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Embryo |
Department of Molecular Animal Breeding and Biotechnology,4 University of Munich, 85764 Oberschleissheim, Germany
Gene Center,5 University of Munich, 81377 Munich, Germany
Institute of Human Genetics,6 Mainz University School of Medicine, 55101 Mainz, Germany
Active demethylation of the paternal genome but not of the maternal genome occurs in fertilized mouse, rat, pig, and bovine zygotes. To study whether this early demethylation wave is important for embryonic development, we have analyzed the global methylation patterns of both in vivo-fertilized and cloned rabbit embryos. Anti-5-methylcytosine immunofluorescence of in vivo-fertilized rabbit embryos revealed that the equally high methylation levels of the paternal and maternal genomes are largely maintained from the zygote up to the 16-cell stage. The lack of detectable methylation changes in rabbit preimplantation embryos suggests that genome-wide demethylation is not an obligatory requirement for epigenetic reprogramming. The methylation patterns of embryos derived from fibroblast and cumulus cell nuclear transfer were similar to those of in vivo-fertilized rabbit embryos. Fluorescence in situ hybridization with chromosome-specific BACs demonstrated significantly increased chromosomal aneuploidy rates in cumulus cell nuclear transfer rabbit embryos and embryos derived from nuclear transfer of rabbit fibroblasts into bovine oocytes compared with in vivo-fertilized rabbit embryos. The incidence of chromosomal abnormalities was correlated with subsequent developmental failure. We propose that postzygotic mitotic errors are one important explanation of why mammalian cloning often fails.
2 Correspondence: Thomas Haaf, Johannes Gutenberg-Universität Mainz, Langenbeckstrasse 1, Bau 601, 55131 Mainz, Germany. FAX: 49 6131 175690; haaf{at}humgen.klinik.uni-mainz.de
3 Current address: Department of Development and Genetics, Evolutionary Biology Center, Uppsala University, Norbyvägen 18A, 75236 Uppsala, Sweden
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