Fig. 4

Pax2.1 and pax6a expression is altered in sox11 morphants.

(A) Fluorescent in situ hybridization on transverse sections from control and sox11 morphants with probes for pax2.1 and pax6a. The expression domain of f pax2.1 was expanded into the optic vesicle of sox11 morphants at 18 hpf (top right, arrows), and there was a modest retraction of pax6a expression compared to controls (top left; number of embryos analyzed: control MO, n = 14; sox11 MO, n = 13). At 36 and 48 hpf, in control retinas pax2.1 expression decreased and was only observed lining the optic nerve (asterisk; left middle and bottom rows); in contrast, pax2.1 expression was expanded and persisted around the open choroid fissure in sox11 morphant retinas (asterisks, right middle and bottom rows). Pax6a expression in the retina of sox11 morphants at 36 and 48 hpf appeared comparable to the control morphant retinas at this stage (number of embryos analyzed: 36 hpf control MO, n = 7; 36 hpf sox11 MO, n = 12; 48 hpf control MO, n = 8; 48 hpf sox11 MO, n = 14). (B) QPCR performed on mRNA from control and sox11 morphant heads at 18 and 24 hpf revealed a significant increase in pax2.1 expression at both 18 and 24 hpf, and a downregulation of pax6a expression at 24 hpf, in sox11 morphants compared to controls. Relative transcript abundance was normalized to atp5h (18 hpf) or gapdh (24 hpf) levels and is presented as the mean fold-change in expression relative to controls (n = 50 embryos per group, 3 independent biological replicates. *p<0.05. D, dorsal; V, ventral; OV, optic vesicle; L, lens; hpf, hours post fertilization; MO, morpholino.