title: Nuclear receptor subfamily 2 group F member 1a (nr2f1a) is required for vascular development in zebrafish.
abstract: Genetic regulators and signaling pathways are important for the formation of blood vessels. Transcription factors controlling vein identity, intersegmental vessels (ISV) growth and caudal vein plexus (CVP) formation in zebrafish are little understood as yet. Here, we show the importance of the nuclear receptor subfamily member 1A (nr2f1a) in zebrafish vascular development. Amino acid sequence alignment and phylogenetic analysis of nr2f1a is highly conserved among the vertebrates. Our in situ hybridization results showed nr2f1a mRNA is expressed in the lateral plate mesoderm at 18 somite stage and in vessels at 24-30 hpf, suggesting its roles in vasculization. Consistent with this morpholino-based knockdown of nr2fla impaired ISV growth and failed to develop fenestrated vascular structure in CVP, suggesting that nr2f1a has important roles in controlling ISV and CVP growth. Consequently, nr2f1a morphants showed pericardial edema and circulation defects. We further demonstrated reduced ISV cells and decreased CVP endothelial cells sprouting in nr2f1a morphants, indicating the growth impairment of ISV and CVP is due to a decrease of cell proliferation and migration, but not results from cell death in endothelial cells after morpholino knockdown. To test molecular mechanisms and signals that are associated with nr2f1a, we examined the expression of vascular markers. We found that a loss of nr2f1a results in a decreased expression of vein/ISV specific markers, flt4, mrc1, vascular markers stabilin and ephrinb2. This indicates the regulatory role of nr2f1a in controlling vascular development. We further showed that nr2f1a likely interact with Notch signaling by examining nr2f1a expression in rbpsuh morphants and DAPT-treatment embryos. Together, we show nr2f1a plays a critical role for vascular development in zebrafish.
authors.affiliation: Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung, Taiwan; Zuoying Branch of Kaohsiung Armed Forces General Hospital, Kaohsiung, Taiwan.
authors.forename: Bao-Jueng
authors.initials: BJ
authors.lastname: Wu
authors.affiliation: Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Chien-Chih
authors.initials: CC
authors.lastname: Chiu
authors.affiliation: Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung, Taiwan; Marine Biotechnology Doctoral Program, National Sun Yat-sen University, Kaohsiung, Taiwan.
authors.forename: Chun-Lin
authors.initials: CL
authors.lastname: Chen
authors.affiliation: Department of Bioagricultural Science, National Chiayi University, Chiayi, Taiwan.
authors.forename: Wen-Der
authors.initials: WD
authors.lastname: Wang
authors.affiliation: Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung, Taiwan.
authors.forename: Jia-Hong
authors.initials: JH
authors.lastname: Wang
authors.affiliation: Marine Biotechnology Doctoral Program, National Sun Yat-sen University, Kaohsiung, Taiwan; Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung, Taiwan.
authors.forename: Zhi-Hong
authors.initials: ZH
authors.lastname: Wen
authors.affiliation: Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Wangta
authors.initials: W
authors.lastname: Liu
authors.affiliation: Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Hsueh-Wei
authors.initials: HW
authors.lastname: Chang
authors.affiliation: Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung, Taiwan; Marine Biotechnology Doctoral Program, National Sun Yat-sen University, Kaohsiung, Taiwan.
authors.forename: Chang-Yi
authors.initials: CY
authors.lastname: Wu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017124:Conserved Sequence; D004268:DNA-Binding Proteins / Q000502:physiology*; D004730:Endothelium, Vascular / Q000254:growth & development / Q000378:metabolism; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D008969:Molecular Sequence Data; D018919:Neovascularization, Physiologic; D014157:Transcription Factors / Q000502:physiology*; D015027:Zebrafish / Q000502:physiology*; D029961:Zebrafish Proteins / Q000502:physiology*
chemical_list: D004268:DNA-Binding Proteins; D014157:Transcription Factors; D029961:Zebrafish Proteins; C080695:nr2f1a protein, zebrafish