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Fig. 2. Pharyngeal pouch formation coincides with epibranchial neurogenesis.
(A-E,G,I) Confocal images of live, homozygous her5:egfp transgenic
embryos; anterior to the left. (A) 16 hpf embryo, lateral view; pouch 1, as
well as an unsegmented endoderm located further posteriorly (arrow), is
labeled. (B) 26 hpf embryo, lateral view; pouches 1-3 are labeled. (C) 18 hpf
embryo, dorsal view; pouches 1 and 2 on each side are joined along the AP axis
by a continuous band of endoderm surrounding mesenchyme of the second arch.
(D) 32 hpf embryo, dorsal view; pouches (1-6) form only in the most lateral
endoderm, while her5:egfp also labels a flattened layer of medial
endoderm (me). (E,G,I) Sequence of confocal images of her5:egfp, in
dorsal view, at 22 (E), 28 (F) and 32 (H) hpf. (E) 22 hpf embryo;
pouch-forming endodermal cells extend filopodial processes laterally (arrow).
(F) An illustration of the embryo shown in E. Arrow indicates movement of GFP+
cells toward the surface ectoderm (se). (G-J) Embryos analyzed for
her5:egfp expression at 28 (G) and 32 (I) hpf were fixed and
subjected to in situ hybridization (H,J) for neurod (blue) combined
with immunolabeling with 
-Hu antibody (brown). 1-6, pharyngeal pouches;
cb, cerebellum; e, eye; gV, trigeminal ganglion; gVII, facial (geniculate)
ganglion; gIX, glossopharyngeal (petrosal) ganglion; gX, vagal (nodose)
ganglion; hb, hindbrain; me, medial endoderm; ot, otic vesicle. Scale bars: 50
µm; in B for A,B; in C for C,D; in F for E-J.
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