Figure 3

Spastin and p60-katanin have non-redundant roles in motor axon guidance.

(A) Immunolabelling of sMN axons in 72-hpf Tg(Hb9:GFP) larvae injected with MO^Ctl (n = 10), 1.3 pmol of MO^p60Kat (n = 10), or co-injected with MO^Ctl or MO^p60Kat and human SPG4 mRNA (MO^Ctl + mRNA^Spast, n = 14; MO^{p60Kat/1.3 pmol} + mRNA^Spast, n = 10) using Zn-5 and GFP antibodies. (B, C) Mean number of misguided/split dorsal nerves (B) and misguided rostral nerves (C) per larva. (D) Immunolabelling of sMN axons in 72-hpf Tg(Hb9:GFP) larvae injected with MO^Ctl (n = 29), MOsp^ATG (n = 30), or co-injected with MO^Ctl or MOsp^ATG and human KATNA1 mRNA (MO^Ctl + mRNA^KATNA1, n = 32; MOsp^ATG + mRNA^KATNA1, n = 29) using Zn-5 and GFP antibodies. (E, F) Mean number of abnormal rostral nerves (i.e., caudally targeted or missing; E) and missing dorsal nerves (F) per larva. (A, D) Dotted lines indicate lateral myosepta. Full arrowheads and full arrows show normal rostral and dorsal nerves, respectively. Empty arrowheads and empty arrows indicate misguided rostral and dorsal projections, respectively. All images are lateral views of the trunk, anterior to the left. Scale bars: 25 µm. (B, C, E, F) Non-blind quantifications were performed on 24 spinal hemisegments around the yolk tube per larva. Analysed larvae were pooled from three independent experiments. Violin Plots; horizontal bars indicate the median ± the 1st and 3rd quartiles. Kruskal–Wallis ANOVA test with Dunn’s post hoc test. P values are displayed on graphs. Source data are available online for this figure.