sema6d mutants display normal eye vesicle development and patterning before optic cup morphogenesis. Whole-mount RNA ISH for patterning genes indicates early development occurs normally in sema6d mutants (B, D, F, H, J) relative to wild-type siblings (A, C, E, G, I). In the bottom right of panels are the number of embryos of the total analyzed that exhibited the normal wild-type expression pattern. A, B, fgf8a expression in 10-ss embryos viewed laterally. C–H, Dorsal whole-mount images of the eye vesicles (dashed outlines) and neural keel of 12 ss processed by whole-mount ISH for the expression of early patterning genes. C, D, Early dorsal (future nasal) tissue with foxg1a. E, F, Early ventral (future temporal) tissue with foxd1. G, H, Early anterior (future ventral) tissue with vax2. I, J, RPE progenitors and neural crest cells express tfec at the appropriate time in wild-type siblings and sema6d mutants. K, The average anteroposterior length of the eye vesicles. L, Ratio (%) of RPE to eye vesicle anteroposterior lengths [bars a:b in J; p values are unpaired t test, df = 22 (K), df = 21 (L), error bars are SD]. Scale bars: 100 μm. A: anterior, C: caudal, ev: eye vesicle, h: hypothalamus, mhb: mid-hindbrain boundary, N: nasal, nc: neural crest, nk: neural keel, P: posterior, R: rostral, som: somites, T: temporal, tb: tailbud.
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