MFNG role in zebrafish is fundamental for heart and valve development. a qRT-PCR analysis of the relative transcript level of MFNG during embryogenesis from 24 to 60 hpf. Data are shown as the mean ± SEM, n = 6. b Schematic representation of MFNG exons with the target sites of MFNG MO-1 and MFNG MO-2, which induce MFNG deficiency in zebrafish embryos by targeting the 3-exon–3-intron junction and the 4-exon–4-intron junction, respectively. c The efficacy of MFNG MO1 was confirmed by qRT-PCR. Data are presented as mean ± SEM, n = 4. d The lethality of ctrl-MO and MFNG-MO embryos from 8 to 48 hpf. The chi-square test was used for statistical analysis. e The heart rate of ctrl-MO and MFNG-MO embryos at 72 hpf. f Bright-field views of live ctrl-MO and MFNG-MO embryos at 72 hpf. Note the pericardial edema (red arrowhead) and tail deformity (black arrowhead). g Compared to the ctrl-MO group, the MFNG-MO group exhibited a significantly higher number of pericardial edema and tail deformities. h, i Lateral views of ctrl-MO embryos and MFNG-MO embryos at 72 hpf, captured in the Tg (myl7: EGFP) background. Zebrafish embryos in the MFNG-MO group displayed absent regular heart looping. The chi-square test was used for statistical analysis. j Sections of zebrafish embryos stained with HE at 120 h post-MO injection. In the MFNG-MO group, the valve leaflets observed in the ctrl-MO group are absent. The red arrow represents the heart valve, and the red oval denotes the atrioventricular boundary
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