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First published online 7 July 2004
doi: 10.1242/dev.01241
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1 Howard Hughes Medical Institute and Department of Cell and Developmental
Biology, University of Pennsylvania School of Medicine, Philadelphia, PA
19104, USA
2 Department of Biology, University of Pennsylvania, Philadelphia, PA 19104,
USA
* Author for correspondence (e-mail: bartolom{at}mail.med.upenn.edu)
Accepted 26 April 2004
Preimplantation development is a period of dynamic epigenetic change that
begins with remodeling of egg and sperm genomes, and ends with implantation.
During this time, parental-specific imprinting marks are maintained to direct
appropriate imprinted gene expression. We previously demonstrated that
H19 imprinting could be lost during preimplantation development under
certain culture conditions. To define the lability of genomic imprints during
this dynamic period and to determine whether loss of imprinting continues at
later stages of development, imprinted gene expression and methylation were
examined after in vitro preimplantation culture. Following culture in
Whitten's medium, the normally silent paternal H19 allele was
aberrantly expressed and undermethylated. However, only a subset of individual
cultured blastocysts (
65%) exhibited biallelic expression, while others
maintained imprinted H19 expression. Loss of H19 imprinting
persisted in mid-gestation conceptuses. Placental tissues displayed activation
of the normally silent allele for H19, Ascl2, Snrpn, Peg3 and
Xist while in the embryo proper imprinted expression for the most
part was preserved. Loss of imprinted expression was associated with a
decrease in methylation at the H19 and Snrpn imprinting
control regions. These results indicate that tissues of trophectoderm origin
are unable to restore genomic imprints and suggest that mechanisms that
safeguard imprinting might be more robust in the embryo than in the
placenta.
Key words: Imprinting, DNA methylation, Placenta, H19, Snrpn, Peg3, Ascl2, Xist, Mouse
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