Single cell transcriptional profiles of endothelial cells sorted from the hearts with ablation of Wnt/beta-catenin signaling activating cardiomyocytes and without ablation [scRNA-seq]

To understand how Wnt/ß-catenin signaling-activated cardiomyocytes (ß-cat ON CMs) promote coronary vessel development, we performed single-cell RNA sequencing of endothelial cells (ECs) sorted from the hearts ablated ß-cat ON CMs and those from control hearts, respectively. Our analyses indicated that ß-cat ON CMs regulates coronary vessel development by promoting arterialization, a step in the transition of endocardial ECs to coronary ECs. Overall design: To induce nitroreductase-metronidazole (NTR-MTZ)-dependent ablation of ß-cat ON CMs, we treated 5 mM MTZ or DMSO to Tg(fli1:EGFP);Tg(cryaa:EGFP, myl7:GAL4db-TCF?C);(5xUAS:NTR-mCherry) fish every other day for 4 weeks from 1 month-old. We isolated live GFP+ cells by FACS sorting from the fish-treated MTZ or DMSO, respectively. Barcoded single-cell cDNA libraries were prepared using the Chromium Next GEM single cell 3'reagent kits v3.1 and sequenced using Illumina NextSeq 500.