Apoptosis in parthenogenetic preimplantation porcine embryos
Apoptosis in parthenogenetic preimplantation porcine embryos
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DOI：
10.1095/biolreprod.103.026005
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发表时间：
2004-06-01
影响因子：
3.6
通讯作者：
Prather, RS
中科院分区：
文献类型：
--
作者：
Hao, YH;Lai, LX;Prather, RS
卵母细胞的孤雌生殖(PA)对于许多与卵母细胞或胚胎相关的技术是必不可少的，例如胞质内单精子注射和核移植克隆。本研究采用末端脱氧核糖核酸转移酶介导的脱氧尿苷5-三磷酸缺口末端标记法(TUNEL)，研究了PA猪胚胎中细胞凋亡的发生和频率，以及符合细胞凋亡一般标准的形态变化。与体外受精(IVF)相比，PA胚胎在体外培养过程中有较高的细胞凋亡率，较低的卵裂率(45%对71%)和较低的囊胚期发育率(16%对29%)。PA胚胎TUNEL阳性信号最早出现在第6天，比IVF胚胎晚1天。PA胚胎的凋亡率从第6天的15%上升到第8天的29%。PA胚胎的平均凋亡率高于IVF胚胎(第5天除外)。尤其是PA胚胎的凋亡率在第6天(15%比6.7%)和第8天(29%比13%)是IVF胚胎的两倍。PA囊胚的平均细胞数显著低于IVF囊胚，而细胞凋亡率显著高于IVF囊胚。PA囊胚单倍体率高(62.5%)。这种倍性可能与高水平的细胞凋亡有关。这些结果可能有助于解释孤雌生殖发育失败的机制，并可能导致改善孤雌生殖发育的方法。
Parthenogenesis (PA) of the oocyte is essential to a number of oocyte- or embryo-related technologies such as intracytoplasmic sperm injection and cloning by nuclear transfer. This study investigated the onset and frequency of apoptosis in PA-porcine embryos and the morphological changes that conform to the general criteria of apoptotic cell death by using a terminal deoxynucleatidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling (TUNEL) assay. PA embryos had a higher degree of apoptotic cell death during in vitro culture, a lower cleavage rate (45% vs. 71%), and a lower development rate to the blastocyst stage (16% vs. 29%), relative to in vitro fertilization (IVF). The earliest positive TUNEL signal in the PA embryos was detected on Day 6, 1 day later than that in IVF embryos. Apoptosis in PA embryos increased from 15% of the embryos on Day 6 to 29% on Day 8. The mean level of apoptosis of the PA embryos was statistically higher than that of IVF embryos, except on Day 5. In particular, apoptosis in PA embryos was twice that of IVF embryos on Day 6 (15% vs. 6.7%) and Day 8 (29% vs. 13%). The mean cell number in PA blastocysts was significantly lower than that of IVF blastocysts, whereas the percentage of apoptosis in PA blastocysts was significantly higher than that of IVF blastocysts. There was it high percentage of haploid (62.5%) PA blastocysts. The ploidy may contribute to a high level of apoptosis. These results may help to explain the mechanism of parthenogenetic developmental failure and may lead to methods that will improve parthenogenetic development.