sacs?/? mutant zebrafish display motor impairment and reduced cerebellar area. (A) Coiling frequency in zebrafish embryos at 30 hpf is decreased in sacs?/? compared with control specimens (sacs?/? n = 113; controls n = 113, in 4 independent experiments). (B) Automated analysis of spontaneous motor activity revealed a reduction in swim distance and velocity in sacs?/? larvae at 120 hpf compared with observations in control siblings (sacs?/? n = 256; controls n = 273, in 5 independent experiments). Statistical analysis (* p ? 0.05, *** p ? 0.001, **** p ? 0.0001) was performed using the Mann-Whitney test. (C) ?Cerebellar? morphology as assessed in vivo by RFP fluorescence in Tg(tagRFP-T:PC:GCaMP5G). The ?cerebellar? area was found to be significantly reduced in sacs?/? compared with control specimens at 120 hpf. (D) Statistical analysis of the data shown in (C), **** p ? 0.00001, calculated by the Mann-Whitney test. The values are expressed as mean ± standard deviation. (E) Dorsal views at 120 hpf of whole-mount larvae labeled with mAB pvalb7 (sacs?/? n = 20; controls n = 20). (F) Pearson?s coefficient of skewness of the distribution of the fluorescence fluctuations, ?F/F0. These statistics are cumulated from 16 recordings in controls and sacs?/? mutants, respectively. (* p ? 0.05 was calculated using the Mann-Whitney test). Abbreviations: n, number of evaluated embryos in total; error bars indicate standard errors of the means.
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