Figure 5

Figure 5 TTLL6 and TTLL11 play non-overlapping roles in spinal motor axon navigation.

(A) Rescue and cross-rescue experiments of TTLL6 morphant phenotypes by co-injection of mouse TTLL6 or TTLL11 transcripts. (B) Reciprocal rescue and cross-rescue experiments of TTLL11 morphant phenotypes by co-injection of mouse TTLL11 or TTLL6 transcripts. (A, B) Upper panels: Overall morphology of 72-hpf control, morphant and rescued larvae. Scale bars: 250 μm. bottom panels: Immunolabelling of sMN axons with Zn-5 and GFP antibodies in 72-hpf Tg(Hb9:GFP) larvae injected with control (MO^Ctl, n = 22), TTLL6 (MO^TTLL6, n = 22), TTLL11 (MO^TTLL11, n = 21) morpholinos or co-injected with MO^TTLL6 or MO^TTLL11 and mouse TTLL6 or TTLL11 mRNA (MO^TTLL6 + mRNA^TTLL6, n = 19; MO^TTLL6 + mRNA^TTLL11, n = 22; MO^TTLL11 + mRNA^TTLL11, n = 17; MO^TTLL11 + mRNA^TTLL6, n = 22). Dotted lines delineate lateral myosepta. Full arrowheads and full arrows, respectively, indicate normal rostral and dorsal nerves. Empty arrowheads and empty arrows, respectively, point at misguided rostral and dorsal projections. Scale bars: 50 μm. (C–H) Quantifications of sMN defects in larvae analysed in (A, B). Mean number of split/misguided dorsal nerves (C), misrouted rostral nerves (D), missing dorsal nerves (E) and defasciculated rostral nerves (F) per larva. (G, H) Percentage of larvae with ectopic sorting of sMN axons (G) and somata (H) from the spinal cord. (C–H) Non-blind quantifications were performed on 24 spinal hemisegments located around the yolk tube per embryo or larva. Analysed larvae were pooled from three independent experiments. (C–F) Violin Plots; horizontal bars indicate the median ± the 1st and 3rd quartiles. One-way ANOVA test with Bonferroni’s post hoc test (D) or Kruskal–Wallis ANOVA test with Dunn’s post hoc test (C, E, F). (G, H) Chi² test. P values are displayed on graphs. Source data are available online for this figure.