Figure 1

terfa knockdown does not affect the development of zebrafish larvae. (A) Experimental design: injection of terfa morpholino (terfa MO) or control morpholino (ctrl MO) at the 1-cell stage. The senescence phenotype was analyzed at 72 h post-fertilization (hpf) by immunofluorescence and at 78 hpf by RT-qPCR. Fin development and function were analyzed by measuring fin length at 144 hpf and zebrafish locomotion and activity at 120 hpf, respectively. (B) Confocal images of immunofluorescence staining to assess ɣH2AX expression in the caudal fin of ctrl morphants (ctrl MO) and terfa morphants (terfa MO) at 72 hpf. The graph shows the quantification of ɣH2AX-positive cells in the caudal fin; data are the mean ± SEM, n = 15 larvae/group; ** p < 0.01 (Mann Whitney test). (C) Relative cdkn2a/b (p15/16), cdkn1a (p21) and tp53 (p53) expression in the caudal fin of terfa MO and ctrl MO was assessed by RT-qPCR at 78 hpf. Ef1a was used as reference gene and data are the mean ± SEM (n = 14 independent experiments); **** p < 0.0001, *** p < 0.001, ** p < 0.01 (Mann Whitney test). (D) Representative images of zebrafish larvae at 144 hpf (left panels) and zoom on the caudal fin folds at 144 hpf (right panels). The graph shows the fin length (mean ± SEM) at 144 hpf in ctrl MO (n = 27) and terfa MO (n = 11) larvae; ns, not significant (Mann Whitney test). (E) Spontaneous locomotion analysis in ctrl and terfa morphants at 120 hpf. Quantification of the mobility period (left panel) and net velocity (middle panel) during locomotion; n = 24 ctrl MO and n = 24 terfa MO; ns, not significant (Mann Whitney test). On the right panel, the red and green trajectories correspond to fast and slow swimming, respectively.