Lin Tao, Lee Jae Eun, Shin Hyeon Yeong, Lee Joo Bin, Kim So Yeon, Jin Dong Il
Division of Animal & Dairy Science, Chungnam National University, Daejeon 34143, Korea.
J Anim Sci Technol. 2019 Jul;61(4):225-233. doi: 10.5187/jast.2019.61.4.225. Epub 2019 Jul 31.
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 cells/number of total cells) or expression levels of apoptosis-related and genes. However, total cell and EdU (5-ethynyl-2'-deoxyuridine)-positive cell numbers in 4N parthenogenetic blastocysts derived from M1 were higher ( < 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 embryos derived from inhibition of PB2 (M2) or PB1 (M3) extrusion.
本研究的目的是使用各种方法生产猪四倍体(4N)孤雌生殖胚胎,并评估其发育潜力。在方法1(M1)中,通过抑制第一极体(PB1)和第二极体(PB2)的排出获得猪4N孤雌生殖胚胎;在方法2(M2)和方法3(M3)中,分别通过对源自抑制PB2或PB1排出的二倍体2细胞期孤雌生殖胚胎进行电融合来获得4N孤雌生殖胚胎。我们发现M1、M2和M3组在卵裂率、囊胚形成率或4N胚胎比例方面没有差异。不同方法也不影响凋亡率(TUNEL阳性细胞数/总细胞数)或凋亡相关基因的表达水平。然而,源自M1的4N孤雌生殖囊胚中的总细胞数和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孤雌生殖胚胎,但抑制PB1和PB2排出(M1)优于对源自抑制PB2(M2)或PB1(M3)排出的二倍体2细胞期孤雌生殖胚胎进行电融合。