oa The role of the epigenetic factor in valvulogenesis in zebrafish in vivo

In this study, we focused on wall shear stress as an epigenetic factor to initiate valve formation. Fast or oscillatory flow is induced in an embryonic zebrafish vessel to mimic the flow in atrioventricular canal (AVC). By creating an environment similar to AVC in non-cardiac region in vivo, we propose to seek a better understanding of the mechanisms associated with the induction of valvulogenesis signaling pathways. Polydimethylsiloxane is injected at the vessel wall of embryonic zebrafish, creating fast or oscillatory flow, therefore modulating local wall shear stress (WSS). Particle image velocimetry (PIV) technique is used to measure blood flow velocity. Whole mount in situ hybridization is performed to detect kruppel-like factor 2a (klf2a) expression. Klf2a is used as a marker gene since it responds to shear stress and is crucial for valve formation. Using PIV, WSS is calculated. In situ hybridization showed klf2a expression in the vessel as a result of either high WSS or oscillatory WSS. Klf2a is not expressed in the vessel during normal embryonic development. With a new gel injection technique, local WSS in the zebrafish vessel has changed, inducing expression of klf2a, an important gene for the initiation of valve formation. Expression of different genes such as TGF-β family that are involved in valve formation will be studied further to establish an innovative model to explore valvulogenesis in vivo.