Penkov L I, Platonov E S, New D A
N.I. Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow.
Int J Dev Biol. 1995 Dec;39(6):985-91.
Parthenogenetic mammalian embryos show reduced placental development and do not develop beyond the 25-somite stage. But non-parthenogenetic embryos in culture, without a functional placenta, can develop to 40 somites or more. We have therefore examined the possibility that parthenogenetic embryos might also show prolonged development in culture. After parthenogenetic activation and diploidization, 23% of CBA and 56-58% of hybrid (CBAxC57BL/6) F1 mouse eggs developed in culture to blastocysts. When transferred to pseudopregnant recipients: 60% of the CBA blastocysts implanted and 26% of these developed to somite stage embryos; 71-72% of the hybrid blastocysts implanted and 11-17% of these developed to somite stage embryos. Improved development of postimplantation embryos explanted into culture at about the 15-20 somite stage was obtained by opening the visceral yolk sac (without exteriorizing the embryo). All the normal (non-parthenogenetic) embryos cultured in this way developed to more than 35 somites and many reached 45-55 somites. Under the same conditions, 11/17 diploid parthenogenetic CBA embryos developed in culture to more than 35 somites and 5 of these reached 45 somites; and 9/28 diploid parthenogenetic (CBAxC57BL/6) F1 embryos developed to 35 somites or more and 5 of these reached 45 somites. The size and protein content of the parthenogenetic embryos after culture was less than that of the normal embryos of equivalent stages. These results raise new possibilities for the analysis of parthenogenesis and genomic imprinting, including studies of the effects of adding to the culture medium specific growth factors and demethylating agents.
孤雌生殖的哺乳动物胚胎显示出胎盘发育受限,且无法发育到超过25体节阶段。但培养中的非孤雌生殖胚胎,在没有功能性胎盘的情况下,可发育到40体节或更多。因此,我们研究了孤雌生殖胚胎在培养中也可能出现发育延长的可能性。在孤雌激活和二倍体化后,23%的CBA小鼠卵和56 - 58%的杂交(CBAxC57BL/6)F1小鼠卵在培养中发育成囊胚。当移植到假孕受体中时:60%的CBA囊胚着床,其中26%发育到体节阶段胚胎;71 - 72%的杂交囊胚着床,其中11 - 17%发育到体节阶段胚胎。通过打开内脏卵黄囊(不将胚胎取出体外),将植入后约15 - 20体节阶段的胚胎植入培养可改善其发育情况。以这种方式培养的所有正常(非孤雌生殖)胚胎都发育到超过35体节,许多达到45 - 55体节。在相同条件下,17个二倍体孤雌生殖CBA胚胎中有11个在培养中发育到超过35体节,其中5个达到45体节;28个二倍体孤雌生殖(CBAxC57BL/6)F1胚胎中有9个发育到35体节或更多,其中5个达到45体节。培养后的孤雌生殖胚胎的大小和蛋白质含量低于同等阶段的正常胚胎。这些结果为孤雌生殖和基因组印记分析带来了新的可能性,包括研究向培养基中添加特定生长因子和去甲基化剂的效果。