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Research Papers
Division of Biology, Beckman Research Institute of the City Hope, Duarte, California 91010, USA.
Abstract
Genomic imprinting determines the monoallelic expression of a small number of genes during at least later stages of development. To obtain information necessary for the elucidation of imprinting mechanisms, we assessed the allele-specific expression and total expression level of four imprinted genes during early stages of development of normal F1 hybrid mice utilizing quantitative allele-specific reverse transcription-PCR (RT-PCR) single-nucleotide primer extension assays. The Igf2r and Snrpn genes were activated by the early 4-cell stage and exhibited biallelic and monoallelic expression, respectively, throughout preimplantation development. Thus, with respect to different imprinted genes, epigenetic systems determining monoallelic expression are not uniform in their time of establishment. Biallelic expression of Igf2r was observed in single blastomeres, discounting the possibility of random allelic inactivation at this stage. The closely linked H19 and Igf2 genes were activated after the blastocyst stage and often exhibited biallelic and monoallelic expression respectively in tissues of pregastrulation postimplantation-stage embryos, rather than reciprocal monoallelic modes as observed at later stages. This raises the possibility that imprinting of H19 is involved only in the maintenance and not in the initiation of monoallelic expression of Igf2. Monoallelic expression of Snrpn was observed in each blastomere at the 4-cell stage, demonstrating that the germ line, which exhibits biallelic expression of imprinted genes, must be derived from cells in which imprinting was once manifest.
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