title: Endocardium Minimally Contributes to Coronary Endothelium in the Embryonic Ventricular Free Walls.
abstract: There is persistent uncertainty regarding the developmental origins of coronary vessels, with 2 principal sources suggested as ventricular endocardium or sinus venosus (SV). These 2 proposed origins implicate fundamentally distinct mechanisms of vessel formation. Resolution of this controversy is critical for deciphering the programs that result in the formation of coronary vessels and has implications for research on therapeutic angiogenesis.
  
  
  
  To resolve the controversy over the developmental origin of coronary vessels.
  
  
  
  We first generated nuclear factor of activated T cells (Nfatc1)-Cre and Nfatc1-Dre lineage tracers for endocardium labeling. We found that Nfatc1 recombinases also label a significant portion of SV endothelial cells in addition to endocardium. Therefore, restricted endocardial lineage tracing requires a specific marker that distinguishes endocardium from SV. By single-cell gene expression analysis, we identified a novel endocardial gene natriuretic peptide receptor 3 (Npr3). Npr3 is expressed in the entirety of the endocardium but not in the SV. Genetic lineage tracing based on Npr3-CreER showed that endocardium contributes to a minority of coronary vessels in the free walls of embryonic heart. Intersectional genetic lineage tracing experiments demonstrated that endocardium minimally contributes to coronary endothelium in the embryonic ventricular free walls.
  
  
  
  Our study suggested that SV, but not endocardium, is the major origin for coronary endothelium in the embryonic ventricular free walls. This work thus resolves the recent controversy over the developmental origin of coronary endothelium, providing the basis for studying coronary vessel formation and regeneration after injury.
authors.affiliation: From the Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences (H.Z., W.P., X.H., L.H., X.T., Q.L., L.Z., B.Z.) and Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences (B.Z.), Chinese Academy of Sciences, Shanghai, China; Cardiovascular Institute, Division of Cardiovascular Medicine, Department of Medicine, and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, CA (G.L., S.M.W.); Broad CIRM Center and Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles (H.M.S.); and School of Life Science and Technology, ShanghaiTech University, Shanghai, China (B.Z.).
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.affiliation: From the Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences (H.Z., W.P., X.H., L.H., X.T., Q.L., L.Z., B.Z.) and Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences (B.Z.), Chinese Academy of Sciences, Shanghai, China; Cardiovascular Institute, Division of Cardiovascular Medicine, Department of Medicine, and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, CA (G.L., S.M.W.); Broad CIRM Center and Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles (H.M.S.); and School of Life Science and Technology, ShanghaiTech University, Shanghai, China (B.Z.).
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Pu
authors.affiliation: From the Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences (H.Z., W.P., X.H., L.H., X.T., Q.L., L.Z., B.Z.) and Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences (B.Z.), Chinese Academy of Sciences, Shanghai, China; Cardiovascular Institute, Division of Cardiovascular Medicine, Department of Medicine, and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, CA (G.L., S.M.W.); Broad CIRM Center and Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles (H.M.S.); and School of Life Science and Technology, ShanghaiTech University, Shanghai, China (B.Z.).
authors.forename: Guang
authors.initials: G
authors.lastname: Li
authors.affiliation: From the Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences (H.Z., W.P., X.H., L.H., X.T., Q.L., L.Z., B.Z.) and Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences (B.Z.), Chinese Academy of Sciences, Shanghai, China; Cardiovascular Institute, Division of Cardiovascular Medicine, Department of Medicine, and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, CA (G.L., S.M.W.); Broad CIRM Center and Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles (H.M.S.); and School of Life Science and Technology, ShanghaiTech University, Shanghai, China (B.Z.).
authors.forename: Xiuzhen
authors.initials: X
authors.lastname: Huang
authors.affiliation: From the Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences (H.Z., W.P., X.H., L.H., X.T., Q.L., L.Z., B.Z.) and Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences (B.Z.), Chinese Academy of Sciences, Shanghai, China; Cardiovascular Institute, Division of Cardiovascular Medicine, Department of Medicine, and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, CA (G.L., S.M.W.); Broad CIRM Center and Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles (H.M.S.); and School of Life Science and Technology, ShanghaiTech University, Shanghai, China (B.Z.).
authors.forename: Lingjuan
authors.initials: L
authors.lastname: He
authors.affiliation: From the Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences (H.Z., W.P., X.H., L.H., X.T., Q.L., L.Z., B.Z.) and Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences (B.Z.), Chinese Academy of Sciences, Shanghai, China; Cardiovascular Institute, Division of Cardiovascular Medicine, Department of Medicine, and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, CA (G.L., S.M.W.); Broad CIRM Center and Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles (H.M.S.); and School of Life Science and Technology, ShanghaiTech University, Shanghai, China (B.Z.).
authors.forename: Xueying
authors.initials: X
authors.lastname: Tian
authors.affiliation: From the Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences (H.Z., W.P., X.H., L.H., X.T., Q.L., L.Z., B.Z.) and Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences (B.Z.), Chinese Academy of Sciences, Shanghai, China; Cardiovascular Institute, Division of Cardiovascular Medicine, Department of Medicine, and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, CA (G.L., S.M.W.); Broad CIRM Center and Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles (H.M.S.); and School of Life Science and Technology, ShanghaiTech University, Shanghai, China (B.Z.).
authors.forename: Qiaozhen
authors.initials: Q
authors.lastname: Liu
authors.affiliation: From the Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences (H.Z., W.P., X.H., L.H., X.T., Q.L., L.Z., B.Z.) and Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences (B.Z.), Chinese Academy of Sciences, Shanghai, China; Cardiovascular Institute, Division of Cardiovascular Medicine, Department of Medicine, and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, CA (G.L., S.M.W.); Broad CIRM Center and Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles (H.M.S.); and School of Life Science and Technology, ShanghaiTech University, Shanghai, China (B.Z.).
authors.forename: Libo
authors.initials: L
authors.lastname: Zhang
authors.affiliation: From the Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences (H.Z., W.P., X.H., L.H., X.T., Q.L., L.Z., B.Z.) and Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences (B.Z.), Chinese Academy of Sciences, Shanghai, China; Cardiovascular Institute, Division of Cardiovascular Medicine, Department of Medicine, and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, CA (G.L., S.M.W.); Broad CIRM Center and Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles (H.M.S.); and School of Life Science and Technology, ShanghaiTech University, Shanghai, China (B.Z.).
authors.forename: Sean M
authors.initials: SM
authors.lastname: Wu
authors.affiliation: From the Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences (H.Z., W.P., X.H., L.H., X.T., Q.L., L.Z., B.Z.) and Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences (B.Z.), Chinese Academy of Sciences, Shanghai, China; Cardiovascular Institute, Division of Cardiovascular Medicine, Department of Medicine, and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, CA (G.L., S.M.W.); Broad CIRM Center and Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles (H.M.S.); and School of Life Science and Technology, ShanghaiTech University, Shanghai, China (B.Z.).
authors.forename: Henry M
authors.initials: HM
authors.lastname: Sucov
authors.affiliation: From the Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences (H.Z., W.P., X.H., L.H., X.T., Q.L., L.Z., B.Z.) and Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences (B.Z.), Chinese Academy of Sciences, Shanghai, China; Cardiovascular Institute, Division of Cardiovascular Medicine, Department of Medicine, and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, CA (G.L., S.M.W.); Broad CIRM Center and Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles (H.M.S.); and School of Life Science and Technology, ShanghaiTech University, Shanghai, China (B.Z.). zhoubin@sibs.ac.cn.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhou
mesh_terms: D000818:Animals; D019070:Cell Lineage; D003331:Coronary Vessels / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D004699:Endocardium / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D006352:Heart Ventricles / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D051379:Mice; D050778:NFATC Transcription Factors / Q000235:genetics / Q000378:metabolism; D017461:Receptors, Atrial Natriuretic Factor / Q000235:genetics / Q000378:metabolism
chemical_list: D050778:NFATC Transcription Factors; C494005:Nfatc1 protein, mouse; D017461:Receptors, Atrial Natriuretic Factor; C073616:atrial natriuretic factor receptor C
keywords: blood vessel; coronary; coronary artery; coronary development; coronary origin; lineage tracing