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Nuclear transfer into mouse zygotes

Nature Genetics volume 24, pages 108–109 (2000)Cite this article

We have previously reported a novel nuclear-transfer technique to clone mice from adult somatic cells1,2 and late-passage embryonic stem cells3. The method entails the injection of a somatic-cell nucleus into an enucleated, unfertilized oocyte using a piezo-electrically driven micromanipulator. Embryonic development is activated by incubation with strontium chloride one to six hours after nuclear transfer.

Here we sought to determine if this cloning method could be adapted to use zygotes as recipients for somatic cell nuclei. In parallel, we have investigated a related issue: the developmental effect of the timing of oocyte activation relative to nuclear transfer. We microinjected nuclei from cumulus cells of adult female B6D2F1 mice into enucleated, unfertilized B6D2F1 oocytes1. Reconstructed oocytes were activated with strontium chloride at one of the following time points: (i) one to three hours after nuclear transfer (post-activation); (ii) immediately after nuclear transfer (immediate activation); or (iii) one hour before nuclear transfer (pre-activation). Additionally, other nuclei were injected into zygotes after removal of both pronuclei (iv), which we estimate to be five to six hours after the zygotes were produced following coital fertilization; this is the earliest point at which pronuclei can be readily visualized for removal. The four experimental conditions were carried out concurrently.

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Figure 1: Fate of oocytes and zygotes after nuclear transfer.
Figure 2: Chromosomes of reconstructed embryos stained with Giemsa just before the first mitotic cell division.

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Acknowledgements

We thank T. Perry for critical and useful comments on the manuscript. R.Y. acknowledges financial support from ProBio America.

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Author notes

  1. Teruhiko Wakayama

    Present address: The Rockefeller University, New York, NY, USA

Authors and Affiliations

  1. Department of Anatomy and Reproductive Biology, University of Hawaii Medical School, Honolulu, Hawaii, USA

    Teruhiko Wakayama, Hiroyuki Tateno & Ryuzo Yanagimachi

  2. The Rockefeller University, New York, New York, USA

    Peter Mombaerts

Authors
  1. Teruhiko Wakayama
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  2. Hiroyuki Tateno
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  3. Peter Mombaerts
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  4. Ryuzo Yanagimachi
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Correspondence to Teruhiko Wakayama.

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Wakayama, T., Tateno, H., Mombaerts, P. et al. Nuclear transfer into mouse zygotes. Nat Genet 24, 108–109 (2000). https://doi.org/10.1038/72749

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