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Effects of in vivo prematuration and in vivo final maturation on developmental capacity and quality of pre-implantation embryos

机译:体内早熟和体内最终成熟对植入前胚胎发育能力和质量的影响

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In current in vitro production (IVP) systems, oocytes lack in vivo dominant and preovulatory follicular development, which may compromise pregnancy and viability of calves born. When an oocyte sets off in vivo on the road toward fertilization, it contains numerous transcripts and proteins necessary to survive the first few cell cycles of embryonic development. It is not yet known during which period of development the oocyte builds up the store, possibly primarily during the major growth phase of the oocyte, which is completed at the time a follicle reaches the size of 3 mm. Here, we investigated to what extent the later phases of follicular development, such as prematuration in the dominant follicle before the LH surge and ensuing final maturation in the preovulatory follicle, contribute to oocyte competence and development into viable blastocysts, Recent studies on in vivo vs in vitro oocyte maturation employed oocytes from an identical preovulatory development by applying ovum pick-up (OPU) twice (before and 24 h after the LH surge) in each cow treated for superovulation with a controlled LH surge. The embryo recovery rates at Day 7 of IVC after IVF were similar: 44% (97/219) for in vivo- vs 41% (87/213) for in vitro-matured oocytes, which shows that the natural environment during final maturation is not essential for the mere in vitro development of the prematured oocyte beyond the 8- to 16-cell stage. However, in vivo maturation appeared to contribute to the oocyte's quality in a more subtle way, as indicated by a significant increase in the proportion of expanded blastocysts and a more physiological degree of chromosome aberrations of the embryos. In blastocysts derived from in vivo-matured oocytes, 21% of the embryos were mixoploid vs 50% from in vitro-matured oocytes, concomitant with a higher number of cells (96 vs 54 per normal blastocyst). The expression pattern of a set of six developmentally important genes was, however, not significantly altered in blastocysts derived from in vivo-matured oocytes. Certain deviations were observed compared with the levels of entirely in vivo-developed control blastocysts, which suggests that the beneficial effects of in vivo maturation are possibly exerted at initial stages of embryonic development. Prematuration in vivo, occurring in a dominant follicle developing from about 8 mm into the preovulatory follicle, is accompanied by changes in protein synthesis of the cumulus oocyte complex (COC). Presumably, the differentially expressed proteins are involved in equipping the oocyte with further developmental competence. Although we have unraveled some important biochemical and cellular biological features of the oocyte, further research on in vivo processes is essential to improve in vitro embryo production in practice.
机译:在当前的体外生产(IVP)系统中,卵母细胞缺乏体内占优势的和排卵前的卵泡发育,这可能会损害妊娠和出生犊牛的生存能力。当卵母细胞在体内受精的过程中启动时,它包含许多转录本和蛋白质,这些转录本和蛋白质可以在胚胎发育的最初几个细胞周期中生存。尚不知道卵母细胞在哪个发育阶段建立储存,可能主要是在卵母细胞的主要生长阶段,卵泡达到3毫米大小时就完成了。在这里,我们调查了卵泡发育的后期阶段,例如LH激增之前优势卵泡的早熟以及排卵前卵泡最终成熟的程度,有助于卵母细胞的能力和发育成存活的胚泡。在体外卵母细胞成熟中,通过对每只经控制的LH浪涌进行超排卵处理的母牛进行两次卵子拾取(OPU)(在LH浪涌之前和之后24小时),采用来自相同排卵前发育的卵母细胞。体外受精后IVC第7天的胚胎恢复率相似:体内成熟率为44%(97/219),体外成熟卵母细胞为41%(87/213),这表明最终成熟过程中的自然环境是对于早于8至16个细胞阶段的成熟卵母细胞的体外发育而言,这不是必需的。然而,体内成熟似乎以更微妙的方式对卵母细胞的质量做出了贡献,这是由膨胀的胚泡比例显着增加和胚胎的染色体畸变更为生理化所表明的。在源自体内成熟卵母细胞的胚泡中,21%的胚胎是混合倍体,而来自体外成熟卵母细胞的胚为50%,伴随着更多的细胞数量(每个正常胚泡96个vs 54个)。但是,在体内成熟的卵母细胞衍生的囊胚中,六个发育重要基因的表达模式没有明显改变。与完全体内发育的对照胚泡的水平相比,观察到某些偏差,这表明体内成熟的有益作用可能在胚胎发育的初始阶段发挥作用。体内早熟发生在占主导地位的卵泡中,该卵泡从约8 mm发育成排卵前卵泡,伴随卵丘卵母细胞复合体(COC)蛋白质合成的变化。据推测,差异表达的蛋白质参与使卵母细胞具有进一步的发育能力。尽管我们已经阐明了卵母细胞的一些重要的生化和细胞生物学特征,但对体内过程的进一步研究对于在实践中提高体外胚胎的产生至关重要。

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