TY - JOUR T1 - Wnt- and Glutamate-receptors orchestrate stem cell dynamics and asymmetric cell division JF - bioRxiv DO - 10.1101/2020.06.16.154609 SP - 2020.06.16.154609 AU - Sergi Junyent AU - Joshua Reeves AU - James L. A. Szczerkowski AU - Clare L. Garcin AU - Tung-Jui Trieu AU - Matthew Wilson AU - Shukry J. Habib Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/07/21/2020.06.16.154609.abstract N2 - Wnt signalling regulates many aspects of cell biology. Wnt-pathway activation and its downstream effects have been extensively studied, but the dynamic analysis of Wnt-ligands on mammalian cellular membranes is obstructed by difficulties of visualization. We overcome this using microbead-tethered Wnts presented to single embryonic stem cells, which undergo Wnt-mediated asymmetric cell division (ACD). Through live imaging and genetic editing, we show that knockout of Wnt co-receptor Lrp5 promotes cytoneme formation and Wnt-recruitment, which requires Lrp6 and β-catenin. Lrp5 facilitates ligand-retention at the membrane, and alongside Lrp6 mediates Wnt-ligand stabilization and positioning. β-catenin or Wnt co-receptor knockout causes misorientation at mitosis, and all but Lrp5 are required for Wnt-orientated ACD. Surprisingly, ionotropic glutamate receptor (iGluR) activity enables initial Wnt-recruitment, positioning, and ultimately oriented ACD. Uniquely, we have scrutinized the early Wnt ligand-membrane interaction, linking roles of Wnt-pathway components and crosstalk with iGluRs in guiding cell fate determination by oriented ACD.Competing Interest StatementThe authors have declared no competing interest. ER -