Transcription profiles of endothelial cells in the rat ductus arteriosus during a perinatal period

PLoS One. 2013 Sep 27;8(9):e73685. doi: 10.1371/journal.pone.0073685. eCollection 2013.

Abstract

Endothelial cells (ECs) lining the blood vessels serve a variety of functions and play a central role in the homeostasis of the circulatory system. Since the ductus arteriosus (DA) has different arterial characteristics from its connecting vessels, we hypothesized that ECs of the DA exhibited a unique gene profile involved in the regulation of DA-specific morphology and function. Using a fluorescence-activated cell sorter, we isolated ECs from pooled tissues from the DA or the descending aorta of Wistar rat fetuses at full-term of gestation (F group) or neonates 30 minutes after birth (N group). Using anti-CD31 and anti-CD45 antibodies as cell surface markers for ECs and hematopoietic derived cells, respectively, cDNAs from the CD31-positive and CD45-negative cells were hybridized to the Affymetrix GeneChip® Rat Gene 1.0 ST Array. Among 26,469 gene-level probe sets, 82 genes in the F group and 81 genes in the N group were expressed at higher levels in DA ECs than in aortic ECs (p<0.05, fold change>2.0). In addition to well-known endothelium-enriched genes such as Tgfb2 and Vegfa, novel DA endothelium-dominant genes including Slc38a1, Capn6, and Lrat were discovered. Enrichment analysis using GeneGo MetaCore software showed that DA endothelium-related biological processes were involved in morphogenesis and development. We identified many overlapping genes in each process including neural crest-related genes (Hoxa1, Hoxa4, and Hand2, etc) and the second heart field-related genes (Tbx1, Isl1, and Fgf10, etc). Moreover, we found that regulation of epithelial-to-mesenchymal transition, cell adhesion, and retinol metabolism are the active pathways involved in the network via potential interactions with many of the identified genes to form DA-specific endothelia. In conclusion, the present study uncovered several significant differences of the transcriptional profile between the DA and aortic ECs. Newly identified DA endothelium-dominant genes may play an important role in DA-specific functional and morphologic characteristics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • DNA Primers
  • Ductus Arteriosus / cytology
  • Ductus Arteriosus / metabolism*
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / metabolism*
  • Female
  • Gene Expression Profiling*
  • Oligonucleotide Array Sequence Analysis
  • Pregnancy
  • Rats
  • Rats, Wistar
  • Real-Time Polymerase Chain Reaction
  • Transcription, Genetic*

Substances

  • DNA Primers

Grants and funding

This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan (U.Y., S.M.), a Grant-in-Aid for Scientific Research on Innovative Areas (U.Y.,), the Yokohama Foundation for Advanced Medical Science (U.Y., S.M.), the “High-Tech Research Center” Project for Private Universities: MEXT (T.K., S.M.), MEXT-Supported Program for the Strategic Research Foundation at Private Universities (T.K., S.M.), the Vehicle Racing Commemorative Foundation (U.Y., S.M.), Miyata Cardiology Research Promotion Funds (U.Y.), the Takeda Science Foundation (U.Y., S.M.), and the Shimabara Science Promotion Foundation (S.M.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.