Figure 4

Defective swimming activity was partially recovered in the SCI larvae with Anp32a overexpression. (A) Experimental flowchart showing the time course of microinjection of either Anp32a mRNA (RNA) or Anp32a morpholinos (MO), SCI performance and behavior test. Embryos at the one-cell stage were microinjected with either wobble-Anp32a-flag mRNA or Anp32a-MO. Following SCI at 48 hpf, we recorded the swimming trajectory route at 24 hpi. (B) To carry out gain-of-function and loss-of function studies, the swimming trajectory route of each fish from three groups was examined, including untreated control (neither injection nor SCI), SCI only (SCI), both wobble-Anp32a-flag mRNA injection and SCI (RNA + SCI), injection of Anp32a-MO and SCI (MO + SCI), and co-injected Anp32a-MO with wobble-Anp32a-flag mRNA injection and SCI (MO + RNA + SCI). (C) Quantification and statistical analysis. Swimming distances (cm) were calculated based on the recorded trajectory route shown in (B). The data in each group were averaged from all examined larvae (untreated control: n = 30; SCI: n = 66; RNA + SCI: n = 35; MO + SCI: n = 39; MO + RNA + SCI: n = 35). One-way ANOVA, followed by Tukey?s multiple comparison test, was used to perform statistical analysis (***, p < 0.001; error bars indicate mean ± SD).