Developmental single-cell atlas of coronary vessel growth and cardiomyocyte interaction in zebrafish

Cardiac morphogenesis requires the intricate coordination of different cell types and molecular cues. Coronary vessel formation is essential for heart development, yet how coronary vessels grow and contribute to ventricular wall formation remains poorly understood. Here, we combine high-resolution imaging and new genetic tools to systematically analyze coronary vasculature development in zebrafish at each millimeter increment from 7 to 30 mm of body length and identify cellular and molecular milestones defining four distinct coronary network developmental stages. Our data show how coronary vessels expand, pattern, specify and act as vascular scaffolds to guide cardiomyocyte growth throughout development. Manipulating coronary network growth by Vegfa signaling perturbation through gain- and loss-of-function approaches disrupts coronary vessel-cardiomyocyte interactions and cardiomyocyte expansion. To gain further resolution into these processes, we build a single-cell RNA sequencing atlas by profiling 37,554 ventricular cells across the different developmental stages and identify new coronary markers, dynamic cellular interactions, and stage-specific endothelial-cardiomyocyte crosstalk. Overall, we present the first comprehensive roadmap of coronary vessel formation and coordinated cellular and molecular interactions with the developing cardiac muscle in zebrafish.