A systems biology approach for understanding the transcriptome wide modulation of cardiac gene expression under oxidative environment during early stage of cardiovascular remodeling in developing embryo

Various internal and external factors (including environmental pollutants) elevates oxidative stress in the developing embryo. Earlier studies unveil that chick embryo developed under oxidative environment due to exposure of reactive oxygen species (ROS) leads to cardiomyocyte hypertrophy associated with modulation of signaling pathways. In this study, we created the oxidative environment with 2,2-azobis(2-amidinopropane) dihydrochloride (AAPH), a free radicals generator to study the effects of oxidative stress on the cardiovascular system during chick embryo development. Here we exposed the chick embryo to AAPH through injecting 100 µL of AAPH (2.6 µmol) into the air chamber of fertilized eggs containing HH 35 chick embryos. The injections were repeated on alternative days for up to 10th day. At HH37 (day 11) the hearts were harvested from all surviving embryos for total RNA isolation. Saline and AAPH exposed embryonic heart RNA was isolated for RNA-Seq using next generation sequencing (NGS) approach.