Parthenogenetic and nuclear transfer rabbit embryo development and apoptosis after activation treatments
Parthenogenetic and nuclear transfer rabbit embryo development and apoptosis after activation treatments
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DOI：
10.1002/mrd.20332
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发表时间：
2005-09-01
影响因子：
2.5
通讯作者：
Chen, DY
中科院分区：
文献类型：
--
作者：
Liu, SZ;Jiang, MX;Chen, DY
以往的研究主要是评价培养条件对植入前胚胎凋亡的影响。为了抑制核移植（NT）胚胎的凋亡，使用假定的凋亡抑制剂处理供体细胞。然而，关于激活处理对胚胎凋亡的影响知之甚少。本研究首先研究了不同的孤雌激活（PA）处理对胚胎发育、囊胚细胞数和凋亡的影响，然后从这些激活处理中选择一种被证明是最有效的激活处理用于激活兔NT胚胎。通过电脉冲激活，30分钟后，用25 μ M的无Ca 2+和Mg 2+的PBS中的D-肌醇1，4，5-三磷酸盐（IP 3）电穿孔，然后暴露于2.0mM的6-二甲基氨基嘌呤（6-DMAP）3小时，有效地激活了兔卵母细胞，并显著提高了囊胚发育率（72.7%）和总细胞数（175 ± 14.1），囊胚细胞凋亡水平（4.3 ± 0.5）显著低于其他各组。将同样的激活方案应用于NT胚胎激活，发现6-DMAP处理3小时可降低囊胚的凋亡水平，提高囊胚率和囊胚细胞数。结果表明，卵母细胞激活不仅影响胚胎发育和质量，而且影响胚胎凋亡。
Previous studies mainly evaluated the effect of culture conditions on preimplantation embryo apoptosis. In order to inhibit apoptosis of nuclear transfer (NT) embryos, putative apoptosis inhibitors were used to treat donor cells. However, little is known about the effect of activation treatments on embryo apoptosis. We firstly investigated the effect of various parthenogenetic activation (PA) treatments on embryo development, blastocyst cell number, and apoptosis, and then one of these activation treatments proved to be most efficient was selected for activation rabbit NT embryos. The activation by electrical pulses and 30 min later, electroporation with 25 mu M D-myoinositol 1,4,5-trisphosphate (IP3) in Ca2+- and Mg2+-free PBS, then exposure to 2.0 mM 6-dimethyl-aminopurine (6-DMAP) for 3 hr effectively activated rabbit oocytes, and resulted in significantly a higher blastocyst development rate (72.7%) and total cell number (175 +/- 14.1), and markedly lower apoptosis level of blastocyst (4.3 +/- 0.5) than all the other groups. When the same activation protocol was applied in NT embryo activation, we found that exposure of the embryos to 6-DMAP for 3 hr could decrease the apoptosis level of blastocyst and increase blastocyst rate and cell number. The results demonstrate that oocyte activation affects not only embryo development and quality but also embryo apoptosis.