Figure 3

ANP32a is required for the neuronal regeneration of zebrafish larvae after SCI. (A) The experimental flowchart showing the time course of microinjection of either Anp32a mRNA or Anp32a morpholinos, SCI performance and observation of spinal cord regeneration. Embryos at the one-cell stage were injected with either wobble-Anp32a-flag mRNA or antisense morpholino oligonucleotides (MO) of Anp32a and SCI at 48 hpf, followed by analysis of the lesion gap at 0, 18 and 24 hpi. (B) Time course image of spinal cord regeneration in control embryos and Anp32a-overexpressed embryos at 0, 18 and 24 hpi. (C) Quantification of temporal regenerative response in SCI-control and wobble-Anp32a-flag mRNA-injected embryos at 18 hpi (left; control: n = 25; RNA + SCI: n = 19) and 24 hpi (right; control: n = 28; RNA + SCI: n = 16). (D) Time course image of spinal cord regeneration in control embryos and Anp32a-morphants at 0, 18 and 24 hpi. (E) Quantification of temporal regenerative response in SCI control embryos and Anp32a-morphants at 18 hpi (left; control: n = 31; MO + SCI: n = 30) and 24 hpi (right; control: n = 32; MO + SCI: n = 34). Scale bar: 100 μm; The Student’s t-test was used to perform statistical analysis (*, p < 0.05; p < 0.01; ***, p < 0.001; error bars indicate mean ± SD).