Rhombomere centers harbor proliferating neural progenitors. (A) Tg[nestin:GFP; Mu4127] embryos at 48 hpf displaying neural progenitors in magenta and the r3 and r5 landmarks in green. (B) Wild-type embryos immunostained with anti-Sox2 (magenta) to visualize the neural progenitors and with anti-HuC (green) to label the differentiated neurons at 48 hpf. (C,D) Wild-type embryos in situ hybridized with fabp7a (green) to stain progenitors and neurog1 or ascl1b (magenta) to label neuronal committed cells at 48 hpf. (a–d) Transverse views of (A–D) through the center of r4. (a) Images displaying a single channel or (b–d) the overlay of both channels. (E–H) Tg[BCP:H2B-GFP] embryos were incubated with EdU for 2h, in situ hybridized with fabp7a, and immunostained with anti-GFP to label boundary cells. The EdU-positive cells are displayed in magenta, the boundaries in blue, and fabp7a expression indicating the center of the rhombomeres in green. (e–h) Transverse views of (E–H) through the r4 center. White arrows in (e–h) indicate fabp7a cells that did not incorporate EdU. (A–H) Dorsal MIP with anterior to the top displaying all channels. White arrowheads indicate the rhombomeric boundaries. Dotted lines in (a–d) indicate the contour of the neural tube. ov, otic vesicle; rl, rhombic lip. Scale bar, 50 μm.
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