Abstract
Neural crest cells exhibit dramatic migration behaviors as they populate their distant targets. Using a line of zebrafish expressing green fluorescent protein (sox10:EGFP) in neural crest cells we developed an assay to analyze and quantify cell migration as a population, and use it here to characterize in detail the subtle defects in cell migration caused by ethanol exposure during early development. The challenge was to quantify changes in the in vivo migration of all Sox10:EGFP expressing cells in the visual field of time-lapse movies. To perform this analysis we used an Optical Flow algorithm for motion detection and combined the analysis with a fit to an affine transformation. Through this analysis we detected and quantified significant differences in the cell migrations of Sox10:EGFP positive cranial neural crest populations in ethanol treated versus untreated embryos. Specifically, treatment affected migration by increasing the left-right asymmetry of the migrating cells and by altering the direction of cell movements. Thus, by applying this novel computational analysis, we were able to quantify the movements of populations of cells, allowing us to detect subtle changes in cell behaviors. Because cranial neural crest cells contribute to the formation of the frontal mass these subtle differences may underlie commonly observed facial asymmetries in normal human populations.
Highlights
The cranial neural crest cells (CNCC) are a population of progenitors that give rise to both craniofacial structures and components of the peripheral nervous system (PNS)
In embryos exposed to 200 mM (Fig. 2C1,2,3, see Video S3), the effects of EtOH on CNCC migration were more severe: the CNCC start migrating anteriorly (Fig. 2 C1,2), though some cells failed to move (Fig. 2C3, arrowhead) and by 20–22 somites the cells had arrested, never reaching the limits of the olfactory placodes (OPs) (Fig. 2C, asterisks)
The resulting morphological craniofacial phenotypes we observed in the EtOH exposed embryos (Boric, Couve, and Whitlock, unpublished) are in agreement with previous studies showing that EtOH treatment results distinct reproducible phenotypes [22,26,27,28,29,30,31]
Summary
The cranial neural crest cells (CNCC) are a population of progenitors that give rise to both craniofacial structures and components of the peripheral nervous system (PNS). These cells migrate from the dorsal neural tube to populate the face. The cell movements that occur during the migration of this population of CNCC are complex and difficult to describe and characterize quantitatively. For this reason we developed an analytical method to characterize cell migration, and use it here to describe the defects of dorsal anterior CNCC migration caused by alcohol exposure
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