Lateral zebrafish hypoblast cells initiate dorsal convergence near mid-gastrulation and exhibit non-polarized morphologies, limited cell-cell contact and indirect migration trajectories. By late gastrulation, mesodermal cells become packed as they engage in planar cell polarity (PCP)-dependent movement. Here, we aimed to understand this transition in cell behavior by examining the relationship between protrusion dynamics and establishment of PCP and directed migration. We found that wild-type cells undergo a reduction in bleb protrusions near late gastrulation accompanied by a VANGL planar cell polarity protein 2 (Vangl2)-regulated increase in filopodia number and polarization. Manipulation of blebs is sufficient to interfere with PCP and directed migration. We show that Vangl2, fibronectin and cadherin 2 function to suppress blebbing. Vangl2 maintains ezrin b (Ezrb) protein levels and higher Ezrb activation rescues defective mediolateral cell alignment and migration paths in vangl2 mutant embryos. Transplantation experiments show that loss of vangl2 disrupts protrusion formation cell-autonomously while fibronectin acts non-autonomously. We propose that dorsal convergence requires the coordinated action of Vangl2, Ezrb and cell-adhesion proteins to inhibit blebs and promote polarized actin-rich protrusive activity and PCP.
Keywords: Ezrin b; Fibronectin; Migration; Polarity; Protrusion; Vangl2.
© 2019. Published by The Company of Biologists Ltd.