Fig. 4

Analysis of the effects of Nlcam on cell migration during OV morphogenesis. (A) Outline of the experimental procedure. Embryos are injected at the one cell stage with either a mixture of H2BmRFP and nlcam RNAs or with eBFP2-Nuc RNA (1). Cells from both donors are transplanted into Rx3::GFP hosts (2). After the onset of GFP expression, embryos are imaged by confocal microscopy (3). (B) Outline of the processing pipeline. Red boxes show the input; grey boxes detail the processing steps; and blue boxes show the final output. (C) Clustering analysis of transplanted cells indicates that nlcam overexpression does not promote homophilic clustering. (D–G) Selected frames from Dataset 1, showing key points during optic vesicle morphogenesis. Between 0 and 64 min (D, E), RPCs are converging towards the midline. Subsequently, RPCs begin to evaginate (F). Approximately 5 h after the onset of imaging (G), the process is essentially complete. The eye field is marked in green; wt cells are blue; nlcam⁺ cells are red. Scale bar is 100 μm. (H-K) Rendered reconstruction of cells within the eye field at the same time points. Trails show the position of the nucleus over the previous 10 time points (∼ 20 min). Green dashed line indicates midline. White dashed lines in H indicate medial (M) and lateral (L) groupings used for analysis. (L–O) Vectorial visualisation of migration. For simplification, the data is collapsed in the DV axis. Arrows show migration direction and displacement of cells over the following 10 time points. Cyan: lateral wt cells. Blue: medial wt cells. Red: lateral nlcam⁺ cells. Orange: medial nlcam⁺ cells.