PUBLICATION
An endothelial-cell-enriched primary culture system to study vascular endothelial growth factor (VEGF A) expression in a teleost, the Japanese eel (Anguilla japonica)
- Authors
- Huang, Y.S., Huang, W.L., Lin, W.F., Chen, M.C., and Jeng, S.R.
- ID
- ZDB-PUB-060703-14
- 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.
Citation
Huang, Y.S., Huang, W.L., Lin, W.F., Chen, M.C., and Jeng, S.R. (2006) An endothelial-cell-enriched primary culture system to study vascular endothelial growth factor (VEGF A) expression in a teleost, the Japanese eel (Anguilla japonica). Comparative biochemistry and physiology. Part A, Molecular & integrative physiology. 145(1):33-46.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping