C da
Faculty of Animal Science and Food Engineering, Department of Veterinary Medicine, University of São Paulo, Pirassununga, SP, Brazil.
Universidade Federal de São Carlos, Centro de Ciências Biológicas e da Saúde, Departamento de Genética e Evolução, São Carlos, SP, Brazil.
Heliyon. 2024 Dec 6;10(24):e40957. doi: 10.1016/j.heliyon.2024.e40957. eCollection 2024 Dec 30.
-produced blastocysts are exposed to different stimuli when compared with ones. This includes the culture of embryos in a sturdy petri-dish, while embryos develop in a soft and dynamic structure. Here we hypothesized that a softer environment could differently modulate the produced embryos To that aim, presumptive zygotes were produced by fertilization and divided into three groups: 1) Cultured in a regular Petri dish - Control (CON); 2) Cultured on top of an alginate hydrogel surface (TOP); 3) Encapsulated inside an alginate hydrogel sphere (ENC) and cultured. We observed a decrease in blastocyst rate in TOP and ENC compared with the CON. Profiling of 383 bovine miRNAs, we found 3 miRNAs involved in cell proliferation being differently modulated by the TOP and ENC groups (miR-1246; miR-1260b, and miR-541). Analyzing global levels of DNA methylation and hydroxymethylation, we observed increased levels of the two marks in the TOP group when compared with the CON and ENC systems. RNA sequencing (RNA-seq) analysis carried out using blastocysts showed alterations in several developmentally important genes among the three groups. In summary, our results indicate that embryo production was possible to achieve up to the blastocyst stage. However, with the experimental conditions used herein, the alginate hydrogels adversely affected the embryo development, which were paralleled by epigenetic and transcriptomic changes.
与对照组相比,所产生的囊胚受到不同的刺激。这包括在坚固的培养皿中培养胚胎,而胚胎是在柔软且动态的结构中发育。在此,我们假设更柔软的环境可能会以不同方式调节所产生的胚胎。为此,通过受精产生推定合子,并将其分为三组:1)在常规培养皿中培养——对照组(CON);2)在藻酸盐水凝胶表面上培养(TOP);3)封装在藻酸盐水凝胶球体内(ENC)并进行培养。我们观察到,与CON组相比,TOP组和ENC组的囊胚率降低。对383个牛miRNA进行分析,我们发现3个参与细胞增殖的miRNA在TOP组和ENC组中受到不同调节(miR - 1246、miR - 1260b和miR - 541)。分析DNA甲基化和羟甲基化的整体水平,我们观察到与CON组和ENC组系统相比,TOP组中这两种标记物的水平有所增加。使用囊胚进行的RNA测序(RNA - seq)分析显示,三组中几个对发育重要的基因发生了改变。总之,我们的结果表明,胚胎发育能够达到囊胚阶段。然而,在此处使用的实验条件下,藻酸盐水凝胶对胚胎发育产生了不利影响,这与表观遗传和转录组变化同时出现。