TRAP1 knock-out in Zebrafish affects early embryogenesis stages and mitochondrial bioenergetics.a Measurement of oxygen consumption rate (OCR) in TRAP1 wild-type and knock-out embryos at 96 hpf. Respiratory complex I and III inhibitors (2 μM rotenone and 5 μM antimycin A, respectively) were added where indicated. Left: representative traces; right: quantification of basal respiration. b–d Succinate-CoQ reductase (SQR) enzymatic activity of succinate dehydrogenase (SDH) in total wild-type or TRAP1 knock-out embryo lysates. Where indicated, embryos were treated with the TRAP1 inhibitor compound 5 (100 µM) for 2 h. e Kinetics of fish growth from 48 hpf to 96 hpf; where indicated, fish were treated with succinate (500 µM or 1 mM). Succinate was added directly in fish water every day from 24 hpf to 96 hpf. The total fish area, excluding the yolk region, was considered. f Quantification of basal mitochondrial respiration in wild-type and TRAP1 knock-out embryos at 96 hpf; where indicated, fish were treated with succinate 1 mM every day from 24 hpf until 96 hpf. Inhibitors of respiratory complex I and III (2 μM rotenone and 5 μM antimycin A, respectively) were added where indicated. Data were normalized to untreated fish and reported as average ±SEM of at least six animals (a, f) or 15 animals (e) and 60 embryos (in b–d) per condition. In a, c, d, e, f data are reported as average ±SEM with an unpaired two-tailed Student’s t test; in (b) data are reported as average ±SEM with one-way ANOVA and Bonferroni’s correction of at least four independent experiments; asterisks indicate significant differences (*p < 0.05, ∗∗p < 0.01, ***p < 0.001).
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