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Fig. 1. Rab3d is required for anterior neurulation. A: Rab3d MO specifically inhibits translation of ectopic Rab3d mRNA, which has 50 UTR region
containing MO target region (5U-ORF-myc). B: Unilateral injection of Rab3d MO (20 ng) inhibits neural plate convergence and folding. a, b: Sox2
expression shows neural plate region. LacZ staining shows injected side. White arrow indicates convergence defect on Rab3d MO injected side.
Black arrow indicates neural folding. c: Cross section followed by immunostaining for b-tubulin. Imaginary midline of the embryo is marked by
white dashed line. Yellow dashed line indicates vertical boundary between neural plate and the underlying mesoderm. Asterisks mark cells with
apical constriction. C: Rab3d MO (20 ng) mediated neurulation defects are accompanied by changes of neural markers. Sox2 and Msx1 expressions
show the defective neural convergence and folding. Otx2 and Krox20 expressions are disrupted on the Rab3d MO injected side. Arrows
indicate MO injected sides. D: Rescue of Rab3d morphants phenotypes by morpholino-insensitive Rab3d mRNA (Sox2 down: reduced Sox2
expression and no NF/CV defects; NF/CV defect: neural folding or convergence defect). E: Rab3d MO (10 ng) was injected to the dorsal-animal
blastomeres of four cell stage embryos. At neurula stage, embryos were collected and subjected to the semi-quantitative RT-PCR analysis. ODC
was used as a loading control. The data are representative of three independent experiments. |
