Understanding the cellular and molecular basis of epithelial migration using the Angiomotin mutant (360G-Wellcome-109100_Z_15_A)

Epithelial morphogenesis is a complex process involving tissue-level integrity and dynamically coordinated morphogenetic change. The visceral endoderm (VE) in the mouse embryo is one such tissue that undergoes morphogenesis required for embryonic development. In angiomotin (amot) mutants, there is a characteristic abnormal furrowing in the anterior VE accompanied by aberrant apical build up of actin. This leads to reduced displacement of anterior VE cells and embryonic lethality. We do not understand the cellular and molecular basis for abnormal cell migration in this mutant, nor what proteins AMOT interacts with in this context. To address these questions, I will initially make detailed quantitative observations of cell behaviour by analysing light sheet and confocal microscopy data, using automated cell segmentation and tracking approaches I am developing. I will explore actin dynamics using FRAP and laser ablation on mutant and wild-type embryos. To understand the molecular pathway through which AMOT regulates epithelial cell behaviour, I will use a phosphoproteomic approach to identify interacting proteins using wild-type and amot mutant ES cell-derived embryoid bodies. Select interacting proteins will be tested for their role using small molecule inhibitors where available, or CRISPR mediated mutagenesis in ES cells and mice.