Fig. 1

Fig. 1 MT3 regulates vegfaa expression during hindbrain vascular development.

a Lateral maximum projections of the hindbrain of Tg(fli1:EGFP) after WISH against vegfaa (magenta) and immunostained against GFP (endothelial marker, white) in CTRMO and MCT8MO zebrafish embryos at 32, 36 and 48 hpf. Yellow arrowhead indicates agglomeration of vegfaa expression around a CtA or the putative location where a CtA is supposed to ingress into the hindbrain. The red box indicates the hindbrain area for every embryonic stage analyzed adapted from (Kimmel et al., 1995). Graphical representation of the number of times, defined as frequency (%), that vegfaa expression can be observed around a specific CtA or where a CtA is supposed to develop at b) 32 hpf, c) 36 hpf, d) 48 hpf between CTRMO and MCT8MO embryos; Fisher’s exact test.n = 22 (32hpf and 48hpf CTRMO), 15 (32hpf MCT8MO), 23 (36hpf CTRMO), 21 (36hpf MCT8MO), 18 (48hpf MCT8MO). e At 48 hpf, Tg(fli1:EGFP) zebrafish embryos co-injected with vegfaa-165 mRNA and MCT8MO (n = 24) are able to partially rescue, in a rhombomere specific manner, some but not all CtAs. CTRMO (n = 10), MCT8MO (n = 14). f Statistical analysis of the number of CtAs present in each experimental group; One-way ANOVA followed by Bonferroni’s multiple comparison analysis. Data are presented as box-and-whisker plot, where the black thick horizontal line represents the median. The first and third quartiles are marked by the lower and upper edges of the boxes, respectively. Error bars represent standard deviations (smallest and higher value). g Frequency (%) of each CtA to develop in each experimental group; Fisher’s exact test. **p < 0.01; ***p < 0.001; ****p < 0.0001. For detailed statistics, see Supplementary Data 2. The red arrowhead indicates the mid-cerebral vein (MCeV), and the green arrowhead indicates the primordial hindbrain channels (PHBC). Numbers in yellow 1 – 8 indicate the CtA in its respective rhombomere. Scale bar: 50 μm. Zebrafish drawings used from Kimmel et al (1995)(Ref. ²⁸) under CC licence number 6041951178663.