title: EphB4 forward-signaling regulates cardiac progenitor development in mouse ES cells.
abstract: Eph receptor (Eph)-ephrin signaling plays an important role in organ development and tissue regeneration. Bidirectional signaling of EphB4-ephrinB2 regulates cardiovascular development. To assess the role of EphB4-ephrinB2 signaling in cardiac lineage development, we utilized two GFP reporter systems in embryonic stem (ES) cells, in which the GFP transgenes were expressed in Nkx2.5(+) cardiac progenitor cells and in α-MHC(+) cardiomyocytes, respectively. We found that both EphB4 and ephrinB2 were expressed in Nkx2.5-GFP(+) cardiac progenitor cells, but not in α-MHC-GFP(+) cardiomyocytes during cardiac lineage differentiation of ES cells. An antagonist of EphB4, TNYL-RAW peptides, that block the binding of EphB4 and ephrinB2, impaired cardiac lineage development in ES cells. Inhibition of EphB4-ephrinB2 signaling at different time points during ES cell differentiation demonstrated that the interaction of EphB4 and ephrinB2 was required for the early stage of cardiac lineage development. Forced expression of human full-length EphB4 or intracellular domain-truncated EphB4 in EphB4-null ES cells was established to investigate the role of EphB4-forward signaling in ES cells. Interestingly, while full-length EphB4 was able to restore the cardiac lineage development in EphB4-null ES cells, the truncated EphB4 that lacks the intracellular domain of tyrosine kinase and PDZ motif failed to rescue the defect of cardiomyocyte development, suggesting that EphB4 intracellular domain is essential for the development of cardiomyocytes. Our study provides evidence that receptor-kinase-dependent EphB4-forward signaling plays a crucial role in the development of cardiac progenitor cells.
authors.affiliation: Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China.
authors.forename: Kang
authors.initials: K
authors.lastname: Chen
authors.forename: Hao
authors.initials: H
authors.lastname: Bai
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Liu
authors.forename: Dixie L
authors.initials: DL
authors.lastname: Hoyle
authors.forename: Wei-Feng
authors.initials: WF
authors.lastname: Shen
authors.forename: Li-Qun
authors.initials: LQ
authors.lastname: Wu
authors.forename: Zack Z
authors.initials: ZZ
authors.lastname: Wang
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology*; D036388:Ephrin-B2 / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology*; D018995:Myosin Heavy Chains / Q000378:metabolism; D011485:Protein Binding; D060888:Real-Time Polymerase Chain Reaction; D036224:Receptor, EphB4 / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D036388:Ephrin-B2; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; C484469:Nkx2-5 protein, mouse; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D036224:Receptor, EphB4; D018995:Myosin Heavy Chains
keywords: CARDIAC PROGENITOR CELLS; CARDIOMYOCYTES; EMBRYONIC STEM (ES) CELLS; EphB4; Nkx 2.5; ephrinB2; α-MHC