The main goal of this study was to identify mRNA transcripts whose content increases during bovine minor embryonic genome activation. We compared the gene expression profile of the bovine 4-cell-stage embryo and MII oocyte using the technique of suppression subtractive hybridization. Differentially expressed amplicons were subcloned, and 60 of them were sequenced. The resulting DNA sequences were compared with GenBank databases using BLAST search. The expression of five differentially expressed genes with an apparent function in cell cycle progression, chromatin remodeling, and splicing or translation initiation was further characterized by a real-time RT-PCR. Centromere protein F, 350/400ka (CENPF), and splicing factor arginine/serine-rich 3 (SRFS3) show an increase in mRNA content during the 2- to 4-cell and late 8-cell stages. For the high mobility group nucleosomal binding domain 2 (HMGN2), the level of mRNA increases in 2- to 4-cell and morula embryos. The transcription of splicing factor SRFS3 is alpha-amanitin sensitive both during 4-cell and late 8-cell stages. The transcription of CENPF and HMGN2 is alpha-amanitin sensitive only at late 8-cell stage and morula, respectively. SRFS3 represents the first described gene with an important function in preimplantation development, which is also expressed during bovine minor genome activation, and it is alpha-amanitin sensitive during this period. All described genes can play an important role in the preimplantation development of bovine embryos.

Institute of Animal Physiology and Genetics Laboratory of Developmental Biology Academy of Sciences of the Czech Republic v v i 277 21 Libechov Czech Republic

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