Fig. 1 | European Journal of Human Genetics

Fig. 1

From: Analysis of cilia dysfunction phenotypes in zebrafish embryos depleted of Origin recognition complex factors

Fig. 1

Loss of ORC1 produces ciliopathy phenotypes in zebrafish embryos. a Live images of 48 hpf zebrafish embryos. NI: uninjected controls, CTRL MO: embryos injected with a control MO, Orc1 MO: translation blocking MO against Orc1. Arrow indicates pericardial oedema. Scale bar: 500 µm. b Orc1 morphants are characterised by an overall short body as it had been shown in the original paper describing the Orc1 MO [5]. c Head size is reduced upon Orc1 depletion. d Eyes are smaller in the absence of Orc1. e Orc1 morphant embryos develop pronounced pericardial oedema. f Fluorescent images of the pronephros of control (NI, CTRL MO) and Orc1 KD embryos. To visualise the pronephros a transgenic line expressing GFP under the wt1b promoter was used (Tg(wt1b:GFP)). Scale bar: 100 µm. g Incidence of cystic pronephric tubules. Two-sided Fisher’s exact test. Number of embryos analysed: NI = 120; CTRL MO = 119; Orc1 MO = 128. h Examples of southpaw (spaw) expression in 20–22 ss embryos showing transcripts in the left or right LPM, the middle picture shows ambiguous expression. Scale bar: 300 µm. i Depletion of Orc1 randomises spaw expression. n = 5–9 experiments. Number of embryos: NI = 231; CTRL MO = 253; Orc1 MO = 114. j The heart of 48 hpf zebrafish embryos is normally looped in an S-shape with the ventricle left and above from the atrium (D-loop). When LR asymmetry is disturbed, hearts stay unlooped or show an inverse loop (L-loop). ISH for cmlc2. Scale bar: 200 µm. k Heart looping is disturbed in Orc1 morphants. n = 4–6 experiments. Number of embryos: NI = 82; CTRL MO = 67; Orc1 MO = 145. l ISH for the endocrine pancreas marker insulin (ins) reveals a normal right-sided localisation of the pancreas (right), which is reversed in asymmetry defects (left). Dashed line: midline of the embryo. Dorsal view. Scale bar: 200 µm. m Wrong-sided pancreas in Orc1 morphants. n = 4–6 experiments. Number of embryos: NI = 82; CTRL MO = 67; Orc1 MO = 145. be Graphs show means ± SEM with circles indicating individual experiments. One-way ANOVA with Sidak’s multiple comparisons test. n = 4–6 independent experiments. Number of embryos analysed: NI = 72; CTRL MO = 86; Orc1 MO = 151. i, k, m Data analysed using two-tailed Fisher’s exact test

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