Vegfr3 and Tbx1 Interact in Cardiac Morphogenesis

Gene inactivation in model organisms has identified numerous genes and signaling pathways involved in mammalian cardiac outflow tract (OFT) development. Human genetics data have implicated the VEGFR3 gene in OFT development, but when and where it is required is unknown. In this study we determined the sensitivity of the developing murine heart to reduced Vegfr3 gene dosage, and we tested whether its requirement is dependent upon TBX1, a known regulator of Vegfr3 expression in cardiac and lymphatic endothelial cells. We found that in the mouse, a single copy of the Vegfr3 gene was sufficient for normal heart development in most cases. Mutation of a single copy of the Tbx1 gene greatly enhanced the sensitivity of heart development to Vegfr3 dosage reduction and led to the formation of cardiac defects. In addition, deletion of Vegfr3 in the Tbx1 expression domain also led to severe cardiac OFT abnormalities. We used RNAscope to reveal the location of Vegfr3 and Tbx1 transcripts in midterm mouse embryos. This revealed co-localization of these transcripts in the endothelium of the aortic sac, caudal pharyngeal arch arteries and proximal OFT, suggesting that these are potential sites of genetic interaction between Vegfr3 and Tbx1 that are critical for murine heart development.