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. 2016 Mar 18;12(3):e1005934. doi: 10.1371/journal.pgen.1005934

Fig 3. Floorplate PCP is neither required nor sufficient for FBMN migration but can support the migration of WT FBMNs.

Fig 3

Confocal images showing dorsal views of 48 hpf hindbrains with anterior to the top. (A,B) Tg(shh:Gal4)-driven expression of Tg(UAS:Xdd1-GFP) (A) and Tg(UAS:Fzd3aΔC-GFP) (B) does not disrupt FBMN (magenta) migration. N = 13 Xdd1-GFP expressing embryos and 26 Fzd3aΔC-GFP expressing embryos. (C,D) Tg(shh:Gal4)-driven floorplate expression of GFP-Vangl2 (blue) in the floorplate of a wild type sibling does not disrupt FBMN migration (magenta) (C) and does not rescue migration in a vangl2 mutant (D). N = 24 vangl2 mutants with GFP-Vangl2 expression in the r4 floorplate, 14 with 5 or more expressing floorplate cells in r4. (C’,D’) Boxed regions from panels C and D respectively, showing a single z-plane where GFP-Vangl2 (blue) is expressed broadly in floorplate cells whose membranes are marked with the Zn5 antibody (yellow) [69]. (E,F) Genetic chimeras. Cascade blue-dextran marks all donor-derived cells (blue), Tg(isl1:mRFP) marks host FBMNs (magenta in E) and Tg(isl1:GFP) marks wild type donor-derived FBMNs (green in F). (E) The presence of wild type floorplate cells (blue) in a vangl2 mutant host embryo does not rescue the migration of host FBMNs. N = 16 embryos with extensive contribution of WT cells to the floorplate. (F) The presence of wild type floorplate cells (blue) in a vangl2 mutant can, however, support the migration of co-transplanted wild type donor derived FBMNs (green, arrows). N = 8 embryos with migrated donor-derived FBMNs/22 embryos with donor-derived FBMNs; N = 76 migrated FBMNs/383 total donor-derived FBMNs. (E’,F’) Single Z-planes of the boxed regions from panels E and F respectively, show that donor-derived cells (blue) are in the Zn-5-positive floorplate (yellow). Scale bars: 50 μm, 5μm in the insets.