2-acetoxyfuranodiene decreased the formation of angiogenic blood vessels during zebrafish embryonic development. (A) Bright field image of a Tg (fli1: EGFP) embryo that was exposed to 0.1% methanol as a control. The embryos developed normally. (B) The fluorescent image of a Tg (fli1: EGFP) embryo at 72 hpf. The green color indicates all the blood vessels. The white arrows show the inter-segmental blood vessels in the trunk of the control embryo, which were the first angiogenic blood vessels that formed at around 48 hpf. The emergence of the second angiogenic blood vessels can be seen on the trunk of the control embryo. (C) A Tg (fli1: EGFP) embryo treated with 15 µM of CM2. The bright field images show that CM2 at 15 µM affected the normal development of zebrafish embryos. The embryos were much smaller in size, the body was curved, and there was no pigmentation. (D) The CM2 treatment disrupted the inter-somatic blood vessel formation in the treated zebrafish embryos, the majority of the inter-somatic blood vessels did not form (white asterisks), and some blood vessels formed but they did not grow and failed to connect to dorsal longitudinal anastomotic vessel. Similarly, the sub-intestinal vein did not form in the transgenic zebrafish embryos that were treated with CM2.
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