April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Glycogen Synthase Kinase-3β Inhibitors Reduce Vegf-induced Neovascular Sprouts In Retinal Explants
Author Affiliations & Notes
  • Satoshi Morooka
    Kyoto University, Kyoto, Japan
  • Ken Ogino
    Kyoto University, Kyoto, Japan
  • Masayuki Nukada
    Kyoto University, Kyoto, Japan
  • Tomoaki Murakami
    Kyoto University, Kyoto, Japan
  • Nagahisa Yoshimura
    Kyoto University, Kyoto, Japan
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 4838. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Satoshi Morooka, Ken Ogino, Masayuki Nukada, Tomoaki Murakami, Nagahisa Yoshimura; Glycogen Synthase Kinase-3β Inhibitors Reduce Vegf-induced Neovascular Sprouts In Retinal Explants. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4838.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: : Glycogen synthase kinase-3β (GSK-3β), a downstream molecule of insulin signaling, plays an important role in glycogen metabolism, although only a few publications regarding angiogenic effects were reported. Here we investigated the effects of GSK-3β inhibitors on vascular endothelial growth factor (VEGF)-induced retinal angiogenesis.

Methods: : Retinas from 7-8 week old C57BL/6J mice were isolated and cultured on Millicell filter with 25 ng/ml VEGF alone or VEGF + each dose of GSK-3β inhibitors. After 96 hours, retinas were fixed and applied to immunohistochemistry with PEACAM and collagen type IV, for the quantification of neovascular sprouts. Human umbilical vein endothelial cells (HUVECs) were cultured with CSC Complete Recombinant Medium (Cell Systems Corporation). Live cell imaging were performed using cytoplasmic staining of HUVECs with CellMaskTM, and confocal images were obtained at 0, 5, 15 min after the treatments. Phalloidin was used for the evaluation of actin cytoskeleton.

Results: : GSK-3β inhibitors, SB216763 25 µM and AR-A014418 250 µM, decreased VEGF-induced neovascular sprouts in ex vivo retinal angiogenic model (p<0.05 and p<0.01, respectively). We additionally performed the timelapse imaging of HUVECs and found that cytoplasm at cell borders was retracted with the filopodia-like structure after the treatment with GSK-3β inhibitors, which might be compatible to their antiangiogenic effects. Immunostaining with phalloidin showed that actin stress fibers were produced 5 min after the treatment with GSK-3β inhibitors.

Conclusions: : These data suggests GSK-3β inhibitors reduced retinal angiogenesis, mediated via cytoskeletal changes and concomitant cellular retraction, at least in part.

Keywords: neovascularization • retinal neovascularization 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×