Waksmundzka M, Czolowska R, Tarkowski A K
Department of Embryology, Institute of Zoology, University of Warsaw, Poland.
Mol Reprod Dev. 1997 Dec;48(4):488-95. doi: 10.1002/(SICI)1098-2795(199712)48:4<488::AID-MRD9>3.0.CO;2-M.
We describe the preimplantation development of mouse embryos that have received the haploid maternal genome derived from early diplotene nuclei of primordial oocytes (PO). Two generations of recipient egg-cells were used. Induction of two meiotic divisions of the PO nucleus and the reduction of the number of chromosomes to the haploid level were achieved in preovulatory oocytes (primary recipients). The developmental potential of the obtained haploid genome was examined in zygotes (secondary recipients). The nuclei of PO obtained from newborn mice were transferred by cell electrofusion to in vitro maturing (IVM) and enucleated preovulatory mouse oocytes. The reconstructed oocytes which had completed maturation, i.e., reached metaphase II, were artificially activated (8% ethanol + CHX). Activated oocytes were used as donors of haploid pronuclei of PO origin which were transferred (by karyoplast fusion) to partially enucleated zygotes containing only the male pronucleus. Thus, reconstituted zygotes were transplanted to the ligated oviducts of the cycling mice and 27% of them developed to the blastocyst stage. Our experiments demonstrate that 1) the nucleus of PO can be induced to premature meiotic divisions in an IVM enucleated preovulatory oocyte; 2) in the presence of a normal male pronucleus, the haploid pronucleus of PO origin can substitute for a female pronucleus during preimplantation development.
我们描述了接受源自原始卵母细胞(PO)早双线期细胞核的单倍体母本基因组的小鼠胚胎的植入前发育情况。使用了两代受体卵细胞。在排卵前的卵母细胞(初级受体)中实现了PO细胞核的两次减数分裂诱导以及染色体数量减少至单倍体水平。在受精卵(次级受体)中检测了所获得单倍体基因组的发育潜力。通过细胞电融合将从新生小鼠获得的PO细胞核转移到体外成熟(IVM)并去核的排卵前小鼠卵母细胞中。完成成熟即达到中期II的重构卵母细胞被人工激活(8%乙醇+放线菌酮)。激活的卵母细胞用作PO来源的单倍体原核的供体,这些单倍体原核被转移(通过核质体融合)到仅含有雄原核的部分去核受精卵中。因此,重构受精卵被移植到处于发情周期的小鼠的结扎输卵管中,其中27%发育到囊胚阶段。我们的实验表明:1)PO细胞核可在IVM去核的排卵前卵母细胞中被诱导进行过早减数分裂;2)在正常雄原核存在的情况下,PO来源的单倍体原核在植入前发育过程中可替代雌原核。