Gas6 stimulates angiogenesis of human retinal endothelial cells and of zebrafish embryos via ERK1/2 signaling
- Authors
- Kim, Y.S., Jung, S.H., Jung, D.H., Choi, S.J., Lee, Y.R., and Kim, J.S.
- ID
- ZDB-PUB-140317-16
- Date
- 2014
- Source
- PLoS One 9(1): e83901 (Journal)
- Registered Authors
- Jung, Seung-Hyun
- Keywords
- none
- MeSH Terms
-
- Animals
- Aorta/drug effects
- Aorta/metabolism
- Cell Movement/drug effects
- Cell Movement/genetics
- Cell Proliferation/drug effects
- Endothelial Cells/drug effects*
- Endothelial Cells/metabolism*
- Gene Knockdown Techniques
- Gene Order
- Human Umbilical Vein Endothelial Cells
- Humans
- Intercellular Signaling Peptides and Proteins/genetics*
- Intercellular Signaling Peptides and Proteins/pharmacology
- MAP Kinase Signaling System/drug effects
- Neovascularization, Physiologic/drug effects
- Neovascularization, Physiologic/genetics*
- Phenotype
- Protein Kinase Inhibitors/pharmacology
- Rats
- Retina/metabolism*
- Zebrafish
- PubMed
- 24409287 Full text @ PLoS One
Aim
To determine if growth arrest-specific 6 (Gas6) plays an important role in the regulation of angiogenesis in human retinal microvascular endothelial cells (HRMECs) and in vessel development of zebrafish.
Methods
Proliferation, wound-healing cell migration, and tube formation were measured in HRMECs treated with recombinant human Gas6 (rhGas6). Sprague-Dawley rat aortas in Matrigels were treated with rhGas6, and microvessel sprouting emanating from arterial rings was analyzed. Transgenic zebrafish embryos (flk:GFP) were microinjected with rhGas6 at 50 hours post-fertilization (hpf), and ectopic sprouting of subintestinal vessels (SIVs) was observed under a confocal microscope. Morpholino oligonucleotides (MOs) were microinjected to knockdown gas6 in zebrafish embryos, and intersegmental vessel impairment was observed. The effect of the extracellular signal-regulated kinase (ERK1/2) inhibitor on the migration of HRMECs and on vessel development in zebrafish embryos was tested.
Results
rhGas6 stimulated proliferation, migration, and tube formation in HRMECs in a dose-dependent manner. In rat aortas, rhGas6 induced vessel outgrowth, and the sprouting length was longer than that of controls. The rhGas6-microinjected zebrafish embryos had significantly increased vessel outgrowth in the SIVs. Recombinant human vascular endothelial growth factor (rhVEGF) served as a positive control. Knockdown of gas6 inhibited angiogenesis in the developing vessels of zebrafish. The ERK1/2 inhibitor inhibited HRMEC migration and intersegmental vessel formation in zebrafish embryos.
Conclusions/Interpretations
These data suggest that Gas6 plays a pivotal role in proliferation, migration, and sprouting of angiogenic endothelial cells in the retina and in zebrafish embryos. Furthermore, Gas6 induced angiogenic processes are induced via phosphorylation of ERK1/2.