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Files in this Data Supplement:
Movie 1. Short-range neural crest cell-cell contacts within migratory streams. This data set was co-injected with Gap43-tagged EGFP (green) and H2B-tagged MRFP (red) to label the cell plasma membranes and nuclei. The 3D reconstruction and rotation of the data 360° around the y-axis shows the r6 stream is a dense stream with many filopodial connections throughout the stream. The cells in the confocal z-stack (30 µm at 1 µm interval) show various connections that were not visible in either the flat projection of the z-stack or the depth coding of the z-stack.
Movie 2. Neural crest cells at the stream fronts display numerous filopodia. A typical hairy neural crest cell displays many long and short filopodia with different lengths. When the data set is rotated around the y-, x-, and an arbitrary axis, the movie shows how the extensions course off in different directions and cover a large area of the micro-environment adjacent to the cell body (the nucleus is approximately 10 µm). The embryo was injected with Gap43-tagged EGFP (confocal z-stack of 30 images at 1 µm interval).
Movie 3. Long-range neural crest cell-cell contacts by bipolar shaped cells within migratory streams. Within a typical r4 migratory stream, many neural crest cells display a bipolar shape with filopodia that extend over long distances. A filopodium may extend and intertwine around and between neighboring migrating neural crest cells before contacting a non-local neural crest cell. The embryo was co-injected with Gap43-tagged EGFP (green) and H2B-MRFP (red) and electroporated to label the cell plasma membranes and nuclei, respectively. This confocal z-stack consists of 25 sections (1 µm intervals), rendered in 3D and rotated 360° around the y-axis using Zeiss AIM software (VisArt).
Movie 4. Neural crest cell filopodia from a trailing cell may track the position of a downstream cell. A typical migrating neural crest cell in high-resolution within a stream moving towards the branchial arches. The neural crest cell displays filopodial and lamellipodial extensions. A long filopodium extension can be seen to contact a downstream migrating neural crest cell and track the movements of the back of the cell. In order to make the perimeters of the trailing and downstream target cells more visually distinct, the outline of each cell was traced from each frame of the time-lapse. The embryo was co-injected with Gap43-tagged EGFP (green) and H2B-MRFP (red), and electroporated to label the cell plasma membranes and nuclei, respectively. The time-lapse represents 2 hours of total time (1 minute intervals, 10 minute intervals shown here).
Movie 5. Neural crest cells maintain long tethered contacts. Migrating neural crest cells within a stream may be in close contact (short-range) between neighboring cells (three cells in the top-middle of the image) or in long-range (>50 µm) contact with non-local neighbors. The 3D rendering shows that the long-range contacts wind around neighboring neural crest cells, as can be seen when the filopod changes shape to avoid a nucleus. When the z-stack is rotated around the y-axis it shows that the filopodial process is continuous. The embryo was co-injected with Gap43-tagged EGFP and H2B-MRFP, and electroporated to label the cell plasma membranes and nuclei, respectively (confocal z-stack of 30 images at 1 µm interval).
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