Fig. 3

Fig. 3 Loss of shp2 Causes Cell Death in Neural Tissue

(A and B) Embryos (28 hpf) stained for the panneural crest marker crestin (A) or dlx2 (B). shp2 MO causes a severe loss of crestin- and dlx2-positive neural crest cells posterior to the otic placode (black arrows in [A] and [B]).

(C) Views of 22 hpf embryos stained for α-activated-Caspase 3. Lack of shp2 causes apoptosis in the head and cells adjacent to the otic placode (arrows), extending down the dorsal region of the trunk (arrowheads).

(D) Lateral views show increased puma expression in morphants at 18 hpf, particularly in the brain (left panels), which by 24 hpf becomes restricted to the neural tube (arrow in right panel).

(E) Depletion of shp2 causes an increase in γ-H2Ax-positive nuclei in 12 somite stage embryos along the dorsal midline (quantified at right).

(F) Lateral views (30x) of Acridine orange-stained embryos (24 hpf); right panels are higher magnification views (80x) of the trunk region (boxed). Cell death in the spinal cord (arrows) and brain is rescued by coexpressing zebrafish bcl-xl mRNA with shp2 MO.

(G) Lateral (left) and Dorsal (right) views of 4 dpf embryos, showing that coexpression of zebrafish bcl-xl mRNA restores pigment cell numbers, but not their migration to the ventral stripe (white arrows). Right panels: Ventral views of 6 dpf embryos stained with Alcian blue, showing rescue of some cartilage elements (arrows).

Error bars correspond to standard deviation of the mean.