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First published online March 23, 2006
doi: 10.1242/10.1242/dev.02317
1 Unité de Biologie du Développement et de la Reproduction, UMR
INRA-ENVA, Institut National de la Recherche Agronomique (INRA), Jouy-en-Josas
78352, France.
2 Laboratoire de Développement des Vertébrés, Institut
Jacques Monod, UMR 7592 CNRS, Université Paris 6 et 7, 2 place Jussieu,
75251 Paris, France.
Authors for correspondence (e-mail:
alice.jouneau{at}jouy.inra.fr;
jeanpaul.renard{at}jouy.inra.fr)
Accepted 9 February 2006
In mammals, cloning by nuclear transfer (NT) into an enucleated oocyte is a very inefficient process, even if it can generate healthy adults. We show that blastocysts derived from embryonic stem (ES) donor cells develop at a high rate, correctly express the pluripotential marker gene Oct4 in ICM cells and display normal growth in vitro. Moreover, the majority of them implant in the uterus of recipient females. We combine embryological studies, gene expression analysis during gastrulation and generation of chimaeric embryos to identify the developmental origin (stage and tissue affected) of NT embryo mortality. The majority died before mid-gestation from defects arising early, either at peri-implantation stages or during the gastrulation period. The first type of defect is a non-cell autonomous defect of the epiblast cells and is rescued by complementation of NT blastocysts with normal ES or ICM cells. The second type of defect affects growth regulation and the shape of the embryo but does not directly impair the initial establishment of the patterning of the embryo. Only chimaeras formed by the aggregation of NT and tetraploid embryos reveal no growth abnormalities at gastrulation. These studies indicate that the trophoblast cell lineage is the primary source of these defects. These embryological studies provide a solid basis for understanding reprogramming errors in NT embryos. In addition, they unveil new aspects of growth regulation while increasing our knowledge on the role of crosstalk between the extra-embryonic and the embryonic regions of the conceptus in the control of growth and morphogenesis.
Key words: Nuclear transfer, Growth, Gastrulation, Mouse, Trophoblast, Chimaera
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