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Files in this Data Supplement:
Movie 1. Pharyngeal arch development in a wild-type animal from 12 to 38 hpf. At the beginning of the movie, H2A.F/Z:GFP (green) labels the nuclei of all cells and cranial NCC can be seen migrating in three streams, two anterior and one posterior to the developing otic vesicle. fli1-GFP expression initiates in NCC as they form the pharyngeal arches. As endodermal pouches develop (seen as black clearings between the labeled arches), the most posterior NCC mass successively subdivides to form five branchial arches. At the end of the movie, seven pharyngeal arches (mandibular, hyoid and five branchial: labeled 1-7) and six pouches (p1-p6) are seen. View is lateral, with anterior to the left and dorsal up.
Movie 2. In this example of pharyngeal arch development in an animal reduced for Fgf8 (fgf8-MO; fli1-GFP), one of the endodermal pouches (arrow) fails to fully form and only four arch segments are made from the posterior NCC mass. At the end of the movie, six pharyngeal arches (1-7 with a fused 4/5 arch) and five pouches (p1-p6 but p4 is missing) are seen.
Movie 3. Red and green channels. Pharyngeal endoderm development in a wild-type embryo from 10 to 30 hpf. The nuclei of all cells are green (H2A.F/Z:GFP) and pharyngeal endoderm cells are red (Alexa568). At the start of the movie, endoderm cells are dispersed throughout the embryo. Within a few hours endoderm cells migrate and coalesce at the midline as the neural keel forms. By 14 hpf, cranial NCC can be seen migrating in three streams, and shortly hereafter fli1-GFP expression initiates in NCC of the pharyngeal arches. At the same time, a few endoderm cells (white arrow) extend cytoplasmic processes and begin to migrate laterally to form the first pouch. As the movie progresses, more posterior pouches and arches develop until eventually four out of seven of the pharyngeal arches (numbered 1-4) and three out of six pouches (p1-p3) can be seen. View is dorsal-lateral, with anterior to the left and dorsal up.
Movie 4. Red channel only. Pharyngeal endoderm development in a wild-type embryo from 10 to 30 hpf. The nuclei of all cells are green (H2A.F/Z:GFP) and pharyngeal endoderm cells are red (Alexa568). At the start of the movie, endoderm cells are dispersed throughout the embryo. Within a few hours endoderm cells migrate and coalesce at the midline as the neural keel forms. By 14 hpf, cranial NCC can be seen migrating in three streams, and shortly hereafter fli1-GFP expression initiates in NCC of the pharyngeal arches. At the same time, a few endoderm cells (white arrow) extend cytoplasmic processes and begin to migrate laterally to form the first pouch. As the movie progresses, more posterior pouches and arches develop until eventually four out of seven of the pharyngeal arches (numbered 1-4) and three out of six pouches (p1-p3) can be seen. View is dorsal-lateral, with anterior to the left and dorsal up.
Movie 5. Red and green channels. Pharyngeal endoderm development in an embryo reduced for both Fgf8 and Fgf3 (fgf8–; fgf3-MO; H2A.F/Z:GFP). Whereas the generation and medial migration of endoderm cells (red) are normal in this embryo, the subsequent lateral migration of pouch endoderm cells is disorganized and delayed. White arrows highlight cytoplasmic processes of migrating pouch endoderm cells that do not properly align along the mediolateral axis. At the end of the movie, pharyngeal endoderm cells remain a single mass and discrete pouches have failed to form. Also note the high levels of cell death in the neural keel, apparent as fragmentation of nuclei (green: H2A.F/Z:GFP) in the top center of Movie 5.
Movie 6. Red channel only. Pharyngeal endoderm development in an embryo reduced for both Fgf8 and Fgf3 (fgf8–; fgf3-MO; H2A.F/Z:GFP). Whereas the generation and medial migration of endoderm cells (red) are normal in this embryo, the subsequent lateral migration of pouch endoderm cells is disorganized and delayed. White arrows highlight cytoplasmic processes of migrating pouch endoderm cells that do not properly align along the mediolateral axis. At the end of the movie, pharyngeal endoderm cells remain a single mass and discrete pouches have failed to form.
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