Figure 6. Photo-morpholino knockdown of Sox10 demonstrates its necessity during the ingression/differentiation process.
(A) and (A') Control (antisense morpholino + sense photo-morpholino injected but not photocleaved) live embryos have robust numbers of ciliated (red) and neural crest–derived microvillous (green) neurons at 38 hpf. In contrast, identically injected embryos subjected to photocleavage at 24 hpf go on to develop ciliated neurons but significantly lack microvillous neurons at 38 hpf (B and B') in comparison to control embryos. Sox10:eGFP: green; OMP:RFP: red. Orientation arrows: D: dorsal; V: ventral; L: lateral. Scale bars: 30 μm. See also Figure 6—figure supplement 1.
Figure 6—figure supplement 1. Shown are more details of the phenotype in Figure 6: (A and A') control (antisense morpholino + sense photo-morpholino injected but not photocleaved) live embryos have robust numbers of ciliated (red) and neural crest–derived microvillous (green) neurons at 38 hpf. In contrast, identically injected embryos subjected to photocleavage at 24 hpf go on to develop ciliated neurons but have slight (B and B') or, much more commonly, large (C and C') decreases in microvillous neuron numbers and organization in comparison to control embryos.
Photocleavage may have been less efficient in some embryos, resulting in a range of phenotypic severity. (D–F') similar results are seen in the same embryos at 54 hpf, now with an even greater difference in microvillous neuron numbers between control and cleaved embryos. Sox10:eGFP: green; OMP:RFP: red. Orientation arrows: D: dorsal; V: ventral; L: lateral. Scale bars: 30 μm.