Characterization of rDNA transcriptional activity in pre-implantation bovine embryos

在最初的胚胎发育过程中的基因表达模式的信息已成为一个重要的工具，了解胚胎技术程序可能会导致基因表达的偏差，可能是有害的进一步胚胎发育。在本项目中，将在牛体外产生的或核移植衍生的胚胎中研究rDNA转录、核仁发育、参与RNA poll转录和加工的蛋白质的相互作用以及在rRNA基因组再激活和沉默期间pre-rRNA对核仁发生的贡献。采用放射自显影和透射电子显微镜，光和超微结构免疫分析，荧光素原位杂交和逆转录-聚合酶链反应，我们将有助于理解rDNA转录和延长nucleologenesis，反映双方，激活和沉默的核糖体基因在牛胚胎。该项目的结果将作为体细胞核发生的替代模型，并将增加检查点的数量，适用于评估和改善生物医学研究或未来农业应用中使用的胚胎的正常性。

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.