Abstract
The first events of cellular differentiation occur before embryo implantation in mammals. They consist in the segregation between the inner cell mass and the trophectoderm, followed by segregation between epiblast and primitive endoderm within the inner cell mass. This differentiation process involves different molecular mechanisms, from cell polarization to temporally regulated expression of lineage-specific transcription factors. Recent data suggest that pre-implantation bovine embryo development can be a good model for human early development. The aim of this project is to investigate mechanisms related to early lineage segregation during bovine embryo development. We propose experiments to test hypotheses related to the first and second events of embryonic cell differentiation. We intend to assess the influence of cell polarization and Hippo signaling activity during inner cell mass and trophectoderm segregation. Then, the roles of NANOG and GATA6 transcription factors on epiblast and primitive endoderm segregation will be tested. Experimental approaches to be used will include use of small molecule inhibitors, CRISPR/Cas9-mediated gene editing and single-cell global gene expression analysis. This research project will bring a deeper understanding about mechanisms involved on the first differentiation choices of the bovine embryo. A deeper knowledge of these mechanisms will aid understanding issues related to in vitro fertilization, nuclear transfer techniques and pluripotent cell culture. (AU)
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