Nuclear receptor subfamily 2 group F member 1a (nr2f1a) is required for vascular development in zebrafish

PLoS One. 2014 Aug 26;9(8):e105939. doi: 10.1371/journal.pone.0105939. eCollection 2014.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Conserved Sequence
  • DNA-Binding Proteins / physiology*
  • Endothelium, Vascular / growth & development
  • Endothelium, Vascular / metabolism
  • Gene Expression
  • Gene Expression Regulation, Developmental
  • Molecular Sequence Data
  • Neovascularization, Physiologic*
  • Transcription Factors / physiology*
  • Zebrafish / physiology*
  • Zebrafish Proteins / physiology*

Substances

  • DNA-Binding Proteins
  • Transcription Factors
  • Zebrafish Proteins
  • nr2f1a protein, zebrafish

Grants and funding

This work was supported by the grants from National Science Council, Taiwan (NSC101-2311-B-110-002, NSC102-2311-B-110-002) and from the NSYSU-KMU Joint Research Project (#NSYSUKMU102-P028) to CYW; the grant from Zuoying Branch of Kaohsiung Armed Forces General Hospital, Taiwan to BJW (ZAFGH 102-03). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.