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Figure 4. Retinol binding protein 1 knockdown results in reduced marker gene expression during eye and brain development. A, Reduced expression of the eye-specific marker genes rax and pax6 was observed at stage 13 by whole mount in situ hybridization after injection of retinol binding protein 1 (Rbp1; black arrowheads) but not control morpholino oligonucleotide (MO). In contrast, the pan- neural marker gene sox3 was not affected. A statistical evaluation is provided. B, Depletion of Rbp1 led to reduced expression of the eye marker genes rax and pax6 (black arrowheads) at stage 23. Control MO-injected and uninjected sides showed normal marker gene expression. Anterior views are shown. A statistical evaluation is given. C, Rbp1 knockdown led to reduced expression of rax at stage 24 in the ventral part of the forebrain (turquoise arrowhead). A statistical evaluation is given. D, At stage 36, expression of rax was reduced in the pineal gland upon bilateral Rbp1 MO injection (blue arrowhead) and is represented statistically. Abbreviations: inj., injected; MO, morpholino oligonucleotide; N, number of evaluated embryos in total; n, number of independent experiments; n.s., not significant; Rbp1, retinol binding protein 1; uninj., uninjected. Statistics: Error bars represent SEM; *P ≤ .05. P values were calculated by a nonparametric, one-tailed Mann-Whitney rank sum test |
