Fig. 4

miR-223-3p resulted in aberrant eye vasculature sprout formation. Normal zebrafish eye vasculature development is characterized by three clearly branched blood vessels (indicated by white arrowheads). Exposure to miR-223-3p mimic restricted the growth of these three vessels. The examined groups were scored as three subclasses: normal, mildly affected, and severely affected. A–C Representative images of Tg (fli1a:roy,mitf:GFP) with vasculature expressing green fluorescent protein; A normal, B mild, C severe. The blood vessels displayed abnormal sprouting in mildly and severely affected eyes. Both miR-223-3p (MiR0%G, MiR 2%G, MiR 5%G) and hyperglycemia conditions (control 2%G, control 5%G) resulted in a significant alteration to the developing of blood vessels compared to control with no glucose incubation (control 0%G). Images were obtained using Lumar 12 stereomicroscope (Zeiss Microscopy) and a Nikon camera at ×100 magnification; scale bar, 10 µm. A total number of larvae measured was n = 10 per group. Statistical analysis was conducted using chi-square. D Bar plots showing the distribution of normal, mild, and severe phenotypes in control and miR-223-3p groups in different hyperglycemic conditions (0% G, 2% G, 5% G). E Representative images of injected miR-223-3p embryos into the one-cell stage, with different dose titration levels (1, and 3 μM) in the absence of glucose. The embryos at three days post fertilization were presented with severe developmental defects with 3 μM titration and were classified into 3 groups (G1, G2, and G3) depending on the severity and percentages of abnormally developed larvae. Compared to the control group, the larvae in 1 μM appeared with mixed effects that were less prominent than 3 μM groups. Representative Images were captured using Lumar V.12 stereomicroscope and Nikon camera at ×25