Fascin1-Dependent Filopodia are Required for Directional Migration of a Subset of Neural Crest Cells

Abstract

Directional migration of neural crest (NC) cells is essential for patterning the vertebrate embryo, including the craniofacial skeleton. Extensive filopodial protrusions in NC cells are thought to sense chemo-attractive/repulsive signals that provide directionality. To test this hypothesis, we generated null mutations in zebrafish fascin1a (fscn1a), which encodes an actin-bundling protein required for filopodia formation. Homozygous fscn1a zygotic null mutants have normal NC filopodia due to unexpected stability of maternal Fscn1a protein throughout NC development and into juvenile stages. In contrast, maternal/zygotic fscn1a null mutant embryos ( fscn1a MZ) have severe loss of NC filopodia. However, only a subset of NC streams display migration defects, associated with selective loss of craniofacial elements and peripheral neurons. We also show that fscn1a-dependent NC migration functions through cxcr4a/cxcl12b chemokine signaling to ensure the fidelity of directional cell migration. These data show that fscn1a-dependent filopodia are required in a subset of NC cells to promote cell migration and NC derivative formation, and that perdurance of long-lived maternal proteins can mask essential zygotic gene functions during NC development.

Author Summary

During vertebrate embryogenesis, neural crest (NC) cells migrate extensively along stereotypical migration routes and differentiate into diverse derivatives, including the craniofacial skeleton and peripheral nervous system. While defects in NC migration underlie many human birth defects and may be coopted during cancer metastasis, the genetic pathways controlling directional NC migration remain incompletely understood. Filopodia protrusions are thought to act as “cellular antennae” that explore the environment for directional cues to ensure NC cells reach their correct location. To test this idea, we generated zebrafish fascin1a (fscn1a) mutants that have severe loss of filopodia. Surprisingly, we found that most NC cells migrate to their correct locations without robust filopodial protrusions. We found that fscn1a embryos have directional migration defects in a subset of NC cells, resulting in loss of specific craniofacial elements and peripheral neurons. Interestingly, these defects were only observed in ∼20% of fscn1a embryos, but were significantly enhanced by partial loss of the chemokine receptor Cxcr4a or disruption of the localized expression of its ligand Cxcl12b. Our data show that subsets of skeletal and neurogenic NC cells require filopodia to migrate and that fscn1a-dependent filopodia cooperate with chemokine signaling to promote directional migration of a subset of NC cells.