P-127: Characterization of Filia, A Maternal Effect Gene, in Bovine Oocytes and Embryos

Background: Genetic analysis in mice has lead to find about maternal effect genes such as Filia. Filia knock out mice have a 50% decrease in fertility. Filia dysfunction causes disorders in pre-implantation development. Mutations in human Filia gene, cause FBHM (Familial Biparental Hydatidiform Mole) in women. Filia protein in mice is homologous to that of rat and human, so this idea has emerged that maybe it is present in the bovine oocytes/embryos. Materials and Methods: To find Filia gene expression, primers were designed with OLIGO5 software and the amplified fragment was sequenced, and then translated to protein. A Real-Time experiment was done to identify the expression pattern in oocytes, and blastocysts. A bioinformatic analysis was done on bovine Filia gene (AC_000166.1) and its 2 kb upstream promoter, in FGENESH online tool, and the consensus transcription start site (TSS) and number of exons were predicted. The partial protein sequence was analysed in PHYRE2 online tool and the consensus domains were predicted. A 3' RACE-PCR analysis was implemented to identify different Filia splice variants. Results: Real-Time Data showed that Filia is expressed in bovine oocytes and pre-implantation embryos with a specific pattern. Three exons and one TSS were predicted in FGENESH and according to PHYRE2 results, the 5' protein sequence consisted of a RNA-binding KHdomain, previously found in mouse Filia protein. The 3' RACE-PCR results showed that just one splice variant was amplified. Conclusion: Filia has a maternal inheritance in bovine, and can be an essential factor in bovine embryo development and female fertility.