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Isolation and Characterization of Single Cells from Zebrafish Embryos
Published on: March 12, 2016
Atlas unicelular del desarrollo del crecimiento coronario y la interacción de los miocardiocitos en el pez cebra
Muhammad Abdul Rouf1,2, Gülsüm Kayman Kürekçi1,2, Shaoqiu Zhang1,2
1Centre de Recherche Azrieli, Centre Hospitalier Universitaire Sainte-Justine, Montréal, QC, Canada.
Abstract:
Cardiac morphogenesis requires the intricate coordination of different cell types and molecular cues. Coronary vessel formation is essential for heart development, yet how coronaries grow and contribute to ventricular wall formation remains poorly understood. Here, we combine high-resolution imaging and new genetic tools to systematically analyze coronary 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 via Vegfa signaling perturbation through gain- and loss-of-function approaches disrupts coronary-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 identified 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.
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