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Protocol for the production of viable bimaternal mouse embryos

Nature Protocols volume 3, pages 197–209 (2008)Cite this article

Abstract

A reliable nuclear transfer method was first reported in 1983; it provided definite evidence that parthenogenetic embryos are lethal at early postimplantation in mammals. Subsequently, nuclear transfer has been extensively used as an important and versatile tool for investigating embryo and somatic-cell cloning and nucleo-cytoplasmic interactions. Further development of this technique has enabled the generation of bimaternal embryos containing two haploid sets of maternal genomes from female germ cells of different origins. By using a 2-d nuclear transfer system for oocyte reconstruction, viable mice can be produced solely from maternal genomes, without the participation of the paternal genome. This oocyte reconstruction system, as described in this protocol, could provide valuable guidelines for exploring the potential endowments of gametes and for conferring novel properties to them.

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Figure 1: Schematic diagram of production process for bimaternal embryos, which contain two sets of haploid genome derived from a non-growing oocyte (green) and an MII-stage fully grown oocyte (red).
Figure 2: Preparation of micromanipulation tools using a microforge.
Figure 3: Hemagglutination test.
Figure 4: Setup of the manipulation chamber.
Figure 5: Creating a perivitelline space in the GV oocytes.
Figure 6: Sequential protocol for the first nuclear transfer.
Figure 7: Sequential protocol for the second nuclear transfer.

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Authors and Affiliations

  1. Department of BioScience, Tokyo University of Agriculture, Setagaya-ku, Tokyo, 156-8502, Japan

    Manabu Kawahara, Yayoi Obata, Toshitaka Tsukadaira, Atsushi Fukuda & Tomohiro Kono

  2. Natural Science Center for Basic Research and Development, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan

    Yusuke Sotomaru

  3. Tsukuba Primate Research Center, National Institute of Biomedical Innovation, 1-1 Hachimandai, Tsukuba, Ibaraki, 305-0843, Japan

    Nobuhiro Shimozawa

  4. The Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge

    Siqin Bao

  5. CB2 1QN, UK

    Siqin Bao

Authors
  1. Manabu Kawahara
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  7. Atsushi Fukuda
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  8. Tomohiro Kono
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Correspondence to Tomohiro Kono.

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Kawahara, M., Obata, Y., Sotomaru, Y. et al. Protocol for the production of viable bimaternal mouse embryos. Nat Protoc 3, 197–209 (2008). https://doi.org/10.1038/nprot.2007.531

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