CYP Pathway Modulators Alter Development and Angiogenesis in Zebrafish Embryos

Abstract

Epoxyeicosatrienoic acids (EETs) are CYP metabolites of arachidonic acid that have vascular actions including vasodilation. Arachidonic acid is also metabolized by CYP hydroxylase enzymes to 20‐hydroxyeicosatetraenoic acid (20‐HETE) that causes vasoconstriction and increases vascular reactivity. These vascular properties make CYP metabolites attractive targets for therapeutics. Thus, we determined the actions of chemical compounds that modulate EETs or 20‐HETE on cardiovascular development. A zebrafish embryo model system was utilized to compare heart and caudal vascular plexus development. Tg(kdr:EGFP) zebrafish embryos were exposed to compounds at 10 hours post fertilization (hpf), and incubated until 48hpf when images were taken, quantified, and analyzed using a 2‐tailed t‐test to determine significance. Heart development was affected significantly with a 20‐HETE antagonist (n=19), eliciting a heart width of 176.7 ± 8.2 μm compared with 114.5 ± 17.1 μm in the control fish (n=4). (p<0.05). A soluble epoxide hydrolase inhibitor (sEHI), EET analogs, and combined EET analog/sEHI all caused a narrowing of the caudal vascular plexus. All compounds affecting the caudal vascular plexus caused a 13–21% narrowing compared with control fish (p<0.05). These data demonstrate that chemical compounds that modulate EETs and 20‐HETE have distinct actions on heart and vascular plexus development in the zebrafish embryos. Therefore, we conclude that 20‐HETE antagonism alters heart and vascular development whereas sEHI and EET analogs significantly alter vascular development in zebrafish.