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Production and development of porcine tetraploid parthenogeneticembryos

机译:猪四倍体孤雌生殖的产生与发展胚胎

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摘要

The aim of this study was to produce porcine tetraploid (4N) parthenogenetic embryos using various methods and evaluate their developmental potential. In method 1 (M1), porcine 4N parthenogenetic embryos were obtained by inhibiting extrusion of both first (PB1) and second (PB2) polar bodies; in methods 2 (M2) and 3 (M3), 4N parthenogenetic embryos were obtained by electrofusion of 2-cell stage diploid parthenogenetic embryos derived from inhibition of PB2 or PB1 extrusion, respectively. We found no differences in the rates of cleavage or blastocyst formation or the proportion of 4N embryos among M1, M2, and M3 groups. The different methods also did not influence apoptosis rates (number of TUNEL-positive cellsumber of total cells) or expression levels of apoptosis-related BAX and BCL2L1 genes. However, total cell and EdU (5-ethynyl-2’-deoxyuridine)-positive cell numbers in 4N parthenogenetic blastocysts derived from M1 were higher (p < 0.05) than those for M2 and M3 groups. Our results suggest that, although porcine 4N parthenogenetic embryos could be produced by a variety of methods, inhibition of PB1 and PB2 extrusion (M1) is superior to electrofusion of 2-cell stage diploid parthenogenetic embryosderived from inhibition of PB2 (M2) or PB1 (M3) extrusion.
机译:这项研究的目的是使用各种方法生产猪四倍体(4N)单性生殖胚胎并评估其发展潜力。在方法1(M1)中,通过抑制第一个(PB1)和第二个(PB2)极体的挤压获得了猪4N单性生殖胚胎;在方法2(M2)和方法3(M3)中,通过电融合2细胞阶段二倍体孤雌生殖胚胎分别获得了4N孤雌生殖胚胎,这些胚胎源自抑制PB2或PB1挤出。我们发现在M1,M2和M3组之间卵裂或胚泡形成的速率或4N胚胎的比例没有差异。不同的方法也没有影响凋亡率(TUNEL阳性细胞数/总细胞数)或凋亡相关的BAX和BCL2L1基因的表达水平。然而,源自M1的4N单性生殖胚泡的总细胞和EdU(5-乙炔基-2'-脱氧尿苷)阳性细胞数高于M2和M3组(p <0.05)。我们的结果表明,尽管可以通过多种方法生产猪4N单性生殖胚胎,但抑制PB1和PB2挤出(M1)优于2细胞阶段二倍体单性生殖胚胎的电融合。源自对PB2(M2)或PB1(M3)挤出的抑制。

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