Characterization of rDNA transcriptional activity in pre-implantation bovine embryos

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Information on the patterns of gene expression during initial embryonic development has become an important tool for understanding how embryo technological procedures may cause deviations in gene expression that can be detrimental for further embryonic development. In the present project rDNA transcription, development of nucleoli, interactions of proteins involved in RNA poll transcription and processing, and contribution of pre-rRNAs to nucleolo genesis during the period of rRNA genome reactivation and silencing will be studied in bovine in vitro produced or nuclear transfer derived embryos. Employing autoradiography and transmission electron microscopy, light and ultrastructural immuno analysis, fluorescein in situ hybridization and reverse transcription-polymerase chain reaction, we will contribute to the understanding of rDNA transcription and protracted nucleologenesis that mirrors both, the activation and silencing of the ribosomal genes in bovine embryo. Results of the project will be relevant as an alternative model to somatic cell nucleologenesis and will increase the number of checkpoints suitable for evaluation and improvement of the normality of embryos used in biomedical research or future agricultural application.