ZFIN ID: ZDB-PUB-060703-14
An endothelial-cell-enriched primary culture system to study vascular endothelial growth factor (VEGF A) expression in a teleost, the Japanese eel (Anguilla japonica)
Huang, Y.S., Huang, W.L., Lin, W.F., Chen, M.C., and Jeng, S.R.
Date: 2006
Source: Comparative biochemistry and physiology. Part A, Molecular & integrative physiology   145(1): 33-46 (Journal)
Registered Authors: Chen, Ming-chyuan
Keywords: bFGF, Capillary endothelial cell, Eel, Fish, Hypoxia, Nitric oxide, Rete mirabile, Sex steroid, VEGF
MeSH Terms:
  • Amino Acid Sequence
  • Anguilla/genetics*
  • Anguilla/metabolism
  • Animals
  • Base Sequence
  • Cells, Cultured
  • Cobalt/pharmacology
  • Culture Media/chemistry
  • Culture Media/pharmacology
  • DNA, Complementary/chemistry
  • DNA, Complementary/genetics
  • Dose-Response Relationship, Drug
  • Endothelial Cells/cytology
  • Endothelial Cells/drug effects
  • Endothelial Cells/metabolism*
  • Estradiol/pharmacology
  • Fibroblast Growth Factor 2/pharmacology
  • Gene Expression/drug effects
  • Immunohistochemistry
  • Lipoproteins, LDL/chemistry
  • Lipoproteins, LDL/metabolism
  • Molecular Sequence Data
  • Phylogeny
  • RNA, Messenger/genetics
  • RNA, Messenger/metabolism
  • Reverse Transcriptase Polymerase Chain Reaction/methods
  • Sequence Analysis, DNA
  • Time Factors
  • Vascular Endothelial Growth Factor A/genetics*
  • Vascular Endothelial Growth Factor A/metabolism
PubMed: 16807025 Full text @ Comp. Biochem. Physiol. A Mol. Integr. Physiol.
ABSTRACT
A partial gene for eel (Anguilla japonica) vascular endothelial growth factor (VEGF) has been cloned and an endothelial-cell-enriched primary culture derived from rete mirabile established to study regulation of the expression of the eel VEGF gene. Cells were cultured in M199 medium containing 0.1% fetal calf serum (FCS) and serum-free M199 medium for long-and short-term experiments, respectively. Cells were separately treated with cobalt ions (Co(2+)), basic fibroblast growth factor (bFGF), and estradiol (E2), which have been demonstrated to stimulate mammalian VEGF A expression, followed by quantification of the VEGF mRNA levels by real-time reverse transcription polymerase chain reaction. Our results show that: (1) the deduced eel VEGF protein encoded by the cloned gene is about 130 amino acids in length, and is closely related to a zebrafish (Danio rerio) VEGF A; (2) the endothelial-cell-enriched rete mirabile primary culture containing mainly (over 70%) the capillary endothelial cells; (3) the expression levels of the eel VEGF transcript were increased by Co(2+), bFGF, and E2 treatments in a dose-and time-dependent manner. Our data demonstrate that an eel partial VEGF gene has been cloned and its regulation of expression in endothelial-cell-enriched rete mirabile cell culture is similar to that in higher vertebrates.
ADDITIONAL INFORMATION