Nuclear transplantation in mouse embryos: assessment of nuclear function.

Abstract

Enucleated mouse zygotes receiving eight-cell nuclei rarely develop beyond the two-cell stage, whereas enucleated two-cell embryos with eight-cell nuclei develop to blastocysts, and a few develop to midgestation. In this study the function of eight-cell nuclei in either the zygote or two-cell recipient was assessed by monitoring several nuclear-dependent events. These included methionine uptake, qualitative changes in protein synthesis, the time of blastocoele formation, and changes in nuclear volume. Although enucleated zygotes with nuclei from eight-cell embryos did not develop past the two-cell stage and had abnormally low levels of methionine uptake, they did show a normal zygote-to-two-cell transition in the types of polypeptides synthesized. Enucleated two-cell embryos with eight-cell nuclei formed blastocysts on schedule with the recipient cell stage and reached levels of methionine uptake equivalent to control embryos. The types of proteins synthesized by two-cell embryos with eight-cell nuclei indicated that the nucleus was developing partly in accord with its own developmental program. Transplanted nuclei did not enlarge to acquire the volume of the recipient cell's nucleus in either the zygote or the two-cell recipient. These results indicate that the difference in development between zygotes and two-cell embryos (each with eight-cell nuclei) may be the result of a greater overlap of function between the two-cell and eight-cell stages, rather than the extent of nuclear reprogramming. Nuclear function was not completely normal in either type of embryo, which may explain their failure to develop to term.